Synthesis and: In vitro anti-proliferative evaluation of naphthalimide-chalcone/pyrazoline conjugates as potential SERMs with computational validation

Article abstract of DOI:10.1039/d0ra01822h

A series of naphthalimide-chalcone/pyrazoline conjugates was prepared and evaluated for their anti-breast cancer potential against estrogen responsive, i.e. MCF-7 (ER+), and triple-negative, i.e. MDA-MB-231 (ER-), cell lines. The structure-activity-relati

Full text of DOI:10.1039/d0ra01822h

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Synthesis and in vitro anti-proliferative evaluation
of naphthalimide–chalcone/pyrazoline conjugates
as potential SERMs with computational validation†
Cite this: RSC Adv., 2020, 10, 15836
Shalini,a Pankaj,a Sourav Taru Saha,b Mandeep Kaur,b Ebenezer Oluwakemi,c
c
a
Parvesh Singhc and Vipan Kumar
*
Paul Awolade,
A series of naphthalimide-chalcone/pyrazoline conjugates was prepared and evaluated for their anti-breast
cancer potential against estrogen responsive, i.e. MCF-7 (ER+), and triple-negative, i.e. MDA-MB-231 (ERꢀ),
cell lines. The structure-activity-relationship (SAR) was deduced based on the influence of linker length,
substituents on the phenyl ring and the generated functionalities, on anti-proliferative activities. Docking
simulations further delineate the type of interactions of the designed molecules with the selected
targets. This report discloses the scope of triazole tethered naphthalimide-chalcone/pyrazoline
conjugates as anti breast cancer agents.
Received 26th February 2020
Accepted 14th April 2020
DOI: 10.1039/d0ra01822h
rsc.li/rsc-advances
several anticancer targets, including thioredoxin reductase, and
tubulin polymerization14,15 and by preferentially killing cancer
cells over non-cancer cells.16 In this context, to improve the
efficacy of chalcones while evading drug-resistance, molecular
hybridization involving dual- or multi-target approach has been
investigated. For instance, Lee et al.,17 synthesized quinoline–
chalcones targeting MDR tumor and metastatic breast cancer
cells, whereas Modzelewska et al.,18 reported a series of bis-
chalcones with exceptional inhibitory potency in the growth of
human breast and colon cancer cells.
Naphthalimides, owing to its at aromatic structure and
DNA-intercalating tendencies, have emerged as target motif in
the area of cancer therapy.19 Some mono or bis-naphthalimide
derivatives have displayed promising activities against tumor
cells, among which, mitonade, amonade, elinade and azo-
nade proceeded to clinical trials.20,21 However, poor thera-
peutic index, poor water solubility, and neurological toxicity
impeded their further advancement in clinical phase.22 Thus,
the ongoing efforts are geared towards modifying the molecular
framework of naphthalimide based scaffolds to improve their
therapeutic properties. A literary survey has disclosed better
results of naphthalimide based scaffolds as anti tumor
including anti-breast cancers.23–26
1H-1,2,3-triazole core, on the other hand, presents desirable
properties such as hydrogen bonding ability, resistance to
oxidation, reduction, and enzymatic hydrolysis in physiological
conditions. Besides, they have shown a broad spectrum of
medicinal potential including anticancer.27–31 Therefore, on this
backdrop and in continuation of our research venture,32–35 the
present manuscript describes the hybridization of the chosen
pharmacophores i.e. substituted chalcones/pyrazolines and 1,8-
naphthalimides using triazole as a linker. Two human breast
cancer cell lines i.e. MCF-7 (ER+) and MDA-MB-231 (ERꢀ) were
Introduction
Cancer, an uncontrolled growth of cells, is the second leading
cause of death worldwide. The most commonly diagnosed
cancers are lung (12.6%), breast (11.9%), colorectal (9.8%), and
prostate (7.7%).1 Breast cancer is the most frequent cancer
among women, affecting 2.1 million women each year. A World
Health Organization (WHO) survey estimated that 627 000
women died from breast cancer worldwide in 2018.2 Treatment
for breast cancer may include surgery, radiation, chemotherapy,
targeted therapy and hormone therapy. The proliferation of
mammary cells is stimulated by the binding of hormone
estrogen to estrogen receptors (ERs, existing in isomeric forms
of ERa and ERb), the later has developed as target for breast
cancer therapy.3 Consequently, selective inhibition of ERa has
led to the emergence of drugs known as Selective Estrogen
Receptor Modulators (SERMs) and is an important strategy in
the treatment of breast cancer.4
Chalcones are a prolic class of avonoids with signicant
occurrence in many edible fruits with medicinal properties.5
Chalcones, with an aromatic enone as a central core, showed
anti-inammatory,6 anti-histaminic,7 anti-oxidant,8 anti-proto-
zoal,9 antibacterial,10 antimalarial,11 antifungal,12 anti-HIV13
activities. Moreover, chalcones have shown potent inhibition of
aDepartment of Chemistry, Guru Nanak Dev University, Amritsar, 143005, India.
E-mail: vipan_org@yahoo.com
bSchool of Molecular and Cell Biology, University of Witwatersrand, Private Bag 3,
Wits-2050, Johannesberg, South Africa. E-mail: mandeep.kaur@wits.ac.za
cSchool of Chemistry and Physics, University of Kwazulu Natal, P/Bag X54001,
Westville, Durban 4000, South Africa
† Electronic supplementary information (ESI) available: Scanned copies of 1H and
13C spectra of few representative compounds i.e. 12b, 12g, 13b, 13d, 13g, 14b, 15b
and Table S1. See DOI: 10.1039/d0ra01822h
15836 | RSC Adv., 2020, 10, 15836–15845
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used to investigate the in vitro anti-proliferative potential of the
synthesized compounds.
Results and discussion
Chemistry
Scheme 1 Reagents and conditions: (a) ethanol amine (n ¼ 2)/prop-
anol amine (n ¼ 3), ethanol, reflux, 1 h (b) mesyl chloride, Et3N, dry
CHCl3, RT, 3 h (c) NaN3, DMF, 60 ꢁC, 2 h.
The rst set of precursors were obtained by treating 1,8-naph-
thalic anhydride 1 with ethanol/propanol amine under reux to
yield 2. Further, compound 2 was treated with mesyl chloride
and Et3N in dry CHCl3 to get the mesylated product 3, followed
by its treatment with NaN3 in DMF to obtain the precursor 4,
(Scheme 1).
For second set of precursors, different aldehydes viz. 6 and 7
were treated with ketone 5, formed by O-propargylation of 4-
hydroxy acetophenone, under the basic condition to form the
Claisen–Schmidt condensation products 8 and 9. The intro-
duction of pyrazoline rings is done by treating 8/9 with hydra-
zine hydrate in acetic acid at 130 ꢁC to afford 10 and 11
respectively, (Scheme 2). The rationale for introducing pyrazo-
line functionality in the designed conjugates is based on its
favourable role in enhancing anti-proliferative potential.36,37
Scheme 2 Reagents and conditions: (a) NaOH (40% w/v), ethanol, RT,
1 h (b) hydrazine hydrate, acetic acid, 130 ꢁC, reflux, 2 h.
Scheme 3 Reagents and condition: (a) CuSO4, sodium ascorbate, EtOH : H2O (90 : 10), RT, 10 h.
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Fig. 1 Representative graphs comparing the percentage growth inhibition of MCF-7 and MDA-MB-231 cells at selected concentrations of test
compounds. 40 mM Plumbagin was used as a positive control. Data are mean ꢂ standard deviation S.D (n ¼ 3), where *p < 0.05, **p < 0.01 and
***p < 0.001 significant difference to untreated control.
Subsequently, the target conjugates viz. 12a–h and 13a–h were
As evident from Table 1, naphthalimide–chalcone conju-
synthesized by azide–alkyne cycloaddition reaction between 4 gates, 12a–h failed to inhibit the growth of both the cell lines
and 8/10 using CuSO4 and sodium ascorbate in absolute even at the highest tested concentration i.e. 100 mM. Replace-
ethanol, (Scheme 3A). The ferrocene based counterparts viz. ment of chalcone core with pyrazoline ring did not result in the
14a–b and 15a–b were synthesized analogously from the cyclo- improvement of their anti-proliferative activities against MDA-
addition reaction between 4 and 9/11, (Scheme 3B). The puri- MB-231 cells as evident from conjugates 13a–h. Nevertheless,
cation of the reaction mixture via column chromatography
using ethyl acetate : hexane (10 : 90) mixture as eluent, afforded
the desired conjugates in good to excellent yields. The struc-
tures of the synthesized molecules were assigned based on the
spectral data and analytical evidences. For example, compound
12b, showed molecular ion peak at m/z 559.1975 [M + H]+ in its
Table 1 The IC50 values (in mM) of tested compounds against MCF-7
and MDA-MB-231 cells
Entries
MCF-7
MDA-MB-231
1
HRMS (ESI) spectrum. Its H NMR spectrum exhibited charac-
12a
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
98.11
62.23
>100
>100
>100
>100
>100
>100
>100
50
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
75
teristic singlets at d 3.83 and 7.76 corresponding to methoxy and
triazole ring proton respectively. The signals at d 188.83, 161.91
and 161.63 corresponding to the carbonyl groups along with
requisite signals in 13C NMR spectrum further conrmed the
assigned structure.
12b
12c
12d
12e
12f
12g
12h
13a
13b
13c
13d
13e
13f
13g
13h
14a
14b
15a
15b
4a
In vitro anti-proliferative evaluation and SAR
The synthesized series of triazole tagged naphthalimide-
chalcone/pyrazoline conjugates was evaluated for their anti-
proliferative activities against estrogen receptor positive and
triple-negative human breast cancer cell lines, MCF-7 (ER+) and
MDA-MB-231 (ERꢀ) respectively, using MTT assay.38 The
percentage growth inhibition of MCF-7 and MDA-MB-231 cells
were recorded at varying concentrations of the synthesized
conjugates using Plumbagin as the positive control, and the
results are depicted in Fig. 1. The IC50 values of the synthesized
compounds, which is the concentration required to inhibit the
growth of 50% of cells, are included in Table 1.
4b
Tamoxifen
Plumbagin
3.5
4.4
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Fig. 2 Generalized SAR of the synthesized compounds.
cytotoxic agent, Plumbagin (IC50¼ 3.5 mM against MCF-7 cells),
but comparable to that of standard drug, Tamoxifen (IC50¼ 50
mM against MCF-7 cells). The graphical representation of
generalized SAR for the synthesized compounds is elucidated in
Fig. 2.
The toxicity of moderately active compounds 13f and 13g was
also tested against normal/non-tumor cell line i.e. Human
Embryonic Kidney (HEK-293). The compounds were non-
cytotoxic with IC50 > 100 mM (Fig. 3 and Table 2).
Fig. 3 Representative graph comparing the percentage growth inhi-
bition of HEK-293 cells at selected concentrations of test compounds
13f and 13g. 40 mM Plumbagin was used as a positive control. Data are
mean ꢂ standard deviation S.D (n ¼ 3), where *p < 0.05, **p < 0.01 and
***p < 0.001 significant difference to untreated control.
Molecular docking and ADMET properties prediction
In order to identify the type of interactions operating between the
synthesized compounds (ligands) and the estrogen receptors
Table 2 The IC50 values (in mM) of tested compounds against HEK-
293 cells
Entries
HEK-293
13f
13g
>100
>100
conjugates 13f and 13g showed moderate cytotoxicity against
MCF-7 cell lines. The conjugate 13f bearing a spacer length of n
¼ 3, pyrazoline ring and mono-methoxy substituted phenyl
ring, showed IC50 value 98.11 mM whereas 13g with n ¼ 3 and di-
methoxy substituted phenyl ring, exhibited an IC50 value 62.23
mM. Further, replacing the aryl core with ferrocene in naph-
thalimide–chalcones, 14a–b as well as naphthalimide–pyrazo-
lines, 15a–b could not improve activity proles. Thus, the SAR
analysis demonstrated that propyl spacer, pyrazoline ring and
electron donating substituents were requisite in order to impart
activity in the designed scaffolds. In contrast, ethyl spacer,
chalcone core and unsubstituted/nitro substituted/ferrocene
tagged conjugates failed to elicit any activity. However, the
observed anti-proliferative activities of the active compounds
against MCF-7 cells is less as compared to the reference
Fig. 4 The docked binding modes of (a) 13f and (b) 13g in the binding
site of ERa (PDB:3ERT).
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Fig. 5 The docked binding modes of (a) 13f and (b) 13g in the binding site of ERb (PDB:3LOS).
Fig. 6 The docked binding modes of 13h in the binding site of (a) ERa and (b) ERb.
(protein), the active compounds viz. 13f and 13g were chosen for interactions with Asp369 and Leu306 while the naphthalene
docking studies with ERa and ERb. Further an inactive compound ring is involved in p-alkyl and p-sigma hydrophobic interac-
13h was also included for comparison purpose.
tions with Ala 318 and Pro365 (Fig. 4b). These results demon-
strate that substituents on the phenyl ring can have drastic
The molecular docking analysis of compound 13f (IC50
¼
98.11 mM; B.E. ¼ ꢀ8.3 kcal molꢀ1), presented hydrophobic effects on the potency of these compounds as ERa antagonists.
interactions with Leu306, Ala307, Leu310, Ala318, Leu372, and
The evaluation of potent compounds in the active region of
Val368 of ERa receptor. The results also revealed that hydro- ERb showed a binding mode for 13f (B.E ¼ ꢀ7.8 kcal molꢀ1) in
phobic forces dominate the interactions between 13f and the which the triazole and pyrazoline NH units donates H-bond to
ERa receptor. Moreover, 1H-1,2,3-triazole ring forms p-alkyl the carbonyl oxygen atoms of Val370 and Asp378 respectively
ꢁ
ꢁ
ꢀ
ꢀ
interaction with Val368 while the protonated nitrogen in the (b.d ¼ 2.54 A and 2.76 A; b.a ¼ 90.5 and 110 respectively). The
triazole ring forms H-bonding interaction with Asp369. The naphthalene ring makes electrostatic contacts with Glu291 and
naphthalene ring also forms hydrophobic interaction with H-bonding interaction with Glu372. Whereas the para-methoxy
residues, Val368 and Leu373 (Fig. 4a). The introduction of a 2,5- substituent forms hydrophobic interactions with Met379 and
dimethoxy substituent in compound 13g (B.E
¼
Ala382, respectively (Fig. 5a). Further, analysis of the docked
ꢀ8.4 kcal molꢀ1) enhanced the anti-proliferative potency (IC50 complex of 13g (B.E ¼ ꢀ8.3 kcal molꢀ1) showed that the
¼ 62.23 mM) compared to 13f (IC50 ¼ 98.11 mM). Interestingly, naphthalene unit interacts with Glu291 via electrostatic contact.
this was reected in the hydrogen bonding and hydrophobic The dimethoxy substituted phenyl ring formed p-alkyl interac-
interactions of compound 13g with Leu372, Ala546, and Arg548 tion with Ala382 while the meta-methoxy unit forms a C–H bond
in the ligand-binding pocket of ERa. The meta-methoxy unit with Met379. Strong conventional H-bonding was also observed
also featured a conventional H-bonding with the amino unit of between the NH group of triazole ring and Val370, (b.d ¼ 2.11,
Leu372 at a bond distance (b.d) of 2.14 A and bond angle (b.a) of b.a ¼ 135ꢁ) (Fig. 5b).
ꢀ
138.73ꢁ. The ortho-methoxy unit on the other hand interacted
The docked binding modes of 13h in the binding site of ERa
with Arg548 and Ala546 via conventional and carbon–hydrogen and ERb have been shown in Fig. 6. The compound featured
ꢁ
ꢀ
ꢀ
ꢀ
bonding (b.d ¼ 2.35 A and 3.35 A; b.a ¼ 154.37 , respectively). three hydrogen bonds with Gly372, Glu371 and Phe289 (3.00 A,
ꢀ
ꢀ
The dimethoxy substituted phenyl ring showed p-alkyl inter- 2.78 A and 3.04 A, respectively), and hydrophobic interactions
action with Leu372. These binding modes helped to lock-in the with Leu374, Cys369, and Glu366 respectively, in the ligand-
ligand within the binding site of the ERa receptor for improved binding pocket of ERa. Compared to 13f and 13g, compound
binding affinity. The H-bonding interaction of triazole linker 13h interacts with amino acid residues in a different active site
with Asp369 is accompanied by p-cation and p-anion of ERb; it showed one hydrogen bond interaction with His377.
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The binding prole is also accompanied by hydrophobic
interactions with Tyr537, Tyr526, Val533, and Lys529, as well as
electrostatic interactions with Glu380 and Cys530, respectively.
The binding prole in the docked complex of compound 13h
with ERa and ERb partly justies the low inhibitory activity
observed (>100 mM).
Furthermore, the predicted physicochemical properties and
ADMET descriptors (Table S1, see ESI†) suggest good drug-like
properties for compounds 13f and 13g. For instance, the
compounds present good gastrointestinal absorption which
correlates with the predicted water solubility. A closer look at the
predicted metabolism interactions towards CYP (Cytochrome
P450) isoforms revealed that the compounds are suitable inhibi-
tors of cytochrome P450 and also CYP2D6/CYP3A4 substrate,
although 13g seems to be resistant to CYP1A2 inhibitor.
General procedure for the synthesis of 4
To a well stirred solution of 1,8-naphthalic anhydride, 1 (1
mmol) in absolute ethanol, ethanol/propanol amine (1.5 mmol)
was added and the reaction mixture was reuxed for 1 h. On
cooling, the reaction mixture was ltered to get solid 2. The
intermediate 2 (1 mmol) was treated with mesyl chloride (1.5
mmol) and Et3N (2 mmol) in dry CHCl3 for 3 h at room
temperature. The resulting product 3 was further reacted with
ꢁ
NaN3 in dry DMF at 60 C for 2 h to afford 4.
General procedure for the synthesis of 8/9 and 10/11
K2CO3 (2 mmol) was added to a stirred solution of 4-hydroxy
acetophenone (1 mmol) in acetone. Aer the generation of
anion, propargyl bromide (1.5 mmol) was added and the reac-
tion mixture was allowed to stir for 5 h to yield 5. The compound
5 (1 mmol) was then dissolved in alkaline ethanol and reacted
with different aldehydes 6/7 (1.2 mmol) at room temperature for
1 h to obtain 8/9. Further, hydrazine hydrate (5 mmol) and
catalytic amount of acetic acid (0.5 mmol) was added to solution
of 8/9 (1 mmol) in ethanol and the reaction mixture was reuxed
for 1 h to afford 10/11.
Conclusion
The current work disclosed the synthesis of naphthalimide-
chalcone/pyrazoline conjugates tethered by triazole ring and
their anti-proliferative effects against two breast cancer cell
lines. The anti-proliferative evaluation identied two
compounds, 13f and 13g to be explicitly targeting estrogen-
responsive breast cancer cell line MCF-7 (ER+) while the
whole series was inactive against estrogen non-responsive/
triple-negative cell line MDA-MB-231 (ERꢀ). The active
compound displayed IC50 value of 62.23 mM which is compa-
rable to the standard anti-breast cancer drug Tamoxifen. In
order to conrm their safety proles, the active compounds
were evaluated against HEK-293 cell line and proved to be non-
cytotoxic. The docking simulations as well as physiochemical
properties were studied to get an insight into the type of inter-
actions and drug likeness of the synthesized compounds. The
only limitation is its moderate activity which needs further
modication to get enhanced. Thus, the designed molecular
framework can be set as therapeutic template to particularly
General procedure for the synthesis of 12/13 and 14/15
To the solution of 4 (1 mmol) and 8/10 (1.2 mmol) in etha-
nol : water (90 : 10) catalytic amount of CuSO4 and sodium
ascorbate was added. The reaction was monitored on TLC and
on its completion, the crude product was extracted using
CHCl3 : H2O. The crude was then puried on column chroma-
tography using ethylacetate : hexane (10 : 90) as eluent to afford
nal set of compounds 12/13. The similar procedure was fol-
lowed using 4 and 9/11 in order to produce the nal hybrids 14/
15 in good yields.
Physical and spectral data
2-(2-(4-((4-Cinnamoylphenoxy)methyl)-1H-1,2,3-triazol-1-yl)
target estrogen responsive breast cancer and thus can ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione1 (12a). Yield-
contribute in the venture of developing SERMs.
86%, bright yellow solid, MP ¼ 148–150 ꢁC, 1H NMR (400 MHz,
CDCl3): d 4.68–4.70 (m, 2H, CH2), 4.78–4.80 (m, 2H, CH2), 5.28
(s, 2H, OCH2), 7.10 (d, J ¼ 8.9 Hz, 2H, ArH), 7.30–7.34 (m, 3H,
ArH), 7.39 (d, J ¼ 15.5 Hz, 1H, olenic H), 7.55 (d, J ¼ 8.6 Hz, 2H,
ArH), 7.65–7.70 (m, 3H, ArH), 7.75 (s, 1H, triazole-H), 7.95 (d, J ¼
8.8 Hz, 2H, ArH), 8.15 (dd, J ¼ 0.9, 8.3 Hz, 2H, ArH), 8.40 (dd, J ¼
Experimental
General information
Stuart Digital Melting Point apparatus (SMP10) and an open 0.8, 7.2 Hz, 2H, ArH). 13C NMR (101 MHz, CDCl3) d 188.69,
capillary was used to determine melting points and are uncor- 163.94, 162.01, 144.02, 134.40, 131.63, 131.56, 131.34, 130.76,
rected. BRUKER AVANCE II (500 and 125 MHz) and JEOL (400 130.33, 129.33, 128.91, 128.35, 128.21, 126.98, 123.40, 121.98,
and 100 MHz) NMR spectrometers were used in order to record 121.93, 114.71, 114.60, 62.12, 48.02, 39.62. HRMS (ESI) calcd for
1H and 13C NMR spectra of the synthesized compounds dis-
solved in CDCl3 (Sigma-Aldrich). Tetramethylsilane (TMS) was
C
32H24N4O4 [M + H]+ 529.1876, found 529.1895.
(E)-2-(2-(4-((4-(3-(4-Methoxyphenyl)acryloyl)phenoxy)-methyl)-
kept as reference and chemical shi values are expressed as 1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-
ꢁ
parts per million (ppm) downeld from TMS. While the dione (12b). Yield-85%, bright yellow solid, MP ¼ 154–156 C,
coupling constants represented by J values are expressed in 1H NMR (400 MHz, CDCl3): d 3.83 (s, 3H, OCH3), 4.64–4.67 (m,
hertz (Hz). Splitting patterns are indicated as s: singlet, d: 2H, CH2), 4.75–4.78 (m, 2H, CH2), 5.24 (s, 2H, OCH2), 6.91 (d, J
doublet, t: triplet, m: multiplet, dd: double doublet, dt: doublet ¼ 8.8 Hz, 2H, ArH), 7.00 (d, J ¼ 9.0 Hz, 2H, Ar H), 7.38 (d, J ¼
of a triplet and br: broad peak. Mass spectra were recorded on 15.6 Hz, 1H, olenic H), 7.57 (d, J ¼ 8.6 Hz, 2H, ArH), 7.67–7.73
a Bruker micrOTOF-QII high resolution mass spectrometer.
(m, 3H, ArH), 7.76 (s, 1H, triazole-H), 7.96 (d, J ¼ 8.9 Hz, 2H,
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ArH), 8.17 (dd, J ¼ 1.0, 8.4 Hz, 2H, ArH), 8.46 (dd, J ¼ 1.1, 7.2 Hz, ArH), 7.65–7.72 (m, 3H, ArH), 7.75 (s, 1H, triazole-H), 7.95 (d, J ¼
2H, ArH). 13C NMR (101 MHz, CDCl3) d 188.83, 164.04, 161.91, 8.8 Hz, 2H, ArH), 8.15 (dd, J ¼ 1.0, 8.5 Hz, 2H, ArH), 8.42 (dd, J ¼
161.63, 143.99, 143.79, 134.53, 131.76, 131.68, 131.67, 130.75, 1.1, 7.3 Hz, 2H, ArH). 13C NMR (101 MHz, CDCl3) d 188.95,
130.21, 128.26, 127.84, 127.08, 123.54, 122.00, 119.57, 114.69, 164.10, 161.92, 161.61, 143.98, 143.75, 134.50, 131.76, 131.69,
114.48, 62.13, 55.49, 48.10, 39.70. HRMS (ESI) calcd for 131.60, 130.85, 130.40, 128.30, 127.91, 127.12, 123.51, 122.10,
C
33H26N4O5 [M + H]+ 559.1981, found 559.1975.
(E)-2-(2-(4-((4-(3-(2,5-Dimethoxyphenyl)acryloyl)phenoxy)- (ESI) calcd for C34H28N4O5 [M + H]+ 573.2138, found 573.2152.
methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]isoquinoline- (E)-2-(3-(4-((4-(3-(2,5-Dimethoxyphenyl)acryloyl)phenoxy)-
119.51, 114.62, 114.45, 62.16, 56.01, 48.10, 39.45, 28.89. HRMS
1,3(2H)-dione (12c). Yield-85%, light brown solid, MP ¼ 146– methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-benzo[de]isoquinoline-
1
ꢁ
148 C, H NMR (400 MHz, CDCl3): d 3.80 (s, 3H, OCH3), 3.86 1,3(2H)-dione (12g). Yield-85%, bright yellow solid, MP ¼ 142–
1
ꢁ
(s, 3H, OCH3), 4.60 (t, J ¼ 6.8 Hz, 2H, CH2), 4.71 (t, J ¼ 7.0 Hz, 144 C, H NMR (400 MHz, CDCl3): d 2.37–2.43 (m, 2H, CH2),
2H, CH2), 5.25 (s, 2H, OCH2), 6.85 (d, J ¼ 8.9 Hz, 1H, ArH), 6.92 3.79 (s, 3H, OCH3), 3.84 (s, 3H, OCH3), 4.25 (t, J ¼ 6.7 Hz, 2H,
(dd, J ¼ 3.1, 9.1 Hz, 1H, ArH), 7.08 (d, J ¼ 8.6 Hz, 2H, ArH), 7.15 CH2), 4.48 (t, J ¼ 7.0 Hz, 2H, CH2), 5.24 (s, 2H, OCH2), 6.84 (d, J
(d, J ¼ 3.0 Hz, 1H, Ar H), 7.58 (d, J ¼ 15.6 Hz, 1H, olenic H), ¼ 9.0 Hz, 1H, ArH), 6.90 (dd, J ¼ 3.0, 9.0 Hz, 1H, ArH), 7.05 (d, J
7.75–7.78 (m, 2H, ArH), 7.89 (s, 1H, triazole-H), 8.01 (d, J ¼ ¼ 8.7 Hz, 2H, ArH), 7.12 (d, J ¼ 2.9 Hz, 1H, ArH), 7.56 (d, J ¼
8.6 Hz, 2H, ArH), 8.05 (d, J ¼ 15.5 Hz, 1H, olenic H), 8.21 (d, J 15.8 Hz, 1H, olenic H), 7.72–7.75 (m, 2H, ArH), 7.87 (s, 1H,
¼ 8.1 Hz, 2H, ArH), 8.58 (d, J ¼ 7.4 Hz, 2H, ArH). 13C NMR (101 triazole-H), 8.00 (d, J ¼ 8.7 Hz, 2H, ArH), 8.02 (d, J ¼ 15.8 Hz, 1H,
MHz, CDCl3) d 189.15, 164.40, 160.95, 153.45, 153.10, 143.65, olenic H), 8.20 (d, J ¼ 8.2 Hz, 2H, ArH), 8.57 (d, J ¼ 7.3 Hz, 2H,
140.40, 135.21, 132.10, 131.95, 131.51, 130.90, 128.15, 127.45, ArH). 13C NMR (101 MHz,) d 189.39, 164.43, 161.95, 153.52,
125.10, 123.40, 122.95, 122.35, 117.15, 114.51, 113.85, 112.50, 153.34, 143.44, 139.53, 134.43, 131.78, 131.67, 131.58, 130.93,
62.15, 56.20, 55.85, 48.21, 38.40. HRMS (ESI) calcd for 128.20, 127.13, 124.67, 123.33, 122.95, 122.35, 117.11, 114.67,
C
34H28N4O6 [M + H]+ 589.2087, found 589.2072.
(E)-2-(2-(4-((4-(3-(4-Nitrophenyl)acryloyl)phenoxy)methyl)- (ESI) calcd for C35H30N4O6 [M + H]+ 603.2243, found 603.2255.
113.80, 112.49, 62.16, 56.19, 55.92, 48.49, 37.57, 28.99. HRMS
1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-
(E)-2-(3-(4-((4-(3-(4-nitrophenyl)acryloyl)phenoxy)methyl)-
dione (12d). Yield-85%, yellow solid, MP ¼ 161–163 ꢁC, 1H NMR 1H-1,2,3-triazol-1-yl)propyl)-1H-benzo[de]isoquinoline-1,3(2H)-
ꢁ
(400 MHz, CDCl3): d 4.65–4.68 (m, 2H, CH2), 4.74–4.76 (m, 2H, dione (12h). Yield-85%, bright yellow solid, MP ¼ 140–142 C,
CH2), 5.25 (s, 2H, OCH2), 6.95 (d, J ¼ 8.8 Hz, 2H, ArH), 7.36 (d, J 1H NMR (400 MHz, CDCl3): d 2.36–2.42 (m, 2H, CH2), 4.26 (t, J ¼
¼ 8.7 Hz, 2H, Ar H), 7.40 (d, J ¼ 15.5 Hz, 1H, olenic H), 7.63 (d, 6.6 Hz, 2H, CH2), 4.45 (t, J ¼ 7.0 Hz, 2H, CH2), 5.25 (s, 2H,
J ¼ 8.8 Hz, 2H, ArH), 7.70–7.74 (m, 3H, ArH), 7.76 (s, 1H, OCH2), 6.94 (d, J ¼ 8.7 Hz, 2H, ArH), 7.35 (d, J ¼ 8.8 Hz, 2H,
triazole-H), 8.10 (d, J ¼ 8.7 Hz, 2H, ArH), 8.18 (dd, J ¼ 0.8, 8.1 Hz, ArH), 7.41 (d, J ¼ 15.4 Hz, 1H, olenic H), 7.64 (d, J ¼ 8.7 Hz, 2H,
2H, ArH), 8.50 (dd, J ¼ 0.7, 7.1 Hz, 2H, ArH). 13C NMR (101 MHz, ArH), 7.71–7.75 (m, 3H, ArH), 7.75 (s, 1H, triazole-H), 8.12 (d, J ¼
CDCl3) d 189.10, 164.10, 161.65, 143.92, 143.70, 141.30, 134.50, 8.8 Hz, 2H, ArH), 8.16 (dd, J ¼ 1.0, 8.0 Hz, 2H, ArH), 8.45 (dd, J ¼
131.91, 131.70, 131.60, 130.66, 130.25, 128.36, 127.95, 125.25, 1.1, 7.2 Hz, 2H, ArH). 13C NMR (101 MHz, CDCl3) d 189.15,
123.85, 123.28, 122.15, 119.20, 114.40, 62.51, 48.15, 39.61. 164.12, 161.64, 143.90, 143.68, 141.28, 134.48, 131.92, 131.75,
HRMS (ESI) calcd for C32H23N5O6 [M + H]+ 574.1726, found 131.61, 130.65, 130.21, 128.35, 127.90, 125.20, 123.81, 123.25,
574.1737.
2-(3-(4-((4-Cinnamoylphenoxy)methyl)-1H-1,2,3-triazol-1-yl)-
propyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (12e). Yield-
122.10, 119.25, 114.41, 62.50, 48.20, 39.60, 28.90. HRMS (ESI)
calcd for C33H25N5O6 [M + H]+ 588.1883, found 588.1896.
2-(2-(4-((4-(1-Acetyl-5-phenyl-4,5-dihydro-1H-pyrazol-3-yl)-
87%, off-white solid, MP ¼ 135–137 ꢁC, 1H NMR (400 MHz, phenoxy)methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]iso-
CDCl3): d 2.35–2.40 (m, 2H, CH2), 4.60–4.64 (m, 2H, CH2), 4.70– quinoline-1,3(2H)-dione (13a). Yield-84%, light brown solid, MP
4.74 (m, 2H, CH2), 5.24 (s, 2H, OCH2), 7.12 (d, J ¼ 8.8 Hz, 2H, ¼ 82–84 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.38 (s, 3H, CH3), 2.94
ArH), 7.31–7.35 (m, 3H, ArH), 7.38 (d, J ¼ 15.6 Hz, 1H, olenic (dd, J ¼ 4.5, 17.4 Hz, 1H, CH2), 3.62 (dd, J ¼ 10.9, 17.3 Hz, 1H,
H), 7.56 (d, J ¼ 8.5 Hz, 2H, ArH), 7.66–7.72 (m, 3H, ArH), 7.76 (s, CH2), 4.65–4.68 (m, 2H, CH2), 4.75–4.78 (m, 2H, CH2), 5.21 (s,
1H, triazole-H), 7.96 (d, J ¼ 8.7 Hz, 2H, ArH), 8.16 (dd, J ¼ 1.0, 2H, OCH2), 5.56 (dd, J ¼ 4.5, 10.5 Hz, 1H, CH), 7.12 (d, J ¼
8.4 Hz, 2H, ArH), 8.42 (dd, J ¼ 1.1, 7.3 Hz, 2H, ArH). 13C NMR 8.8 Hz, 2H, ArH), 7.31–7.35 (m, 3H, ArH), 7.54 (d, J ¼ 8.5 Hz, 2H,
(101 MHz, CDCl3) d 188.65, 163.95, 162.10, 144.10, 134.42, ArH), 7.65–7.69 (m, 2H, ArH), 7.75 (s, 1H, triazole-H), 7.94 (d, J ¼
131.65, 131.57, 131.35, 130.72, 130.30, 129.35, 128.90, 128.40, 8.8 Hz, 2H, ArH), 8.16 (dd, J ¼ 1.1, 8.4 Hz, 2H, ArH), 8.39 (dd, J ¼
128.21, 126.95, 123.41, 121.95, 121.90, 114.71, 114.56, 62.20, 1.0, 7.3 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.75, 164.10,
48.15, 38.90, 28.85. HRMS (ESI) calcd for C33H26N4O4 [M + H]+ 159.95, 159.10, 153.65, 134.60, 134.25, 131.70, 131.65, 128.31,
543.2032, found 543.2045.
128.20, 127.10, 126.95, 124.61, 123.45, 122.05, 115.10, 114.30,
(E)-2-(3-(4-((4-(3-(4-Methoxyphenyl)acryloyl)phenoxy)methyl)- 62.10, 55.40, 48.10, 41.75, 39.60, 22.05. HRMS (ESI) calcd for
1H-1,2,3-triazol-1-yl)propyl)-1H-benzo[de]isoquinoline-1,3(2H)-
C
34H28N6O4 [M + H]+ 585.2250, found 585.2272.
2-(2-(4-((4-(1-Acetyl-5-(4-methoxyphenyl)-4,5-dihydro-1H-pyr-
1
dione (12f). Yield-86%, off white solid, MP ¼ 151–153 ꢁC, H
NMR (400 MHz, CDCl3): d 2.37–2.43 (m, 2H, CH2), 3.80 (s, 3H, azol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo
OCH3), 4.61–4.65 (m, 2H, CH2), 4.72–4.75 (m, 2H, CH2), 5.21 (s, [de]isoquinoline-1,3(2H)-dione (13b). Yield-85%, bright yellow
2H, OCH2), 6.89 (d, J ¼ 8.7 Hz, 2H, ArH), 6.95 (d, J ¼ 8.9 Hz, 2H, solid, MP ¼ 85–87 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.37 (s, 3H,
ArH), 7.35 (d, J ¼ 15.5 Hz, 1H, olenic H), 7.51 (d, J ¼ 8.5 Hz, 2H, CH3), 3.09 (dd, J ¼ 4.4, 17.6 Hz, 1H, CH2), 3.66 (dd, J ¼ 11.7,
15842 | RSC Adv., 2020, 10, 15836–15845
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17.6 Hz, 1H, CH2), 3.74 (s, 3H, OCH3), 4.64–4.67 (m, 2H, CH2), 114.35, 62.12, 55.41, 48.15, 41.70, 39.42, 29.15, 22.09. HRMS
4.74–4.77 (m, 2H, CH2), 5.20 (s, 2H, OCH2), 5.51 (dd, J ¼ 4.4, (ESI) calcd for C35H30N6O4 [M + H]+ 599.2407, found 599.2419.
11.6 Hz, 1H, CH), 6.81 (d, J ¼ 8.7 Hz, 2H, ArH), 6.97 (d, J ¼
2-(3-(4-((4-(1-Acetyl-5-(4-methoxyphenyl)-4,5-dihydro-1H-pyr-
8.9 Hz, 2H, ArH), 7.14 (d, J ¼ 8.6 Hz, 2H, ArH), 7.64 (d, J ¼ azol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-
8.9 Hz, 2H, ArH), 7.69–7.73 (m, 2H, ArH), 7.76 (s, 1H, triazole-H), benzo[de]isoquinoline-1,3(2H)-dione (13f). Yield-85%, light
8.19 (dd, J ¼ 1.0, 8.4 Hz, 2H, ArH), 8.50 (dd, J ¼ 1.1, 7.3 Hz, 2H, brown solid, MP ¼ 82–84 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.36–
ArH). 13C NMR (101 MHz,) d 168.74, 164.06, 159.99, 159.06, 2.40 (m, 5H, CH3 + CH2), 3.10 (dd, J ¼ 4.4, 17.5 Hz, 1H, CH2),
153.68, 134.52, 134.27, 131.72, 131.68, 128.31, 128.25, 127.08, 3.65 (dd, J ¼ 11.6, 17.5 Hz, 1H, CH2), 3.75 (s, 3H, OCH3), 4.63–
126.99, 124.64, 123.45, 122.08, 115.13, 114.30, 62.13, 59.39, 4.66 (m, 2H, CH2), 4.75–4.78 (m, 2H, CH2), 5.21 (s, 2H, OCH2),
55.35, 48.05, 42.44, 39.72, 22.05. HRMS (ESI) calcd for 5.50 (dd, J ¼ 4.5, 11.3 Hz, 1H, CH), 6.82 (d, J ¼ 8.6 Hz, 2H, ArH),
C
35H30N6O5 [M + H]+ 615.2356, found 615.2368.
6.96 (d, J ¼ 8.8 Hz, 2H, ArH), 7.15 (d, J ¼ 8.5 Hz, 2H, ArH), 7.65
2-(2-(4-((4-(1-Acetyl-5-(2,5-dimethoxyphenyl)-4,5-dihydro-1H- (d, J ¼ 8.9 Hz, 2H, ArH), 7.70–7.75 (m, 2H, ArH), 7.77 (s, 1H,
pyrazol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-
triazole-H), 8.20 (dd, J ¼ 1.1, 8.5 Hz, 2H, ArH), 8.48 (dd, J ¼ 1.2,
benzo[de]isoquinoline-1,3(2H)-dione (13c). Yield-84%, yellow 7.4 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.70, 164.10, 159.90,
solid, MP ¼ 78–80 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.38 (s, 3H, 159.10, 153.65, 134.53, 134.25, 131.75, 131.65, 128.35, 128.20,
CH3), 2.94 (dd, J ¼ 4.4, 17.5 Hz, 1H, CH2), 3.60 (dd, J ¼ 11.6, 127.10, 126.99, 124.65, 123.45, 122.10, 115.20, 114.30, 62.15,
17.5 Hz, 1H, CH2), 3.69 (s, 3H, OCH3), 3.80 (s, 3H, OCH3), 4.25 (t, 59.40, 55.30, 48.12, 42.40, 39.70, 29.10, 22.04. HRMS (ESI) calcd
J ¼ 6.7 Hz, 2H, CH2), 4.47 (t, J ¼ 7.0 Hz, 2H, CH2), 5.20 (s, 2H, for C36H32N6O5 [M + H]+ 629.2512, found 629.2525.
OCH2), 5.76 (dd, J ¼ 4.5, 11.5 Hz, 1H, CH), 6.54 (d, J ¼ 3.0 Hz,
2-(3-(4-((4-(1-Acetyl-5-(2,5-dimethoxyphenyl)-4,5-dihydro-1H-
1H, ArH), 6.71 (dd, J ¼ 2.9, 8.7 Hz, 1H, ArH), 6.76 (d, J ¼ 8.7 Hz, pyrazol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-
1H, ArH), 6.98 (d, J ¼ 8.6 Hz, 2H, ArH), 7.64 (d, J ¼ 8.7 Hz, 2H, benzo[de]isoquinoline-1,3(2H)-dione (13g). Yield-88%, light
ArH), 7.74–7.78 (m, 2H, ArH), 7.85 (s, 1H, triazole-H), 8.23 (d, J ¼ brown solid, MP ¼ 76–78 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.38–
8.1 Hz, 2H, ArH), 8.60 (d, J ¼ 7.3 Hz, 2H, ArH). 13C NMR (101 2.42 (m, 5H, CH2 + CH3), 2.95 (dd, J ¼ 4.5, 17.6 Hz, 1H, CH2),
MHz,) d 168.76, 164.40, 159.85, 154.50, 153.71, 150.25, 143.60, 3.61–3.66 (m, 1H, CH2), 3.68 (s, 3H, OCH3), 3.79 (s, 3H, OCH3),
134.40, 131.70, 131.56, 130.61, 128.25, 128.20, 127.15, 124.75, 4.26 (t, J ¼ 6.6 Hz, 2H, CH2), 4.48 (t, J ¼ 7.0 Hz, 2H, CH2), 5.19 (s,
123.20, 122.35, 114.95, 112.45, 112.15, 111.85, 62.10, 56.12, 2H, OCH2), 5.77 (dd, J ¼ 4.4, 11.6 Hz, 1H, CH), 6.55 (d, J ¼
55.75, 55.40, 48.45, 41.65, 38.01, 22.05. HRMS (ESI) calcd for 2.9 Hz, 1H, ArH), 6.70 (dd, J ¼ 2.9, 8.8 Hz, 1H, ArH), 6.78 (d, J ¼
C
36H32N6O6 [M + H]+ 645.2461, found 645.2475.
2-(2-(4-((4-(1-Acetyl-5-(4-nitrophenyl)-4,5-dihydro-1H-pyr-
8.8 Hz, 1H, ArH), 6.97 (d, J ¼ 8.7 Hz, 2H, ArH), 7.63 (d, J ¼
8.7 Hz, 2H, ArH), 7.73–7.77 (m, 2H, ArH), 7.84 (s, 1H, triazole-H),
8.22 (d, J ¼ 8.1 Hz, 2H, ArH), 8.58 (d, J ¼ 7.3 Hz, 2H, ArH).13C
azol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo
[de]isoquinoline-1,3(2H)-dione (13d). Yield-86%, light brown NMR (101 MHz,) d 168.75, 164.44, 159.88, 154.55, 153.72,
solid, MP ¼ 94–96 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.40 (s, 3H, 150.23, 143.62, 134.45, 131.68, 131.59, 130.62, 128.27, 128.23,
CH3), 3.09 (dd, J ¼ 4.9, 17.0 Hz, 1H, CH2), 3.77 (dd, J ¼ 12.0, 127.14, 124.79, 123.21, 122.36, 114.98, 112.41, 112.14, 111.87,
17.6 Hz, 1H, CH2), 4.64–4.67 (m, 2H, CH2), 4.74–4.77 (m, 2H, 62.09, 56.11, 55.71, 55.42, 48.48, 41.70, 37.58, 29.02, 22.04.
CH2), 5.21 (s, 2H, OCH2), 5.61 (dd, J ¼ 4.8, 11.9 Hz, 1H, CH), 6.99 HRMS (ESI) calcd for C37H34N6O6 [M + H]+ 659.2618, found
(d, J ¼ 8.8 Hz, 2H, ArH), 7.38 (d, J ¼ 8.7 Hz, 2H, ArH), 7.64 (d, J ¼ 659.2627.
8.8 Hz, 2H, ArH), 7.71–7.75 (m, 2H, ArH), 7.77 (s, 1H, triazole-H),
2-(3-(4-((4-(1-Acetyl-5-(4-nitrophenyl)-4,5-dihydro-1H-pyr-
8.16 (d, J ¼ 8.7 Hz, 2H, ArH), 8.21 (dd, J ¼ 0.7, 8.2 Hz, 2H, ArH), azol-3-yl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-
8.51 (dd, J ¼ 0.8, 7.2 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.99, benzo[de]isoquinoline-1,3(2H)-dione (13h). Yield-85%, bright
164.10, 160.26, 153.52, 149.03, 147.40, 143.81, 134.62, 131.74, yellow solid, MP ¼ 86–88 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.35–
131.72, 128.37, 128.31, 127.12, 126.77, 124.40, 123.93, 123.51, 2.40 (m, 5H, CH2 + CH3), 3.10 (dd, J ¼ 5.2, 16.9 Hz, 1H, CH2),
122.04, 115.22, 62.09, 59.40, 48.05, 42.21, 39.74, 21.96. HRMS 3.76 (dd, J ¼ 11.9, 16.8 Hz, 1H, CH2), 4.63–4.66 (m, 2H, CH2),
(ESI) calcd for C34H27N7O6 [M + H]+ 630.2101, found 630.2115. 4.75–4.78 (m, 2H, CH2), 5.23 (s, 2H, OCH2), 5.60 (dd, J ¼ 5.1,
2-(3-(4-((4-(1-Acetyl-5-phenyl-4,5-dihydro-1H-pyrazol-3-yl)- 11.8 Hz, 1H, CH), 6.95 (d, J ¼ 8.7 Hz, 2H, ArH), 7.35 (d, J ¼
phenoxy)methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-benzo[de]iso-
8.4 Hz, 2H, ArH), 7.64 (d, J ¼ 8.8 Hz, 2H, ArH), 7.70–7.76 (m, 2H,
quinoline-1,3(2H)-dione (13e). Yield-87%, light brown solid, MP ArH), 7.76 (s, 1H, triazole-H), 8.15 (d, J ¼ 8.3 Hz, 2H, ArH), 8.20
¼ 78–80 ꢁC, 1H NMR (400 MHz, CDCl3): d 2.35–2.40 (m, 5H, CH3 (dd, J ¼ 0.8, 8.1 Hz, 2H, ArH), 8.52 (dd, J ¼ 0.9, 7.3 Hz, 2H, ArH).
+ CH2), 2.95 (dd, J ¼ 4.8, 17.9 Hz, 1H, CH2), 3.61 (dd, J ¼ 11.1, 13C NMR (101 MHz,) d 168.95, 164.20, 160.20, 153.50, 149.10,
17.8 Hz, 1H, CH2), 4.65–4.68 (m, 2H, CH2), 4.74–4.77 (m, 2H, 147.38, 143.80, 134.62, 131.75, 131.70, 128.40, 128.30, 127.20,
CH2), 5.25 (s, 2H, OCH2), 5.56 (dd, J ¼ 4.7, 11.5 Hz, 1H, CH), 7.12 126.71, 124.45, 123.90, 123.50, 122.10, 115.20, 62.10, 59.40,
(d, J ¼ 8.7 Hz, 2H, ArH), 7.32–7.36 (m, 3H, ArH), 7.55 (d, J ¼ 48.10, 42.25, 38.01, 29.10, 21.95. HRMS (ESI) calcd for
8.5 Hz, 2H, ArH), 7.65–7.68 (m, 2H, ArH), 7.76 (s, 1H, triazole-H),
7.95 (d, J ¼ 8.7 Hz, 2H, ArH), 8.15 (dd, J ¼ 0.9, 8.5 Hz, 2H, ArH),
C
35H29N7O6 [M + H]+ 644.2257, found 644.2265.
(E)-2-(2-(4-((4-(3-Ferrocenyl-acryloyl)phenoxy)methyl)-1H-
8.40 (dd, J ¼ 0.8, 7.5 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.72, 1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione
164.20, 159.90, 159.20, 153.60, 134.62, 134.26, 131.75, 131.66, (14a). Yield-85%, red solid, MP ¼ 220–222 ꢁC, 1H NMR (400
128.30, 128.21, 127.12, 126.91, 124.60, 123.41, 122.06, 115.12, MHz, CDCl3): d 4.18 (s, 5H, FeH), 4.47 (s, 2H, FeH), 4.60 (s, 2H,
FeH), 4.69 (t, J ¼ 6.5 Hz, 2H, CH2), 4.80 (t, J ¼ 7.0 Hz, 2H, CH2),
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5.26 (s, 2H, OCH2), 7.03 (d, J ¼ 8.5 Hz, 2H, ArH), 7.13 (d, J ¼ cultured in DMEM supplemented with 5% FBS and 1% peni-
15.1 Hz, 1H, olenic H), 7.72–7.76 (m, 3H, ArH + olenic H), 7.78 cillin–streptomycin, both incubated at 37 ꢁC and 5% carbon
(s, 1H, triazole-H), 7.97 (d, J ¼ 8.5 Hz, 2H, ArH), 8.22 (dd, J ¼ 0.9, dioxide.
8.1 Hz, 2H, ArH), 8.50 (dd, J ¼ 0.8, 7.3 Hz, 2H, ArH). 13C NMR
(101 MHz, CDCl3) d 188.20, 164.45, 161.72, 146.15, 143.41, MTT assay
134.45, 131.89, 131.65, 131.50, 130.65, 128.20, 127.25, 123.30,
Cells were seeded in 96 well plates at a density of 5000 cells per
well in triplicate in media. Aer 24 hours, the test compounds
diluted in complete Dulbecco's media Eagle's medium (DMEM)
were added to each well. Cells were treated with a range of
122.30, 118.80, 114.61, 79.54, 71.30, 69.92, 69.10, 62.31, 48.45,
39.12. HRMS (ESI) calcd for C36H28FeN4O4 [M + H]+ 637.1538,
found 637.1549.
(E)-2-(2-(4-((4-(3-Ferrocenyl-acryloyl)phenoxy)methyl)-1H-
different concentrations of the compounds (1, 5, 10, 20, 50, 100
1,2,3-triazol-1-yl)propyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione
mM) for 24 hours at 37 ꢁC and 5% carbon dioxide. Subsequently,
(14b). Yield-85%, brick red solid, MP ¼ 210–212 ꢁC, 1H NMR (400
sterile 5 ml of 5 mg mLꢀ1 MTT (Sigma-Aldrich) dissolved in PBS
MHz, CDCl3): d 2.37–2.44 (m, 2H, CH2), 4.15 (s, 5H, FeH), 4.26 (t, J
was added to each well and incubated with cells for 2 hours.
Solubilization solution (10% SDS, 10 mM HCl) of equal volume
to the wells was then added to each well, which was incubated
with cells for 16 hours at 37 ꢁC. The optical density of each well
was read at 570 nm using a microtiter plate reader (Thermo
Fisher Scientic Multiskan GO Microplate Reader, SkanIt™
soware).
¼ 6.7 Hz, 2H, CH2), 4.45 (t, J ¼ 1.8 Hz, 2H, FeH), 4.49 (t, J ¼
7.1 Hz, 2H, CH2), 4.56 (t, J ¼ 1.8 Hz, 2H, FeH), 5.24 (s, 2H, OCH2),
7.05 (d, J ¼ 8.8 Hz, 2H, ArH), 7.11 (d, J ¼ 15.3 Hz, 1H, olenic H),
7.69–7.77 (m, 3H, ArH + olenic H), 7.87 (s, 1H, triazole-H), 7.98
(d, J ¼ 8.8 Hz, 2H, ArH), 8.21 (dd, J ¼ 0.7, 8.2 Hz, 2H, ArH), 8.58
(dd, J ¼ 0.8, 7.2 Hz, 2H, ArH). 13C NMR (101 MHz, CDCl3)
d 188.18, 164.44, 161.78, 146.01, 143.46, 134.44, 131.87, 131.67,
131.59, 130.69, 128.21, 127.14, 123.29, 122.36, 118.85, 114.64,
79.43, 71.32, 69.85, 69.02, 62.16, 48.49, 37.57, 29.01. HRMS (ESI)
calcd for C37H30FeN4O4 [M + H]+ 651.1694, found 651.1685.
2-(2-(4-((4-(1-Acetyl-5-ferrocenyl-4,5-dihydro-1H-pyrazol-3-yl)-
phenoxy)methyl)-1H-1,2,3-triazol-1-yl)ethyl)-1H-benzo[de]iso-
quinoline-1,3(2H)-dione (15a). Yield-85%, light brown solid, MP
Statistical analysis
The statistical analysis was performed using Excel®, and IC50
values were estimated using GraphpadPrism5 soware (Hearne
Scientic Soware). The experiments were performed in
duplicate, and the statistical signicance was calculated using
the student's t-test. A p-value of less than 0.05 was used to
estimate the signicance of the observations. A Z-factor was
calculated for each 96-well plate and assays having Z-factor
above >0.5 were included in the statistical analysis.39
1
ꢁ
¼ 198–200 C, H NMR (400 MHz, CDCl3): d 2.26 (s, 3H, CH3),
3.45 (dd, J ¼ 4.8, 17.3 Hz, 1H, CH2), 3.56 (dd, J ¼ 11.6, 17.6 Hz, 1H,
CH2), 3.94–3.96 (m, 2H, CH2), 4.07–4.12 (m, 7H, FeH), 4.20–4.24
(m, 2H, CH2), 4.47 (s, 2H, FeH), 5.25 (s, 2H, OCH2), 5.45 (dd, J ¼
4.6, 11.5 Hz, 1H, CH), 7.05 (d, J ¼ 8.1 Hz, 2H, ArH), 7.73–7.78 (m,
4H, ArH), 7.86 (s, 1H, triazole-H), 8.21 (d, J ¼ 7.7 Hz, 2H, ArH), 8.56
Molecular docking
(d, J ¼ 6.8 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.82, 164.42, Molecular docking was carried out using Auto dock Vina40 and
159.95, 153.81, 143.60, 134.40, 131.75, 131.60, 128.30, 128.20, the crystal structures of human estrogen receptor alpha (ERa,
ꢀ
127.14, 124.70, 123.26, 122.35, 115.16, 87.42, 70.30, 68.69, 68.27, PDB code 3ERT, 1.9 A resolution) and estrogen receptor beta
ꢀ
62.10, 55.30, 48.45, 39.65, 37.56, 22.20. HRMS (ESI) calcd for (ERb, PDB code 3OLS, 2.2 A resolution). The compatibility
C38H32FeN6O4 [M + H]+ 693.1912, found 693.1925.
between the residue and the moderate active ligands was
2-(2-(4-((4-(1-Acetyl-5-ferrocenyl-4,5-dihydro-1H-pyrazol-3-yl)- further checked using Protein–ligand Interaction Proler so-
phenoxy)methyl)-1H-1,2,3-triazol-1-yl)propyl)-1H-benzo[de]iso-
ware. SwissADME41 and pkCSM42 web-based applications were
quinoline-1,3(2H)-dione (15b). Yield-85%, light brown solid, MP used to predict the ADMET properties of active compounds.
1
ꢁ
¼ 190–192 C, H NMR (400 MHz, CDCl3): d 2.27 (s, 3H, CH3),
2.39–2.42 (m, 2H, CH2), 3.47 (d, J ¼ 15.5 Hz, 1H, CH2), 3.59 (dd, J ¼
11.6, 16.3 Hz, 1H, CH2), 3.97 (s, 1H, CH2), 4.07–4.11 (m, 7H, FeH),
4.26 (s, 2H, CH2), 4.46–4.49 (m, 3H, FeH + CH2), 5.23 (s, 2H, OCH2),
5.43 (d, J ¼ 8.5 Hz, 1H, CH), 7.05 (d, J ¼ 8.0 Hz, 2H, ArH), 7.72–7.77
(m, 4H, ArH), 7.87 (s, 1H, triazole-H), 8.22 (d, J ¼ 7.8 Hz, 2H, ArH),
8.58 (d, J ¼ 6.8 Hz, 2H, ArH). 13C NMR (101 MHz,) d 168.81, 164.46,
159.99, 153.83, 143.61, 134.47, 131.69, 131.60, 128.25, 128.23,
127.16, 124.72, 123.28, 122.36, 115.17, 87.45, 70.35, 68.66, 68.27,
62.13, 55.29, 48.50, 39.68, 37.58, 29.03, 22.11. HRMS (ESI) calcd for
C39H34FeN6O4 [M + H]+ 707.1991, found 707.1985.
Abbreviations
ER
Estrogen receptor
SERMs
SAR
Selective estrogen receptor modulators
Structure activity relationship
Conflicts of interest
The authors declare no conict of interest.
Materials and methods
Cell culturing
Acknowledgements
MCF7 cells were cultured in DMEM media with 10% FBS, and Department of Science and Technology (DST), New Delhi, India,
1% penicillin–streptomycin and MDA-MB-231 cells were under INSPIRE programme (Shalini IF160180) has been
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