Theoretical and experimental investigations into structural, electronic, molecular and biological properties of 4-(3-chlorophenyl)-1-(3-chloropropyl) piperazin-1-ium chloride

Article abstract of DOI:10.1016/j.molstruc.2018.05.019

A convenient and facile synthesis of 4-(3-chlorophenyl)-1-(3-chloropropyl)piperazin-1-ium chloride is accomplished by stirring 1-(3-Chlorophenyl)piperazine hydrochloride with 3-chloropropanal by reductive amination in ethanol. The resulting compound was characterized using spectral data analysis augmented by X-ray. Single crystal analysis depicted that the synthesized compound crystallizes in monoclinic crystal system with P 21/c point group. The structural and electronic properties of the title compound have been calculated at DFT/B3LYP/6-311G (d,p) level of theory. Theoretically obtained parameters were well compared to the experimentally obtained results, showing excellent agreement. Molecular electrostatic potential surface, frontier orbital analysis and vibrational analysis were also carried out. HOMO-LUMO energy gap was calculated which allowed the calculation of relative properties like chemical hardness, chemical inertness, chemical potential, nucleophilicity and electrophilicity index of the synthesized products. Pass prediction was carried out which revealed that the target compound can be active against Prostate specific membrane protein, bearing Pa value of 0.411. Based on Pass, molecular docking studies of compound was carried out against Prostate specific membrane protein. The title compound depicted a binding free energy of ?6.3 kcal/mol and is seen to be involved in key bonding interactions which include both alkyl and mixed pi-alkyl hydrophobic interactions: Alkyl hydrophobic interactions: (LEU 259; 4.64 ?, LEU 261; 3.79 ?), mixed Pi/alkyl hydrophobic interactions: (HIS 552; 4.96 ?, HIS 553; 4.64, 5.31 ? LEU; 3.45 ?) respectively with Prostate specific membrane protein. Target compound was screened for their cytotoxic potential with various cancer cell lines being most effective against prostate. In short, this study reveals the synthesis of a 4-(3-Chlorophenyl)-1-(3-chloropropyl)piperazin-1-ium chloride and exposes its structural, electronic and biological properties, paving way for further research in the field of drug development.

Full text of DOI:10.1016/j.molstruc.2018.05.019

Journal of Molecular Structure 1168 (2018) 242e249
Contents lists available at ScienceDirect
Journal of Molecular Structure
journal homepage: http://www.elsevier.com/locate/molstruc
Theoretical and experimental investigations into structural, electronic,
molecular and biological properties of 4-(3-chlorophenyl)-1-(3-
chloropropyl) piperazin-1-ium chloride
, b,
Muzzaffar A. Bhat a *, Shabir H. Lone c, Ray J. Butcher d, Sanjay K. Srivastava a
a School of Studies in Chemistry, Jiwaji University, Gwalior, 474011, India
b Department of Chemistry, Islamic University of Science and Technology, Awantipora, Kashmir, 192122, India
c Govt Degree College Khanabal, Anantnag, Kashmir, 192101, India
d Department of Chemistry, Howard University, Washington, DC, 20059, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
A convenient and facile synthesis of 4-(3-chlorophenyl)-1-(3-chloropropyl)piperazin-1-ium chloride is
accomplished by stirring 1-(3-Chlorophenyl)piperazine hydrochloride with 3-chloropropanal by reduc-
tive amination in ethanol. The resulting compound was characterized using spectral data analysis
augmented by X-ray. Single crystal analysis depicted that the synthesized compound crystallizes in
monoclinic crystal system with P 21/c point group. The structural and electronic properties of the title
compound have been calculated at DFT/B3LYP/6-311G (d,p) level of theory. Theoretically obtained pa-
rameters were well compared to the experimentally obtained results, showing excellent agreement.
Molecular electrostatic potential surface, frontier orbital analysis and vibrational analysis were also
carried out. HOMO-LUMO energy gap was calculated which allowed the calculation of relative properties
like chemical hardness, chemical inertness, chemical potential, nucleophilicity and electrophilicity index
of the synthesized products. Pass prediction was carried out which revealed that the target compound
can be active against Prostate specific membrane protein, bearing Pa value of 0.411. Based on Pass,
molecular docking studies of compound was carried out against Prostate specific membrane protein. The
title compound depicted a binding free energy of ꢀ6.3 kcal/mol and is seen to be involved in key bonding
interactions which include both alkyl and mixed pi-alkyl hydrophobic interactions: Alkyl hydrophobic
interactions: (LEU 259; 4.64 Å, LEU 261; 3.79 Å), mixed Pi/alkyl hydrophobic interactions: (HIS 552; 4.96
Å, HIS 553; 4.64, 5.31 Å LEU; 3.45 Å) respectively with Prostate specific membrane protein. Target
compound was screened for their cytotoxic potential with various cancer cell lines being most effective
against prostate. In short, this study reveals the synthesis of a 4-(3-Chlorophenyl)-1-(3-chloropropyl)
piperazin-1-ium chloride and exposes its structural, electronic and biological properties, paving way for
further research in the field of drug development.
Received 23 February 2018
Received in revised form
13 April 2018
Accepted 7 May 2018
Available online 9 May 2018
Keywords:
Synthesis
X-ray
DFT
Pass
Docking
© 2018 Elsevier B.V. All rights reserved.
1. Introduction
reductive amination and nucleophilic substitution reactions. Syn-
thetic piperazines are important in biomedical applications as ion
N,N0-disubstituted piperazines have found application as li-
gands in metal complexes and form the basis of various natural
products that exhibit favorable pharmacological properties. Piper-
azine is conveniently substituted at the N and N0 positions via
channel and anticancer agents [1,2]. The substitutional flexibility of
piperazines also makes them tunable ligands. The behavior of
piperazine ligands in metal complexes is also of interest because
they can act either as chelating or bridging ligands. Although the
cyclohexane-type ring is relatively flexible, the chelate bite of the
nitrogen atoms requires adoption of a boat conformation leading to
significant ring strain. Therefore, the piperazine is at least as likely
to engage in bridging behavior between metal centers despite the
well known thermodynamic preference for chelation [3e11]. Many,
but not all, chelated piperazine complexes feature ancillary
* Corresponding author. Department of Chemistry, IUST, Awantipora Kashmir,
192122, India.
E-mail addresses: muzzaffarbhat9@gmail.com, muzzaffarbhat9@yahoo.com
(M.A. Bhat).
https://doi.org/10.1016/j.molstruc.2018.05.019
0022-2860/© 2018 Elsevier B.V. All rights reserved.

M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
243
coordination from pendant groups. A variety of bridging piperazine
complexes has been reported, including metal dimers and tetra-
mers, polymeric chains, and 2-D and 3-D frameworks [3,12e22].
Based on the above mentioned versatile properties and our
previous work on the development of pharmacologically potent
scaffolds [23e28], we explored the structural, molecular, electronic
and vibrational properties of the synthesized compound both
experimentally and theoretically which depicted a notable agree-
ment to each other. Furthermore to explore the predicted biological
properties of this compound, molecular docking studies with
prostate specific membrane protein was carried out.
Fig. 1. Single crystal structure of 4-(3-chlorophenyl)-1-(3-chloropropyl)piperazin-1-
ium chloride.
2. Experimental
Table 1
2.1. Materials and methods
Crystal data and structure refinement for the title compound.
3-chloropananal, 1-(3-chlorophenyl) piperazine hydrochloride,
sodium triacetoxy borohydride (NaBH(OAc)3), deuterated chloro-
form were procured from SigmaeAldrich (USA) and used as
received. FT-IR spectra of the compound was recorded on Agilent
FT-IR spectrometer in KBr discs (4000e400 cmꢀ1). 1H and 13C NMR
spectra were recorded on a Bruker Spectrospin DPX-400 NMR
spectrometer at 400.13 and 100.47 respectively using TMS as an
internal standard. The diffraction data on single crystals of title
compound was collected on a Bruker AXS SMART Apex CCD
Identification code
shelx
Empirical formula
Formula weight
Temperature
Wavelength
Crystal system
Space group
C13H19Cl3N2
309.65
100(2) K
0.71073 Å
Monoclinic
P 21/c
a ¼ 10.9608(9) Å
b ¼ 9.5199(8) Å
c ¼ 14.0262(11) Å
1457.1(2) Å3
4
Unit cell dimensions
a
b
g
¼ 90ꢂ
¼ 95.3980(10)ꢂ
¼ 90ꢂ
Volume
Z
diffractometer using MoeKa (0.71073 Å) radiations at 100 K. The
software SADABS [29] was used for absorption correction and
SHELXTL for space group, structure determination and refinements
[30,31]. All non-hydrogen atoms were refined anisotropically. All
the computations are carried out using GAUSSIAN 09 software [32].
The DFT modeling method, using the hybrid B3LYP [33] functional
was used to calculate theoretical parameters for title compound
with the basis set combination 6e311 G (d,p) [34]. Geometry opti-
mization was carried out until global minima were achieved. The
SRB cytotoxic assay was used to screen the compounds for cell
cytotoxicity [35,36]. Various human cancer cell lines, human leu-
kemia (THP-1, at a density of 7 ꢁ 103), human lung carcinoma cell
Density (calculated)
Absorption coefficient
F(000)
Crystal size
Theta range for data
collection
1.412 Mg/m3
0.613 mm-1
648
0.550 ꢁ 0.320 ꢁ 0.300 mm3
2.589e32.106ꢂ.
Index ranges
ꢀ16 ꢃ h<¼16, ꢀ14 ꢃ k<¼14, ꢀ20
ꢃ l<¼20
Reflections collected
32810
Independent reflections 4808 [R(int) ¼ 0.0247]
Completeness to
theta ¼ 25.500ꢂ
99.9%
Absorption correction
Refinement method
Data/restraints/
None
line (A-549, at a density of 8 ꢁ 103 cells per mL per 100
mL per well),
Full-matrix least-squares on F2
4808/0/167
human prostate cancer cell line (PC-3, at a density of 8 ꢁ 103 cells
parameters
Goodness-of-fit on F2
Final R indices
per mL per 100 mL per well) and human colon cancer cell line (HCT-
1.075
116, at a density of 1 ꢁ 104 cells per mL per 100
mL per well) used in
this study were purchased from European collection of cell culture
R1 ¼ 0.0296, wR2 ¼ 0.0811
[I > 2sigma(I)]
R indices (all data)
Extinction coefficient
R1 ¼ 0.0328, wR2 ¼ 0.0833
(ECACC) USA) and seeded in flat-bottomed 96-well plates.
n/a
Largest diff. peak and hole 0.688 and ꢀ0.326 e.Å-3
2.2. Synthesis of 4-(3-chlorophenyl)-1-(3-chloropropyl) piperazin-
1-ium chloride
To a stirring solution of 1-(3-chlorophenyl)piperazine hydro-
chloride in dichloro methane, 3-chloropropanal in the same solvent
was added drop wise under dry nitrogen and the resulting solution
stirred for 0.25 h. Thereafter sodium triacetoxy borohydride (NaB-
H(OAc)3) was added in pinches and the resulting solution was
stirred overnight. Cream coloured solid precipitate obtained was
further purified and recrystallized using ethanol. Cylindrical crys-
tals were obtained after allowing the solvent to evaporate slowly at
room temperature.
conformation [NeCeCeC ¼ ꢀ177.25 (8)ꢂ and CeCeCeCl ¼ 174.23
(7)ꢂ]. In the crystal, charge-assisted NeH/Cl hydrogen bonds link
the cation and anion into ion pairs. Numerous weak CeH/Cl in-
teractions link the ion pairs into a three-dimensional network.
Short Cl/Cl contacts [3.2419 (4) Å] are also observed (see: Scheme
1)
Theoretical studies using Density Functional Theory (DFT)
involving the well-known Becke three-parameter Lee-Yang-Parr
function (B3LYP) and 6-311G (d, p) basis set for the synthesized
compound was carried out Geometric optimization of the title
compound was carried out. Theoretically determined bond lengths
and bond angles were well compared with those determined
experimentally (X-ray) and depicted an excellent agreement
(Table 2). Theoretical calculations allowed the estimation of MEPS,
FMO's, HUMO-LUMO energy gap and related parameters which
depicted the potential kinetic stability and reactivity of the target
compound.
3. Results and discussion
The molecule crystallizes in monoclinic crystal systems with P
21/c point group (Fig. 1). The crystal refinement of the molecule is
shown in Table 1. The piperazine ring adopts a chair conformation
with the exocyclic NeC bonds in equatorial orientations. The
dihedral angle between the piperazine ring (all atoms) and the
benzene ring is 28.47 (5)ꢂ. The chloropropyl group has an extended

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M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
Fig. 2. MEP surface of target compound determined by DFT/B3LYP and 6-311G (d, p).
Scheme 1. Synthesis of 4-(3-chlorophenyl)-1-(3-chloropropyl)piperazin-1-ium
chloride.
3.1. Molecular electrostatic potential surface (MEPS) analysis
Molecular electrostatic potential surface is used to calculate the
molecular reactivity towards charged reactants and depict the
hydrogen bond interactions. These surfaces help to understand the
size shape and variation of electron density while correlating it
with dipole moment, partial charges, electronegativity, and
chemical reactivity sites located in the molecule. Computational
organic chemists use MEP surfaces for analyzing drug-receptor and
enzyme-substrate interactions along with H-bonding interactions
[37e41]. Fig. 2 shows the MEP surface of the target compound that
was calculated using DFT/B3LYP/6311G (d, p) basis set. The pictorial
representation with rainbow colour scheme of electrostatic po-
tential for the target compound lies in the range of ꢀ7.415e-2
to þ7.415e-2. MEP surface for target compound in the region sur-
rounding the chlorine atoms seems to be darkest red that is one
with highest electron density.
3.2. Frontier molecular analysis
Frontier molecular orbitals (FMO) (Fig. 3) determines the elec-
tric optical properties, electronic transitions and kinetic stability
Fig. 3. HOMO-LUMO surfaces of the title compound simulated using DFT/BL3YP/
6e311 G (d, p) level of theory.
Table 2
Comparison of theoretical and experimentally obtained bond lengths, bond angles and torsional angles for 4-(3-chlorophenyl)-1-(3-chloropropyl)
piperazin-1-ium chloride.
Bond length X-Ray DFT
Bond angle X-ray DFT
Torsional angles X-Ray DFT
N(1)eC(1) 1.40 1.47
N(2)eC(11) 1.50 1.47
C(3)eCl(1) 1.74 1.76
N(1)eC(7) 1.47 1.46
N(1)eC(10) 1.47 1.46
C(1)eC(2) 1.40 1.54
C(2)eC(3) 1.38 1.35
C(7)eC(8) 1.52 1.54
C(9)eC(10) 1.51 1.54
C(11)eC(12) 1.51 1.54
C(13)eCl(2) 1.76 1.79
C(4)-H(4A) 0.99 1.07
C(9)eH(9) 0.99 1.07
C(5)eH(5) 0.95 1.07
C(12)eH(12) 0.99 1.07
C(1)-N(1)-C(7) 119.02 109.47
C(1)-N(1)-C(10) 117.42 109.47
N(1)-C(1)-C(2) 118.65 120
N(1)-C(1)-C(6) 122.71 120
C(1)-C(2)-C(3) 119.62 120
C(2)-C(3)-Cl(1) 118.44 120
C(8)-N(2)-C(9) 110.04 109.47
C(8)-N(2)-C(11) 112.66 109.47
N(2)-C(11)-C(12) 118.0109.47
C(11)-C(12)-C(13)107.62 109.47
C(12)-C(13)-Cl(2) 110.47 109.47
N(2)-C(9)-H(9) 109.5109.47
N(1)-C(7)-H(7) 109.8109.47
C(1)-C(6)-H(6) 119.9120
N(1)-C(1)-C(2)-C(3) 157.5179.9
N(1)-C(1)-C(6)-C(5) 111.74 179.9
N(2)-C(9)-C(10)-N(1) ꢀ69.8 -60
N(2)-C(8)-C(7)-N(1) 0.4e0.6
N(2)-C(11)-C(12)-C(13) 178.8180
C(4)-C(3)-Cl(1) 118.77 120

M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
245
Table 3
like softness, hardness (h), electronegativity, chemical potential (m),
Calculated energy values for target compound using B3LYP/6-311G (d, p)
basis set.
electron-affinity and ionization energy for the synthesized com-
pound was carried out. These properties are calculated as follows
[43e45]:
Parameter
Target compound
Energy (au)
Dipole moment (Debye)
EHOMO (eV)
ELUMO (eV)
EHOMO-LUMO (eV)
EHOMO-1 (eV)
ꢀ1996.72
8.1447
ðI ꢀ AÞ
ꢀðI þ AÞ
ðI þ AÞ
ꢀ0.21635
ꢀ0.02277
0.19358
ꢀ0.22068
ꢀ0.01524
0.20544
0.09678
ꢀ0.11956
0.11956
0.07385
h
¼
;
m
¼
;
c
¼
2
2
2
where I and A represent ionization potential and electron affinity of
the compound, which are actually obtained from HOMO and LUMO
energies as I ¼ -EHOMO and A ¼ -ELUMO as per Janak theorem [46] and
Perdew et al. [47]. A large HOMO-LUMO gap represents a hard
molecule while as a small gap indicates a soft or more reactive/less
stable molecule. As can be seen the HOMO-LUMO energy gap in the
E
E
LUMOþ1 (eV)
(HOMO-1)-(LUMOþ1) (eV)
Hardness (
Chemical Potential (
Electronegativity (
Electrophilicity index (
h)
m
)
)
c
u
)
target compound is 0.19358 eV. Similarly chemical potential ‘m’ that
measures the escaping ability of electrons from an equilibrium
system for target compound is ꢀ0.11956 eV. Similarly the global
[42]. FMO of target compound was calculated using DFT/BL3YP/6-
311G (d, p) level of theory. As can be seen from Fig. 3 HOMO are
located over the whole skeleton of the molecule except the chlor-
opropyl group whereas LUMO are fully spread over chloropropyl
ring and partially over piperazine ring and terminal CeCl atoms.
The HOMO-LUMO energy gap is 0.19358 eV for the title compound.
(Table 3). Using this HOMO-LUMO energy gap chemical descriptors
electrophilicity index (u), a global reactivity index that is related to
chemical hardness and chemical potential as introduced by Parr
et al. [45] represents the measure of the stabilization in energy
achieved when the system acquires an additional electronic charge
from the environment and is given by
sponding value for target compound is 0.07385 eV. All these
u
¼
m2=2
h. The corre-
Fig. 4. IR of 1 [a) Theoretical, b) Experimental].

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M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
Fig. 5. 1H NMR of target compound [a) Theoretical, b) Experimental].
parameters have been calculated for the target compound using
B3LYP/6-311G (d, p) basis set and are depicted in Table 3.
3.4. Nuclear magnetic resonance (NMR) analysis
In the 1H NMR spectra of target compound, two triplets
appearing at 2.95e2.98 and 2.46e2.50 ppm are assigned to CleCH2
and CH2eN protons respectively, while multiplet at 1.88e1.95 ppm
is due to eCH2e. protons. Piperazine ring protons appear as two
triplets at 2.52e2.55 and 3.28e3.46 ppm. Theoretically calculated
1H NMR showed a very good correlation with the experimental one
and displayed CleCH2 and CH2eN protons at 2.55e2.9 and
1.8e1.9 ppm respectively, while as eCH2e protons around 2.1 ppm.
Piperazine ring protons appear at 1.95e2.05 and 3.05e3.10 ppm.
Aromatic protons (AreH) are as expected and are positioned at
6.5e7.5 ppm as two doublets, singlet and triplet. However aromatic
protons displayed a greater shift as compared to calculated data
and appeared around 5.6e6.4 ppm (Fig. 5).
3.3. Fourier-transform infrared (FTIR) analysis
Theoretical IR of target compound depicted a total of 105 normal
modes of vibration. However in the desired region between 400
and 4000 cmꢀ1 only 86 modes were observed and this region was
compared to the experimentally obtained values. The theoretical
and experimental IR spectra are presented in Fig. 4. Although a
good agreement between the experimental and theoretical IR is
observed, yet the slight variation observed is attributed to the fact
that theoretically the calculations were done in gas phase while as
experimentally the IR was carried out in solid KBr pellets. Experi-
mental FT-IR of target compound displayed peaks in the region
3120e3300, 2915e2808, 1612e1008 and 912e500 with prominent
wave numbers being 3120, 3049, 3008, 2981, 2929, 2866, 2808,
1614, 1573, 1492, 1477, 1446, 1392, 1389, 1328, 1315, 1282, 1180,
1062, 1035, 1008, 966, 912, 891, 862, 779, 767, 651, 607, 520. Bands
in the region 3120-2808 cmꢀ1 were assignable to CeH stretching's
of aromatic ring and CH3 stretching's of the methyl groups, a broad
band between 2800 and 2250 cmꢀ1 is the positive indication of
quaternary piperazinium salt and bands observed between 1622
and 1008 were assigned to C]C stretching vibrations in aromatic
ring and bending HeC]C modes in aromatic ring. Further bands in
the region between 1000 and 500 cmꢀ1 were assigned to the dis-
tortions in torsion angles.
In the 13C{1H} NMR spectrum of target compound, signals at
58.13, 47.55 and 25.83 ppm are assigned to CleCH2, CH2eN and
eCH2e carbons respectively. Piperazine ring carbons absorb at
53.19 and 48.82 ppm. In the theoretically calculated 13C NMR
spectra, CleCH2, CH2eN and eCH2e carbon signals appear at 45, 42
and 18 ppm respectively, while as piperazine carbons appear at 42
and 37 ppm.
Aromatic carbons absorb at 152.54, 114.01, 130.21, 115.90, 135.15,
119.40 ppm in the experimental 13C NMR spectra, while as in
theoretical spectra aromatic carbons are less deshielded and appear
at 136, 99, 117, 103, 135, 102 ppm (Fig. 6).

M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
247
Fig. 6. 13C NMR of target compound [a) Theoretical, b) Experimental].
3.5. Drug likeness and molecular docking
generally Pa þ Pi s 1 as these probabilities are calculated inde-
pendently [49]. PASS analysis results for the target compound have
been enlisted in Table 4. A thorough analysis of PASS results shows
that the target compound is likely to be antineoplastic (Multiple
myeloma) with corresponding Pa value of 0.411. To know the exact
possible mechanistic pathway as how the target compound acts we
carried out the cytotoxic evaluation and docking studies of the
target molecule against the predicted target Prostate specific
membrane protein.
PASS (Prediction of activity spectra) is an easily available online
tool to predict the possible pharmacological properties of target
compound in relation to its structure [48]. PASS assesses the drug
likeness and toxicities of molecules like teratogenicity, carcinoge-
nicity embroyogenecity etc. The average accuracy of prediction is
about 95% according to leave-one-out-cross validation (LOOCV)
estimation and the probabilities, Pa (Probable activity) and Pi
(Probable inactivity), are values that vary from 0.000 to 1.000, and
X-ray crystal structure of Prostate-specific membrane protein
Table 4
PASS predicts results of target compound with Pa > 0.411.
Pa
Pi
Activity
Pa
Pi
Activity
0.843
0.824
0.731
0.692
0.675
0.670
0.668
0.640
0.638
0.620
0.652
0.626
0.639
0.664
0.598
0.588
0.007
0.025
0.032
0.004
0.010
0.010
0.012
0.020
0.020
0.005
0.037
0.016
0.034
0.078
0.015
0.008
Glycosylphosphatidylinositol phospholipase D inhibitor
Phobic disorders treatment
Antineurotic
Monoamine uptake inhibitor
CYP2D6 substrate
CYP2D substrate
CYP2A6 substrate
CYP2B6 substrate
27-Hydroxycholesterol 7alpha-monooxygenase inhibitor
Leukopoiesis inhibitor
Antiischemic. Cerebral
CYP2A substrate
CYP3A4 substrate
Gluconate 2-dehydrogenase (acceptor) inhibitor
CYP1A2 substrate
0.597
0.589
0.579
0.610
0.579
0.575
0.597
0.561
0.588
0.546
0.566
0.511
0.524
0.517
0.486
0.411
0.022
0.014
0.007
0.038
0.019
0.015
0.040
0.014
0.051
0.015
0.037
0.005
0.029
0.035
0.005
0.021
CYP2C9 substrate
CYP2C19 substrate
CYP2C18 substrate
Anaphylatoxin receptor antagonist
X-methyl-His dipeptidase inhibitor
Sigma receptor agonist
CYP3A substrate
CYP2E1 substrate
5-O-(4-coumaroyl)-D-quinate 30-monooxygenase inhibitor
CYP2E substrate
Acetylcholine neuromuscular blocking agent
5 Hydroxytryptamine 2B agonist
CYP2A8 substrate
CYP2C8 substrate
5 Hydroxytryptamine 1A antagonist
Antineoplastic (multiple myeloma)
Chemosensitizer

248
M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
Table 6
(PDB-ID 1Z8L) was obtained from the Protein Data Bank web site
(http://www.rrscb.org/pdb). Water molecules and heteroatoms
were removed. The preparation of Target, Ligand, Grid and Docking
Parameter files for the target compound was done employing
AutoDock 4.2 using the Lamarkian Genetic Algorithm [50]. The free
IC50 values of the target compound (mM).
Compound
Leukemia
(THP-1)
Colon
(HCT-
116)
Prostate
(PC-3)
Lung
(A-
549)
Target
compound
17
19
16
18
energy of binding (
(Fig. 7.) calculated using AutoDock 4.2 are listed in Table 5. As can be
seen the best free energy of binding ( Gb) of target compound was
DGb and along with the binding interactions
D
found to be ꢀ6.3 kcal/mol against prostate-specific membrane
protein. It is seen that various types of bonding interactions be-
tween the amino acid residues of prostate-specific membrane
protein and target compound take place (Table 5). Which include
both alkyl and mixed pi alkyl hydrophobic interactions: Alkyl hy-
drophobic interactions: (LEU 259; 4.64 Å, LEU 261; 3.79 Å). Mixed
Pi/alkyl hydrophobic interactions: (HIS 552; 4.96 Å, HIS 553; 4.64,
5.31 Å LEU; 3.45 Å) respectively.
3.6. SRB cytotoxicity assay
The synthesized target compound was screened for cytotoxic
potential using Sulforhodamine B (SRB) cytotoxicity assay against
leukemia (THP-1), colon (HCT-116), prostate (PC-3) and human
lung (A-549) cancer cell lines. Target compound displayed a
different cytotoxic potential towards the cancer cell lines, with the
leukemia and prostate cell being most sensitive (Table 6). Target
compound displayed excellent cytotoxicity towards leukemia (PC-
3, IC50 of 16 mm).
4. Conclusion
Synthesis of 4-(3-chlorophenyl)-1-(3-chloropropyl)piperazin-
1-ium chloride is accomplished by stirring of 1-(3-chlorophenyl)
piperazine hydrochloride with 3-chloropropanal by reductive
amination in dichloro methane. Single crystal analysis depicted
that the synthesized compound crystallizes in monoclinic crystal
system with P 21/c point group. The structural and electronic
properties of the title compound was calculated using DFT/B3LYP/
6-311G (d.p) level of theory. Theoretically obtained parameters
depicted excellent agreement to the experimentally obtained re-
sults. HOMO-LUMO energy gap was calculated which allowed the
calculation of relative properties like chemical hardness. chemical
inertness. Chemical potential. nucleophilicity and electrophillicity
index of the synthesized product. Pass prediction was carried which
revealed that target compound can be highly active against
Prostate-specific membrane protein bearing Pa value of 0.411. To
validate the pass results and to explore the protein-compound in-
teractions molecular docking studies of target compound was car-
ried out against Prostate-specific membrane protein. Target
compound depicted a binding free energy of ꢀ6.3 kcal/mol. Target
compound displayed various types of interactions which include
both alkyl and mixed pi alkyl hydrophobic interactions: Alkyl hy-
drophobic interactions: (LEU 259; 4.64 Å, LEU 261; 3.79 Å). Mixed
Pi/alkyl hydrophobic interactions: (HIS 552; 4.96 Å, HIS 553; 4.64,
5.31 Å LEU; 3.45Å) respectively with Prostate-specific membrane
protein. Target compound was screened for their cytotoxic poten-
tial with various cancer cell lines being most effective against
prostate.
Fig. 7. Interactions of target compound with amino acids of Prostate-specific mem-
brane protein.
Conflicts of interest
Table 5
Approximate mean Binding free energy and bonding interactions of target
compound.
The authors declare no conflict of interest.
The authors declare no competing financial interests.
Compound
No of
Rotational
bonds
Binding energy
Gb)Kcal/Mol
Bonding
Interactions
(
D
Acknowledgment
The authors are very thankful for 1H and 13C NMR studies to IIIM
Srinagar. RJB wishes to acknowledge NSF award 1205608. Part-
nership for Reduced Dimensional Materials for partial funding of
this research.
Target
compound
5
ꢀ6.3
Alkyl hydrophobic
interactions: (LEU 259;
4.64 Å, LEU 261; 3.79
Å).
Mixed Pi/alkyl
hydrophobic
interactions: (HIS 552;
4.96 Å, HIS 553; 4.64,
5.31 Å LEU; 3.45 Å)
Appendix A. Supplementary data
Supplementary data related to this article can be found at

M.A. Bhat et al. / Journal of Molecular Structure 1168 (2018) 242e249
249
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