Synthesis, Characterization and Antimicrobial Evaluation of Biphenyl derivatives against C. albicans and E. coli
Shivali1, Priyankul Palia1*, Sunny Dhiman1, Inder Kumar2, Pankaj Kumar1, Vikrant Dalwal1
1School of Pharmacy, Abhilashi University, Chail-Chowk, Mandi, Himachal Pradesh, India.
2Minerva College of Pharmacy, Indora, Distt. Kangra, HP, India.
*Corresponding Author E-mail: priyankulpalia23@gmail.com
ABSTRACT:
Biphenyls are polynuclear aromatic hydrocarbons with a molecular formula (C12 H10) having more than one aromatic nucleus. Biphenyl derivatives substituted with an aromatic or heteroaromatic radical have therapeutic use in pharmaceutical compositions intended for use in humans or cosmetic compositions. The present study included the synthesis of biphenyls derivatives and their antifungal and antimicrobial activities. Four different compounds (SCPD1-SCPD4) were synthesized via different synthesis methods. All the compounds were characterized by different methods like TLC, IR, and NMR. All the peaks and spectra revealed the presence of synthesized compounds. Among all the synthesized compounds, SCPD3 has shown maximum activity against gram-negative E. coli, and SCPD4 has shown maximum activity against C. albicans when compared with standard drug ketoconazole. The series of biphenyl derivatives may be a potent alternative to some other antibacterial and antifungal agents.
KEYWORDS: Biphenyls, Anti-inflammatory, Anti-microbial, Biphenyl derivatives, Topical, Synthesis.
INTRODUCTION:
Biphenyls are polynuclear aromatic hydrocarbons with a molecular formula (C12H10) having more than one aromatic hydrocarbons. The two nuclei are attached at one point. Biphenyls are also named as Diphenyl or 1, 1’ Biphenyl or limonene. It is a solid compound that forms colorless to yellowish crystals. Biphenyl is insoluble in water but is soluble in organic solvents. Literature findings have also shown its various therapeutic uses as an anti-inflammatory, analgesic, anti-pyretic, anti-microbial, anti-protozoal, anti-viral, and anti-fungal1-3. Biphenyl derivatives substituted with an aromatic or heteroaromatic radical have therapeutic use in pharmaceutical compositions intended for use in human or veterinary medicine, or in cosmetic compositions.
The compounds according to the invention have pronounced activity in the fields of cell differentiation and proliferation and find applications more particularly in the topical and systemic treatment of dermatological complaints associated with a keratinization disorder, dermatological (or other) complaints with an inflammatory and/or immune allergic component, and dermal or epidermal proliferation, whether they are benign or malignant4-7. These compounds can also be used in the treatment of connective tissue degenerative diseases, for controlling the aging of the skin, whether this is light-induced or chronological aging, and for treating cicatrization disorders. They moreover find an application in the ophthalmological field, in particular in the treatment of homeopathy. The compounds can also be used in cosmetic compositions for body and hair hygiene. Honokiol (HNK), a naturally occurring biphenyl, has potent anti-neoplastic and anti-angiogenic properties. HNK exhibited potent antiproliferative activity against breast cancer cell lines and enhanced the activity of other drugs used for the treatment of breast cancer. Honokiol (HNK) is a naturally occurring biphenyl that can be extracted either from the root, stem bark, or seed cone of several Magnolia species. HNK has long been known to have anti-thrombotic, anti-bacterial, and anxiolytic effects8,9. The objective of the present work involves the synthesis of new derivatives having biphenyl as a basic skeleton since this ring system is present in some of the existing antibacterial and antifungals and the structural confirmation of the synthesized compounds by spectroscopic IR and NMR analysis and to carry out biological activities of newly synthesized compounds.
MATERIAL AND METHODS:
The solvent used for the study were of reagent grade and were purified before their use. All solvents, and chemicals, biphenyl, acetic anhydride, pyridine, aluminium chloride, were procured from CDH Pvt. Ltd New Delhi. Glacial acetic acid, hydrochloric acid, bromine, and all other solvents were purchased from SD fines Pvt. Ltd New Delhi.
Synthesis of biphenyl derivatives:
This scheme involves Friedal Crafts acylation of biphenyl with acetic anhydride in the presence of aluminum chloride as Lewis’s acid catalyst to give p-Phenylacetophenone. Bromination of p-Phenyl acetophenone was carried out to give desired product 1-Biphenyl-4-yl-2-bromoethanone10-13. This 1-Biphenyl-4-yl-2-bromoethanone was substituted with different aryl or heteroaryl amines(R) to give different analogs in presence of pyridine (fig. 1).
Synthesis of p-Phenylacetophenone (SCPD1):
In a 250ml three-necked flask provided with a dropping funnel, a mechanical stirrer, and a reflux condenser, placed 77g (0.5mol) of biphenyl, 150gm (1.125mol) of finely powdered anhydrous aluminium chloride, and 350 ml of anhydrous carbon disulphide, charged the dropping funnel with 51gm (47.5ml, 0.5mol) of pure acetic anhydride and closed the mouth of the funnel with a calcium chloride guard tube. The mixture was heated on a water bath until gentle reflux commenced and acetic anhydride was added during one hour; the addition product made its appearance as a curdy mass when about three-quarters of the acetic anhydride had been added. The reaction mixture was refluxed gently for a further hour, allowed to cool and poured the reaction product slowly and with stirring on to crushed ice to which hydrochloric acid had been added. The product was filtered and washed with water to remove traces of hydrochloric acid and dried. It was recrystallized from methanol (Fig. 1)15,16.
Fig.1: Synthesis of biphenyl derivatives
Synthesis of p-Phenylphenacylbromide (SCPD2):
p-Phenylacetophenone (36g, 0.183mol) was suspended in 200ml of glacial acetic acid in a 500ml flask and warmed gently on a water bath until a clear solution resulted, then cooled as far as possible without the formation of crystals. To this solution 29.5g (9.5ml, 0.184mol) of bromine was added and the temperature was maintained below 45ºC during the addition. The brominated product separated from the solution when about three-quarters of the bromine had been added. After 2 hours, cooled the flask in a bath of ice and salt, filtered the product, washed with a little water, cooled the flask in a bath of ice and salt, filtered the product, washed with a little cold glacial acetic acid, followed by small volumes of water until all the acid has been removed17,18. Recrystallised from hot rectified spirit (600-700ml) (Fig. 1).
Synthesis of 1-Biphenyl-4-yl-2-(4-chloro-phenylamino)-ethanone (SCPD3):
In a 250ml RBF, 1-(biphenyl-4-yl)-2-bromoethanone (0.01mol) was dissolved in 16.11ml (0.2mol) of pyridine. To this solution, 1.2gm (0.01 mol) of p-chloro aniline was added. The resulting reaction mixture was then kept on stirring for 5 hrs till the TLC of the reaction mixture showed completion of the reaction, stirring was continued up to the completion of the reaction. The reaction product was poured slowly and with stirring on crushed ice. The product separated was filtered, dried, and was recrystallized from methanol19.
Synthesis of 1-Biphenyl-4-yl-2-(4-nitro-phenylamino)-ethanone (SCPD4):
In a 250ml RBF, 1-(biphenyl-4-yl)-2-bromoethanone (0.01mol) was dissolved in 16.11ml (0.2mol) of pyridine. To this solution, 1.0gm (0.01mol) of p-nitro aniline was added. The resulting reaction mixture was then kept on stirring for 5hours till the TLC of the reaction mixture showed completion of the reaction, stirring was continued up to the completion of the reaction. The reaction product was poured slowly and with stirring on crushed ice20-22. The product separated was filtered, dried, and was recrystallized from methanol (Fig. 1).
Chemical characterization:
Physicochemical characterization of all synthesized compounds was done by assessment of their melting points, Rf values determined by using TLC, and Infra-Red (IR) and Proton Nuclear Magnetic Resonance (1H NMR) spectra. The single spots on TLC as well as their sharp melting points assessed the purity of all synthesized compounds. TLC was prepared with silica gel G and activated at 110oC for 30 min. The plates were developed by exposure to iodine vapors. The melting points were determined on the melting point apparatus. The structural confirmation of all synthesized final compounds with their intermediates was done with the help of FT-IR and 1H NMR spectroscopy. Proton (1H) nuclear magnetic resonance (1H-NMR) spectra were obtained using Brucker AC-400 F, 400 MHz spectrometer and are reported in parts per million (ppm), downfield from tetramethylsilane (TMS) as internal standard. The spin multiplicities are indicated by the symbols: s (singlet), d (doublets), t (triplet), q (quatret), m (multiplet), and br (broad). Infrared (IR) spectra were obtained with Perkin Elmer 882 Spectrum and RXI17,20, 21.
2.3. In vitro Antibacterial screening:
The antibacterial activity of the synthesized Biphenyl derivatives was carried out using the cup plate method. The compounds were tested against one strain of bacteria E. coli (NCIM-121). Stock solutions of the synthesized compounds and the Ciprofloxacin were prepared in DMF. Dilution of the stock solution of synthesized compound was done using DMF and three different concentrations 50, 100, and 250mg/ml were prepared from micro test tubes. The stock solution of standard was diluted in DMF to give a concentration of 250 mg/ml.100ml of organism suspension was seeded on culture plates. Then in each of the agar plate cavities/ well (9mm) were prepared using a cork borer. 100ml of each of the dilutions of compound (50, 100, and 250 mg/ml) were transferred to the wells, and Ciprofloxacin (250 mg/ml) was added to the remaining well. The plates were incubated at 30oC. The plates were observed for growth at the end of 24 to 48 hours 4, 6.
In vitro Antifungal screening:
The antifungal activity of the synthesized Biphenyl derivatives was carried out using the cup plate method. The compounds were tested against one stain of fungi Candida albicans ATCC-10231. Stock solutions of the synthesized compounds and standard drug i.e., Ketoconazole were prepared in DMF. Dilution of the stock solution of synthesized compound was done using DMF and three different concentrations 50, 100, and 250 mg/ml were prepared from micro test tubes. The stock solution of standard was diluted in DMF to give a concentration of 250mg/ml. 100ml of organism suspension was seeded on culture plates. Then in each of the agar plate cavities/ well (9mm) were prepared using a cork borer. 100ml of each of the dilutions of compound (50, 100, and 250mg/ml) were transferred to the wells, and Ciprofloxacin (250mg/ml) was added to the remaining well. The plates were incubated at 30oC. The plates were observed for growth at the end of 24 to 48 hours4,6.
RESULTS AND DISCUSSION:
Characterization of synthesized compounds:
Characterization of p-Phenylacetophenone (SCPD1):
Yield: 82%, M.P.: 114-116˚C, RF: 0.65 [benzene: ethyl acetate:4: 1], IR (KBr) in cm-1: 3071.6 (Ar.-CH stretching), 2915.7(Ali. -CH Stretching), 1679 (C=O), 1600(C=C stretching), 1H-NMR (DMSO-d6):d2.6(s, 3H, -CH3), d 7.3-8.0 (m, 9H, (Ar H), -CO-C6H4- C6H5)
Characterization of 1- biphenyl-4-yl-2- bromoethanone (SCPD2):
Yield: 82%, M.P.: 122-124°C, RF: 0.65 [benzene], IR (KBr) in cm-1:3399(N-H), 3058(Ar. C-H), 1662 (C=O), 1297(C-N),1598(Ar.C=C), 1H-NMR (DMSO-d6): d4.4(s, 2H, -CH2Br), d 7.2-8.0 (m, 9H, (Ar H), -CO-C6H4-C6H5)
Characterization of 1-Biphenyl-4-yl-2-(4-chloro-phenylamino)-ethanone (SCPD3):
Yield: 73%, M.P.:157-159°C, RF:0.56 [benzene], IR (KBr) in cm-1: 3151(N-H), 2916(Ar. C-H),2847(Ali. -CH Stretching), 1681 (C=O), 1263 (C-N),1598 (Ar. C=C), 748 (C-Cl), 1H-NMR (DMSO-d6 ppm): d4.7 (s, 2H, -CH2), d7.8(s,1H, NH), ),d 7.0-8.0(m, 13 H, (Ar H), CH2CO-C6H5-C6H4- C6H4)
Characterization of 1-Biphenyl-4-yl-2-(4-nitro-phenylamino)-ethanone (SCPD4):
Yield: 70%, M.P.:145-148°C, RF:0.52 [benzene], IR (KBr) in cm-1: 3399 (N-H), 3058(Ar. C-H), 1662 (C=O), 1297 (C-N),1598 (Ar. C=C), 1H-NMR (DMSO-d6 ppm):d4.8 (s, 2H, -CH2), d6.6(s,1H, NH),), d 6.7-8.3(m, 13 H, (Ar H), CH2CO-C6H5-C6H4- C6H4)
Biological evaluation:
The compounds were evaluated for in vitro antibacterial activity against E. coli, and Ciprofloxacin was used as standard also evaluated for in vitro antifungal activity against C. albicans and Ketoconazole was used as standard. The zone of inhibition of the synthesized compounds against E. coli is presented in Table 1. Amongst, all the synthesized compounds SCPD3 have shown maximum activity against gram-negative bacteria E. coli, compared to standard drug. For antifungal activities, compound SCPD4 has shown maximum activity against C. albicans, compared to standard drug.
Table 1: In vitro antifungal and antibacterial activity of the synthesized compound against C. albicans and E. coli
|
S. No. |
Derivatives |
Zone of inhibition (mm) At different concentrations (µg/ml) |
|||||
|
C. albicans |
E. coli |
||||||
|
50µg/ml |
100µg/ml |
250µg/ml |
50µg/ml |
100µg/ml |
250µg/ml |
||
|
1 |
SCPD1 |
2 |
7 |
9 |
3 |
6 |
9 |
|
2 |
SCPD2 |
3 |
9 |
10 |
4 |
7 |
10 |
|
3 |
SCPD3 |
4 |
8 |
10 |
4 |
7 |
11 |
|
4 |
SCPD4 |
4 |
9 |
11 |
4 |
6 |
11 |
|
5 |
Ketoconazole |
5 |
10 |
12 |
*** |
*** |
*** |
|
6 |
Ciprofloxacin |
*** |
*** |
*** |
5 |
7 |
12 |
|
|
|
|
Fig. 2: Comparison of antibacterial activity of synthesized biphenyl derivatives with standard drug |
Fig. 3: Comparison of antifungal activity of synthesized biphenyl derivatives standard drug |
CONCLUSION:
In the present study, four biphenyl compounds were synthesized by different methods. All four compounds (SCPD1-SCPD4) were identified and characterized using TLC, IR, and NMR. The peaks, values and spectral analysis showed the presence of synthesized compounds. All the compounds were evaluated for their biological evaluations (antifungal and antibacterial activities) concerning standards. Among all the synthesized compounds SCPD3 has shown maximum antibacterial activity against gram-negative E. coli, when compared with standard drug and SCPD4 has shown maximum antifungal activity when compared with standard drug. So, these new therapeutic agents could be considered as lead molecule for the future development of drugs which could be used as antibacterial and antifungal agents.
ACKNOWLEDGEMENT:
The authors are grateful to the Abhilashi University, Mandi, HP, India for providing various facilities in connection with this work.
CONFLICT OF INTEREST:
The authors declare no conflict of interest, financial or otherwise.
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Received on 19.07.2023 Modified on 07.02.2024
Accepted on 26.06.2024 ©Asian Pharma Press All Right Reserved
Asian J. Pharm. Tech. 2024; 14(3):208-212.
DOI: 10.52711/2231-5713.2024.00034