BIOCIDAL EFFICACY OF SYNTHETIC COPOLYESTERS HAVING CHALCONE MOIETY

BIOCIDAL EFFICACY OF SYNTHETIC COPOLYESTERS HAVING CHALCONE MOIETY

Rajesh Kumari ¹ and Tom John ^{* 2}

Department of Dravyaguna Vigyan¹, Shiv Shakti Ayurvedic College, Bhikhi (Mansa), Punjab, India. Neuropharmacology Section ², National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.

ABSTRACT: Two chalcone diols (2E)-1-(4-hydroxyphenyl)-3(4-hydroxy, 3-ethoxy phenyl) prop-en-1-one and (2E)-1-(4-hydroxy-3-methoxy phenyl)-3(4-hydroxy, 3-ethoxy phenyl) prop-en-1-one were synthesized by acid catalyzed ClaisenSchmidt reaction. The synthesized chalcones were made to undergo polymerization using three aliphatic dichlorides namely glutaryl, succinyl and oxalyl chlorides. The polymers were random copolyesters and characterized by solubility and viscosity measurement. The structure of the copolyester was established by FTIR, ¹H-NMR and ¹³C-NMR. The biocidal efficacy was done by Disc diffusion method and found to be nominal against the standards used. Thus these copolyesters may be considered as drug carriers

Chalcones,

Polycondensation, Copolyesters, Antibacterial, Antifungal

INTRODUCTION: Chalcones (α, β-Unsaturated ketones) are biogenetic precursors of flavonoids ¹. Chemically, chalcones are 1,3-diaryl-2-propen-1-ones in which two aromatic rings are joined by a three carbon bridge having a cabonyl moiety and α, β unsaturation ². Conventionally, chalcones are synthesized by Claisen-Schmidt condensation of arylaldehydes and acetophenones. The traditional methods for the synthesis of chalcones involves the use of strongbases such as NaOH, KOH, Ba (OH) hydrotalcites, LiHMDS, calcined NaNO3/natural phosphate. There are also some reports of acid-catalyzed aldol condensations, e.g. AlCl₃, BF₃, dry HCl, ZrH₂/NiCl₂ and RuCl₃ (for cyclic and acyclic ketones).

Chalcones and its derivatives have attracted increasing attention due to numerous pharmacological applications ³. Literature survey reveals that numerous chalcones and their derivatives exhibit a broad spectrum of pharmacological activities such as anti-amoebic ⁴, antibacterial, antifungal ⁵, anticancer ⁶, anticonvulsant ⁷, anti-inflammatory, antifungal, antibacterial ⁸, Antileishmanial ⁹, Anti-malarial ¹⁰, Anti-malarial, Anti-tubercular ¹¹. Senthamizh Selvi and coworkers have synthesized certain chalcone based random copolyesters and reported their biocidal behavior ¹².

Perundevi and coworkers have synthesized certain bischalcone based random copolyesters and reported their antibacterial behavior ¹³.

The objective of our study is to synthesize four copolyesters by incorporating the chalcone moieties in the copolyester main chain by polycondensation process, then characterizing them analytically and to study their anti bacterial and anti fungal activity.

 Experimental section:

Aldrich samples of 4-hydroxy acetophenone and 3-ethoxy, 4-hydroxy benzaldehyde, 3-methoxy, 4-hydroxy acetophenone and were used as received.  Ethanol (Merck) was used as non-solvent for the copolyesters and as solvent for the preparation of the two chalcone diols. Aldrich samples of succinyl chloride, oxalyl chloride, glutaryl and isopthaloyl chloride were used as such in the synthesis of the 5 copolyesters. Spectral grade DMSO-d6 was used as internal standard for recording NMR spectra.

 Synthesis of chalcone diols:

The monomer diols namely (2E)-1-(4-hydroxyphenyl)-3(4-hydroxy, 3-ethoxy phenyl) prop-en-1-one and (2E)-1-(4-hydroxy-3-methoxy phenyl)-3(4-hydroxy, 3-ethoxy phenyl) prop-en-1-one were synthesized by the process reported by Jasmine Francis and et al ¹⁴.

 Synthesis of HHEP:

Dry HCl gas was passed through a well cooled and stirred solution of 4-hydroxy acetophenone (50 mmol) and 3-ethoxy-4-hydroxy benzaldehyde (50mmol) in 100 ml of dry ethanol taken in a 250 ml round bottom flask. The yellow colored crystals of HHEP which got separated was washed with double distilled water and recrystallized from hot methanol. Yield - 90%, M.P. – 174℃.

Synthesis of MHEP:

Dry HCl gas was passed through a well cooled and stirred solution of 4-hydroxy-3-methoxy acetophenone (50 mmol) and 3-ethoxy-4-hydroxy benzaldehyde (50mmol) in 100 ml of dry ethanol taken in a 250 ml round bottom flask.  The yellow coloured crystals of MHEP which got separated was washed with double distilled water and recrystallised from hot methanol. Yield - 74%, M.P. –194ºC

 Synthesis of Copolyesters:

The procedure ¹³ for the synthesis of a typical aliphatic diacid chloride-based copolyester is given here. The diol HHEP (1mmol) was dissolved in 10 ml of dry DMF and taken in a 100 ml round bottomed flask. To this 0.2ml of succinyl chloride and 0.2 ml of oxalyl chloride were added with constant stirring and the temperature was maintained at 120ºC with continuous stirring for 3 hours. At end the reaction mixture was cooled to room temperature and poured into 100 ml of methanol when the copolyester was precipitated. It was filtered, washed with dry methanol and dried in vaccum. The other four copolyesters were also prepared by similar method. The diols and the diacid chlorides used and the copolyester code of the four copolyesters are presented in Table 1.

 Antimicrobial Activity (Agar Disc Diffusion Method):

Antibacterial activity of sample was determined by disc diffusion method ¹⁵ on Muller Hinton agar (MHA) medium. The Muller Hinton Agar medium was weighed as 3.8gmsand dissolved in 100ml of distilled water and add 1gm of agar. Then the medium is kept for sterilization. After sterilization the media was poured in to sterile petriplates and were allowed to solidify for thirty minutes. After the medium was solidified, the inoculums were spread on the solid plates with sterile swab moistened with the bacterial suspension. Add 20 µl of sample [concentration: 1000 µg, 500 µg, 250 µg, 12 5 µg & 62.5 µg], negative control add 20 µl of DMSO and positive control add 10 µl (10 µg) streptomycin on respective disc and placed on MHA plates. These plates were incubated for 24 hrs at 37ºC. Then the microbial growth was determined by measuring the diameter of zone of inhibition.

 Antifungal Activity Assay:

Antifungal activity of copolyester sample was determined by antifungal susceptibility test. Prepare PDA Broth and inoculate the culture. Then it is kept in shaker for a day. The potato dextrose agar was weighed as 3.9gms and dissolved in 100ml of distilled water to which 1gm of agar was added. Then the medium was kept for sterilization. After sterilization the media was poured in to sterile Petri plates and were allowed to solidify for twenty minutes.

After solidification, the inoculums were spread on the solid plates with sterile swab moistened with the fungal suspension. The discs were placed in PDA plate and 20µl of sample [concentration: 500µg, 250µg, 125µg, 62.5µg] were added. The plates were kept at room temperature. Then the microbial growth was determined by measuring the diameter of zone of inhibition.

 RESULTS AND DISCUSSION:

Solubility of all the five copolyesters synthesized were determined in various solvents qualitatively. The inherent viscosity (ηinh) of the copolyesters were determined in DMAc solution at a concentration 0.1 gdL-1 using Ubbelohde viscometer in which the pure solvent had a flow rate of 104 seconds at 30°C. FT-IR spectra of the entire random copolyesters were recorded using Shimadzu FT-IR instrument. The 1H and 13C-NMR spectra were recorded with BRUKER AV III 500 MHz NMR instrument in DMSO-d6 solvent.

 Solubility:

The copolyesters stated here are found to be soluble in highly polar solvents such as DMAc and dimethyl formamide, partially soluble in moderately polar solvents like tetrahydrofuran and acetone but thoroughly insoluble in least polar solvents like benzene and hexane ¹⁶. Copolyesters with methoxy substituent in the benzene ring of the chalcone moiety had better solubility which may be attributed to their competence to disrupt the macromolecular chain. Similar explanation was offered by Perundevi and coworkers ¹⁶ in a series of copolyester.

 Viscosity Measurements:

The ηinh value of all the five copolyesters was determined in DMAc solution at 30℃ using Ubbelohde viscometer. 25mg of each of pure dry copolyester sample was dissolved in 25ml of DMAc, kept aside for some time with occasional shaking. Then they were left undisturbed for 24 hours. The ηinh was calculated from the flow time measurements ¹⁶. The inherent viscosity values were found to be in the range of 0.73–1.10dL/g and are presented in Table 1. The data shows that these copolyesters are reasonably of high molecular weight.

 Spectral Studies:

FT-IR spectrum of the five copolyesters were recorded using Shimadzu FT-IR instrument. The FT-IR spectrum of all the five copolyesters revealed characteristic absorption in the range of 1742–1764cm-1 due to ester C=O stretching frequency. Similar observations were made by Samuel and Coworkers ¹⁷ in a series of copolyesters.

The NMR spectra were recorded with BRUKER AV III 500 MHz NMR instrument in DMSO-d6 solvent to categorize the repeating structural units present in the copolyester chain. The aromatic protons are observed in the range of 7.2–8.1ppm. The vinyllic protons attached to the carbonyl carbon are observed in the range of 6.7–6.9ppm. The methoxy protons in the chalcone moiety are represented in the range of 3.1–3.4ppm. The methylene protons are observed in the range of 1.3–3.3 ppm. Similar remarks were made Perundevi and coworkers ¹⁶ in a series of copolyesters derived from bischalcones.

The signals in the range of 170–205ppm and 165–175ppm in the 13C-NMR spectra of the copolyesters are due to the carbonyl carbon of the α, β-unsaturated ketone and ester groups, respectively, which indicates the formation of copolyester.

Bactericidal Study:

The antibacterial activity ¹⁸ of the five copolyesters PSGH, PSOH, PSIH, PSOM and PSGM were assayed against (Staphylococcus aureus, Bacillus subtillus, Klebsilla pnemoniae, Pseudomonas aeruginosa & amp; Escherichia coli by disc diffusion method ¹⁹. Streptomycin inhibited the escalation of Staphylococcus aureus by 20mm, Enterrococcus faecalis.

By 19mm, Bacillus cereus by 26mm, and Escherichia coli by 17mm. From table 2 it is apparent that the four copolyesters were found to be bactericidal in nature. With increase of concentration of the copolyester material it was observed that the inhibition effect increased.

 Fungicidal Study:

The antifungal activity ²⁰ of the synthesized copolyesters PSGH, PSOH, PSIH, PSOM and PSGM were assayed against A. fumigates, Aspergillus flavus, T. mentogrophyte by disc diffusion method. Streptomycin subdued the growth of Candida albicans by 14mm, Aspergillus flavus by 13mm, Penicillium by 10mm, and T. Mentogrophyte by 10mm. From Table 3 it is obvious that the five copolysters were found to be fungicidal in nature.

SCHEME 1: SYNTHESIS OF MONOMER DIOL HHEP

SCHEME 2: SYNTHESIS OF MONOMER DIOL MHEP

TABLE 1: MONOMERS USED AND THE COPOLYESTER CODE OF THE FOUR COPOLYESTERS TOGETHER WITH PERCENTAGE OF YIELD AND INHERENT VISCOSITIES (ηinh)

TABLE 2: INHIBITION EFFECTS OF THE FIVE COPOLYESTERS ON THE GROWTH OF STAPHYLOCOCCUS AUREUS, KLEBSELLIA PNEUMONIA, BACILLUS SUBTILIS, E.COLI AND PSEUDOMONAS AERUGINOSA

TABLE 3: INHIBITION EFFECTS OF THE FIVE COPOLYESTERS ON THE GROWTH OF A.FLAVUS, A. FUMIGATES, AND T.MENTAGROPHYTE

CONCLUSION: All the five copolyesters synthesized using a diacid chloride-I (Succinyl chloride), diacid chloride-II (Glutaryl chloride, oxalyl chloride and isopthaloyl chloride) and a chalcone diol. The chalcone diols are varied. The chalcone diols used are (2E)-1-(4-hydroxyphenyl)-3(4-hydroxy,3-ethoxy phenyl) prop-en-1-one (HHEP) and (2E) – 1 - (4-hydroxy-3-methoxy phenyl)-3(4-hydroxy,3-ethoxy phenyl) prop-en-1-one (MHEP). The copolyesters synthesized here are highly soluble in polar organic solvents. These random copolyesters are characterized by solubility studies, viscosity measurements and spectral data. The copolyesters exhibited significant bactericidal activity against pathogenic bacteria. Some of them are also found to be fungicidal in nature.

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    How to cite this article:

    Kumari R and John T: Biocidal Efficacy of Synthetic Copolyesters Having Chalcone Moiety. Int J Pharm Sci Res 2016; 7(9): 3611-16.doi: 10.13040/IJPSR.0975-8232.7(9).3611-16.

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