European Journal of Medicinal Chemistry 44 (2009) 2347–2353

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European Journal of Medicinal Chemistry

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Original article

Synthesis, antimicrobial and antiviral evaluation of substituted imidazole

derivatives
Deepika Sharma a, Balasubramanian Narasimhan b, *, Pradeep Kumar a, Vikramjeet Judge a,
Rakesh Narang a, Erik De Clercq c, Jan Balzarini c
a
Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science and Technology, Hisar 125001, India
b
Faculty of Pharmaceutical Sciences, Maharshi Dayanand University, Rohtak 124001, Haryana, India
c
Laboratory of Virology and Chemotherapy, Rega Institute for Medical Research, Minderbroedersstraat 10, B-3000 Leuven, Belgium

a r t i c l e i n f o a b s t r a c t

Article history: In the present study, we have synthesized 2-(substituted phenyl)-1H-imidazole (1–12) and (substituted
Received 30 July 2008 phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanone (13–26) analogues and screened them for
Received in revised form 22 August 2008 their antimicrobial activity against Gram positive, Gram negative and fungal species. The results of
Accepted 25 August 2008
antibacterial study indicated that compounds 15, 17 and 24 showed appreciable antibacterial activity and
Available online 11 September 2008
compound 26 emerged as the most potential antifungal agent. The results of SAR studies indicated that
the presence of electron withdrawing groups is necessary for the antimicrobial activity of the synthe-
Keywords:
sized compounds. The results of the present study indicated that compounds 15, 17 and 24 might be of
Substituted imidazoles
Antibacterial activity interest for the identification of new antimicrobial molecules as their antibacterial activity is equivalent
Antifungal activity to the standard drug norfloxacin. Further, the antiviral screening of (substituted phenyl)-[2-(substituted
Antiviral activity phenyl)-imidazol-1-yl]-methanones (13–26) against a panel of viral strains indicated that compounds 16
and 19 can be selected as lead compounds for the development of novel antiviral agents.
Ó 2008 Elsevier Masson SAS. All rights reserved.

1. Introduction actions, which are distinct from those of well known classes of
antimicrobial agents to which many clinically relevant pathogens
The incidence of invasive microbial infections caused by oppor- are now resistant [5].
tunistic pathogens, often characterized by high mortality rates, has The herpes virus family contains eight known human viruses,
been increasing over the past two decades. Patients who become amongst them are herpes simplex virus-1 (HSV-1) and herpes
severely immunocompromised because of underlying diseases such simplex virus-2 (HSV-2) that cause mucocutaneous infections
as leukemia or recently acquired immunodeficiency syndrome or resulting in cold sores (HSV-1) and genital lesions (HSV-2),
patients who undergo cancer chemotherapy or organ trans- respectively. Much research has been focused on HSV-1 and HSV-2
plantation are particularly susceptible to opportunistic microbial as these viruses have a high incidence rate and a high prevalence
infection [1]. Almost all of the major classes of antibiotics have [6]. Vesicular stomatitis virus (VSV), a member of the Rhabdoviridae
encountered resistance in clinical applications [2]. The emergence of family, is an enveloped single-stranded RNA virus that causes an
bacterial resistance to b-lactam antibiotics, macrolides, quinolones economically important disease in cattle, horses and swine [7].
and vancomycin is becoming a major worldwide health problem [3]. The alphaviruses are a genus of approximately 27 arthropod-
A matter of concern in the treatment of microbial infections is transmitted plus-strand RNA viruses found in the Togaviridae family
the limited number of efficacious antimicrobial drugs. Many of the which is responsible for a wide range of diseases and many of them
currently available drugs are toxic, enable recurrence because they are important human and animal pathogens. Several examples of
are bacteriostatic/fungistatic and not bactericidal/fungicidal or lead alphaviruses are Sindbis, Semliki Forest, and Venezuelan equine
to the development of resistance due in part to the prolonged encephalitis viruses. Infection can result in fever, rash, arthralgia or
periods of administration [4]. There is a real perceived need for the arthritis, lassitude, headache, and myalgia. The prototypic alpha-
discovery of new compounds that are endowed with antibacterial virus is Sindbis virus (SINV), which is transmitted to humans
and antifungal activities, possibly acting through mechanism of through mosquito bites [8]. Respiratory syncytial virus (RSV),
a paramyxovirus, is an important cause of respiratory tract infection
in infants, young children and adults [9].
* Corresponding author. Tel.: þ91 1262 272535; fax: þ91 1262 274133. Previous antiviral research on herpes simplex viruses has
E-mail address: [email protected] (B. Narasimhan). primarily focused on the development of nucleoside analogues,

0223-5234/$ – see front matter Ó 2008 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2008.08.010
2348 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353

such as acyclovir (Zovirax), valacyclovir, famciclovir, and penciclo- Dahiya and Pathak [35] resulted in resinous products. Therefore,
vir. Recently, immunomodulators (imiquimod and resiquimod), coupling was carried out using sodium acetate along with stirring
nonnucleoside viral polymerase inhibitors (4-hydroxyquinoline-3- in cold conditions for the initial 3 h followed by 48 h stirring
carboxamides), and viral helicase inhibitors (thiazolylphenyl and at room temperature which resulted in a solid product. For the
thiazolylamide) have received considerable attention. Though synthesis of (substituted phenyl)-[2-(substituted phenyl)-imida-
numerous strategies and considerable efforts have been spent in zol-1-yl]-methanones (13–26), the key intermediates (1–12) were
search of the next generation antiviral therapy, it has proved reacted with substituted benzoyl chloride which was prepared by
difficult to outperform acyclovir [6,10]. the reaction of substituted benzoic acid with thionyl chloride. The
The incorporation of the imidazole nucleus is an important physicochemical characteristics of the synthesized compounds are
synthetic strategy in drug discovery. The high therapeutic proper- presented in Table 1.
ties of the imidazole related drugs have encouraged the medicinal The structures of compounds 1–26 were assigned by IR and 1H
chemists to synthesize a large number of novel chemotherapeutic NMR spectroscopic data, which are consistent with the proposed
agents. Imidazole drugs have broadened scope in remedying molecular structures. The appearance of medium out-of-plane
various dispositions in clinical medicines. Medicinal properties deformation bands (C–C bending) at 725–680 cm1 indicated the
include anticancer [11], b-lactamase inhibitors [12], 20-HETE syn- presence of 1,3-disubstituted benzene ring in compounds 3 and 18.
thase inhibitors [13], carboxypeptidase inhibitors [14], hemeox- Similarly, the appearance of the C–C out-of-plane band at 790–
ygenase inhibitors [15], antiaging agents [16], anticoagulants [17], 810 cm1 indicated the presence of 1,4-disubstituted benzene ring
anti-inflammatory [18], antibacterial [19], antifungal [20], antiviral in compounds 2, 6, 13, 17, 18 and 25. The presence of 2-substituted
[21], antitubercular [22], antidiabetic [23] and antimalarial [24], all benzene in structures of compounds 17, 23, and 25 was confirmed
are unique characteristics known for imidazole derivatives. by strong out-of-plane deformation bands (C–H bending) around
In continuation with our research program concerning the 740 cm1 which were visible from their IR spectra. Moreover,
synthesis and antimicrobial evaluation of medicinally important the presence of NO2 group in compounds 4, 6, 13, 17 and 18
compounds [25–33], in the present study we have synthesized was indicated by the appearance of asymmetric and symmetric
2-(substituted phenyl)-1H-imidazoles (1–12). Further recent liter- NO2 stretching bands at 1550–1510 cm1 and 1365–1335 cm1,
ature revealed that N1-substitution of imidazoles improves the respectively. The appearance of medium bands at 3696.2 cm1 in
antibacterial activity [34]. This created interest among us to the IR spectra of compound 17 indicated the presence of a free OH
synthesize (substituted phenyl)-[2-(substituted phenyl)-imidazol- in the carboxylic acid group of 2-substituted benzene. Further, the
1-yl]-methanones (13–26) and screen them for their antimicrobial appearance of strong C]O stretching bands at 1695–1670 cm1 in
and antiviral activity. the IR spectra of (substituted phenyl)-[2-(substituted phenyl)-
imidazol-1-yl]-methanones (13–26) demonstrated the presence of
2. Chemistry tertiary amide linkage between substituted benzoic acid and
imidazole nucleus. Compounds 23 and 25 showed C–Br stretching
Synthesis of compounds 1–26 followed the general pathway at 522.8 cm1 and 533.8 cm1, which confirms the presence of 2-
that is depicted in Scheme 1. Compounds 1–12 are the key inter- bromo substituted benzene ring. The presence of aliphatic CH
mediates for the synthesis of compounds 13–26. The key inter- stretching at 2923.2 cm1 and CH stretching of aralkyl ether at
mediates, 2-(substituted phenyl)-1H-imidazoles (1–12), were 3054.5 cm1 in compound 25 confirms the existence of OCH3 group
prepared by the condensation of imidazoles with corresponding stretching.
substituted aryldiazonium chlorides which in turn were prepared Compounds 13 and 17 showed multiplet signal at d 7.69–
by the diazotization of substituted anilines. However, based on our 7.77 ppm corresponding to the protons of substituted benzene at C2
experience, the application of cupric chloride to the condensation of imidazole. In contrast, compounds 18 and 23 showed the
of aryldiazonium chloride with benzimidazole as suggested by multiplet signals at d 6.98–7.42 ppm for the protons of substituted

R1 H
R1 -lC+N2 N R1 R2
H2N R2 R2
N
NaNO2/HCl N
R3
0-10°C 48 h
R5 R3 R5 R3 N
H
R4 R4 R5 R4
1-12

COOH SOCl2 COCl

24 h
X X

R1 R2
N
R3
N
R5 R4
O

X
13-26

Scheme 1. Synthetic scheme for the synthesis of 2-(substituted phenyl)-1H-imidazoles and (substituted phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanones.
 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353 2349

Table 1
Physicochemical characteristics of synthesized 2-(substituted phenyl)-1H-imidazoles and (substituted phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanones
R1 R2
N
R3
R1 R2 N
N R5 R4
R3
O
N
H
R5 R4 X
1-12 13-26
Compound R1 R2 R3 R4 R5 X Mol. formula Mol. wt Mp ( C) Rf %Yield
1 Cl H H H H – C9H7N2Cl 178.66 100 0.80a 28.67
2 H H Cl H H – C9H7N2Cl 178.66 >250 0.86a 59.13
3 H Cl H H H – C9H7N3O2 189.23 196–199 0.41a 12.45
4 H NO2 H H H – C9H7N3O2 189.23 201–203 0.81a 53.13
5 NO2 H H H H – C9H7N2Cl 178.66 99–101 0.80a 44.31
6 H H NO2 H H – C9H8N2 144.17 51–53 0.60a 34.65
7 H H H H H – C9H7N3O2 189.23 >250 0.20a 47.07
8 COOH H H H H – C10H8N2O2 188.24 >250 0.12a 41.79
9 H H OCH3 H H – C10H10N2O2 190.25 109–111 0.29a 57.89
10 CH3 CH3 H H H – C11H12N2 172.23 219–221 0.71b 43.70
11 CH3 H H CH3 CH3 – C11H12N2 172.23 209–211 0.53b 74.32
12 H H Br H H – C9H7BrN2 123.07 >250 0.2b 53.61
13 H NO2 H H H 4-NO2 C16H10N4O5 338.28 139–141 0.13a 35.73
14 NO2 H H H H 4-NO2 C16H10N4O5 338.28 149–151 0.14a 41.09
15 Cl H H H H 4-NO2 C16H10N3O3Cl 327.73 221–223 0.11a 56.07
16 H H Cl H H 4-NO2 C16H10N3O3Cl 327.73 219–221 0.33a 65.72
17 COOH H H H H 4-NO2 C17H11N3O5 337.29 194–196 0.34a 19.56
18 H Cl H H H 4-NO2 C16H10ClN3O3 327.73 214–216 0.11a 76.54
19 H H NO2 H H 4-NO2 C16H10N4O5 338.28 209–211 0.66a 59.80
20 H H OCH3 H H 4-NO2 C17H13N3O4 323.31 229–231 0.10a 78.65
21 NO2 H H H H 2-Br C16H10BrN3O3 372.18 119–121 0.85a 60.98
22 H NO2 H H H 2-Br C16H10BrN3O3 372.18 114–116 0.73a 58.65
23 Cl H H H H 2-Br C16H10BrClN2O 361.63 149–151 0.85a 64.23
24 H H Cl H H 2-Br C16H10BrClN2O 361.63 139–141 0.82a 38.65
25 H H OCH3 H H 2-Br C17H17BrN2O2 357.21 139–141 0.82a 75.86
26 H H NO2 H H 2-Br C16H10BrN3O3 372.18 109–111 0.78a 49.78
a
Toluene:chloroform (7:3).
b
Benzene.

benzene at C2 of imidazole. This may be due to the interchange of presented in Table 2. In general the compounds showed improved
electron withdrawing groups in compounds 13 and 17 to electron antibacterial activity when compared to their antifungal activity.
donating groups in compounds 18 and 23. Compounds 13, 17 and 18 The deduced patterns of antimicrobial activity of substituted
showed signals at d 8.07–8.39 ppm due to the presence of the p- imidazoles are in the following order: antibacterial > antifungal.
NO2 acyl substituent at N1 of imidazole but in compounds 23 and All of the synthesized substituted imidazole derivatives showed
25 a downfield shift at d 7.50–7.59 ppm due to the substitution of appreciable in vitro antimicrobial activity against the tested
the electron withdrawing NO2 with bromine was observed. In microorganisms. Compounds 15, 17, 20, 21, 24 and 26 were found to
compounds 13 and 18 signal corresponding to C4 of imidazole was be highly active against S. aureus showing a bacterial inhibition at
observed at d 7.33 ppm but in compounds 17 and 23 the signal of C4 2  103 mM/ml. The synthesized derivatives 15, 17, 20, 21 and 24
of imidazole was seen at d 7.15 ppm. This effect may probably be showed high antibacterial potential at 2  103 mM/ml against B.
due to the presence of the 3-substituted benzene in compounds 13 subtilis. The antibacterial potential against E. coli was exhibited by
and 18 and 2-substituted benzene in compounds 17 and 23. The compounds 15, 17, 22 and 24 at concentrations ranging from
appearance of signal at d 3.90 ppm confirmed the presence of OCH3 2  103 mM/ml to 18  103 mM/ml. Compounds 14, 16, 21 and 26
group in compound 25. were found to be effective against A. niger having MIC of
4  103 mM/ml. Compounds 22, 25 and 26 were found to possess
3. Results and discussion high antifungal potential against C. albicans when compared to
other derivatives.
3.1. Antimicrobial activity The most active antibacterial agents, 15, 17 and 24, with MIC
value of 2  103 mM/ml have activities equivalent to that of the
The synthesized compounds were screened for their in vitro standard drug norfloxacin (MIC ¼ 2  103 mM/ml). Further modifi-
antimicrobial activities against two Gram-positive bacteria – cation of the above structures may provide compounds with better
Staphylococcus aureus, Bacillus subtilis; Gram-negative bacterium – antibacterial potential than the reference compound, norfloxacin.
Escherichia coli and fungal strains – Aspergillus niger and Candida Compound 21 emerged as the most effective antibacterial agent
albicans by the tube dilution method [36] using norfloxacin and against all of the tested microorganisms, except for E. coli and C.
fluconazole as control drugs for antibacterial and antifungal activ- albicans, based on its MIC values but it failed to prove effective against
ities, respectively. The results of the antimicrobial studies are the tested strains on the basis of its MBC and MFC values (Table 3).
2350 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353

Table 2 Table 3
Antimicrobial activity of synthesized 2-(substituted phenyl)-1H-imidazoles and MBC/MFC values of synthesized 2-(substituted phenyl)-1H-imidazoles and
(substituted phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanones (substituted phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanones

Compound Minimum inhibitory concentration (1  103 mM/ml) Compound Minimum bactericidal/fungicidal concentration (1  103 mM/ml)

S. aureus B. subtilis E. coli A. niger C. albicans S. aureus B. subtilis E. coli A. niger C. albicans
1 35 702 702 35 702 1 561 561 561 280 561
2 4 8 8 702 35 2 35 702 702 561 280
3 4 17 702 35 35 3 35 140 561 280 280
4 4 4 4 33 33 4 33 33 33 264 264
5 4 4 4 33 33 5 33 33 33 264 264
6 4 16 4 33 33 6 33 132 33 264 264
7 5 5 5 86 43 7 43 43 43 694 347
8 4 4 4 33 66 8 531 33 265 265 526
9 4 4 32 65 65 9 32 32 263 526 526
10 4 4 4 17 72 10 36 36 36 144 289
11 4 4 72 9 36 11 36 36 578 144 289
12 3 3 3 14 28 12 28 28 28 112 224
13 18 36 9 9 18 13 147 295 36 73 295
14 36 36 36 4 36 14 295 295 295 73 295
15 2 2 2 9 38 15 19 19 19 76 152
16 38 19 38 4 19 16 152 305 305 76 152
17 2 2 2 9 18 17 18 18 18 148 256
18 38 38 38 19 38 18 305 305 305 152 304
19 73 36 73 18 18 19 295 295 295 147 147
20 2 2 18 9 18 20 18 18 147 147 147
21 2 2 33 4 33 21 268 268 268 67 268
22 4 33 2 16 16 22 33 268 16 67 268
23 69 69 69 8 17 23 268 268 268 138 138
24 2 2 2 8 17 24 17 17 17 69 138
25 16 16 33 16 16 25 134 628 628 134 134
26 2 4 4 4 16 26 16 33 33 67 134

Std 2a 2a 2a 1b 1b Std 19a 19a 19a 40b 40b

a a
Norfloxacin. Norfloxacin.
b b
Fluconazole. Fluconazole.

In general, the MFC and MBC values of the synthesized imid- et al. [38] who stated that the p-substituted phenyl causes an
azole derivatives were 3-fold higher than the MIC values which increase in activity than the unsubstituted phenyl ring in the
indicated that the synthesized compounds were bacteriostatic and case of oxazoles.
fungistatic in action (a drug is considered to be bacteriostatic/ (iv) The high antimicrobial activity of compounds 13–26 in
fungistatic when its MFC and MBC values are 3-fold higher than its comparison to compounds 1–12 may be attributed to the
MIC value) [37]. presence of carbonyl group in the former. This is supported
by the fact that removal of carbonyl group in bis-imidazole
derivatives resulted in loss of their antifungal activity [39].
3.2. Structure activity relationship (SAR) studies (v) Compounds 15, 17 and 24 were found to be the most effective
antibacterial agents, whereas compound 26 emerged as the
From the results of the antimicrobial activity of the synthesized most effective antifungal agent. This indicated that there are
substituted imidazole derivatives, the following structure activity different structural requirements for binding of drug to
relationships can be derived: bacterial or fungal targets, respectively [40].
(vi) In general, the most active compounds contain an electron
(i) N1-Substituted imidazole derivatives exhibited high antimi- withdrawing substituent on phenyl ring at second position of
crobial activity in comparison to N1-non-substituted imidazole. The role of electron withdrawing group in
imidazole derivatives. This was similar to the literature improving antimicrobial activities is supported by the studies
reports [1,34] where little or no activity was found for of Sharma et al. [41].
the non-N1-substituted derivatives of benzimidazoles and (vii) In contrast to point (vi) mentioned above, the presence of
imidazoles. electron donating methoxy group also showed significant
(ii) Compound 17, having COOH substitution i.e. 2-[1-(4-nitro- activity against S. aureus, B. subtilis and C. albicans. The role of
benzoyl)-1H-imidazol-2-yl]-benzoic acid, showed profound methoxy group in improving antifungal activity of imidazoles
antibacterial activity without any significant activity against is supported by the studies of Emami et al. [20].
tested fungal strains. The significant antibacterial activity (viii) As far as antifungal activity against A. niger is concerned,
(2  103 mM/ml) may be attributed to the presence of the the N1-substituted phenyl ring have both 4-NO2 and 2-Br
COOH group which is also present in the standard drug substituents as electron withdrawing groups (14, 16, 21
norfloxacin. The lower antifungal activity is supported by the and 26) whereas in case of C. albicans the N1-substituted
work of Goker et al. [1] who reported that the presence of phenyl ring has only a bromine substituent as an electron
trifluoromethyl, carboxylic, ester and amide groups does not withdrawing moiety. The results indicated that the presence
improve antifungal activity. of the electron withdrawing NO2 substituent on the N1-
(iii) In general, it was observed that most of the compounds with substituted phenyl ring may not be favourable for the
substituted phenyl rings showed better antimicrobial activity binding of compounds 22, 25 and 26 with the fungal target in
than the ones with a non-substituted phenyl ring (compound case of C. albicans. The above SAR results are summarized in
7). This was similar to the results observed by Tekiner-Gulbas Fig. 1.
 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353 2351

Presence of COOH increased the Table 5

antibacterial activity Cytotoxicity and antiviral activity of (substituted phenyl)-[2-(substituted phenyl)-
N imidazol-1-yl]-methanones in HeLa cell cultures
R Presence of electron withdrawing groups
improved the antibacterial activity Compound Minimum EC50b (mg/ml)
N
Presence of H decreased the antibacterial cytotoxic
Vesicular Coxsackie Respiratory
O activity concentrationa
stomatitis virus virus B4 syncytial virus
Necessary for enhanced antibacterial activity (mg/ml)
13 >100 >100 >100 >100
X The antifungal activity against C. albicans is
14 >100 >100 >100 >100
increased if X is 2-Br group
15 >100 >100 >100 >100
16 >100 >100 45 >100
Fig. 1. SAR for antimicrobial activity of (substituted phenyl)-[2-(substituted phenyl)- 17 >100 >100 >100 >100
imidazol-1-yl]-methanones. 18 >100 >100 >100 >100
19 100 >20 >20 >20
20 >100 >100 >100 >100
3.3. Antiviral activity 21 >100 >100 >100 >100
22 >100 >100 >100 >100
The results of antiviral screening of (substituted phenyl)-[2- 23 >100 >100 >100 >100
24 >100 >100 >100 >100
(substituted phenyl)-imidazol-1-yl]-methanones (13–26) against
25 >100 >100 >100 >100
herpes simplex virus-1 (KOS) (HSV-1 KOS), herpes simplex virus-2 26 >100 >100 >100 >100
(G) (HSV-2G), vaccinia virus (VV), vesicular stomatitis virus (VSV), DS-5000 >100 >100 9 0.8
herpes simplex virus-1 TK KOS ACVr (HSV-1 TK KOS ACVr) in HEL (S)-DHPA >250 >250 >250 >250
cell cultures, vesicular stomatitis virus (VSV), Coxsackie virus B4 Ribavirin >250 29 146 10
a
(CV-B4), respiratory syncytial virus (RSV) in HeLa cell cultures and Required to cause a microscopically detectable alteration of normal cell
parainfluenza-3 virus (PI-3V), reovirus-1 (RV-1), Sindbis virus (SV), morphology.
b
Required to reduce virus-induced cytopathogenicity by 50%.
Coxsackie virus B4 (CV-B4), Punta Toro virus (PTV) in Vero cell
cultures were determined using a CPE reduction assay [42].
Compound 19 emerged as the best antiviral agent against HSV-1 vaccinia virus at a concentration of 2 mg/ml and 4 mg/ml, respec-
KOS (EC50 ¼ 59 mg/ml, Table 4), HSV-2G (EC50 ¼ 50 mg/ml, Table 4) tively, which was almost 50 times and 25 times less than that of
and HSV-1 TK KOS ACVr (EC50 ¼ 45 mg/ml, Table 4) among the their cytotoxic concentration (>100 mg/ml) which made them to be
tested (substituted phenyl)-[2-(substituted phenyl)-imidazol-1- considered as compounds having a marked therapeutic index and
yl]-methanones (13–26), but it was much less active than the be selected as lead compounds for the synthesis of newer antiviral
reference compounds brivudin, cidofovir and ganciclovir. Against agents. All other compounds tested in HEL cell culture were active
VSV none of the tested compounds were active in HEL cell culture. at a concentration which caused microscopically detectable alter-
Further, compounds 16 and 19 (having a p-substituted electron ation of normal cell morphology.
withdrawing group at both 1 and 2 positions of imidazole) emerged In HeLa cell cultures (Table 5) none of the compounds proved
as the most promising antiviral agents against vaccinia virus tested active against any of the tested strains of viruses (VSV, CV-B4, RSV)
in HEL cell culture with an EC50 value of 2 and 4 mg/ml, respectively except compound 19 which showed appreciable antiviral activity
(Table 4) and their anti-VV activity was equivalent to that of the against the all aforesaid viruses in HeLa cell culture at an
standard drugs brivudin (EC50 ¼ 5 mg/ml) and cidofovir (EC50 ¼ EC50 > 20 mg/ml, which was  5 times lower than its cytotoxic
7 mg/ml). Compounds 16 and 19 inhibited the viral replication of concentration (100 mg/ml).
In Vero cell culture no activity was noted with any of the
compounds against parainfluenza-3 virus, reovirus-1, Sindbis virus,
Table 4
Cytotoxicity and antiviral activity of (substituted phenyl)-[2-(substituted phenyl)-
imidazol-1-yl]-methanones in HEL cell cultures Table 6
Cytotoxicity and antiviral activity of (substituted phenyl)-[2-(substituted phenyl)-
Compound Minimum EC50b (mg/ml) imidazol-1-yl]-methanones in VERO cell cultures
cytotoxic
Herpes Herpes Vaccinia Vesicular Herpes
concentrationa Compound Minimum EC50b (mg/ml)
simplex simplex virus stomatitis simplex
(mg/ml) cytotoxic
virus-1 virus-2 virus virus-1 TK Parainfluenza- Reovirus- Sindbis Coxsackie Punta
concentrationa
(KOS) (G) KOS ACVr 3 virus 1 virus virus B4 Toro
(mg/ml)
13 >100 >100 >100 >100 >100 >100 virus
14 >100 >100 >100 >100 >100 >100 13 >100 >100 >100 >100 >100 100
15 >100 >100 >100 >100 >100 >100 14 >100 >100 >100 >100 >100 >100
16 >100 >100 >100 2 >100 >100 15 >100 >100 >100 >100 >100 >100
17 >100 >100 >100 >100 >100 >100 16 >100 >100 >100 >100 >100 >100
18 >100 >100 >100 45 >100 >100 17 >100 >100 >100 >100 >100 >100
19 >100 59 50 4 >100 45 18 >100 >100 >100 >100 >100 >100
20 >100 >100 >100 >100 >100 >100 19 >100 >100 >100 >100 >100 >100
21 >100 >100 >100 >100 >100 >100 20 >100 >100 >100 >100 >100 >100
22 >100 >100 >100 >100 >100 >100 21 >100 >100 >100 >100 >100 >100
23 >100 >100 >100 >100 >100 >100 22 >100 >100 >100 >100 >100 >100
24 >100 >100 >100 >100 >100 >100 23 >100 >100 >100 >100 >100 >100
25 >100 >100 >100 >100 >100 >100 24 >100 >100 >100 >100 >100 >100
26 >100 >100 >100 >100 >100 >100 25 >100 >100 >100 >100 >100 >100
Brivudin >250 0.04 29 5 >250 96 26 >100 >100 >100 >100 >100 >100
Ribavirin >250 >250 >250 >250 146 >250 DS-5000 >100 >100 >100 59 >100 100
Cidofovir >250 3 2 7 >250 2 (S)-DHPA >250 >250 >250 >250 >250 >250
Ganciclovir >100 0.06 0.06 >100 >100 1 Ribavirin >250 45 >250 >250 >250 146
a a
Required to cause a microscopically detectable alteration of normal cell Required to cause a microscopically detectable alteration of normal cell
morphology. morphology.
b b
Required to reduce virus-induced cytopathogenicity by 50%. Required to reduce virus-induced cytopathogenicity by 50%.
2352 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353

Coxsackie virus B4, Punta Toro virus at an EC50 value of 100 mg/ml 7.20–7.28 (m, 1H, CH of C4 of ArH), 7.34–7.37 (d, 1H, CH of C6 of
(Table 6). Of some importance, no cytotoxicity was noted with any ArH), 7.50 (s, 1H, CH of C2 of ArH); IR (KBr pellets) cm1: 3488.02 (NH
of the compounds in Vero cells. str, imidazole), 1507.27 (C–C str, Ar), 1660.60 (C–H str, Ar), 738.69
(C–Cl str).
4. Conclusion Analytical data for compound 3: mp ( C) – 197; yield – 12.45%; 1H
NMR (CDCl3): d 6.55–6.57 (d, 2H, CH of C4 and C5 of imidazole),
During an effort to discover new 2-(substituted phenyl)-1H- 8.15–8.17 (d, 2H, CH of C3 and C5 of ArH), 7.54–7.56 (d, 2H, CH of C2
imidazoles and (substituted phenyl)-[2-(substituted phenyl)- and C6 of ArH); IR (KBr pellets) cm1: 3481.27 (NH str, imidazole),
imidazol-1-yl]-methanone analogues with antimicrobial activity we 1519.80 (C–C str, Ar), 1653.52 (C–H str, Ar), 1342.25 (NO2 sym, str).
found that compounds 15, 17 and 24 showed appreciable antibac- Analytical data for compound 6: mp ( C) – >250; yield – 59.13%;
1
terial activity. Compounds 14, 16, 21 and 26 were found to be H NMR (CDCl3): d 7.21–7.26 (d, 2H, CH of C4 and C5 of imidazole),
significantly active against A. niger. Compounds 22, 25 and 26 were 7.34–7.36 (d, 2H, CH of C3 and C5 of ArH), 7.45–7.47 (d, 2H, CH of C2
found to have antifungal activity against C. albicans. The results of and C6 of ArH); IR (KBr pellets) cm1: 3489.95 (NH str, imidazole),
SAR studies indicated that the presence of electron withdrawing 1521.73 (C–C str, Ar), 1653.85 (C–H str, Ar), 749.29 (C–Cl str).
groups is necessary for the antimicrobial activity of the synthesized Analytical data for compound 13: mp ( C) – 139–141; yield –
compounds. Further, the results of the present study indicated that 35.73%; 1H NMR (CDCl3): d 7.69–7.77 (m, 3H, CH of C5 of imidazole
compounds 15, 17, 24 and 26 might be of interest for the develop- and CH of C5 and C6 of m-ArNO2), 7.33 (d, 1H, CH of C4 of imidazole),
ment of new antimicrobial molecules. Especially, it could be noticed 8.19–8.39 (m, 6H, CH of ArNO2); IR (KBr pellets) cm1: 1604.4 (three
that most of the compounds were more active as antibacterials than ring stretching bands, imidazole), 1540.8 (NO2 asym str, aromatic
as antifungals, which could be of some guidance to design new lead NO2), 1349.6 (NO2 sym str, aromatic NO2), 715.8 (C–C out-of-plane
antibacterial compounds. In fact, the most active antibacterial bending, 1,3-disubstituted benzene ring), 800.3 (CH out-of-plane
compounds (15, 17 and 24) showed antibacterial activity equivalent bending, 1,4-disubstituted benzene), 1694.0 (C]O str).
to that of the standard drug norfloxacin in both MIC and MBC Analytical data for compound 17: mp ( C) – 194–196; yield –
determinations. Further, the antiviral screening of (substituted 19.56%; 1H NMR (CDCl3): d 7.69–7.71 (m, 4H, CH of ArCOOH), 7.15 (d,
phenyl)-[2-(substituted phenyl)-imidazol-1-yl]-methanones (13– 1H, CH of C4 of imidazole), 8.07–8.38 (m, 4H, CH of ArNO2), 7.85 (d,
26) against a panel of viral strains indicated that compounds 16 and 1H, CH of C5 of imidazole); IR (KBr pellets) cm1: 1604.3 (three ring
19 could be selected as lead compounds for the development of stretching bands, imidazole), 1540.6 (asym str, aromatic NO2),
novel antiviral agents active against pox viruses. 1349.8 (sym str, aromatic NO2), 930.7 (OH out-of-plane bending,
COOH), 1290.0 (OH in plane bending, COOH), 3062.1 (CH str,
5. Experimental benzene), 799.6 (CH out-of-plane bending, 1,4-disubstituted
benzene), 3696.2 (OH str, COOH), 1693.2 (C]O str).
Melting points were determined in open capillary tubes on Analytical data for compound 18: mp ( C) – 214–216; yield –
a Sonar melting point apparatus and are uncorrected. Reaction 76.54%; 1H NMR (CDCl3): d 8.21–8.26 (m, 4H, CH of ArNO2), 7.33 (d,
progress was monitored by thin layer chromatography on silica gel 1H, CH of C4 of imidazole), 6.98–7.31 (m, 4H, CH of ArCl); IR (KBr
sheets (Merck silica gel-G) and the purity of the compounds was pellets) cm1: 1601.7 (three ring stretching bands, imidazole), 714.2
ascertained by single spot on TLC sheet. 1H nuclear magnetic (C–C out-of-plane bending, 1,3-disubstituted benzene ring), 1536.2
resonance (1H NMR) spectra were recorded on a Bruker Avance II (NO2 asym str, aromatic NO2), 1348.3 (NO2 sym str, aromatic NO2),
400 NMR spectrometer using appropriate deuterated solvents and 797.0 (CH out-of-plane bending, 1,4-disubstituted benzene), 1694.4
are expressed in parts per million (d, ppm) downfield from tetra- (C]O str).
methylsilane (internal standard). Infrared (IR) spectra were recor- Analytical data for compound 23: mp ( C) – 149–151; yield –
ded on a Shimadzu FTIR spectrometer. 64.23%; 1H NMR (CDCl3): d 7.16–7.42 (m, 4H, CH of ArCl), 7.10 (d, 1H,
CH of C4 of imidazole), 7.50–7.70 (m, 4H, CH of ArBr), 7.87 (d, 1H, CH
5.1. General procedure for the synthesis of 2-(substituted of C5 of imidazole); IR (KBr pellets) cm1: 1586.4 (three ring
phenyl)-1H-imidazoles (1–12) stretching bands, imidazole), 1683.8 (C]O str, Aryl COOH), 3010.30
(CH str, aromatic), 522.2 (C–Br str, aromatic), 684.0 (C–Cl str), 740.7
Substituted anilines (0.13 mol) in hydrochloric acid/water (CH out-of-plane bending, 1,2 disubstituted benzene ring), 791.1
mixture (1:1) were diazotized using solution of sodium nitrite at 0– (CH out-of-plane bending, 1,4-disubstituted benzene).
10  C. To the diazotized mixture, imidazole (0.004 mol) was added Analytical data for compound 25: mp ( C) – 144–146; yield –
with vigorous shaking. A solution of sodium acetate (40 g in 75.86%; 1H NMR (CDCl3): d 7.40–7.44 (m, 4H, CH of ArOCH3), 7.15 (d,
100 ml) was added dropwise to the above mixture by maintaining 1H, CH of C4 of imidazole), 7.59–7.79 (m, 4H, CH of ArBr), 7.87 (d, 1H,
the temperature at 5–10  C. The above solution was stirred initially CH of C5 of imidazole), 3.90 (s, 3H, OCH3); IR (KBr pellets) cm1:
for 3 h at cold condition followed by continuation of stirring at 1587.6 (three ring stretching bands, imidazole), 2923.2 (CH str,
room temperature for 48 h. The product obtained was filtered, CH3), 3054.5 (CH str, ArOCH3), 533.8 (C–Br str, aromatic), 685.4 (C–
dried and recrystallized from alcohol. Cl str), 741.4 (CH out-of-plane bending, 1,2 disubstituted benzene
ring), 810.9 (CH out-of-plane bending, 1,4-disubstituted benzene
5.2. General procedure for the synthesis of (substituted phenyl)-[2- ring), 1684.1 (C]O str).
(substituted phenyl)-imidazol-1-yl]-methanones (13–26)
5.3. Evaluation of antimicrobial activity
A solution of 2-(substituted phenyl)-1H-imidazoles (1–12)
(0.002 mol) in diethyl ether (50 ml) was added to a solution of 5.3.1. Determination of MIC
corresponding substituted benzoyl chloride (0.002 mol) in diethyl The antimicrobial activity was performed against Gram-positive
ether (50 ml). The above mixture was stirred for 24 h at room bacteria: S. aureus, B. subtilis, Gram-negative bacterium: E. coli and
temperature. The resultant product was isolated by evaporation of fungal strains: C. albicans and A. niger. The standard and test
ether and purified by recrystallization with methanol. samples were dissolved in DMSO to give a concentration of 100 mg/
Analytical data for compound 2: mp ( C) – 100; yield – 44.31%; ml. The minimum inhibitory concentration (MIC) was determined
1
H NMR (CDCl3): d 7.00–7.03 (d, 2H, CH of C4 and C5 of imidazole), by tube dilution method. Dilutions of test and standard compounds
 D. Sharma et al. / European Journal of Medicinal Chemistry 44 (2009) 2347–2353 2353

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