- Research article
- Open Access
Synthesis and investigation of antimicrobial properties of pyrrolidine appended calixarene
© The Author(s). 2017
- Received: 16 August 2016
- Accepted: 24 January 2017
- Published: 9 February 2017
Calixarenes are widely used macromolecules in supramolecular chemistry synthesized by simple phenol formaldehyde condensation reaction in the presence of base. This article describes the synthesis, antibacterial, and antifungal properties of calixarene appended with pyrrolidine at the upper rim (CAP3).
Antimicrobial activity of CAP3 was determined by Kirby-Bauer well agar diffusion method in Mueller-Hinton agar (MHA) medium of growth against Escherichia coli (ATCC 8739), Staphylococcus aureus (ATCC 10231) and Streptococcus viridans (ATCC 12392), Candida albicans (ATCC 32333), Aspergillus niger (ATCC 16404), and Aspergillus flavus (ATCC 90906). Structure–activity relationship was also used to evaluate the active site of CAP3.
Bacterial strains have shown minimum inhibitory concentration (MIC) values in 1.17–2.34 mg/mL range; whereas, the fungal strains have shown MIC values at the range of 0.58-2.34 mg/mL. It is observed that CAP3 has excellent antifungal action for A. niger as well as antibacterial action for S. viridans having MIC values of 0.58 and 1.17 mg/mL, respectively. The SAR (structure–activity relationship) study of compound reveals that there is a significant antimicrobial activity shown by CAP3 probably due to the pyrrolidine substituents at phenyl rings of calixarene.
The results indicate that CAP3 is an effective material against the microbial strains.
- Structure–activity relationship
- Antimicrobial activity
Because of diverse bioactive and potential pharmaceutical characteristics, pyrrolidine and related heterocyclic compounds have possessed specific position among diverse organic classes (Pinto and Abdala 2000). From the last few decades, these molecules have got considerable attention in pharmaceutical industry due to the extensive clinical requests (Mohammat et al. 2011; Feng et al. 2012). It is because of the outstanding antiviral and antibiotic properties of these molecules caused by heterocyclic core, such as pyrrolidine-2,4-dione (natural tetramic acids) (Gallardo et al. 2004; Gitterman et al. 1964; Chen et al. 2008; Gitterman 1965; la Croix et al. 1975; Janardhanan and Husain 1984; Suzuki et al. 1967; Mourer et al. 2009). Arun et al. has prepared and estimated the antimicrobial activity of dispirooxindole-pyrrolidine derivatives via 1,3-dipolar cycloaddition of an azomethine ylide obtained from sarcosine and isatin. Dandia et al. prepared spiro[acenaphthylene-1,2′-pyrrolidine] in the course of 1,3-dipolar cycloaddition reaction of azomethine ylide generated from sarcosin and di/tri ketone. The cycloadducts ketocarbazalo spiro N-methyl pyrrolidines have demonstrated highly remarkable antimicrobial activity even in very low concentrations (Dandia et al. 2013). However, importance in drug delivery and potentially enhanced neuroprotection and anticancer properties often found in novel compounds. In the last few decades, calixarene molecules are found to have multiple uses in basic as well as applied sciences. They are also a class of important macrocyclic molecules like cyclodextrins, crown ethers, and porphyrins, which act as host molecules. Structure of calixarenes possesses the lower and upper rims with central annulus part joined via methylene groups, while hydroxyl functionality at meta-position (Memon and Bhatti 2015; Fátima and Fernandes 2009). Because of easy functionalization at the upper and lower rims, calixarenes are widely used in various fields like molecular recognition, nanotechnology, catalysis, sensing, drug delivery, and to mimic the biological processes (Nimsea and Kim 2013). The molecular recognition property makes them a versatile tool for complexation of organic and biomolecules. In this regard, the pioneer work is done by Cornforth et al. against tuberculosis (TB), which will prove to explore the calixarenes’ for examining their biological properties. Furthermore, several pharmacological properties including antibacterial, antifungal, antiviral, and anticancer activities (Colston et al. 2004) of calixarenes have also been reported. Thus, herein, we report the synthesis and investigation of calixarene (CAP3) containing pyrrolidine moiety and divulge the remarkable screening results against its antibacterial and antifungal activities.
For synthesis, all reagents were of analytical grade and used without further purification. Mueller-Hinton agar was purchase from Sigma-Aldrich; antimicrobial growth was performed locally prepared at laminar flow cabinet. Preparation of medium was done on vertical autoclave (i.e., Robus Technologies). Antimicrobial growth was carried out on Thermo Scientific incubator. Gallenkamp melting point apparatus was used for the melting point determination and was uncorrected. Elemental analysis was carried out using a CHNS instrument model Flash EA 1112 elemental analyzer. FT-IR spectra were obtained by applying KBr pellet method in the spectral range 400–4000 cm−1 on a Thermo Nicollet 5700 FT-IR spectrometer. TLC was carried out on pre-coated silica gel plates (SiO2, Merck PF254).
Synthesis of compound
2.8 mmol of compound (2) was dissolved in 45 mL of THF with of 37% formaldehyde (12 mmol) added drop wise to reaction mixture at room temperature. By the addition of pyrrolidine (12 mmol), the reaction mixture start to become yellowish wax type. The solution was stirred for 24 h and filtered, washed with water and crystallized in methanol to furnished (3).
Distribution of culture strains and medium
G +ve bacteria
G -ve bacteria
Various methods were employed for the determination of the antimicrobial activity. Antimicrobial activity of CAP3 was tested by using Kirby-Bauer well diffusion method (Perret and Coleman 2011). Different concentrations were used for compound CAP3 ranging from 75.0, 37.5, 18.7, 9.37, 4.68, 2.34, 1.17, and 0.58 mg/mL in dimethylsulphoxide (DMSO) (Zgoda and Porte 2001). A well was developed in the sterile Petri dish with medium plates were prepared with the help of cork borer (6 mm in diameter) and (50 μL) solution of each concentration was poured into the wells. Under sterilized condition, wells were filled with diluted solution; micropipette was used for filling purpose. This work was done at room temperature and stored them for further use. Sterilized Petri dishes were incubated for 48 h at 37 °C. At the same time, DMSO as −ve control was employed, and the solution of CAP3 was also prepared in the same solvent. While the relative antifungal and antibacterial activities in millimeter were measured for evaluation. The results were studied in triplicate. MIC data have also been appraised (Dumazet et al. 1997; Mistry and Desai 2014).
Zones of inhibition for antimicrobial activities of CAP3
MIC value (mg/mL)
Zones of inhibition (mm)
G +ve bacteria
G -ve bacteria
It shows that CAP3 exhibits good antibacterial action against Gram +ve bacteria comparative to Gram -ve. It also shows better antifungal activity against A. niger than all other selected fungal strains due to morphological change in their structures.
Structure–activity relationship hypothesis
Minimum inhibitory concentration
In the present study, pyrrolidine appended calixarene (CAP3) was synthesized and explored for its antimicrobial activity using Kirby-Bauer well agar diffusion method. The study revealed that CAP3 show good antimicrobial activity against selected bacterial and fungal strains. The MIC values calculated are in the range 1.17–2.34 and 0.58–2.34 mg/mL for bacterial and fungal strains, respectively. The convenient synthetic scheme and low toxicity as well as cost effectiveness of the starting materials make CAP3 a promising candidate for the pharmaceutical formulations against the tested microorganisms. Consequently, the study shows a good impact for controlling the infections created by different microorganism in future.
All authors read and approved the final manuscript.
The authors declare that they have no competing interests.
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