Benzyl and sulfonyl derivatives of n-(2,6-dimethylphenyl)-2(piperazin-1-yl)acetamide (t2288): biological screening and fingerprint applications

Article abstract of DOI:10.13005/ojc/350117

A series of five N-(2,6-dimethylphenyl)-2-(piperazin-1-yl)acetamide (T2288) sulfonamides 6a-e and its five alkylated piperazine derivatives 8a-e have been synthesized, characterized and screened for antibacterial, antifungal and anthelmintic activity. Some of the compounds showed significant biological activities. Molecular docking to crystal structures of target proteins revealed that, active compounds show similar binding poses as that of standards, indicating good correlation of the binding energy with observed in vitro data for the active compounds. Finally, the study of latent fingerprint analysis showed that the compound 6c exhibits good stickiness and finger rhythm without dense dust. The resulting compound can be used to detect fingerprints on all types of flat surfaces and hence easily accepted for detecting hidden fingerprints. This research can offer an excellent setting that can lead to the discovery of potential antibacterial, antifungal, anthelmintic and fingerprint agents.

Full text of DOI:10.13005/ojc/350117

ISSN: 0970-020 X
CODEN: OJCHEG
2019, Vol. 35, No.(1):
Pg. 157-166
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Access, Peer Reviewed Research Journal
www.orientjchem.org
Benzyl and Sulfonyl Derivatives of N-(2,6-dimethylphenyl)-2-
(piperazin-1-yl)acetamide (T2288): Biological Screening and
Fingerprint applications
GHOUSE KHAN1,2, SWAMY SREENIVASA1*, SHIVARAJA GOVINDAIAH1
and VIVEK CHANDRAMOHAN3
1Department of Studies and Research in Organic Chemistry, Tumkur University, Tumkur-572103,
Karnataka, India.
2IDSG Government First Grade College, Chikkmagaluru-577101, Karnataka, India.
3Department of Biotechnology, Siddaganga Institute of Technology, Tumkur-572103,
Karnataka, India.
*Corresponding author E-mail: drsreenivasa@yahoo.co.in
http://dx.doi.org/10.13005/ojc/350117
(Received: September 22, 2018; Accepted: December 29, 2018)
ABSTRACT
A series of five N-(2,6-dimethylphenyl)-2-(piperazin-1-yl)acetamide (T2288) sulfonamides 6a-e
and its five alkylated piperazine derivatives 8a-e have been synthesized, characterized and screened for
antibacterial, antifungal and anthelmintic activity.Some of the compounds showed significant biological
activities. Molecular docking to crystal structures of target proteins revealed that, active compounds
show similar binding poses as that of standards, indicating good correlation of the binding energy with
observed in vitro data for the active compounds.Finally, the study of latent fingerprint analysis showed
that the compound 6c exhibits good stickiness and finger rhythm without dense dust. The resulting
compound can be used to detect fingerprints on all types of flat surfaces and hence easily accepted for
detecting hidden fingerprints.This research can offer an excellent setting that can lead to the discovery
of potential antibacterial, antifungal, anthelmintic and fingerprint agents.
Keywords: T2288, Sulphonamide, Molecular Docking, Piperazine, Finger Print, Biological screening.
effectiveness on global surveillance of antimicrobial
resistance1. Infections caused by most of the
microorganisms including Staphylococcus aureus,
Escherichia coli, Proteus vulgaris, Pseudomonas
aeruginosa, Salmonella typhi, Salmonella paratyphi
and some fungi acquiring resistant day by day to the
presently available antimicrobial agents and are the
INTRODUCTION
In recent years, the situation of antimicrobial
resistance to a number of available antimicrobial
agents is increasing drastically. According to World
Health Organization report, from different parts of the
world, indicates an overall decrease in the antibiotic
This is an
Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC- BY).
Published by Oriental Scientific Publishing Company © 2018

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most prevalent of fatal infectious diseases2–4. The
ExpERIMENTAL
problem is still very severe in case of Mycobacterium
tuberculosis and hence needs a new potent
antimicrobial drug with new mechanism of action5.
In this context, chemical modification of approved
drug molecules is a well-known approach affording
the new chemical entities (NCE) with enhanced
therapeutic characteristics. In this view, Piperazine
derivatives emerged as good candidates as they
showed broad spectrum of pharmacological activities
like antidepressant6, anticancer7, antibacterial8,
anthelmintic9, antimycobacterial10, antifungal11,
anticonvulsant12 and antimalarial13. The presence
of these heterocycles can be viewed in several well-
known drugs, like fluphenazine as antipsychotic14,15
cyclizine as antihistamine16, trimetazidine as
antianginal17.
,
Scheme 1. Synthesis of Target compounds. Reagents and
conditions: (a) 2-chloroacetyl chloride, NaOH, DCM, 0oC,
3 h; (b) N-Boc piperazine, TEA, CH3CN, 25oC, 4 h; (c) TFA,
DCM, 0oC, 3 h; (d) RSO2Cl 5a-e, DCM, TEA, 25oC, 8 h; (e)
R1CH2Cl 7a-e, K2CO3, MeCN, rt, 10 hours
Many scientific reports reveals that
sulfonyl group when linked to piperazine derivatives
possess wide pharmacological activities like
MATERIALS AND METHODS
anti-inflammatory18, antidiabetic19, anticancer20-23
antibacterial24-25, antiprotozoal26, antifungal27-28
,
,
Melting points reported were determined by
open capillary method. Structures were confirmed
using infrared spectroscopy, nuclear magnetic
spectroscopy like proton and carbon-13 finally by
LC-MS data. FT-IR Spectra was recorded in Jasco
FT-IR spectrometer, 1H and 13C NMR spectra were
recorded at 399.65 MHz and 100.40 MHz using
CDCl3.The chemical shift was reported in ppm.The
LC-MS were recorded using Waters Alliance 2795
separation module andWaters Micromass LCT mass
detector. All chemicals were purchased from Merck
India, Spectrochem and Sigma-Aldrich.Solvent and
chemicals used are of AR grade.All bacterial strains
were purchased from CSIR - National Chemical
Laboratory (NCL), Pune, India.
nonpeptidic vasopressin receptor antagonists29
and translation initiation inhibitors30-31. In addition,
many acetamide derivatives have been reported to
exhibit various biological activities such as anthelmintic,
anticonvulsant, antioxidant, anti-inflammatory,
anti-arthritic, anticancer, anti-bacterial and
antifungal32-36. Some acetamide derivatives were
found to be used in the development of enzymes
that prevent the replication of HIV virus37,38
.
Certain acetamide compounds potentially inhibit
the enzyme farnesyltransferase, a major strategy
for the development of novel potential anticancer
drugs39. Some Arylacetamides reported as
antimicrobial agents such as herbicides, antifungal
and disinfectants40-41.A number of thioacetamide and
sulfonyl acetamide derivatives of glycosyl were found
to possess antitubercular activity42-44. These three
active pharmacophores viz, piperazine, sulfonyl
and acetamide groups, if they combined together
would produce new molecular templates hope to
exhibit enhanced biological activities.So we inspired
and synthesized hybrid derivatives and test their
biological activities.
Synthesis of N-(2,6-dimethylphenyl)-2-(piperazin-
1-yl)acetamide (4)
Intermediate 4 was synthesized as per the
reported literature54. m.p 118oC and yield 72%.
SpectralcharacterizationofN-(2,6-dimethylphenyl)-
2-(piperazin-1-yl)acetamide (4)
1H NMR (400 MHz, CDCl3, TMS): δ=8.64
(s, 1H, NH), 7.02-7.03 (m, 3H, Ar-H), 3.08 (s, 2H,
CH2), 2.84-2.87 (t, 4H, CH2), 2.56-2.57 (t, 4H, CH2),
2.16 (s, 6H, CH3);13C NMR (100 MHz, CDCl3, TMS):
δ=168.39, 134.89, 133.73, 128.75, 126.99, 76.92,
62.38, 46.22, 18.55. Mass: m/z 248.1.

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General procedure for the synthesis of
compounds 6a-e
133.29, 129.12, 128.29, 127.405, 61.22, 52.77,
45.95, 18.56. IR (KBr): δ = 1665.8 (CO), 1321.4
(S = O asymmetric), 1169.3 (S = O symmetric).
LCMS m/z: cal. 421.12; found 422.21(M+1).
An equimolar mixture of N-(2,6-dimethy
lphenyl)-2-(piperazin-1-yl)acetamide 4 (0.001 mol)
and different substituted benzene sulfonyl chlorides
5a-e (0.001 mol) in methylene dichloride were stirred
for about 8 h in presence of triethylamine at 25oC.
The completion of the reaction was checked byTLC.
Then it was poured into ice-cold water and extracted
with MDC. The organic layer was washed with brine
solution, dried over anhydrous sodium sulphate and
concentratedunderreducedpressuretogetpiperazine
sulphonamide derivatives 6a-e in high yield.
N-(2,6-dimethylphenyl)-2-(4-((4-nitrophenyl)
sulfonyl)piperazin-1-yl)acetamide 6c
Colour: white solid,Yield: 89, m.p:176-177
oC. 1H NMR (400 MHz, CDCl3, TMS): δ = 8.38-8.36
(d, J = 8Hz, 2H; Ar-H), 8.21 (s, 1H, NH), 7.95-7.92
(d, J = 12Hz, 2H, Ar-H), 7.07-7.02 (m, 3H, Ar-H), 3.22
(s, 2H, CH2), 3.16 (s, 4H, CH2), 2.79 (s, 4H, CH2),
2.12 (s, 6H, CH3).13C NMR (100 MHz, CDCl3, TMS):
δ = 167.35, 150.36, 141.7, 134.14, 133.18, 128.87,
127.47, 124.45, 61.19, 52.75, 45.95, 18.53.IR (KBr):
δ = 1659.4 (CO), 1312.1(S = O asymmetric), 1163.5
(S = O symmetric). LCMS m/z: cal. 432.15; found
433.24 (M+1).
General procedure for the synthesis of
compounds 8a-e
To a solution of N-(2,6-dimethylphenyl)-2-
(piperazin-1-yl)acetamide 4 in acetonitrile, different
substituted benzyl chlorides 7a-e (0.001 mol) were
slowly added drop wise in presence of K2CO3 (0.003
mol). Reaction mixture was stirred for 10 h at 25oC
and completion of reaction was checked by TLC.
Reaction mixture was poured into ice-cold water
and extracted with methylene dichloride.The organic
layers was washed with brine solution, dried over
anhydrous sodium sulphate and concentrated to get
alkylated piperazine derivatives 8a-e in good yield.
2-(4-((4-bromophenyl)sulfonyl)piperazin-1-yl)-N-
(2,6-dimethylphenyl) acetamide 6d
Colour:white solid,Yield:90, m.p:184-185
oC. 1H NMR (400 MHz, CDCl3, TMS):δ = 8.27 (s, 1H,
NH), 7.65-7.63 (d, J = 8Hz, 2H, Ar-H), 7.57-7.55
(d, J = 8Hz, 2H, Ar-H), 7.05-7.00 (m, 3H, Ar-H),
3.15 (s, 2H, CH2), 3.04 (s, 4H, CH2), 2.71-2.69
(t, J = 8Hz, 4H, CH2), 2.11 (s, 6H, CH3). 13C NMR
(100 MHz, CDCl3,TMS):δ = 167.73, 135.05, 134.55,
133.37, 132.50, 129.80, 128.26, 128.14, 127.36,
61.19, 52.72, 45.94, 18.58.IR (KBr):δ = 1650.0 (CO),
1315.9 (S = O asymmetric), 1159.7 (S = O symmetric).
LCMS m/z: cal. 465.07; found 468.16 (M+1).
N-(2,6-dimethylphenyl)-2-(4-((4-methoxyphenyl)
sulfonyl)piperazin-1-yl)acetamide 6a
Colour: white solid,Yield: 91, m.p:180-181
oC. 1H NMR (400 MHz, CDCl3, TMS): δ = 8.28 (s,
1H, NH), 7.68-7.66 (t, J = 8Hz, 2H, Ar-H), 7.07-6.97
(m, 5H, Ar-H), 3.84 (s, 3H, OCH3), 3.17 (s, 2H,
CH2), 3.06 (s, 4H, CH2), 2.75-2.72 (t, J = 12Hz, 4H,
CH2), 2.13 (s, 6H, CH3). 13C NMR (100 MHz, CDCl3,
TMS): δ = 167.78, 163.22, 135.02, 133.32, 129.89,
128.27, 127.37, 126.92, 114.34, 61.20, 55.62, 52.80,
46.06, 18.5. IR (KBr): δ = 1662.2 (CO), 1322.2
(S = O asymmetric), 1170.3 (S = O symmetric).
LCMS m/z: cal. 417.17; found 418.25 (M+1).
N-(2,6-dimethylphenyl)-2-(4-tosylpiperazin-1-yl)
acetamide 6e
Colour:white solid,Yield:87, m.p:170-171
oC. 1H NMR (400 MHz, CDCl3, TMS):δ = 8.29 (s, 1H,
NH), 7.62-7.61 (d, J = 4Hz, 2H, Ar-H), 7.32-7.31
(d, J = 4Hz, 2H, Ar-H), 7.07-7.05 (m, 3H, Ar-H),
3.167-3.162 (d, J = 2Hz, 2H, CH2), 3.05 (s, 4H, CH2),
2.73-2.72 (s, 4H, CH2), 2.12 (s, 6H, CH3). 13C NMR
(100 MHz, CDCl3,TMS):δ = 167.73, 143.97, 135.05,
134.55, 133.37, 132.47, 129.20, 128.26, 127.77,
127.36, 61.19, 52.72, 45.94, 18.58. IR (KBr): δ =
1649.1 (C = O), 1342.5 (S = O asymmetric), 1140.1
(S = O symmetric), 2918.89 (Ar-H). LCMS m/z: cal.
401.18; found 402.26 (M+1).
2-(4-((4-chlorophenyl)sulfonyl)piperazin-1-yl)-N-
(2,6-dimethylphenyl)acetamide 6b
Colour :white solid,Yield:90, m.p:188-189
oC. 1H NMR (400 MHz, CDCl3, TMS):δ = 8.26 (s, 1H,
NH), 7.68-7.66 (d, J = 8Hz, 2H, Ar-H), 7.51-7.49
(d, J = 8Hz, 2H, Ar-H), 7.07-7.02 (m, 3H, Ar-H), 3.18
(s, 2H, CH2), 3.08 (s, 4H, CH2), 2.76-2.73 (t, J = 12Hz,
4H, CH2), 2.13 (s, 6H, CH3). 13C NMR (100MHz,
CDCl3, TMS): δ = 167.60, 139.67, 135.03, 134.05,
2-(4-(4-bromobenzyl)piperazin-1-yl)-N-(2,6-
dimethylphenyl) acetamide 8a
Colour: white solid,Yield: 92, m.p:130-131oC.

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1H NMR (400 MHz, CDCl3, TMS): δ = 8.65 (s, 1H,
NH), 7.43-7.41 (d, J = 8Hz, 2H, Ar-H), 7.21-7.18
(d, J = 12Hz, 2H, Ar-H), 7.08-7.04 (m, 3H, Ar-H), 3.47
(s, 2H, CH2), 3.17 (s, 2H, CH2), 2.70 (s, 4H, CH2), 2.52
(s, 4H, CH2), 2.19 (s, 6H, CH3). 13C NMR (100 MHz,
CDCl3, TMS): δ = 168.47, 136.72, 135.23, 134.97,
133.62, 132.66, 131.41, 130.76, 128.27, 127.18,
121.06, 61.58, 53.67, 18.62. IR (KBr): δ = 1652.2 (C
= O). LCMS m/z: cal. 415.13; found 416.68 (M+1).
N-(2,6-dimethylphenyl)-2-(4-(2-fluorobenzyl)
piperazin-1-yl)acetamide 8e
Colour: white solid, Yield: 93, m.p: 162-
163oC. 1H NMR (400 MHz, CDCl3, TMS): δ = 8.65
(s, 1H, NH), 7.39-7.35 (m, 1H, Ar-H), 7.24-7.23
(d, J = 2Hz, 1H, Ar-H), 7.13-7.00 (m, 5H, Ar-H), 3.61
(s, 2H, CH2), 3.18 (s, 2H, CH2), 2.72 (s, 4H, CH2),
2.58 (s, 4H, CH2), 2.20 (s, 6H, CH3). 13C NMR (100
MHz, CDCl3, TMS): δ = 168.51, 162.16, 160.16,
134.97, 133.62, 131.53, 128.97, 128.89, 127.15,
124.15, 124.35, 115.40, 61.63, 55.03, 52.90, 18.62.
IR (KBr): δ = 1635 (C = O). LCMS m/z: cal. 355.21;
found 356.71 (M+1).
N-(2,6-dimethylphenyl)-2-(4-(4-fluorobenzyl)
piperazin-1-yl) acetamide 8b
Colour: white solid,Yield: 94, m.p:150-151
oC. 1H NMR (400 MHz, CDCl3, TMS):δ = 8.66 (s, 1H,
NH), 7.29-7.24 (d, J = 20Hz, 2H, Ar-H), 7.09-7.08
(m, 3H, Ar-H), 7.02-6.97 (t, 2H, Ar-H), 3.48 (s, 2H,
CH2), 3.19 (s, 2H, CH2), 2.71 (s, 4H, CH2), 2.51
(s, 4H, CH2), 2.21 (s, 6H, CH3). 13C NMR (100 MHz,
CDCl3, TMS): δ = 168.49, 163.25, 160.82, 134.96,
133.62, 130.54, 128.27, 127.15, 115.18, 62.03,
61.65, 53.79, 18.61. IR (KBr): δ = 1648.2 (C = O).
LCMS m/z: cal. 355.21; found 356.71 (M+1).
Biological Screening
Antibacterial activity
Compounds 6a-e and 8a-e were evaluated
for antibacterial activity. Agar well diffusion method
was adopted for antibacterial activity using known
literature procedure45.Experiment was performed in
triplicate the mean values of zone of inhibition were
given in Table 1.
Antifungal Activity
2-(4-(3,4-dichlorobenzyl)piperazin-1-yl)-N-(2,6-
dimethylphenyl) acetamide 8c
Antifungalactivitiesofallpiperazinederivatives
towards two mold fungi were studied, viz. Candida
albicans (human pathogen) Aspergillus flavus (mold).
Poisoned food technique method was used to assess
the antifungal activity of the synthesized compounds46,
Nystatin (10µg/disc) was used as standard fungicide.
The experiments were performed in triplicate, average
values were reported in Table 2.
Colour: white solid, Yield: 93, m.p:142-
143oC. 1H NMR (400 MHz, CDCl3, TMS): δ = 8.64
(s, 1H, NH), 7.43-7.42 (d, J = 4Hz, 1H, Ar-H), 7.38-
7.36 (d, J = 8Hz, 1H, Ar-H), 7.24 (s, 1H, Ar-H), 7.17-
7.14 (m, 1H, Ar-H), 7.09-7.05 (m, 3H, Ar-H), 3.46
(s, 2H, CH2), 3.19 (s, 2H, CH2), 2.72 (s, 4H, NCH2),
2.51 (s, 4H, NCH2), 2.21 (s, 6H, CH3). 13C NMR (100
MHz, CDCl3, TMS): δ = 168.41, 138.48, 134.96,
133.60, 132.36, 131.03, 130.24, 128.28, 128.19,
127.17, 61.63, 53.14, 18.65; IR (KBr): δ = 1639.4
(C = O). LCMS m/z: cal. 405.14; found 406.68 (M+1).
Anthelmintic activity
Anthelmintic activity of compounds 6a-e
and 8a-e were evaluated using Pheretima posthuma
(Indian Earthworm), anthelmintic activity of the
compounds were evaluated as per the standard
experimental method reported47.Timetakenforparalysis
and time taken for death were documented in minutes
and the observed results were given in Table 3.
2-(4-(2-chloro-6-fluorobenzyl)piperazin-1-yl)-N-
(2,6- dimethylphenyl) acetamide 8d
Colour: white solid,Yield: 91, m.p:130-131
oC. 1H NMR (400 MHz, CDCl3, TMS): δ = 8.67 (s,
1H, NH), 7.24-7.18 (m, 2H, Ar-H), 7.09-7.07 (m, J
= 2Hz, 3H, Ar-H), 6.98 (m, 1H, Ar-H), 3.72 (s, 2H,
CH2), 3.17 (s, 2H, CH2), 2.69-2.64 (m, 8H, CH2), 2.21
(s, 6H, CH3). 13C NMR (100 MHz, CDCl3, TMS): δ =
168.55, 163.29, 160.81, 136.66, 134.98, 133.65,
129.28, 125.44, 123.70, 114.08, 61.59, 53.78, 52.85,
52.24,18.6. IR (KBr): δ = 1655 (C = O). LCMS m/z:
cal. 389.17; found 390.70 (M+1).
Molecular docking
pharmacokinetic properties prediction
The drug-specificity of the compounds
was determined in accordance with Lipinski’s rule
of five48-49. Newly synthesized compounds were
subject to the QICPROP50 computation software.
Schrodinger software modules are designed to detect
pharmacokinetic properties such as absorption,
delivery, metabolism and division (ADME). The

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ligands for the installation were developed using
LigPrep, Schrödinger. Energy was minimized by the
OPLS 2005 force field51.Protein was prepared using
Maestro's protein preparation wizard52.Docking was
performed using GLIDE, Schrödinger. The GLIDE
XP (extra nozzle) method was used for installation
calculations53.
and 6c displayed highest activity against E. coli and
Staphylococcus aureus compared to Ciprofloxacin.
Table 1: Antibacterial activity Zone of Inhibition (mm)
of 6a-e and 8a-e
Samples Treatment Escherichia coli Staphylococcus aureus
(µg/ml)
(Mean SE)
(Mean SE)
6a
6b
6c
6d
6e
8a
8b
8c
8d
8e
50
50
50
50
50
50
50
50
50
50
15.44 0.55
16.01 0.34
17.62 0.62
16.12 0.27
17.91 0.24
12.92 0.30
26.89 0.23
25.45 0.37
15.93 0.76
14.10 0.41
31.00 0.43
6.43 0.30
7.14 0.33
8.41 0.15
7.06 0.41
8.89 0.42
12.98 0.16
13.35 0.20
12.96 0.16
12.21 0.25
11.80 0.29
15.00 0.39
RESULTS AND DISCUSSION
Chemistry
The basic acetamide piperazine skeleton i.e.
N-(2,6-dimethylphenyl)-2-(piperazin-1-yl)acetamide
4 was synthesized as per the reported literature54
1
and confirmed by IR, H and 13C NMR. Scaffold
N-(2,6- dimethylphenyl)-2-(piperazin-1- yl) acetamide
4 on reaction with different substituted benzene
sulfonyl chlorides 5a-e gave pure piperazine
sulfonamides 6a-e and with substituted benzyl
chlorides 7a-e gave alkylated piperazine derivatives
8a-eingoodyield.ProtonNMRofthetitlecompounds
6a-e were characterised by their resonance of
aromatic protons in the range of 7.0-7.6 ppm,
aliphatic protons at 2.1-3.2 ppm and NH proton of
amide group resonate at chemical shift of 8.2 ppm.
In case of 8a-e series, aromatic protons appear in
the range of 6.9-7.4 ppm, aliphatic protons resonate
in between 2.2-3.7 ppm. Interestingly NH proton of
amide series shifted to downfield at 8.6 ppm due to
electron withdrawing nature of halogen substituents
in the phenyl ring. In Carbon- 13CNMR, characteristic
carbonyl carbon of 6a-e and 8a-e resonate at 167 and
168 ppm respectively. Aromatic as well as aliphatic
carbons of both the series appeared in the range 115-
135 ppm and 50-60 ppm respectively, mass spectra
of all the compounds are in good agreement.
Ciprofloxacin 10
Antifungal activity
Compounds 6a-e and 8a-e showed more or
less inhibitory action against Candida albicans and
Aspergillus flavus as compared to standard Nystatin.
Initially different concentrations of 5 µg, 15 µg, 25 µg,
35 µg and 50 µg/mL have been tested. At 50µg/mL
was considered as minimum inhibitory concentration
(MIC) for both the fungal strains.
Table 2: Antifungal activity Zone of Inhibition (mm)
of 6a-e and 8a-e
Samples Treatment
(µg/ml)
Candida albicans Aspergillus flavus
(Mean SE)
(Mean SE)
6a
6b
50
50
50
50
50
50
50
50
50
50
10
10.75 0.24
10.12 0.42
15.03 0.22
15.74 0.11
11.05 0.17
23.25 0.17
23.97 0.12
25.67 0.25
14.10 0.62
25.52 0.29
29.33 0.13
16.13 0.63
17.56 0.52
12.87 0.13
13.09 0.13
11.14 0.26
18.44 0.17
19.09 0.43
19.12 0.26
12.23 0.23
18.20 0.11
21.33 0.23
6c
6d
6e
8a
8b
8c
8d
Biological Screening
Antibacterial Activity
8e
Nystatin
Compounds 6a-e and 8a-e showed
significant inhibitory action against tested Escherichia
coli and Staphylococcus aureus strains. Initially
different concentrations of 5 µg, 15 µg, 25 µg,
35 µg and 50µg/mL have been tried. At 50µg/
mL, no bacteria growth was observed hence this
concentration was considered as minimum inhibitory
concentration (MIC). Data obtained are presented
in Table 1, shows that the derivatives are active at
and above 50 µg/50 mL. Compounds 8b, 8c, 6e
Data presented in Table 2 showed that the
derivatives were active at and above 50 µg/50 µL.
Amongst all, derivatives 8c, 8e, 8b and 8a showed
significant activity against Candida albicans and
Aspergillus flavus. All other molecules showed
moderate activity against tested microorganisms
compared to standard Nystatin.
Anthelmintic Activity
Compounds 6a-e and 8a-e tested for

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Table 4. Lipinski’s rule of five factors of compounds
6a–e and 8a-e
anthelmintic activity against Pheretima posthuma
(Indian earthworm) showed moderate results
compared to standard piperazine citrate. Earthworms
belonging to control group showed paralysis time
of 142.33 0.49 min and death time of 167.17 0.87
minutes. The paralysis and death time for test
samples 6a-e and 8a-e at the concentrations of
50mg/mLwerereportedinTable3.Standardpiperazine
citrate exhibited 25.17 0.48 and 47.00 0.58 min time
for paralysis and death at concentration of 50 mg/
mL.Examination of anthelmintic activity revealed that
compound 8d, 6a and 6e showed significant activity
against Pheretima posthuma.
Factors of Lipinski’s rule of five
Comp. mol_MW Donor HB Accpt HB QPlogPo/w Rule of Five
(<500)
(< 5)
(< 10)
(< 5)
6a
6b
6c
6d
6e
8a
8b
8c
8d
8e
417.522
421.941
432.493
466.392
401.523
416.36
1
1
1
1
1
1
1
1
1
1
9.75
9
1.536
1.778
0.847
1.855
1.646
3.026
2.694
3.525
3.175
2.804
0
0
0
0
0
0
0
0
0
0
10
9
9
6.5
6.5
6.5
6.5
6.5
355.454
406.354
389.899
355.454
Table 3: Anthelmintic activity of compounds 6a-e and 8a-e
Test Samples Concentration Time taken for
Time taken
The tested compounds 8a-e have more
(mg/mL)
paralysis (min) for death(min)
than 82 percentage of human oral absorption
whereas 6a-e have more than 74 percentage of
human oral absorption except compound 6c which
has less than 60%.The aqueous solubility (QPlogS)
parameter and IC50 values of HERG K+ channel
blockage (QPlogHERG) of all the tested compounds
possess permissible parameters.
Control
-
142.33 0.49
167.17 0.87
6a
50
50
50
50
50
50
50
50
50
50
50
28.01 0.86
29.27 0.60
35.81 1.26
29.00 1.05
28.00 1.06
30.23 1.48
36.31 0.67
37.97 0.48
28.31 1.26
30.34 1.06
25.17 0.48
28.07 1.16
29.34 1.03
36.47 0.60
29.27 0.95
28.41 0.27
30.43 0.87
37.33 1.33
39.89 0.76
31.27 0.48
37.97 0.58
47.00 0.58
6b
6c
6d
6e
8a
8b
8c
Docking Studies
8d
The molecular docking studies were
8e
Piperazine citrate
carried out using Lead IT for antibacterial, antifungal
and anthelmintic activity. Docking energy of the
synthesized molecules along with standards was
tabulated in Table 5. The Molecular Docking result
shows that compounds 8a-e have higher docking
energy compared with antibacterial standard
ciprofloxacin. Antifungal standard Nystatin has
the highest docking score compared to all the
compounds, but in case of anthelmintic activity
most of the compounds have docking score which
are more comparable to piperazine citrate. All
the molecules were docked in the active sites of
respective proteins with effectively.The fitting of the
most potent compounds 8b, 8c, 6c and 6e against
bacteria, 8a, 8b, 8c and 8e against fungi, the
anthelmintic activity of 6c, 8b and 8c are discussed
below. Since conventional hydrogen bonding
is important, our molecular docking discussion
is restricted to hydrogen bonding even though
compounds show interactions like hydrophobic, ri-ri
stacked ri-alkyl interactions.
Molecular Docking
prediction of pharmacokinetic properties
The 2D structures of compounds 6a-e and
8a-e were subjected to a computational program
using Qikprop module of Schrödinger software for the
in silico determination of pharmacokinetic properties.
The statistical parameters of the pharmacokinetic
properties of compounds were tabulated in Table 4.
The pharmacokinetic properties predictions
of compounds 6a-e and 8a-e indicate that all the
compounds were endowed with drug like properties.
The results revealed that there is no violation in
agreement with the rule of five.The molecular weight
of compounds ranges from 355 to 466 a.m.u. The
number of hydrogen bond donor is one whereas
the hydrogen bond acceptor values vary from 6.5
to 10. In addition, the partition coefficient values of
all compounds are less than five.

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SREENIVASA et al., Orient. J. Chem., Vol. 35(1), 157-166 (2019)
Table 5: Docking energy in Kcal/mole of Target
molecules
conventional hydrogen bond with bromo and fluro
substituents separately in the compounds 8a and 8b
respectively. OD1 group of Asp110 residue accepts
hydrogen from NH group of compound 8c to form a
conventional hydrogen bond. Compound 8e donate
hydrogen bond to Asp412 residue. These are the
main significant interactions observed in the active
pocket of antifungal protein and our molecules.
Comp.
Antibacterial
Antifungal
Anthelmintic
PDBID:1SAO
PDB ID:3ACX
PDBID:1IYK
6a
-6.115
-7.212
-11.532
-7.176
-10.773
-7.921
-12.121
-11.463
-7.248
-8.511
-9.185
-
-5.658
-5.458
-7.093
-7.755
-5.545
-18.706
-16.845
-19.471
-7.939
-17.664
-
-12.873
-7.149
-7.139
-7.257
-12.873
-7.283
-7.811
-8.332
-13.225
-7.969
-
6b
6c
6d
6e
8a
8b
8c
8d
8e
Ciprofloxacin
Nystatin
-23.499
-
-
Piperizine Citrate
-
-9.254
Binding mode of potent Antibacterial compounds
In binding pose of nitro substituted
piperazine sulfonamide 6c, the prominent interactions
observed are as follows one hydrogen bond formed
by donating hydrogen from NH group to OD1 group
of Asp48 residue. The amino acid residues Asn168,
Arg171 and Tyr248 forms common hydrogen
interaction with O21 group of methyl piperazine
sulfonamide 6e. O11 atom of Alkylated piperazine
derivative 8b have hydrogen bond interaction with
Amino acid residues Arg171 and Tyr248. There
are each one hydrogen bond established between
Asp48 amino acid and compounds 8c. The docking
pose of the proteins and ligands are shown in
Figure1.
Fig. 2. Binding pose of the active compound’s 8a, 8b, 8c
and 8e with protein 1IYK
Binding mode of potent Anthelmintic compounds
In the binding pocket of the protein tubulin
shows two hydrogen bonding interaction between
O20, O21 groups of piperazine sulfonamide 6c and
NH, HE22 groups of Gln11.Compound 8b with fluro
and chloro substituents forms three hydrogen bond
interaction with Phe141, Gly146 and Glu183.Three
hydrogen interactions were observed between
alkylated piperazine derivative 8c with Phe141,
Gly146 and Glu183 as shown in Fig. 3. These
interactions may responsible for the high docking
score of the active compounds.
Fig. 1. Docking poses of compounds 6c, 6e, 8b and 8c with
protein 3ACx
Fig. 3. Binding pose of the active compound’s 6a, 6e and 8d
Binding mode of potent Antifungal compounds
Molecular docking of the compounds into
with protein 1SA0
the active site of N-myristoyltranferase shown in
Fig. 2 reveals an O10 group of His227 forms one
Finger print Analysis
Inrecentyears,manyfluorescentcompounds

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SREENIVASA et al., Orient. J. Chem., Vol. 35(1), 157-166 (2019)
havebeenusedasalabelingmaterialforfingerprinting
in forensics55. Usually, LFGs are often not seen or
investigated in places of crime. Extraordinary
fluorescent powders were used to improve LFG
in crime scene, and have been well-known for the
last few decades56. The powder dusting method is
the most widely used method for the development
of LFPs in forensic science.57,58 However, these
powders were effective in detecting LFPs under
normal conditions. Powder-dusting method using
these powders is still challenged by severe
problems, such as low sensitivity, low contrast, high
background interference, and high auto fluorescence
labeling agent on various porous and non-porous
surfaces such as, magazine covers (porous), glass
slide, spray bottle cap, compact disk, black plastic
bottle, coin, granite, spoon and highlighter pen
surface (Figure 5).
interactions59-61
.
Revelation of eccrine LFps using synthesized
compounds
All eccrine LFPs were collected by a donor
Fig. 5. Fingerprint visualization of 6c optimized on (a, b)
magazine cover, (c) glass slide, (d) spray bottle cap, (e)
compact disk, (f) black plastic bottle, (g) coin, (h) granite,
(i) spoon and (j) highlighter pen surface.
using cleaned and dried hands. Later, the fingers
were pressed at low pressure on various surfaces
with infiltrating and non-infiltrating materials at
370C temperature to obtain LFP. Each compound is
carefully applied to the eccrine LFPs to improve the
LFP and the excess compound powder was removed
by light brushing method. The obtained fingerprints
were photographed with normal white light with a digital
camera Nikon D3100/AF_S Nikkor50 mm f/2.8G ED.
From the images it is clear that Minutiae
ridges have been detected in all surfaces without
any color barrier, indicating that the improvement of
eccrine finger prints by powder dusting method with
compound 6c used as labeling agents on forensic
relevant porous and non-porous surfaces.
CONCLUSION
In this study two new series of compounds
weresynthesizedandevaluatedfortheirantibacterial,
antifungal, anthelmintic activity. Amongst all the
tested compounds 6c, 6e, 8b and 8c showed
noteworthy antibacterial activity against E. coli
and Staphylococcus aureus. Compounds 8a, 8b,
8c and 8e showed significant antifungal activity
and other alkylated piperazine derivatives 6a, 6e
and 8d showed good anthelmintic activity. It can
be concluded that acetamide derivatives linked
to piperazine show considerable antibacterial,
antifungal, and anthelmintic activity.Additionally, the
molecular docking and drug-likeliness shows that
these compounds have good docking energies.This
result implies that some of these compounds are
new potent antimicrobial and anthelmintic agents.
Compound 6c, which has been tested for latent
fingerprints detection, has shown that fingerprints
are better than other substances and may be used
for forensic application.
Fig. 4. Fingerprint visualization of 8a-e and 6a-e samples
on aluminum foil surface under normal light illumination
Figure 4 shows the development of eccrine
LFPs of synthesized compounds on aluminum foil
surface to examine their labeling efficacy in forensic
applications. This visualization of eccrine LFPs on
aluminum foil surface was developed under normal
white light illumination. From the Figure it is found
that among the entire samples, sample 6c shows
the clear ridge details after powder dusting method.
Hence, sample 6c was optimized and used as

165
University for providing docking software facility.
SREENIVASA et al., Orient. J. Chem., Vol. 35(1), 157-166 (2019)
ACKNOWLEDGEMENT
The authors are grateful to Tumkur
University and College of Science for the laboratory
facilities and also authors acknowledge Annamalai
Conflict of Interest
Authors declare no conflict of interest.
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