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Open AccessArticle

In Vitro Anti-Candida Activity of Certain New 3-(1H-Imidazol-1-yl)propan-1-one Oxime Esters

Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
Medicinal and Pharmaceutical Chemistry Department, Pharmaceutical and Drug Industries Research Division, National Research Centre, Dokki, Giza 12622, Egypt
Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
Department of Microbiology, Faculty of Pharmacy, Alexandria University, Alexandria 21500, Egypt
Author to whom correspondence should be addressed.
Molecules 2013, 18(10), 12208-12221;
Received: 1 September 2013 / Revised: 19 September 2013 / Accepted: 23 September 2013 / Published: 30 September 2013
(This article belongs to the Section Medicinal Chemistry)


Anti-Candida activities of certain new oximes 4ad and their respective aromatic esters 5al are reported. The tested compounds 4ad and 5al exhibited better anti-Candida profiles than fluconazole. Compound 5j, namely (E)-3-(1H-imidazol-1-yl)-1-phenylpropan-1-one O-4-chlorobenzoyl oxime emerged as the most active congener, with a MIC value of 0.0054 µmol/mL being more potent than both fluconazole (MIC > 1.6325 µmol/mL) and miconazole (MIC value = 0.0188 µmol/mL) as a new anti-Candida albicans agent.
Keywords: synthesis; Mannich reaction; azoles; oxime esters; anti-Candida synthesis; Mannich reaction; azoles; oxime esters; anti-Candida

1. Introduction

Fungal infections have recently emerged as a growing threat to human health, especially in patients with weakened or compromised immune systems [1,2]. The organisms most often responsible for invasive fungal infection are Candida and Aspergillus species [3]. Candida infections are adverse in their manifestations, varying from superficial skin problems, chronic infection of the nails, mouth, throat or vagina to frequently fatal systemic diseases that involve the lungs, heart, gastrointestinal tract, central nervous system and other organs [4]. These infections are considered to be opportunistic in nature, since some aspect of the host’s defense system is impaired in some way. In spite of the large number of the available antifungal agents, the medical need is still largely unmet and therefore, efforts to discover new antifungal agents are a must. This is largely due to the perceived threat of emerging new pathogenic fungi and resistance of many strains to existing therapy [5,6,7].
Five major classes of the clinically used antifungal drugs are available, namely polyenes (such as amphotericin B and nystatin), echinocandins (such as caspofungin), allylamines (such as naftifine and terbinafine), fluoropyrimidines (such as 5-fluorocytosine) and azoles (such as miconazole, fluconazole and oxiconazole) (Figure 1) [8,9,10]. Azole antifungal drugs remain the mainstay of therapy for candidal life-threatening fungal infections due to their safety profile and high therapeutic index [11]. The mechanism of action of azole antifungals relies on their ability to inhibit Cyt-P450 dependent sterol 14α-demethylase through binding to the heme cofactor of the cytochrome CYP51 leading to inhibition of sterols synthesis in fungi [4,12].
Figure 1. Azole antifungal agents used in clinical therapy.
Figure 1. Azole antifungal agents used in clinical therapy.
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An evaluation of the literature exposed that some potent clinically used azole antifungals are derived from oxime-containing scaffolds [13]. Additionally, most of the available imidazole-containing antifungal agents have a two carbon atom spacer between the imidazole pharmacophore and an aromatic moiety, whereas limited information is available about imidazole-containing antifungals having a three-carbon atom linker between the imidazole pharmacophore and the aromatic moiety [14,15]. Moreover, Walker et al. reported that some aryl and aralkyl esters of 2-(1H-imidazol-1-yl)-1-phenylethanols displayed more anti-Candida albicans activity than miconazole [16].
Based upon the aforementioned premises, we became interested in the development of new imidazole-containing drug-like anti-Candida agents incorporating oxime functionality, exemplified by compounds 4ad as well as their respective aromatic esters, compounds 5al.

2. Results and Discussion

2.1. Chemistry

The pivotal ketones 3ad were prepared using the synthetic strategy outlined in Scheme 1. Thus, the appropriate acetophenone 1ad was reacted with dimethylamine hydrochloride and paraformaldehyde in the presence of a catalytic amount of concentrated hydrochloric acid to yield Mannich base hydrochlorides 2a–d. Imidazole was alkylated with the appropriate Mannich base 2ad to give ketones 3ad in good yields (Scheme 1).
Scheme 1. Synthesis of the ketones 3ad.
Scheme 1. Synthesis of the ketones 3ad.
Molecules 18 12208 g003
Reagents and conditions: (i) HN(CH3)2.HCl, (CH2O)n, conc. HCl, ethanol, reflux, 2 h; (ii) imidazole, water, reflux, 5 h.
Ketones 3ad were allowed to react with hydroxylamine hydrochloride in the presence of potassium hydroxide to yield oximes 4ad. X-ray crystallography is a decisive analytical tool which can confirm the configuration of the produced oximes 4ad. Accordingly, the assigned (E)-configuration of compounds 4ad was established via single crystal X-ray structure of the oxime 4a (Figure 2) [17].
The produced oximes 4ad were subjected to esterification with the appropriate carboxylic acid derivatives using ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) in the presence of 4-dimethylaminopyridine (DMAP) to yield the target compounds 5al (Scheme 2). The chemical structures of oximes 4ad and the title compounds 5al were confirmed via IR, 1H-NMR, 13C-NMR and mass spectral data.
Figure 2. ORTEP diagram of the title compound 4a drawn at 50% ellipsoids for non-hydrogen atoms.
Figure 2. ORTEP diagram of the title compound 4a drawn at 50% ellipsoids for non-hydrogen atoms.
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Scheme 2. Synthesis of the target compounds 5al.
Scheme 2. Synthesis of the target compounds 5al.
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Reagents and conditions: (i) H2NOH·HCl, KOH, ethanol, reflux, 18 h.; (ii) an appropriate carboxylic acid, EDCI·HCl, DMAP, DCM, rt, 18 h.

2.2. In Vitro Anti-Candida Activity and SARs

Fluconazole is the gold standard azole antifungal and used clinically as the first line of treatment for fungal infections, especially those caused by C. albicans. However, its extensive medical use has led to the emergence of resistance [18]. The in vitro anti-Candida activity of the synthesized imidazole-containing oximes 4ad and their respective aromatic esters 5al was evaluated against two clinical isolates of Candida, C. albicans and C. tropicalis, which are resistant to fluconazole (MIC > 1.6325 µmol/mL). The test compounds 4ad and 5al incorporate a three-carbon atom bridge between the imidazole pharmacophore and the aromatic moiety to gain insight about anti-Candida activity of this type of compounds. The anti-Candida activities, expressed as diameter of the inhibition zone (DIZ) and minimum inhibition concentration (MIC) for the oximes 4ad, the target compounds 5al as well as for the reference drugs fluconazole and miconazole, are summarized in Table 1.
Table 1. Anti-Candida activity of oximes 4ad and the target compounds 5alagainst Candida albicans and Candida tropicalis.
Table 1. Anti-Candida activity of oximes 4ad and the target compounds 5alagainst Candida albicans and Candida tropicalis.
Compound NoCandida albicansCandida tropicalis
DIZ ± SD *MIC (µmol/mL) **DIZ ± SD *MIC (µmol/mL) **
4a21 ± 1.00.580720 ± 0.50.5807
4b13 ± 0.60.50198 ± 1.00.5019
4c9 ± 1.150.50998 ± 1.00.2549
4d8 ± 1.00.54568 ± 1.00.5456
5a11 ± 1.20.39198 ± 1.00.7837
5b18 ± 1.10.080512 ± 1.00.6439
5c16 ± 0.40.325716 ± 1.20.3257
5d18 ± 0.90.169914 ± 0.50.3398
5e16 ± 0.90.370814 ± 0.60.3708
5f7 ± 1.00.37528 ± 1.00.1876
5g13 ± 0.60.645414 ± 1.00.3227
5h24 ± 1.10.011217 ± 1.00.3582
5i17 ± 1.10.305314 ± 0.50.3053
5j25 ± 1.00.005425 ± 1.20.1767
5k20 ± 0.90.022114 ± 0.50.7069
5l12 ± 1.00.706912 ± 0.70.3535
Fluconazole15 ± 0.5>1.632516 ± 0.5>1.6325
Miconazole38 ± 1.10.018824 ± 0.50.0024
* The arithmetic mean of the inhibition zone diameters in mean ± standard deviation in mm. ** The lowest concentration of the compound that produced 80% microbial growth inhibition (μmol/mL).
The preliminary anti-Candida potential of the test compounds 4ad and 5al was evaluated using the DIZ assay and the results are presented in Table 1. The test compounds displayed a promising anti-Candida activity (DIZ = 7–25 mm) where compound 5j was the most active congener (DIZ = 25 ± 1 and 25 ± 1.2 mm against C. albicans and C. tropicalis, respectively).
The oxime 4a exhibited good anti-Candida activity (MIC value = 0.5807 µmol/mL) toward both C. albicans and C. tropicalis, being more potent than fluconazole (MIC value > 1.6325 µmol/mL) but weaker than miconazole (MIC value = 0.0188 and 0.0024 µmol/mL for C. albicans and C. tropicalis, respectively). Substitution of the aromatic ring of 4a with substituents endowed with different electronic and steric properties like chloro, methoxy and/or methyl groups gave compounds 4b, 4c and 4d, respectively, aiming to enhance its anti-Candida activity. Unfortunately, the anti-Candida activity of 4a did not improve significantly, except for compound 4c toward C. tropicalis (MIC value = 0.2549 µmol/mL).
De Vita et al. reported that the presence of a second aromatic ring could enhance the antifungal activity of azoles [19]. Consequently, the respective aromatic esters 5al of the oximes 4ad were prepared and biologically evaluated as new anti-Candida agents. Esterification of the hydroxyl group of 4a with benzoic acid gave compound 5a which displayed better anti-Candida activity (MIC value = 0.3919 µmol/mL) than that of 4a toward C. albicans. Moreover, esterification of the hydroxyl group of the oximes 4bd with 4-chlorobenzoic acid gave the respective esters 5bd. Compounds 5bd showed better anti-Candida profile than their respective oximes 4bd, where compound 5b is the most active congener with a MIC value of 0.0805 µmol/mL toward C. albicans.
Substitution of the aromatic ester functionality of 5a with fluoride, methyl and/or trifluoromethyl groups gave compounds 5eg which exhibited anti-Candida albicans activity comparable with that of 5a, except for compound 5g (MIC value = 0.6454 µmol/mL) which was weaker than 5a. Compounds 5eg displayed better anti-Candida tropicalis profiles than that of 5a where compound 5f is the most active candidate with a MIC value of 0.1876 µmol/mL.
Substitution of the aromatic ester functionality of 5a with a group endowed with negative inductive effect and positive mesomeric properties like a methoxy group gave compound 5h, which showed a comparable anti-Candida albicans profile (MIC value = 0.0112 µmol/mL) with that of miconazole (MIC value = 0.0188 µmol/mL) and was about 145-fold more potent than fluconazole (MIC > 1.6325 µmol/mL). This result encouraged us to synthesize the trimethoxy analogue of 5a, compound 5i. Unfortunately, 5i exhibited weaker anti-Candida albicans (MIC value = 0.3053 µmol/mL) than that of 5h.
Compound 5j emerged as the most active congener in the whole series of synthesized compounds against both C. albicans and C. tropicalis. Compound 5j, the 4-chloro analogue of 5a, exhibited about 3.5-fold and 300-fold more potency than miconazole and the gold standard azole antifungal, fluconazole, respectively, as a drug-like anti-Candida albicans agent. The positional isomers of compound 5j, compounds 5k and 5l, displayed weaker anti-Candida activity than that of 5j.

3. Experimental

3.1. Chemistry

3.1.1. General

Melting points were determined on a Gallenkamp melting point apparatus, and are uncorrected. Infrared (IR) spectra were recorded as KBr disks using the Perkin Elmer FT-IR Spectrum BX apparatus. NMR spectra were carried out on a Bruker NMR spectrometer operating at 500 MHz for 1H and 125.76 MHz for 13C at the Research Center, College of Pharmacy, King Saud University, Saudi Arabia. TMS was used as internal standard and chemical shift values were recorded in ppm on δ scale. The 1H-NMR data were represented as follows: chemical shifts, multiplicity (s. singlet, d. doublet, t. triplet, m. multiplet, br. broad) and number of protons. The 13C-NMR data were represented as chemical shifts and type of carbon. Mass spectra were measured on Agilent Triple Quadrupole 6410 QQQ LC/MS with an electrospray ionization (ESI) source. Silica gel thin layer chromatography (TLC) plates from Merck (silica gel precoated aluminium plates with a 245 nm fluorescent indicator) were used for thin layer chromatography. Visualization was performed by illumination with UV light source (254 nm). Column chromatography was carried out on silica gel 60 (0.063–0.200 mm) obtained from Merck and chloroform/methanol (9:0.5) was used as a solvent system.

3.1.2. General Procedure for Preparation of the Ketones 3ad

The appropriate acetophenone 1ad (200 mmol), dimethylamine hydrochloride (270 mmol) and paraformaldehyde (90 mmol) were heated to reflux in absolute ethanol (35 mL) in the presence of catalytic amount of concentrated hydrochloric acid (0.5 mL). Reflux of the reaction mixture was continued under stirring for two hours, cooled and acetone (200 mL) was added. The formed Mannich base hydrochlorides 2ad were precipitated, filtered off and dried. Subsequently, compounds 2ad (100 mmol) were dissolved in water (100 mL) and imidazole (200 mmol) was added. The reaction mixture was heated to reflux for five hours, cooled and the precipitated solids were collected by filtration to give ketones 3ad which were pure enough to be used in the next step.
3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one (3a). Synthesis of 3a was previously reported [14].
1-(4-Chlorophenyl)-3-(1H-imidazol-1-yl)propan-1-one (3b). Synthesis of 3b was previously reported [15].
3-(1H-Imidazol-1-yl)-1-(4-methoxyphenyl)propan-1-one (3c). Synthesis of 3c was previously reported [20].
3-(1H-Imidazol-1-yl)-1-(4-methylphenyl)propan-1-one (3d). Synthesis of 3d was previously reported [21].

3.1.3. General Procedure for Preparation of the Oximes 4ad

A mixture of the appropriate ketone 3ad (10 mmol), hydroxylamine hydrochloride (20 mmol), and KOH (20 mmol) in ethanol (10 mL) was heated to reflux under stirring for 18 h. The reaction mixture was allowed to cool to room temperature and the insoluble solids were filtered off. The filtrate was concentrated under vacuum and the residue was poured onto ice-cold water (15 mL). The precipitated solids were collected by filtration and dried to give oximes 4ad which were subsequently subjected to the esterification step without any further purification.
(1E)-N-Hydroxy-3-(1H-imidazol-1-yl)-1-phenylpropan-1-imine (4a). [17] Yield 70%; colourless solid mp. 155–157 °C (ethanol); IR (KBr): ν (cm−1) 3508 (OH), 3149, 3002, 2703, 1644 (C=N), 1600, 1573, 1221, 758; 1H-NMR (CDCl3): δ 3.31 (t, J = 7.1 Hz, 2H, -CH2-CH2-N), 4.28 (t, J = 7.1 Hz, 2H, -CH2-CH2-N), 6.96 (s, 1H, -N-CH=CH-N=), 7.07 (s, 1H, -N-CH=CH-N=), 7.29–7.49 (m 5H, Ar-H), 7.58 (s, 1H, -N-CH=N-); 13C-NMR (CDCl3): δ 28.3 (-CH2-CH2-N), 41.8 (-CH2-CH2-N), 119.1 (-N-CH=CH-N=), 126.1, 128.8, 128.9 (-N-CH=CH-N=, Ar-CH), 135.1, 137.0 (Ar-C), 139.5 (-N-CH=N-), 155.4 (C=N-OH); MS m/z (ESI): 216.0 [M + 1]+.
(1E)-1-(4-Chlorophenyl)-N-hydroxy-3-(1H-imidazol-1-yl)propan-1-imine (4b). The synthesis and characterization of 4b were previously reported [22].
(1E)-N-Hydroxy-3-(1H-imidazol-1-yl)-1-(4-methoxyphenyl)propan-1-imine (4c). Yield 65%; pale yellow solid mp. 136–138 °C (ethanol); IR (KBr): ν (cm−1) 3512 (OH), 3135, 3026, 2632, 1648 (C=N), 1680, 1566, 1228, 752; 1H-NMR (CDCl3): δ 3.26 (t, J = 6.5 Hz, 2H, -CH2-CH2-N), 3.83 (s, 3H, OCH3), 4.28 (t, J = 7.1 Hz, 2H, -CH2-CH2-N), 6.89 (d, J = 7.5 Hz, 2H, Ar-H), 6.97 (s, 1H, -N-CH=CH-N=), 7.08 (s, 1H, -N-CH=CH-N=), 7.44 (d, J = 7.5 Hz, 2H, Ar-H), 7.58 (s, 1H, -N-CH=N-); 13C-NMR (CDCl3): δ 28.9 (-CH2-CH2-N), 43.6 (-CH2-CH2-N), 55.4 (OCH3), 114.0 (Ar-CH), 119.1 (-N-CH=CH-N=), 127.4, 127.9, 129.1 (-N-CH=CH-N=, Ar-CH, Ar-C), 137.1 (-N-CH=N-), 155.4 (C=N-OH), 160.5 (Ar-C); MS m/z (ESI): 246.0 [M + 1]+.
(1E)-N-Hydroxy-3-(1H-imidazol-1-yl)-1-(4-methylphenyl)propan-1-imine (4d). Yield 65%; white solid mp. 147–149 °C (ethanol); IR (KBr): ν (cm−1) 3509 (OH), 3119, 2702, 1639 (C=N), 1679, 1512, 1230, 738; 1H-NMR (CDCl3): δ 2.27 (s, 3H, CH3), 3.18 (t, J = 7.0 Hz, 2H, -CH2-CH2-N), 4.18 (t, J = 7.0 Hz, 2H, -CH2-CH2-N), 6.88 (s, 1H, -N-CH=CH-N=), 6.99 (s, 1H, -N-CH=CH-N=), 7.08 (d, J = 7.8 Hz, 2H, Ar-H), 7.30 (d, J = 7.9 Hz, 2H, Ar-H), 7.49 (s, 1H, -N-CH=N-); 13C-NMR (CDCl3): δ 21.3 (CH3), 28.9 (-CH2-CH2-N), 43.6 (-CH2-CH2-N), 119.1 (-N-CH=CH-N=), 125.9, 129.1, 129.4 (-N-CH=CH-N=, Ar-CH), 132.6, 137.1, 139.4 (-N-CH=N-, Ar-C), 154.7 (C=N-OH); MS m/z (ESI): 230.0 [M + 1]+.

3.1.4. General Procedure for the Synthesis of the Target Oxime Esters 5al

A solution of the appropriate carboxylic acid (7 mmol) and EDCI·HCl (7.3 mmol) was stirred in DCM (75 mL) in the presence of DMAP (400 mg). The appropriate oxime 4ad (6.9 mmol) was added to the stirred reaction mixture and stirring was continued for further 18 h at room temperature. The reaction mixture was washed successively with water (2 × 20 mL), 10% NaHCO3 solution (2 × 15 mL), and water (2 × 15 mL). The organic layer was separated, dried (Na2SO4) and evaporated under reduced pressure and the residue was purified either by recrystallisation (for solids) or by column chromatography (for oils).
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-benzoyl oxime (5a). Yield 41%; colourless viscous oil; IR (KBr): ν (cm−1) 3115, 2943, 1746 (C=O), 1650 (C=N), 1510, 1243, 735; 1H-NMR (CDCl3): δ (ppm) = 3.38 (t, J = 7.1 Hz, 2H, -CH2-CH2-N), 4.21 (t, J = 7.1 Hz, 2H, -CH2-CH2-N), 6.84 (s, 1H,-N-CH=CH-N=), 6.95 (s, 1H, -N-CH=CH-N=), 7.36-7.46 (m, 6H, -N-CH=N-, Ar-H), 7.55-7.60 (m, 2H, Ar-H), 7.96 (d, J = 7.6 Hz, 2H, Ar-H.); 13C-NMR (CDCl3): δ 31.0 (-CH2-CH2-N), 43.7 (-CH2-CH2-N), 118.8 (-N-CH=CH-N=), 127.3, 128.6, 128.8, 129.1, 129.6, 130.0, 131.3 (-N-CH=CH-N=, Ar-CH, Ar-C), 133.0, 133.8, 136.9 (-N-CH=N-, Ar-CH, Ar-C), 163.4 (C=N), 163.5 (C=O); MS m/z (ESI): 320.1 [M + 1]+.
(E)-1-(4-Chlorophenyl)-3-(1H-imidazol-1-yl)propan-1-one O-4-chlorobenzoyl oxime (5b). Yield 56%; white solid mp. 132–134 °C (isopropanol); IR (KBr): ν (cm−1) 3107, 1744 (C=O), 1650 (C=N), 1560, 1513, 1261, 748; 1H-NMR (CDCl3): δ (ppm) = 3.43 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 4.29 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 6.89 (s, 1H, -N-CH=CH-N=), 7.02 (s, 1H, -N-CH=CH-N=), 7.41 (d, J = 8.5 Hz, 2H, Ar-H.), 7.49–7.51 (m, 3H, -N-CH=N-, Ar-H), 7.60 (d, J = 8.5 Hz, 2H, Ar-H.), 7.94 (d, J = 8.5 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 30.8 (-CH2-CH2-N), 43.7 (-CH2-CH2-N), 118.8 (-N-CH=CH-N=), 126.9, 128.5, 129.2, 129.4, 130.0, 130.9, 131.2, (-N-CH=CH-N=, Ar-CH, Ar-C), 136.9, 137.8, 140.4 (-N-CH=N-, Ar-C), 162.5 (C=N), 162.6 (C=O); MS m/z (ESI): 388.0 [M ]+.
(E)-3-(1H-Imidazol-1-yl)-1-(4-methoxyphenyl)propan-1-one O-4-chlorobenzoyl oxime (5c). Yield 70%; white solid mp. 131–133 °C (isopropanol); IR (KBr): ν (cm−1) 3123, 2366, 1758 (C=O), 1684 (C=N), 1564, 1514, 1252, 747; 1H-NMR (CDCl3): δ (ppm) = 3.44 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 3.87 (OCH3), 4.32 (t, J = 7.0 Hz, 2H, -CH2-CH2-N), 6.92 (s, 1H, -N-CH=CH-N=), 6.96 (d, J = 8.8 Hz, 2H, Ar-H.), 7.09 (s, 1H, -N-CH=CH-N=), 7.49 (d, J = 8.6 Hz, 2H, Ar-H.), 7.67 (d, J = 9.0 Hz, 2H, Ar-H.), 7.77 (s, 1H, -N-CH=N-), 7.94 (d, J = 8.6 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 30.5 (-CH2-CH2-N), 44.2 (-CH2-CH2-N), 55.5 (OCH3), 114.5 (Ar-CH), 119.1 (-N-CH=CH-N=), 128.5, 128.9, 129.2, 130.9, 131.2, (-N-CH=CH-N=, Ar-CH, Ar-C), 136.8, 140.2 (-N-CH=N-, Ar-C), 162.3, 162.8, 162.9 (C=N, C=O, Ar-C); MS m/z (ESI): 384.2 [M + 1]+
(E)-3-(1H-Imidazol-1-yl)-1-(4-methylphenyl)propan-1-one O-4-chlorobenzoyl oxime (5d). Yield 58%; white solid mp. 142–144 °C (isopropanol); IR (KBr): ν (cm−1) 3065, 1744 (C=O), 1654 (C=N), 1646, 1559, 1508, 1254, 749; 1H-NMR (CDCl3): δ (ppm) = 2.42 (s, 3H, CH3), 3.46 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 4.31 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 6.91 (s, 1H, -N-CH=CH-N=), 7.09 (s, 1H, -N-CH=CH-N=), 7.27 (d, J = 7.9 Hz, 2H, Ar-H.), 7.49 (d, J = 8.5 Hz, 2H, Ar-H.), 7.62 (d, J = 8.0 Hz, 2H, Ar-H.), 7.79 (s, 1H, -N-CH=N-), 7.94 (d, J = 8.5 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 21.5 (CH3), 30.6 (-CH2-CH2-N), 44.2 (-CH2-CH2-N), 119.1 (-N-CH=CH-N=), 127.2, 128.4, 128.5, 129.2, 129.9, 130.9, 131.2, (-N-CH=CH-N=, Ar-CH, Ar-C), 136.8, 140.2, 142.1 (-N-CH=N-, Ar-C.), 162.7 (C=N), 163.3 (C=O); MS m/z (ESI): 368.2 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-4-fluorobenzoyl oxime (5e). Yield 62%; pale yellow solid mp. 114–116 °C (isopropanol); IR (KBr): ν (cm−1) 3115, 2848, 1746 (C=O), 1660 (C=N), 1571, 1249, 739; 1H-NMR (CDCl3): δ (ppm) = 3.46 (t, J = 7.0 Hz, 2H, -CH2-CH2-N), 4.29 (t, J = 7.0 Hz, 2H, -CH2-CH2-N), 6.91 (s, 1H, -N-CH=CH-N=), 7.02 (s, 1H, -N-CH=CH-N=), 7.18–7.21 (m, 2H, Ar-H), 7.43–7.52 (m, 4H, -N-CH=N-, Ar-H), 7.68 (d, J = 7.4 Hz, 2H, Ar-H.), 8.03–8.05 (m, 2H, Ar-H); 3C-NMR (CDCl3): δ 30.9 (-CH2-CH2-N), 43.8 (-CH2-CH2-N), 116.1 (d, J = 22.1 Hz, Ar-CH), 118.8 (-N-CH=CH-N=), 124.9, (d, J = 2.6 Hz, Ar-C), 127.3, 129.1, 129.8, 131.4 (-N-CH=CH-N=, Ar-CH), 132.2 (d, J = 9.4 Hz, Ar-CH), 132.8, 136.9 (-N-CH=N-, Ar-C), 162.5 (C=N), 163.5 (C=O), 167.1 (d, J = 254.0 Hz, Ar-C); MS m/z (ESI): 338.2 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-4-methylbenzoyl oxime (5f). Yield 62%; pale yellow solid mp. 125–127 °C (isopropanol); IR (KBr): ν (cm−1) 3115, 2964, 1736 (C=O), 1647 (C=N), 1605, 1506, 1248, 750; 1H-NMR (CDCl3): δ (ppm) = 2.37 (s, 3H, CH3), 3.36 (t, 2H, J = 7.1 Hz, -CH2-CH2-N), 4.21 (t, 2H, J = 7.0 Hz -CH2-CH2-N), 6.84 (s, 1H, -N-CH=CH-N=), 6.94 (s, 1H, -N-CH=CH-N=), 7.23 (d, J = 7.8 Hz, 2H, Ar-H), 7.35–7.41 (m, 4H, -N-CH=N-, Ar-H), 7.59 (d, J = 7.0 Hz, 2H, Ar-H.), 7.85 (d, J = 8.0 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 21.8 (CH3), 31.1 (-CH2-CH2-N), 43.7 (-CH2-CH2-N), 118.8 (-N-CH=CH-N=), 125.8, 127.2, 129.0, 129.5, 129.6, 130.1, 131.2, 133.1 (-N-CH=CH-N=, Ar-CH, Ar-C), 136.9 (-N-CH=N-), 144.7 (Ar-C), 163.2 (C=N), 163.5 (C=O); MS m/z (ESI): 334.0 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-4-(trifluoromethyl)benzoyl oxime (5g). Yield 39%; white solid mp. 125–127 °C (isopropanol); IR (KBr): ν (cm−1) 3050, 2360, 1750 (C=O), 1653 (C=N), 1559, 1507, 1264, 737; 1H-NMR (CDCl3): δ (ppm) = 3.47–3.49 (m, 2H, -CH2-CH2-N), 4.29–4.32 (m, 2H, -CH2-CH2-N), 6.91 (s, 1H, -N-CH=CH-N=), 7.05 (s, 1H, -N-CH=CH-N=), 7.36–7.55 (m, 4H, -N-CH=N-, Ar-H), 7.71–7.72 (m, 2H, Ar-H), 7.79 (d, J = 8.0 Hz, 2H, Ar-H.), 8.13 (d, J = 8.0 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 30.8 (-CH2-CH2-N), 43.8 (-CH2-CH2-N), 118.8 (-N-CH=CH-N=), 125.8 (d, J = 3.4 Hz, CF3), 125.9, 127.3, 128.6, 129.2, 129.8, 130.0, 131.6, 132.6 (-N-CH=CH-N=, Ar-CH, Ar-C), 135.6, 136.9 (-N-CH=N-, Ar-C), 162.4 (C=N), 164.0 (C=O); MS m/z (ESI): 388.1 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-4-methoxybenzoyl oxime (5h). Yield 40%; off white solid mp. 108–110 °C (isopropanol); IR (KBr): ν (cm−1) 3117, 2968, 1735 (C=O), 1650 (C=N), 1602, 1507, 1248, 765; 1H-NMR (CDCl3): δ (ppm) = 3.46 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 3.91 (s, 3H, OCH3), 4.30 (t, J = 6.9 Hz, 2H, -CH2-CH2-N), 6.94 (s, 1H, -N-CH=CH-N=), 7.01 (d, J = 8.8 Hz, 2H, Ar-H), 7.10 (s, 1H, -N-CH=CH-N=), 7.45–7.50 (m, 4H, -N-CH=N-, Ar-H), 7.67 (d, J = 7.0 Hz, 2H, Ar-H.), 8.01 (d, J = 8.8 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 31.0 (-CH2-CH2-N), 43.7 (-CH2-CH2-N), 55.6 (OCH3), 114.1 (Ar-CH), 118.8 (-N-CH=CH-N=), 120.7, 127.2, 129.0, 130.0, 131.2, 131.7, 133.1 (-N-CH=CH-N=, Ar-CH, Ar-C), 136.9 (-N-CH=N-), 162.9 (C=N), 163.2 (Ar-C), 164.0 (C=O); MS m/z (ESI): 350.0 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-3,4,5-trimethoxybenzoyl oxime (5i). Yield 53%; white solid mp. 135–137 °C (isopropanol); IR (KBr): ν (cm−1) 3103, 2938, 1742 (C=O), 1645 (C=N), 1593, 1503, 1231, 749; 1H-NMR (CDCl3): δ (ppm) = 3.45 (t, J = 6.8 Hz, 2H, -CH2-CH2-N), 3.92 (s, 6H, 2 × OCH3), 3.94 (s, 3H, OCH3), 4.28 (t, J = 6.8 Hz, 2H, -CH2-CH2-N), 6.91 (s, 1H, -N-CH=CH-N=), 7.02 (s, 1H, -N-CH=CH-N=), 7.27–7.50 (m, 6H, -N-CH=N-, Ar-H), 7.67 (d, J = 7.0 Hz, 2H, Ar-H.);13C-NMR (CDCl3): δ 30.9 (-CH2-CH2-N), 43.6 (-CH2-CH2-N), 56.5 (2 × OCH3), 61.0 (OCH3), 106.9 (Ar-CH), 118.6 (-N-CH=CH-N=), 123.5, 127.3, 129.1, 130.1, 131.3, 132.9 (-N-CH=CH-N=, Ar-CH, Ar-C), 136.8 (-N-CH=N-), 142.9, 153.2 (Ar-C), 163.2 (C=N), 163.6 (C=O); MS m/z (ESI): 410.1 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-4-chlorobenzoyl oxime (5j). Yield 54%; colourless crystals mp. 126–128 °C (isopropanol); IR (KBr): ν (cm−1) 3115, 2970, 1743 (C=O), 1648 (C=N), 1508, 1249, 736; 1H-NMR (CDCl3): δ (ppm) = 3.44 (t, J = 6.7 Hz, 2H, -CH2-CH2-N), 4.27 (t, J = 6.7 Hz, 2H, -CH2-CH2-N), 6.90 (s, 1H, -N-CH=CH-N=), 7.02 (s, 1H, -N-CH=CH-N=), 7.45–7.50 (m, 6H, -N-CH=N-, Ar-H), 7.68 (d, J = 8.4 Hz, 2H, Ar-H.), 7.95 (d, J = 8.4 Hz, 2H, Ar-H); 13C-NMR (CDCl3): δ 30.9 (-CH2-CH2-N), 43.7 (-CH2-CH2-N), 118.7 (-N-CH=CH-N=), 127.1, 127.3, 129.1, 129.2, 130.1, 130.9, 131.4, 132.8 (-N-CH=CH-N=, Ar-CH, Ar-C), 136.9 (-N-CH=N-), 140.2 (Ar-C.), 162.7 (C=N), 163.7 (C=O); MS m/z (ESI): 354.1 [M + 1]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-3-chlorobenzoyl oxime (5k). Yield 61%; pale yellow viscous oil; IR (KBr): ν (cm−1) 3113, 1751 (C=O), 1654 (C=N), 1510, 1282, 739; 1H-NMR (DMSO-d6): δ (ppm) = 3.52 (br. s, 2H, -CH2-CH2-N), 4.30 (br. s, 2H, -CH2-CH2-N), 6.80 (s, 1H, -N-CH=CH-N=), 7.17 (s, 1H, -N-CH=CH-N=), 7.51–7.82 (m, 8H, -N-CH=N-, Ar-H), 8.00 (d, J = 1.5 Hz, 2H, Ar-H.); 13C-NMR (DMSO-d6): δ 30.1 (-CH2-CH2-N), 43.0 (-CH2-CH2-N), 119.3 (-N-CH=CH-N=), 127.3, 128.1, 128.5, 128.9, 130.3, 130.9, 131.1, 132.9, 133.6, 133.7 (-N-CH=CH-N=, Ar-CH, Ar-C), 137.1 (-N-CH=N-), 161.7 (C=N), 164.8 (C=O); MS m/z (ESI): 354.1 [M]+.
(E)-3-(1H-Imidazol-1-yl)-1-phenylpropan-1-one O-2-chlorobenzoyl oxime (5l). Yield 60%; white solid mp. 118–120 °C (isopropanol); IR (KBr): ν (cm−1) 3054, 1763 (C=O), 1658 (C=N), 1640, 1511, 1265, 739; 1H-NMR (DMSO-d6): δ (ppm) = 3.45 (br. s, 2H, -CH2-CH2-N), 4.25 (br. s, 2H, -CH2-CH2-N), 6.80 (s, 1H, -N-CH=CH-N=), 7.08 (s, 1H, -N-CH=CH-N=), 7.51-7.67 (m, 7H, -N-CH=N-, Ar-H), 7.75 (d, J = 6.9 Hz, 2H, Ar-H.), 7.90 (d, J = 7.2 Hz, 1H, Ar-H.); 13C-NMR (DMSO-d6): δ 30.2 (-CH2-CH2-N), 43.0 (-CH2-CH2-N), 119.2 (-N-CH=CH-N=), 127.3, 127.6, 128.5, 128.8, 128.9, 130.9, 131.1, 131.3, 131.9, 132.8, 133.7 (-N-CH=CH-N=, Ar-CH, Ar-C), 137.1 (-N-CH=N-), 162.2 (C=N), 164.6 (C=O); MS m/z (ESI): 354.1 [M]+.

3.2. Anti-Candida Activity

3.2.1. Anti-Candida Agents

Miconazole was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and fluconazole from Shouguang-Fukang Pharmaceutical Ltd. (Shandong, China). The antifungal discs (containing 25 µg fluconazole and/or 10 µg miconazole) were purchased from ROSCO (Neo-Sensitabs, Taastrup, Denmark).
Dimethyl sulfoxide (100%) was used to dissolve stock solutions of miconazole, fluconazole and/or the synthesized compounds 4ad and 5al to obtain an initial concentration of 1000 µg/mL. These stock solutions were then diluted to the desired concentration with sterile distilled water. Miconazole and fluconazole antifungal discs were stored at –80 °C until used.

3.2.2. Media

Liquid RPMI 1640 medium supplemented with L-glutamine was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and was added to 2% sodium bicarbonate and 0.165 M morpholine- propane sulfonic acid (MOPS) from Dojindo Laboratories (Kumamoto, Japan) then adjusted to pH 7.0. Sabouraud Dextrose Agar (SDA) and Brain Heart Infusion Broth (BHI) from Difco Laboratories (Detroit, MI, USA). Potato dextrose agar (PDA) was purchased from Eiken Chemical Co. Ltd. (Tokyo, Japan).

3.2.3. Organisms

Two clinical isolates of Candida species were obtained from King Khaled Hospital, Riyadh, Saudi Arabia. One was identified as C. albicans and the other as C. tropicalis. The yeasts were stored at –70 °C in BHI with glycerol 5% until tested.

3.2.4. Preparation of Inocula

Preparation of inocula for the broth microdilution testing was performed in accordance with CLSI documents M27-A2 [23] with RPMI 1640 medium. Yeast isolates were subcultured at 35 °C for 48 h on PDA plates. Candida cells were then recovered and suspended in 5 mL of sterile saline. The turbidity of each suspension was adjusted to a 0.5 McFarland standard (corresponding to 1–3 × 106 to 5–3 × 106 CFU/mL) at a wavelength of 530 nm according to the reported method [23]. Each suspension was diluted 1,000-fold with sterile RPMI 1640 medium to give a final inoculum of 1–3 × 103 to 5–3× 103 CFU/mL.

3.2.5. Disk Diffusion Assay

The disk diffusion assay was performed as described previously [24]. Colonies obtained from the Candida strains under test were suspended in sterile saline and adjusted to a 0.5 McFarland standard (corresponding to 5 × 106 CFU/mL). An aliquot of 100 µL of each yeast suspension was spread uniformly onto SDA plates. Six mm Whatmann filter paper disks were impregnated with 1000 µg of the synthesized compounds 4ad and 5al and were allowed to dry. Then they were placed onto the surface of the inoculated agar plates together with the standard antifungal discs which were then incubated at 35 °C. Diameters of inhibition zones were measured at 24 h.

3.2.6. Antifungal Susceptibility Studies

The MIC of the reference standards and/or the synthesized compounds 4ad and 5al were determined with a microdilution test (M27-A2 Protocol), according to the reference method of the CLSI. The previously prepared yeast inocula (100 µL) were added to each well of 96-well flat-bottom microdilution plates; each well contained 100 µL of twofold serial dilutions of the standard or the synthesized compounds 4ad and 5al ranging from 1 µg/mL to 500 µg/mL in RPMI 1640 medium. Readings were measured at 490 nm with a microplate ELISA reader after each plate was incubated at 35 °C for 48 h. The MICs for the reference standards and/or the synthesized compounds were determined with 80% growth inhibition at the end point relative to the turbidity of the growth control.

4. Conclusions

Anti-Candida activities of certain new imidazole-containing oximes 4ad and their respective aromatic esters 5al have been reported. The synthesized compounds 4ad and 5al exhibited anti-Candida activity better than that of the gold standard antifungal drug, fluconazole. Compound 5j emerged as the most active congener among the all synthesized compounds, being about 3.5-fold and 300-fold more potent than miconazole and fluconazole, respectively. Compound 5j could be considered as a prodrug and could serve as a new lead for anti-Candida agents.


The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding of this research through the Research Group Project no. RGP-VPP-196.

Conflicts of Interest

The authors have declared that there is no conflict of interests.


  1. Imwidthaya, P.; Poungvarin, N. Cryptococcosis in AIDS. Postgrad. Med. J. 2000, 76, 85–88. [Google Scholar] [CrossRef]
  2. Sutton, D.A.; Fothergill, A.W.; Rinaldi, M.G. Guide to Clinically Significant Fungi, 1st ed.; Williams and Wilkins: Baltimore, MD, USA, 1998. [Google Scholar]
  3. Saral, R. Candida and Aspergillus infections in immunocompromised patients: An overview. Rev. Infec. Dis. 1991, 13, 487–492. [Google Scholar] [CrossRef]
  4. Odds, F.; Brown, A.J.P.; Gow, N.A.R. Antifungal agents: Mechanisms of action. Trends Microbiol. 2003, 11, 272–279. [Google Scholar] [CrossRef]
  5. Joseph-Horne, T.; Hollomon, D.W. Molecular mechanisms of azole resistance in fungi. FEMS Microbiol. Lett. 1997, 149, 141–149. [Google Scholar] [CrossRef]
  6. Vanden Bossche, H.; Dromer, F.; Improvisi, I.; Lozano-Chiu, M.; Rex, J.H.; Sanglards, D. Antifungal drug resistance in pathogenic fungi. Med. Mycol. 1998, 36, 119–128. [Google Scholar]
  7. Marichal, P.; Vanden Bossche, H. Mechanisms of resistance to azole antifungals. Acta Biochim. Pol. 1995, 42, 509–516. [Google Scholar]
  8. De Luca, L. Naturally occurring and synthetic imidazoles: Their chemistry and their biological activities. Curr. Med. Chem. 2006, 13, 1–23. [Google Scholar]
  9. Pennisi, M.; Antonelli, M. Clinical aspects of invasive candidiasis in critically III patients. Drugs 2009, 69, 21–28. [Google Scholar] [CrossRef]
  10. Chen, S.C.; Playford, E.G.; Sorrell, T.C. Antifungal therapy in invasive fungal infections. Curr. Opin. Pharmacol. 2010, 10, 522–530. [Google Scholar] [CrossRef]
  11. Wang, W.; Sheng, C.; Che, X.; Ji, H.; Miao, Z.; Yao, J.; Zhang, W. Design, synthesis, and antifungalactivity of novel conformationally restricted triazole derivatives. Arch. Pharm. 2009, 342, 732–739. [Google Scholar] [CrossRef]
  12. Hamdan, J.S.; Hahn, R.C. Antifungal drugs for systemic mycosis: An overview of mechanism of action and resistance. Anti-Infective Agents Med. Chem. 2006, 5, 403–412. [Google Scholar] [CrossRef]
  13. Rossello, A.; Bertini, S.; Lapucci, A.; Macchia, M.; Martinelli, A.; Rapposelli, S.; Herreros, E.; Macchia, B. Synthesis, Antifungal activity, and molecular modeling studies of new inverted oxime ethers of oxiconazole. J. Med. Chem. 2002, 45, 4903–4912. [Google Scholar] [CrossRef]
  14. Aboul-Enein, M.N.; El-Azzouny, A.A.; Attia, M.I.; Saleh, O.A.; Kansoh, A.L. Synthesis and Anti-Candida Potential of Certain Novel 1-[(3-Substituted-3-phenyl)propyl]-1H-imidazoles. Arch. Pharm. 2011, 344, 794–801. [Google Scholar] [CrossRef]
  15. Roman, G.; Mares, M.; Nastasa, V. A novel antifungal agent with broad spectrum: 1-(4-Biphenylyl)-3-(1H-imidazol-1-yl)-1-propanone. Arch. Pharm. 2013, 346, 110–118. [Google Scholar] [CrossRef]
  16. Walker, K.A.; Hirschfeld, D.R.; Marx, M. Antimycotic imidazoles. 2. Synthesis and antifungal properties of esters of 1-[2-hydroxy(mercapto)-2-phenylethyl]-1H-imidazoles. J. Med. Chem. 1978, 21, 1335–1338. [Google Scholar] [CrossRef]
  17. Fun, H.-K.; Quah, C.K.; Attia, M.I.; Almutairi, M.S.; Ghoneim, S.W. (E)-N-[3-(Imidazol-1-yl)-1-phenylpropylidene]hydroxylamine. Acta Crystallogr. E-Struct. Rep. 2012, E68, o627. [Google Scholar]
  18. Pfaller, M.A.; Andes, D.; Diekema, D.J.; Espinel-Ingroff, A.; Sheehan, D. Wild-type MIC distributions, epidemiological cutoff values and species-specific clinical breakpoints for fluconazole and Candida: Time for harmonization of CLSI and EUCAST broth microdilution methods. Drug Resist. Update. 2010, 13, 6180–6195. [Google Scholar]
  19. De Vita, D.; Scipione, L.; Tortorella, S.; Mellini, P.; Di Rienzo, B.; Simonetti, G.; D’Auria, F.D.; Panella, S.; Cirilli, R.; Di Santo, R.; et al. Synthesis and antifungal activity of a new series of 2-(1H-imidazol-1-yl)-1-phenylethanol derivatives. Eur. J. Med. Chem. 2012, 49, 334–342. [Google Scholar] [CrossRef]
  20. Wan, J.; Peng, Z.-Z.; Li, X.-M.; Ouyang, P.-K.; Zhang, S.-S. 3-(1H-Imidazol-1-yl)-1-(4-methoxyphenyl)propan-1-one. Acta Crystallogr. E-Struct. Rep. 2005, E 61, o2585–o2586. [Google Scholar]
  21. Selvamurugan, V.; Aidhen, I.S. Simple Synthetic Equivalents for the β-(N,N-Disubstituted)ethylamino Acyl Cation Synthon and their Applications. Synthesis 2001, 2239–2246. [Google Scholar] [CrossRef]
  22. Attia, M.I.; Ghabbour, H.A.; Almutairi, M.S.; Ghoneim, S.W.; Abdel-Aziz, H.A.; Fun, H.-K. Synthesis and X-ray crystal structure of (1E)-1-(4-chlorophenyl)-N-hydroxy-3-(1H-imidazol-1-yl)propan-1-imine. J. Chem. 2013. [Google Scholar] [CrossRef]
  23. Clinical and Laboratory Standards Institute (CLSI), Reference method for broth dilution antifungal susceptibility testing of yeasts. In Approved Standard M27-A2, 2nd ed.; CLSI: Villanova, PA, USA, 2002.
  24. Clinical and Laboratory Standards Institute (CLSI), Performance Standards for Antimicrobial Disk Susceptibility Tests. In Approved Standard, M2-A8, 8thed.; CLSI: Wayne, PA, USA, 2005.
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