Multi-drugs resistance has addressed a growing trouble in the treatment of many infectious diseases caused by several kinds of bacteria and fungi. The discovery and development of effective antibacterial and antifungal agents with novel chemical structures become urgent tasks for infectious disease research programs.
In the current work, the powder obtained from crushing the seeds of Red Delicious apple was extracted by solvents of different polarities including water, methanol, chloroform, and n-hexane. The extraction was carried out via three extraction methods named kinetic maceration, ultrasound-expedited and microwave-expedited extraction techniques. For each one, the extraction was performed in three styles, which are non-serial, serial risingly- and fallingly-arranged in polarity.
The resultant extracts were submitted to the phytochemical screening tests for seeking the existence of many primary and secondary plant metabolites. The investigated phytochemicals were flavonoids, tannins, terpenoids, carbohydrates, alkaloids, emodins, phenols, steroids, anthocyanins, betacyanins, amino acids, proteins, saponins, glycosides, coumarins, anthraquinones, and fixed oils.
The results acquired from the phytochemical analysis documented that coumarins could be detected in the obtained methanol and chloroform extracts. According to these results, the chloroform extract obtained from a non-serial ultrasound-expedited extraction method was selected to isolate its coumarin components.
Since the processes of separation and purification completed, four novel furanocoumarins have been acquired. Their chemical structures were illustrated by analyzing their FTIR, 1H-NMR and 13C-NMR spectra and corresponding their spectroscopic data with those found in the literature.
Two in vitro antimicrobial studies were verified for the isolated products via a broth dilution method; the antibacterial activity versus the following standard bacterial strains: Pseudomonas aeruginosa, Klebsiella pneumonia, Haemophilus influenzae and Escherichia coli utilizing Ciprofloxacin as a reference, and the antifungal activity versus the following standard fungal strains: Candida albicans and Aspergillus niger using Nystatin as a reference. The results indicated that the isolated furanocoumarins had a promising antimicrobial activity against the test pathogens with the superior activity attributed to compound E1 ((E)-12-(2'-Chlorovinyl)bergapten) . Also, the isolated products displayed encouraging bactericidal and fungicidal activities based on their MBC/MIC and MFC/MIC values.
In the literature, there is a large number of reports which documented that the enhanced lipophilicity of the antimicrobial agent may improve its penetration into pathogenic microorganisms resulting in a better antimicrobial effect. To examine this assumption on the isolated furanocoumarins, one of them, which is compound E3 (12-(2'-chloropropan-2'-yl)-8-hydroxybergapten) was chemically modified in such a way to increase its lipophilicity. The results of testing the antimicrobial activity of this semisynthetic product revealed improvement in the activity versus the same test pathogens. Accordingly, it is concluded that it is possible to increase the antimicrobial activity of the isolated natural furanocoumarins by enhancing their lipophilicity. This may be accomplished by either introducing a non-polar functional group or modifying the currently available polar active group.