Alkaloid and benzopyran compounds of Melicope latifolia fruit exhibit anti-hepatitis C virus activities

Abstract

Background New agents for developing alternative or complementary medicine to treat the hepatitis C virus (HCV) are still needed due to high rates of HCV infection globally and the current limitations of available treatments. Treatment of HCV with a combination of direct acting antivirals have been shown to be approximately 90% effective but will be limited in the future due to the emergence of drug resistance and high cost. The leaves of Melicope latifolia have previously been reported to have anti-HCV activity and are a potential source of bioactive compounds for future novel drug development. This study aimed to evaluate the efficacy of the extract of M. latifolia fruit to treat HCV and to isolate its active compounds. Method M. latifolia fruit was extracted using methanol and purified using vacuum liquid chromatography (VLC) and Radial Chromatography. The anti-HCV activity was analyzed using cell culture lines Huh7it-1 and JFH1 (genotype 2a). Time-of-addition and immunoblotting studies were performed to identify the mode of action of the isolated active compounds. The structures of the active compounds were determined using nuclear magnetic resonance (NMR) spectra, UV, IR, and Mass Spectra. Results Six known compounds were isolated from M. latifolia fruit: O-methyloktadrenolon, alloevodionol, isopimpinellin, alloxanthoxyletin, methylevodionol, and N-methylflindersine. N-methylflidersine was the most active compound with IC50 value of 3.8 μg/ml while methylevodionol, isopimpinellin, and alloevodionol were less active. O-methyloktadrenolon and alloxanthoxyletin were moderately active with IC50 values of 10.9 and 21.72 μg/ml, respectively. N-methylflidersine decreased level of HCV NS3 protein expression in the cells. Conclusion The alkaloid compound, N-methylflindersine which was isolated from M. latifolia possesses anti-HCV activity through post-entry inhibition and suppressed NS3 protein expression.

Phytochemical, Metabolite Compound, and Antioxidant Activity of Clinacanthus nutans Leaf Extract from Indonesia

“…Alloevodionol contained in Melicope sp. also showed antioxidant, antiproliferative and anti-hepatitis C virus activity (Johnson et al, 2010;Widyawaruyanti et al, 2021;Le et al, 2021. Boesenbergin is a compound from Boesenbergia rotunda with antioxidant and cytotoxic e ects on various cancer cell lines and anti-in ammatory activity on RAW 264.7 (Isa et al, 2012) . Meanwhile, 6-Gingerdiol, a Gingerol metabolite from Zingiberaceae, is reported to ameliorate cisplatin-induced oxidative stress by decreasing lipid peroxide markers and increasing antioxidant enzymes (Kuhad et al, 2006) .…”

A Review of the Ethnomedicinal, Phytochemical, and Anticancer Properties of Melicope Species

“…This metabolite will lead to their biological activities. [40,43] Six compounds from the class of alkaloid and benzopyran were successfully isolated and identified from the dried fruit of M. latifolia. They are O-methyloktadrenolon, alloevodionol, isopimpinellin, alloxanthoxyletin, methylevodionol, and N-methylflindersine.…”

“…They are O-methyloktadrenolon, alloevodionol, isopimpinellin, alloxanthoxyletin, methylevodionol, and N-methylflindersine. [43] The isolation and extraction work on dry leaves of methanol extract had successfully characterized five spiroke- [39] In another study, the isolation and extraction of M. latifolia successfully isolated two monoterpenoids and four coumarins. The monoterpenoids are melifoliones A and melifoliones B while the four coumarin derivatives are bergapten, 5,7,8trimethoxycoumarin, 6,7,8-trimethoxycoumarin, and bruceol.…”

An in vitro study of an Artocarpus heterophyllus substance as a hepatitis C antiviral and its combination with current anti-HCV drugs

“…The cytotoxicity of the samples to the cells was assessed using a 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay as described previously [21,25]. Huh7it-1 cells (2.4 × 10 4 ) placed in a 96 well plate were combined with sample at various concentrations and incubated for 48 h. After incubation, the medium was discarded and 150 μL of medium containing MTT (15 μL) was added and incubated for a further 4 h. Then 100 μL of DMSO was added to dissolve the precipitate that formed from the MTT reaction.…”

“…Equal amounts of proteins were separated using SDS polyacrylamide gel electrophoresis and transferred onto a polyvinylidene difluoride (PVDF) membrane (Millipore, Bed-ford, MA, USA). The membranes were first probed with primary antibodies: a HCV NS3 mouse monoclonal antibody (clone H23; Abcam, Cambridge, MA, USA) and a βactin antibody (MBL, Nagoya, Japan) followed by a secondary antibody, HRP-conjugated goat anti-mouse immunoglobulin (MBL) [21,25]. Target proteins were visualized using an enhanced chemiluminescence detection system (Biorad; GE Healthcare, UK).…”