International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019 | Volume 9| Issue 3| Page 89-97

Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

Synthesis, Characterization, and in silico Studies of

Novel Alkanoylated 4-Methylphenyl sulphonamoyl
Carboxylic Acids as Potential Antimicrobial and
Antioxidant Agents
Egbujor, Melford. C1*, Okoro, Uchechukwu. C2, Okafor, Sunday. N3, Nwankwo, N.E4
1
Department of Industrial Chemistry, Renaissance University, Ugbawka, Enugu, Nigeria.
2
Synthetic Organic Chemistry Division, Department of Pure and Industrial Chemistry, University of Nigeria, Nsukka,
Nigeria.
3
Department of Pharmaceutical and Medicinal Chemistry, University of Nigeria, Nsukka, Nigeria.
4
Natural Science Unit, School of General Studies, University of Nigeria, Nsukka, Nigeria.
ABSTRACT
The synthesis of new alkanoylated 4-methylphenyl sulphonamoyl carboxylic acids, their molecular docking,
and antimicrobial and antioxidant activities were reported. The process involved the mild reaction of acetic
anhydride and sodium acetate with 4-methylsulphonamoyl carboxylic acids. The characterization of the
compounds was done using FTIR, 1H-NMR, 13C-NMR, and elemental analysis. They were tested for their
antimicrobial activities against selected human pathogens such as Pseudomonas aeruginosa, Salmonella typhi,
Candida albicans, Aspergillusniger, Staphylococcus aureus, Escherichia coli, and Bacillus subtilis. The antioxidant
study of the compounds was evaluated in vitro by the inhibition of generated stable 2,2-diphenyl-1-
picrylhydrazyl (DPPH) free radical. The in silico study was carried out in order to study five different prevalent
disease conditions, namely: bacterial infections, fungal infections, oxidative stress, trypanosomiasis, and
malaria. The findings demonstrated that compounds 2b and 2c exhibited the best in vitro antibacterial and
antifungal activities compared to ofloxacin and fluconazole. The antioxidant study revealed that compound 2a
was the most excellent antioxidant agent. The molecular docking study revealed that compounds 2a, 2b, 2c, 2e,
and 2f exhibited excellent antibacterial, antifungal, antioxidant, antitrypanosomal, and antimalaria activities
compared to the corresponding standard drugs such as Penicillin, Ketoconazole, α-Tocopherol, Melarsoprol,
and Chloroquine, respectively. The physicochemical evaluations showed that all the compounds were drug-like
according to “Lipinski’s rule of five”.The synthesized compounds were found to be potent antibacterial,
antifungal, antioxidant, and antitrypanosomal agents.
Key Words: Alkanoylation; 4-methylphenyl sulphonamoyl carboxylic acids; Antibacterial activity; antifungal
activity; Antioxidant activity, molecular docking.
eIJPPR 2019; 9(3):89-97
HOW TO CITE THIS ARTICLE: Egbujor, Melford. C, Okoro, Uchechukwu. C, Okafor, Sunday. N, Nwankwo, N.E (2019). “Synthesis,
Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as Potential Antimicrobial and
Antioxidant Agents ”, International Journal of Pharmaceutical and Phytopharmacological Research, 9(3), pp. 89-97.

INTRODUCTION groups from oxoacid [1]. Alkanoylation process leads to

the prevention of rearrangement reactions that would have
Alkanoylation is the process of incorporating an acyl been possible with alkylation [2]. It has also been found
group (R-C=O-) into a compound. This reaction method to protect the amino group of sulphonamide during
is also known as acylation, in which the acylating agent aminolysis in order to ensure regioselectivity [3, 4].
provides the needed acyl group. Acid anhydrides and acyl Fundamentally, sulphonamides are commonly employed
halides are generally utilized as acylating agents. Acyl in the management of acute systematic or local infections.
moiety is often gotten by removing one or more hydroxyl

Corresponding author: Egbujor, Melford. C

Address: Department of Industrial Chemistry, Renaissance University, Ugbawka, Enugu, Nigeria.
E-mail:  [email protected]
Relevant conflicts of interest/financial disclosures: The authors declare that the research was conducted in the absence of any
commercial or financial relationships that could be construed as a potential conflict of interest.
Received: 20 January 2019; Revised: 10 June 2019; Accepted: 14 June 2019

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

Many critical disease conditions including, coccidiosis, elemental analysis was carried out with elemental
mastitis, metritis, respiratory infections, and analyzer (Exeter Analytical Inc.model:CE440).
toxoplasmosis are also treatable with sulphonamoyl Precipitation of the tittle compounds was in analytical
compounds [5, 6]. Early and prompt administered of these grade and the reactions were monitored using TLC. The
drugs in the course of chronic infections give better antimicrobial studies were carried out at the Department
results. However, in severe cases, the first doses are to be of Microbiology, University of Nigeria, Nsukka while the
intravenously administered to minimize the time lag antioxidant studies were carried out at the Biochemistry
between dose and effect [7]. Compounds having Department, University of Nigeria, Nsukka
sulphonamoyl functional group are crucial in medicinal
chemistry for the design of drugs with a broad spectrum SYNTHESIS OF ALKANOYLATED 4-METHYLPHENYL
of biological activities. Also in organic chemistry study SULPHONAMIDES
and application, sulfonamides have numerous
2g of various 4-methyphenyl sulphonamoyl carboxylic
importances. Industrially, sulphonamides are useful in the
acids was weighed into a beaker, 9ml of concentrated
production of some health products, food colorants, and
hydrochloric acid and 25ml of distilled water were added
others. Considering the inherent ability of sulfonamides to
to the beaker followed by vigorous stirring to ensure
cure high profile diseases such as arthritis, cancer, and
homogeneous mixture. 16.0g of sodium carbonate was
osteoporosis, their syntheses are very crucial [8]. In spite
dissolved in 50ml distilled water in a separate beaker.
of the fact that the incorporation of acyl group into
Then, 13.5ml of acetic anhydride was added in small
sulphonamide compounds enhances their biological
portion over an interval of 1 hour to the 4-methyphenyl
activities and improves their drug potency, only a few
sulphonamoyl carboxylic acid solution after which it was
research have been reported on the successful acylation
poured into the sodium acetate solution. The reaction
and biological evaluation of these important
mixture was stirred thoroughly with a glass rod and
pharmaceutical compounds. Moreover, a little or no
immersed in an ice bath for 2 hours and filtered to afford
attention has been given to the synthesis of alkanoylated
the various alkanoylated 4-methyphenyl sulphonamoyl
sulphonamides despite the fact that the acyl group
carboxylic acids (2a-2f) in good to excellent yields (70-
component has been found to exist in major biochemical
97%).
molecules, and are essential in the formation of molecules 90
like DNA, proteins, carbohydrates, and lipids [9, 10]. 2-{Acetyl[(4-methylphenyl)sulfonyl]amino}propanoic
Biochemically, acyl group-containing compounds such as acid (2a)
Acetyl-CoA are required in several biosynthetic processes Yield=2.03g (91.4%), mp=108-109 °C, IR (KBr) cm-1:
[11]. Considering the numerous pharmacological 3312 (O-H of COOH); 3094 (N-H); 3001 (C-H); 1923
importances of acyl and sulphonamide functionalities, it (C-H aromatic); 1703, 1691(C=O); 1491, 1402 (C=C);
was found necessary to synthesize alkanoylated 4- 1311, 1292 (2S=O);1170 (SO2-NH); 1115 (C-N); 741
methylphenyl sulponamoyl carboxylic acids and evaluate (Ar-H). 1HNMR (CD3CN, 400MHZ) δ: 7.968 (d, J= 8.Hz,
their antimicrobial and antioxidant activities, as well as 2H, ArH), 7.523 (d, J= 8.4 Hz, 2H, ArH), 2.732 (s, 3H,
molecular docking, in order to maximize the synergistic CH3-C=O), 2.492 (s,3H, CH3-Ar), 1.985-1.968 (m, IH,
actions arising from their successful coupling and this CH), 1958 (d, J=1.2Hz, 3H, CH3-CH). 13CNMR (CD3N,
underscores the novelty of this study. 400MHZ) δ: 171.253, 170.244 (C=O); 147.937, 141.130,
130.613, 129.474, 126.355, 117.356 (aromatic carbons);
MATERIALS AND METHODS
58.141, 51.344, 49.553 43.915 (aliphatic carbons).
Chemistry Anal.calcd(%)for C12H15NO5S (285.32): C, 50.48; H,
Reagents were purchased from Sigma Aldrich. Melting 5.27; N, 4.92; S, 11.23. Found: C, 50.52; H, 5.31; N,
points of the synthesized compounds were determined 4.93; S, 11.26.
using electrothermal melting point apparatus and were
2-{Acetyl[(4-methylphenyl)sulfonyl]amino}-3-
uncorrected. Infrared spectra data were recorded on 8400s
sulfanylpropanoic acid (2b)
Fourier Transform Infrared (FTIR) (ABU,Zaria, Kaduna
Yield=1.95g (89.8%), mp=181-182 °C, IR (KBr) cm-1:
State, Nigeria). Nuclear Magnetic Resonance (1H-NMR
3308 (O-H of COOH), 3004 (N-H), 2919 (C-H alphatic);
and 13C-NMR) were run on 400MHz using NMR
2960 (S-H); 2088 (C-H aromatic); 1793, 1622 (C=O);
spectrophotometer at Sandeep VermaLabouratory,
1660, 1656 (C=C); 12987,1193 (S=O); 1155 (SO2NH);
Department of Chemistry, Indian Institute of
1137 (C-N); 785 (Ar-H). IHNMR (DMSO, 400MHz) δ:
Technology, Kanpur. Chemical shifts were reported in δ
7.930 (m, 2H, ArH),7.713 (m, 2H, ArH), 7.645-7.627(m,
scale (neat) using tetramethylsilane as a standard. The
IH, ArH), 7.598-7.571 (m, IH, ArH); 6.577 (s, IH, NH);

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

3.354 (s, 2H, NH2); 2.487 (s,3H, CH3-C=O). 13CNMR C13H17NO6S (315.35): C, 49.48; H, 5.39; N, 4.45; S,
(DMSO, 400MHz) δ: 172.212, 170.122 (C=O); 137.176, 10.15. Found: C, 49.52; H, 5.43; N, 4.48; S, 10.21.
137.063, 133.822, 133.689. 133.625, 132.692 (aromatic
carbons); 40.637, 40.425, 40.212, 39.988 (aliphatic 2-{Acetyl[(4-methylphenyl)sulfonyl]amino}-4-
carbons). Anal.calcd (%) for C1`2H15NO5S2 (317.39): C, methylpentanoic acid(2f)
45.38; H, 4.74; N, 4.42; S, 20.18. Found: C, 45.41; H, Yield=2.29g (95.8%); mp.217-218 °C, IR (KBr) cm-1:
4.76; N, 4.39; S, 20.22. 3273 (OH of COOH); 3073 (N-H); 2970 (C-H aliphatic);
1992 (C-Haromatic); 1871, 1695 (C=O); 1670, 1641
2-{Acetyl[(4-methylphenyl)sulfonyl]amino}-4- (C=C); 1336, 1135 (S=O); 1121 (SO2-NH); 1088 (C-N);
(methylsulfanyl)butanoic acid(2c) 741 (Ar-H). 1HNMR (C6D6, 400MHz) δ: 7.411 (m, 2H,
Yield=2.17g (93.7%), mp.217-218 °C, IR(KBr) Cm-1: ArH); 7.043 (m, 2H, ArH); 3.261 (s,IH, CH-CO2H);
3448(O-H of COOH); 3251(N-H); 2923(C-H), 1919 (C-H 2.147 (s,3H, CH3-C=O); 2.860 (s,3H,CH3-Ar); 1.747 (s,
aromatic), 1719, 1671 (C=O), 1654,1623 (C=C), 1232, 2H, 2CH); 1.277 (s, 6H, 2CH3-CH). 13C-NMR
1165 (S=O); 1089 (SO2NH), 1040 (C-N); 822 (Ar-H). (C6D6,400MHz) δ:171.244, 170.142(C=O); 138.557,
1
HNMR (CDCL3 /C6D6, 400MHz) δ: 7.172 (m, 2H, ArH); 133.883, 133.688 133.056, 132.821, 132.578 (aromatic
7.175 (m, 2H, ArH). 13(NMR (CDCL3/C6D6, 400MHz) carbons); 78.349, 72.463, 69.789, 65.892, 56.453, 45.152
δ:176.135, 170.123 (C=O); 137.412, 133.830, 133.648, (aliphatic carbons). Anal.calcd (%) for C15H21NO5S
128.458, 127.980, 127.737 (aromatic carbons); 77.464, (327.41): C, 54.99, H, 6.42, N, 4.29, S, 9.78. Found: C,
77.145, 76.811, 71.342, 68.566, 63.457 (aliphatic 54.97, H, 6.46, N, 4.31, S, 9.83.
carbons). Anal.calcd (%) for C14H19NO5S2 (345.43): C,
48.65; H, 5.51; N, 4.06; S, 18.54. Found: C, 48.68; H, BIOLOGICAL STUDIES
5.48; N, 4.10; S, 18.49.
ANTIMICROBIAL STUDIES
2-{Acetyl[(4-methylphenyl)sulfonyl]amino}--3-
hydroxypropanoic acid(2d) Preparation of media
Yield=2.31g (96.4%); mp.201-202 °C, IR (KBr) cm-1: Nutrient agar: 27.5g of nutrient agar powder was
3653 (free OH); 3297 (OH of COOH); 3073 (C-H dissolved in 1000ml of distilled water and was allowed to
soak for 15mins. The agar suspension was melted by 91
aliphatic); 1998 (C-H aromatic); 1820, 1735 (C=O);
1649, 1605 (C=C); 1329, 1240 (S=O); 1172(SO2-NH), boiling in a water bath. 20ml of the molten nutrient agar
1078 (C-N); 733(Ar-H). 1HNMR (DMSO/CDCL3) was dispensed into a bijou bottle, cocked, and sterilized in
400MHz) δ: 10.245 (m, 2H, OH); 7.495-7.475 (d, J= an autoclave at 121oC for 15mins. The sterile molten
8.0Hz, 2H, ArH); 7.033-7.012 (d, J= 91.2 Hz, 2H, ArH); nutrient agar was stored at 42oc until the time of use.
2.210 (S, 3H, CH3-C=O); 2.176 (S, 3H, CH3Ar). 13CNMR
Potato Dextrose Agar (PDA): 47g of PDA powder was
(DMSO/CDCl3, 400 MHz) δ: 171.133, 170.122 (C=O);
dissolved in 1000ml of distilled water and was allowed
141.859, 140.318, 129.497, 128.890, 126.864, 125.756,
soaking for 15mins. The agar suspension was melted by
(aromatic carbons); 78.693, 78.367, 78.040, 39.282
boiling in a water bath. 20ml of the molten PDA was
(aliphatic carbons). Anal.calcd (%) for C12H15NO6S
dispensed into a bijou bottle, cocked, and sterilized in an
(301.33): C, 47.80; H, 4.98; N, 4.66; S, 10.63. Found: C,
autoclave at 121oC for 15mins. The sterile molten potato
47.76; H, 4.96; N4.70, S, 10.59.
dextrose agar was stored at 42oc until use.
2-{Acetyl[(4-methylphenyl)sulfonyl]amino}-3-
The used test microorganisms: The antimicrobial
hydroxybutanoic acid(2e)
screening was done according to the agar dilution method
Yield=2.30g (96.0%); mp.217-218 °C, IR (KBr) cm-1:
[12]. The test microorganisms used (Staphylococcus
3387 (O-H of COOH); 3093 (N-H); 2932 (C-H aliphatic);
aureus, Escherichia coli, Bacillus subtilis, Pseudomonas
1995 (C-H aromatic); 1708, 1691 (C=O); 1650, 1640
aeruginosa, Salmonella typhi, Candida albicans, and
(C=C); 1381, 1294 (S=O); 1180 (SO2-NH); 1131 (C-N);
Aspergillusniger) were clinical isolates obtained from the
746 (Ar-H); IHNMR (DMSO, 400MHz) δ: 7.772-7.755
department of pharmaceutical microbiology and
(d, J= 6.8Hz, 2H, ArH); 7.564-7.548 (d, J= 6.4Hz, 2H,
biotechnology laboratory, University of Nigeria, Nsukka.
ArH); 4.099 (s, IH, OH); 3.892 (s, IH, COOH); 2.489 (s,
3H, CH3-C=O); 0.957-0.944 (d, J= 5.2Hz,3H, CH3-CH) Standardization of the test organism suspension: The
13
(NMR (DMSO, 400MHz) δ:171.222; 170.233 (C=O); organisms were standardized using 0.5 McFarland turbid
140.918, 132.895, 129.444, 126.970, 124,688, 120.233 equivalents.
(aromatic carbons); 67.675. 61.612, 40.061, 39.848,
39.272 (aliphatic carbons). Anal.calcd (%)for

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

Preparation of different concentrations of the solution was added to 2 ml of each sample solution and
synthesized compounds used: 5mg/ml of the stock ascorbic acid. The reaction mixture was vortexed
concentration of the compounds was prepared by thoroughly and left in the dark at room temperature for 30
dissolving 10g of the extract in 2ml of 50% DMSO. minutes. The absorbance of the mixture was measured (in
1.0mg/ml, 0.9mg/ml, 0.8mg/ml, 0.7mg/ml, 0.6mg/ml, triplicate) spectrophotometrically at 517 nm against the
0.5mg/ml, 0.4mg/ml, 0.3mg/ml, 0.2mg/ml, and 0.1mg/ml, corresponding blank solution. The percentage of DPPH
concentrations were obtained using C1V1=C2V2 formula. radical scavenging activity was calculated by using the
following formula:
Where C1 (initial concentration) =5mg/ml
V1 (Initial volume) = X DPPH radical scavenging activity (%) =
C2 (final concentration) = 1.0mg/ml
V2 (final volume) = 20ml, 𝐀𝐛𝐬𝐜𝐨𝐧t𝐫𝐨𝐥 − 𝐀𝐛𝐬𝐬𝐚𝐦𝐩𝐥𝐞
× 𝟏𝟎𝟎 Eq 2
𝐀𝐛𝐬𝐜𝐨𝐧t𝐫𝐨𝐥
Control test (standard): The standard antibiotic used
was Ofloxacin and Fluconazole. where Abscontrol was the absorbance of DPPH radical and
n-hexane/methanol, Abssample was the absorbance of
Experimental: 4.0ml of sample suspension of stock DPPH radical and sample/standard.
concentration 50mg/ml was transferred to the sterile Petri
dish, 16.0ml of double strength sterile molten agar was In silico methodology
transferred to the same plate to mix uniformly and thus, Physicochemical properties
1mg/ml concentration was obtained. The other The physicochemical properties of the synthesized
concentrations 0.9mg/ml, 0.8mg/ml, 0.7mg/ml, 0.6mg/ml, compounds were generated in silico. They included
0.5mg/ml, 0.4mg/ml, 0.3mg/ml, 0.2mg/ml, 0.1mg/ml, molecular weight (MW), number of hydrogen bond
were obtained using the same C1V1=C2V2 formula. The acceptor (HBA), number of hydrogen bond donor (HBD),
molten agar plates with different concentrations of the number of rotatable bond (NoRB), octanol/water partition
sample were allowed to gel. The plates were divided into coefficient logP(o/w), aqueous solubility (SlogP), and
seven equal parts with a permanent marker. The test total polar surface area (TPSA). These parameters were
microorganisms were streaked on the segments and 92
computed by descriptors calculator in Molecular
labeled. The culture plates were incubated in an inverted Operating Environment (MOE, 2018). The drug-likeness
position at 37 oC for 24 hours and at 25 oC for 48 hours. was evaluated using Lipinski’s rule of five.
After the incubation period , the plates were observed for
sensitivity and resistivity of the organisms to the agents Molecular docking
and the observations were recorded. The plates were Five different disease conditions were studied, namely:
further incubated for another 24 hours at 37 oC, and 48 trypanosomiasis, malaria, bacterial and fungal infections,
hours at 25 oC to determine whether the activity was and oxidative stress. A drug target was chosen for each of
bacteriostatic or bactericidal. The observations were also the disease conditions for molecular docking studies. The
recorded. drug targets for anti-trypanosomiasis: T. bruce farnesyl
diphosphate synthase complexed with minodronate (PDB
ANTIOXIDANT STUDIES code: 2EWG); antimalarial: plasmepsin II, a hemoglobin-
degrading enzyme from Plasmodium falciparum, in
Antioxidant activity by DPPH method complex with pepstatin A (PDB code: 1SME);
The antioxidant activity was determined according to antibacterial: E. coli DNA gyrase in complex with 1-
Blois Method [13]. The antioxidant behavior of the ethyl-3-[8-methyl-5-(2-methyl-pyridin-4-yl)-isoquinolin-
synthesized compounds was measured in vitro by the 3yl]urea (PDB code: 5MMN); antifungal: urate oxidase
inhibition of generated stable 2,2-diphenyl-1- from Aspergillus flavus complexed with uracil (PDB
picrylhydrazyl (DPPH) free radical. The DPPH solution code: 1WS3), and antioxidant: human peroxiredoxin 5
was prepared by dissolving 1.9 mg of DPPH in 100 ml of (PDB code: 1HD2).
methanol. Three different concentrations (50, 100, and The 3-dimensional structures of these drug targets were
200 µg/ml) of the DPPH solution were prepared. 2 mg of downloaded from the Protein Data Bank (PDB),
each synthesized compound was weighed out and (http://www.pdb.org) database. The drug targets were
dissolved in 10 ml of an appropriate solvent. The stock loaded in Molecular Operating Environment (MOE) and
solution (200 µg/ml) was diluted further to get 100 and 50 prepared using the QickPrep in MOE. The MMFF94
µg/ml of each sample. The standard solution of ascorbic force field was used for energy minimization of the ligand
acid was prepared in a similar manner. 1 ml of DPPH molecules. The prepared compounds were then subjected

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

to interact with each of the receptors through molecular The already existing 4-methylphenyl sulphonamoyl
docking. The protocol facilitates flexible compound carboxylic acids were subjected to alkanoylation reaction
docking for various compound conformers within the using acetic anhydride in the presence of sodium acetate.
rigid receptor. The best conformation for each compound This reaction step was carried out for the purpose of
was chosen and the interaction was visualized in protecting the amine functional group and incorporation
Discovery studio. of the biologically active acyl group into 4-methylphenyl
sulphonamoyl carboxylic acids in order to achieve
RESULTS AND DISCUSSION enhanced biological activities and improved drug
potency.
SYNTHESIS

O
O O
O O
S
S N CH3
NH (CH3CO)2O R
R H3C O
O CH3CO2Na
H3C OH
OH 2a-2f
1
R= CH3 for 2a, CH2SH for 2b, CH2CH2SCH3 for 2c, CH2OH for 2d, CH2CH3OH for
2e and CH2CH(CH3)CH3 for 2f

Scheme 1: synthesis of alkanoylated 4-methylphenyl sulphonamoyl carboxylic acids

REACTION MECHANISM the first step, the compound dissolves in water with one
equivalent of hydrochloric acid. The amino group of 4- 93
In compliance with Lumiere-Barbier method, the amino methylphenyl sulphonamide reacts in aqueous solution as
group can be alkanoylated in aqueous solutions [14]. In follows:

O O
O O
S S
NH H-OH NH3+
.. 2
H3C
H3C
Scheme 2: Reaction of the amino group in aqueous solution

Then, 1.2 equivalent of acetic anhydride is added after methylphenyl sulphonamide attacks acetic anhydride
which 1.2 equivalent of aqueous sodium acetate solution followed by deprotonation of the ammonium ion as
is added to the solution. The amino group of the 4- follows (Scheme 3):

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

- O
O O Me O
O O O O S
S N O
NH +
.. 2 O H
H3C H
H3C

B:

Then acetate acts as a living group of the substrate:

O
O O
O
O O Me O- O S -
+
S N H3C O
N O H
H H3C
H3C alkanoylated 4-methylphenylsulphonamide

Scheme 3: reaction mechanism for the synthesis of alkanoylated 4-methylphenyl sulphonamoyl carboxylic acids

O
O
S
NH

H3C R
O O O
1
S OH
NH O
O
H3C CH3 O
S
(CH3CO)2 1a O CH3
+ OH
N

CH3CO2Na CH3
H3C
O O O
2a OH
S
NH
O
O 94
H3C O
S
O
(CH3CO)2 + 1b OH
SH N CH3

CH3CO2Na
O O H3C
2b O SH
S
CH3 OH
NH O
S O
H3C O
S
(CH3CO)2 + 1c O
OH
N CH3
CH3CO2Na CH3
H3C 2c S
O O O
S OH
O
NH O
O
H3C S
O N CH3
(CH3CO)2 + 1d
OH
OH
2d
CH3CO2Na H3C
O O O OH
S OH
NH CH3 O
O
H3C O
O
(CH3CO)2 + 1e
OH
OH S
N CH3
2e CH3
CH3CO2Na
H3C
O O O OH
S H3C OH
NH O
O
H3C CH3
O
O S
(CH3CO)2 + 1f N CH3
OH CH3
CH3CO2Na 2f
H3C CH3
O
OH
R= CH3 for 2a, CH2SH for 2b, CH2CH2SCH3 for 2c, CH2OH for 2d, CH2CH3OH for 2e and CH2CH(CH3)CH3 for 2f

Scheme 4: a synthetic pathway for alkanoylated 4-methylphenyl sulphonamoyl carboxylic acid.

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

ANTIMICROBIAL ACTIVITY RESULTS

Table 1: Antimicrobial activities of compounds (2a-2f)

minimum inhibitory concentration (MIC)(mg/ml)
COMPOUNDS E.coli S.typhi S. aureus B. sub Ps. aerug C. albicans A. niger
2a 0.8 0.7 + 0.8 + 0.8 +
2b 0.6 0.9 0.7 0.4 0.9 0.6 0.9
2c 0.7 0.7 0.9 0.5 0.7 + +
2e 0.9 1.0 + + + 0.9 +
2f 0.9 0.7 + 0.8 + 0.7 +
Ofloxacin 0.005 0.010 0.010 0.020 0.020 + +
Fluconazole + + + + + 0.020 0.005
Key: + = no inhibition, Ofloxacine and Fluconazole are the antibacterial and antifungal standard drugs

The antimicrobial studies (Table 1) revealed that the title implies that these alkanoylated 4-methylphenyl
compounds had good antimicrobial (antibacterial and sulphonamoyl carboxylic acids can serve as good
antifungal) activities when compared to the commercial antibacterial and antifungal agents.
standard drugs. It was also observed that compounds 2b
and 2c possess the best antibacterial activities while ANTIOXIDANT STUDIES
compound 2b had the best antifungal activity. This

Table 2: Antioxidant activities results

200 µg/ml 100 µg/ml 50 µg/ml
Sample % inhibition Std % inhibition Std % inhibition Std
Ascorbic acid 96.83 0.001 97.68 0.001 97.31 0.001
290a 93.41 0.000 87.67 0.001 84.92 0.001 95
290b 75.03 0.002 71.79 0.002 77.05 0.001
290e 79.12 0.001 82.17 0.001 79.55 0.001
290f 79.67 0.000 ND ND 75.34 0.001

The in vitro antioxidant studies (Table 2) showed that antioxidant agent and therefore could be subjected to
some of the tested compounds had antioxidant activities. further derivatization to improve the drug-likeness.
Compound 2a showed impressive antioxidant activities.
This implies that compound 2a was the most potent Drug-likeness Studies

Table 3: Physicochemical properties

comp HBA HBD NoRB logP(o/w) SlogP TPSA Weight lip_violation
290a 5 2 5 1.22 1.01 91.75 285.32 0
290b 5 2 5 1.40 0.87 130.55 303.36 0
290c 5 2 8 1.87 1.74 91.75 345.44 0
290e 6 3 6 0.64 0.37 111.98 315.35 0
290f 5 2 7 2.63 2.03 91.75 327.40 0

The physicochemical properties to evaluate the drug- value of ≤ 500, hydrogen bond donor (HBD)≤ 10 and
likeness of the synthesized compounds are shown in partition coefficient (Log P) value ≤ 5. Violation of more
Table 3. Lipinski’s rule of five (Ro5) is used for the than one parameter may pose a challenge to the
assessment of the drug-likeness of a molecule. According bioavailability of the molecule in case of the oral
to this rule, a molecule must have a molecular weight formulation. From the results in Table 3, the synthesized

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

compounds are in agreement with (Ro5). The TPSA, a cross the BBB and penetrate the CNS [16]. Therefore,
reflection of the ligand hydrophilicity, is vital in protein- they can be very valuable in treating CNS related diseases
ligand interaction. NoRB ≤ 10 and TPSA ≤ 140 Å2 such as cerebral malaria, Alzheimer’s diseases.
would have a high probability of good oral bioavailability
in rats [15]. Also, a compound with TPSA of ≤ 90 Å2 can Results of Molecular Docking Studies

Table 4: In silicoAntitrypanosomal, Antimalarial, Antibacterial, Antifungal, and Antioxidant Activities.

Compound Trypanosomiasis Malaria Antibacterial Antifungal Antioxidant
2EWG 1SME 5MMN 1WS3 1HD2
2a -13.35 -10.83 -9.63 -11.79 -12.66
2b -13.68 -11.48 -10.17 -10.62 -11.81
2c -13.68 -11.46 -9.46 -9.49 -11.33
2e -14.54 -11.28 -10.54 -10.48 -12.28
2f -13.04 -11.78 -11.22 -9.49 -11.45
Standard drug -19.36 -10.08 -10.89 -10.38 -9.34
Standard drugs for 2EWG - Melarsoprol; 1SME - Chloroquine; 5MMN - Penicillin; 1WS3 - Ketoconazole; 1HD2 - α-Tocopherol

Molecular docking respectively possess more excellent antifungal activities

The calculated free binding energy after molecular that the commercial standard drug with binding energy -
docking is given in Table 4. Our compounds showed 10.38 Kcal/mole. This implies that compound 2a, 2b, and
strong binding affinities with all the receptors used in this 2e can serve as better antifungal agents than standard
study. Among all the compounds tested on 2EWG, 2a and commercial drugs and therefore should be considered
2f gave the lowest binding energy (highest binding accordingly.
affinity) of -13.35 and 13.04 kcal/mol, respectively. In silico Antioxidant Activities: The in silico antioxidant
However, the standard drug for the treatment of studies (Table 4) revealed that only compounds 2a, 2b,
96
trypanosomiasis (melarsoprol) showed the highest 2c, 2e and 2f (-12.66, 12.81, 11.33, 12.28, and 11.45
binding affinity with 2EWG (-19.36 kcal/mol). Likewise, Kcal/mol) had comparable binding energy with α-
compound 2f showed the highest binding affinity (-11.78 tocopherol (-9.34 Kcal/mol). Compounds 2a, 2b, 2c, 2e,
kcal/mol) with the Plasmodium falciparum pepstatin A and 2f are more promising than the standard drug.
receptor (1SME) when compared to the standard drug In Silico antitrypanosomal activities: The in silico
(chloroquine) for malaria treatment, whose binding antitrypanosomal studies (Table 4) revealed that all the
affinity is -10.08 kcal/mol. For the DNA gyrase receptor, compounds possessed antitrypanosomal activities but
compound 2f had a more binding affinity with it (-11.93 none of the tested compounds were as effective as the
kcal/mol) than with penicillin (-10.89 kcal/mol). The standard drug.
receptor for antifungal study, 1WS3, had the highest In Silico antimalarial activities: The in silico
binding affinity with compound 2a (-11.79 kcal/mol), antimalarial studies (Table 4) revealed that all the
which was better than that of ketoconazole (-10.38 compounds 2a, 2b, 2c, 2e, and 2f with binding energies
kcal/mol). Finally, compounds 2a, 2b, 2c, 2e, and 2f of -10.83, -11.48, -11.46, and -11.28, -11.78 kcal/mol,
outperformed α-tocopherol in their binding affinities with respectively against Plasmodium Falciparuim as an
1HD2 standard drug (-9.34 kcal/mol). excellent inhibitor of dihydrofolate reductase. These
In silico antibacterial activities: The in silico alkanoylated4-methylphenyl sulphonamoyl carboxylic
antibacterial studies (Table 4) revealed that compound 2a acids when compared to the most potent standard
with binding energy of -11.22 Kcal/mol possessed more antimalaria agent, Chloroquine (-10.08 Kcal/mol),
excellent antibacterial activities than the commercial showed better binding energies with compound 2f being
standard drug with a binding energy of -10.89 Kcal/mole. the most excellent compound and therefore can be used as
This implies that compound 2a can serve as better antimalarial agents.
antibacterial agent than standard commercial drugs and
therefore should be considered accordingly. CONCLUSION
In silico antifungal activities: The in silico antifungal
studies (Table 4) revealed that compounds 2a, 2b, and 2e, In conclusion, convenient synthesis of alkanoylated 4-
with binding energies -11.79, 10.62 and 10.48Kcal/mol methylphenyl sulphonamoyl carboxylic acids (2a-2f) was
successful. The assigned structures were in perfect

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International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | June 2019| Volume 9| Issue 3| Page 89-97
Egbujor, Melford. C, Synthesis, Characterization, and in silico Studies of Novel Alkanoylated 4-Methylphenyl sulphonamoyl Carboxylic Acids as
Potential Antimicrobial and Antioxidant Agents

agreement with the spectral data. The antimicrobial, [4] Egbujor, Melford. C, Okoro, Uchechukwu. C,
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The authors have no financial involvement with any Agar and broth dilution methods to determine the
organization or entity. There was no financial aid received minimal inhibitory concentration (MIC) of
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