Synthesis, Characterization and Antimicrobial Activity of 7’Methyl 3 Phenyl 2, 3 Dihydro 4’ Furo Chromene

 

S. Janet Beula*, Dr. R. Suthakaran, M. Bhanu Prakash, G. Sreeya, P. Pavan Kalyan, P. Shiva Prasad

Vijaya College of Pharmacy, Munuganoor, RR District, India.

*Corresponding Author E-mail: pharmjanet123@gmail.com

 

ABSTRACT:

Synthetic medicines developed are popular even among younger generations. The coumarine derivative drugs claim efficient cure of various disorders. Recently, coumarine compound analogues and their derivatives have attracted strong interest in medicinal chemistry due to their biological and pharmacological properties. The structure have offered a high value of diversity that is proven useful for the development of new medicinal drugs and improved potency, less toxicity and good pharmacological activity. The synthesized compounds were screened for their in-vitro growth inhibiting activity against different strains of bacteria Staphylococcus aureus and Escherichia coli at 50, 100, 150 and200 µg/ml concentrations by using agar well diffusion technique. The results were compared with the standard antibiotics Ciprofloxacin (100 µg/ ml). The newly synthesized compound produced significant action against the bacterial strains at 100 µg/ml concentration.

 

KEYWORDS: Melting point, TLC, H1NMR, IR, Mass spectroscopy, Antimicrobial Evaluation and Agar well diffusion technique.

 

 

 

INTRODUCTION:

Coumarin is the parent organic compound of a class of naturally occurring phytochemicals found in many plant species. Coumarin comes under the benzopyrone family of compounds, all of which consist of a benzene ring joined to a pyrone ring1. Coumarins have attracted considerable attention of medicinal chemists and pharmacologists in recent years as they been demonstrated to bear various pharmacological activities like anti-microbial, antioxidant, antiinflammatory and analgesic, anti-cancer, ulcerogenic, antimalarial, antihyperlipidemic, tyrosinase inhibitor, anticonvulsant, anti-parkinsonian, antihepatitis, anticoagulant, cholinesterase inhibitor, vasorelaxant.

 

Coumarins comprise a very large class of compounds found throughout the plant kingdom2-4. Coumarin is also known as 2H-lbenzopyran-2-one, 1, 2-benzopyrone5-9. Coumarins are competitive inhibitors of vitamin K in the biosynthesis of prothrombin10-11. In the present study we have planned to prepare 7’methyl 3 phenyl 2, 3 dihydro 4’ furo chromene and elucidate the structure by physical, chemical, spectral analysis and evaluate the antimicrobial activity15-17.

 

Materials:

All the chemicals used in the synthesis were of laboratory grade (Sri Venkateswara Scientifics, Parsigutta and Hyderabad). The melting points were determined in open capillary on Veego (VMP-D) electronic apparatus and the errors were rectified. The IR spectra of the synthesized compound were recorded on Perkin Elmer BX2 FT-IR spectrophotometer in potassium bromide (anhydrous IR grade) pellets. NMR spectrum was recorded in DMSO using Bruker AV 500 ultra-shield NMR instrument. A UV spectrum was recorded on a Specords 100 diode array spectrometer. Progress of the reactions was monitored using TLC, which was performed on glass slides (2x1.5cm) coated with silica gel-G using ethyl acetate: hexane (1:9) as the solvent system and the spots were visualized by exposure to iodine vapors or under UV light.

 

Methods:

(STEP – 1): Synthesis of 4- hydroxyl 7 methyl coumarin7, 8:

A mixture of m – cresol (15ml), Malonic acid (10ml) and ConH2SO4 (20ml) was taken in a beaker. So that the temperature of reaction mixture did not raise above the 100C the reaction on complete addition mixture was kept at ambient temperature for 18 hr and then poured with vigorous stirring to the mixture of ice and water. The precipitated was filter off and washed with cold water then dried under reduced pressure to afford the crude solid mass it was recrystallised from aqueous alcohol gives 4- hydroxyl 7 methyl coumarin.

 

(STEP – 2): Synthesis of β -Nitro styrene14:

A solution of benzaldehyde (10ml) and nitro methane (7.5ml) in methanol 100ml was cooled to 5ºC. Sodium hydroxide solution (7.5gm) in methanol (70ml) was added drop wise added at 5-10ºC and the reaction mixture was stirred for further 10 minutes. The resulting cold solution was run in to 30ml dilute HCL, when the product precipitated out wich was recrystallised from ethanol to give β -Nitro styrene.

 

(Step – 3): Synthesis of 7’Methyl 3 Phenyl 2, 3 Dihydro 4’ Furo Chromene:

An equimolar proportion of I and II in benzene was refluxed for 3 hours. The resulting solid was recrystallized from methanol.

 

SCHEMATIC REPRESENTATION

(Step 1)

 

 

(Step 2)

 

 

 

(Step 3)

 

7’Methyl 3 Phenyl 2,3 dihydro 4’ furo chromene

 

Antimicrobial activity18-20:

The antimicrobial activity of synthesized compound was determined by well plate or agar diffusion technique.

 

Preparation of media:

Wash the hands an wear gloves, sterilize all equipments then take 1gm of poptone bacteriological powder, 1gm beef extract, 2.5gm agar and 0.5gm sodium chloride then make the volume 100ml with distilled water. Then heat todissolve completely and sterilize by using autoclave.

 

Screening for antimicrobial activity:

The in vitro antimicrobial activity was carried out against 24 hours old cultures of bacteria. The different strains of bacteria Staphylococcus aureus Gram positive bacteria and Escherichia coli Gram negative bacteria are used. Pure cultures of the test microorganisms were procured from the compounds were tested at the concentrations of 50, 100, 150 and 200μg/ml and solutions were prepared by dissolving in distilled water. The petri dishes used for antibacterial screening were incubated at 37±1ºC for 48 hours. The results were compared to Ciprofloxacin (100μg/ml) for antibacterial activity, respectively by measuring zone of inhibition in mm.

 

 

Figure-1

RESULT AND DISCUSSION:

Physical and Chemical Method:

The TLC of the synthesized compound was shown in Figure-1. The synthesized compound, Rf value was found to be 0.84 in Ethyl acetate: Hexane (1:9). It is light yellow crystals. Melting point: 150-170oC. Molecular formula: C15H16O2N4S.

 

Ultraviolet Spectrum:

About 200mg of synthesized compound was dissolved in 10ml of anhydrous methanol. From this solution 0.1ml of solution was taken and makes up to 10ml with methanol. This was again made up to 10ml with methanol by taking 1ml of the above sample. This sample was scanned from 200nm-800nm. The synthesized compound melting point (150-170oC) showed an intense λ max at 226nm and absorbance is 0.862. Figure-2 represents the λmax of the synthesized compound.

 

Figure-2

 

 

 

Figure-3

 

 

Figure-4

 

Infrared Spectrum:

The IR spectrum of synthesized compound is shown in Fig: 3. The interpretation of various peaks obtained in the IR spectra of synthesized compound an enlisted below. The presence of peaks at 3200 indicates aromatic C-H, the value of 1500 indicates the presence of aromatic C=C in the compound. The presence of peaks at 1400 indicates aliphatic C-H group present in the structure. The value of 1600 indicates the presence of ketone C=O group present in the compound and the presence of peaks at 1100 indicates presence of ether C-O group in the compound.

 

NMR Spectrum:

The 1H NMR spectrum of synthesized compound is shown in Figure- 4. The interpretation of various signals obtained in the 1H NMR spectrum of synthesized compound is an enlisted below. The presence of a signals between δ 4-4.5ppm 1H NMR spectrum confirmed the presence of five proton (furon H). The signals between δ 6-7 accounts for the presence of (Ar-H) group of five protons. The signals between δ 1.5-2.5 assigned for the presence of three protons (CH3). The signals between δ 7.5-8 assigned for the presence of three protons (Coumarin-H).

 

 

Figure-5

 

TABLE-1

 

Zone of inhibition(mm)

Zone of inhibition(mm)

Test Compound

Gram positive

Gram negative

 

Staphylococcus aureus

E.coli

 

50 μg/ml

100 μg/ml

150 μg/ml

200 μg/ml

50 μg/ml

100 μg/ml

150 μg/ml

200 μg/ml

7’Methyl 3 Phenyl 2, 3 Dihydro 4’ Furo Chromene.

9.5

9.7

11.2

13.9

9.2

10.5

11.1

13.5

Ciprofloxacin(100μg/ml)

14.5

14.2

 

 

Mass spectrum:

The Mass spectrum of synthesized compound is shown in figure- 5. ESI-MS spectrum of synthesized compound exhibited a peak (M+ + 1•) at m/z 281. The molecular formula of synthesized compound is C15H16O2N4S. The molecular weight of compound is 280. It was determined by ESI-MS measurement.

 

Antimicrobial activity:

The antibacterial screening results were represented in Table 1. Table no 1 showing evaluation of in vitro antibacterial activity of 7’Methyl 3 Phenyl 2, 3 Dihydro 4’ Furo Chromene.

 

Ciprofloxacin (100μg/ml) was used as positive reference standard antibiotic.

According to table, the minimum inhibitory concentration (MIC) revealed that the 200 μg/ml drug was showing good antimicrobial activity against gram negative and gram-positive bacteria.

 

 

Figure – 5 Antibacterial activity of the synthesized compound and Ciprofloxacin antibiotic against Staphylococcus aureus

 

Figure – 6 Antibacterial activity of the synthesized compound and Ciprofloxacin antibiotic against E.coli

 

CONCLUSION:

The synthesized compound, melting point: 180ºC-188ºC, slight brown crystals on the U.V. studies indicated a coumarine nucleus in the compound. The 1H NMR spectrum of synthesized compound showed the presence of 16-hydrogen or proton present in the compound. Finally, the IR spectrum confirms the functional groups present in the compound and mass confirmed the molecular weight of the compound. Based on the Rf values, 1H NMR, IR and MASS studies the structure of synthesized compound has been characterized as 7’Methyl 3 Phenyl 2, 3 Dihydro 4’ Furo Chromene. A perusal of the literature reveals reports coumarine nucleus having antimicrobial activity. Our synthesized compound was having coumarine basic nucleus. So we are concluding according to result and discussion our compound also having antimicrobial activity.

 

ACKNOWLEDGEMENT:

I express my sincere thanks to Dr. R. Suthakaran and Management of Vijaya College of Pharmacy an encouragement throughout my research work and also thankful to NIPER, Hyderabad for helping the spectral analysis of our compound.

 

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Received on 26.02.2020            Modified on  21.03.2020           

Accepted on 16.04.2020   ©Asian Pharma Press All Right Reserved

Asian J. Pharm. Tech. 2020; 10(2):85-89.

DOI: 10.5958/2231-5713.2020.00016.1