Phytochemistry of Red Betel leaf (Piper crocatum) and its antifungal activity against Lanosterol 14 alpha demethylase in Candida albicans in vitro and in silico Tessa Siswina (a), Mia Miranti (b), Dadan Sumiarsa (c), Dikdik Kurnia (c*)
(a) Universitas Padjadjaran, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Sumedang, 45363, Indonesia- Poltekkes Kemenkes Pontianak, Department of Midwifery, Pontianak, 78241, Indonesia
(b) Universitas Padjadjaran,Department of Biology, Faculty of Mathematics and Natural Sciences, Sumedang, 45363, Indonesia
(c) Universitas Padjadjaran, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Sumedang, 45363, Indonesia
(*) correspondence : dikdik.kurnia[at]unpad.ac.id
Abstract
90% of Indonesian women and 31.8% of unmarried adolescents aged 15-24 years experience vaginal discharge due to Candida albicans, and 45% of recurrent vaginal discharge. In the Asia Pacific, the incidence of azole resistance reaches 83%, so non-pharmacological treatment in vaginal discharge is urgently needed. Red betel leaf (P. crocatum) is an antifungal plant. The mechanism of antifungal agents by inhibiting lanosterol 14 alpha-demethylase (CYP51) is a promising target to be developed. Lanosterol plays an important role in the biosynthesis of ergosterol as a key enzyme that damages membranes and kills fungal cells. The aims of this study were to identify the phytochemicals of P. crocatum, evaluate the antifungal activity against C. albicans, and predict its molecular interactions in vitro and in silico. Phytochemical tests were carried out using standard procedures, in vitro antifungal analysis by disc diffusion and microdilution methods, and in silico tests by docking protein ligands. The results showed that the P. crocatum methanol extract had secondary metabolites in the form of flavonoids, steroids, alkaloids, phenolics, tannins, and saponins. The antifungal activity was indicated by the zone of inhibition by ketoconazole, P. crocatum, metronidazole, and aquadest of 20.7, 8.4, 0 and 0 mm in fraction 5, respectively- 18.5, 9.3, 0, and 0 mm in fraction 4- 17.5, 8.1, 0, and 0 mm in fraction 2- 16.7, 8.3, 0, and 0 mm in fraction 3. Meanwhile, the binding energies of ketoconazole (positive control), metronidazole, selinene, eugenyl acetate, eugenol, and safrole to lanosterol 14 alpha-demethylase were -10.61, -5.55, -7.76, -5.77, -5.61, and -5.56 Kcal/mol at His377, respectively. Selinene has a stronger bond than other compounds in Red Betel leaf. Although not as good as the positive control (ketokonazole), it has the natural antibiotics potentially in vaginal discharge caused by C. albicans by inhibiting lanosterol 14 alpha-demethylase.
