title: Khaya grandifoliola C.DC: a potential source of active ingredients against hepatitis C virus in vitro.
abstract: In this study, we examined the antiviral properties of Khaya grandifoliola C.DC (Meliaceae) on the hepatitis C virus (HCV) life cycle in vitro and identified some of the chemical constituents contained in the fraction with the most antiviral activity. Dried bark powder was extracted by maceration in a methylene chloride/methanol (MCM) system (50:50; v/v) and separated on silica gel by flash chromatography. Infection and replication rates in Huh-7 cells were investigated by luciferase reporter assay and indirect immunofluorescence assay using subgenomic replicons, HCV pseudotyped particles, and cell-culture-derived HCV (HCVcc), respectively. Cell viability was assessed by MTT assay, and cellular gene expression was analysed by qRT-PCR. The chemical composition of the fraction with the most antiviral activity was analysed by coupled gas chromatography and mass spectrometry (GC-MS). Five fractions of different polarities (F0-F100) were obtained from the MCM extract. One fraction (KgF25) showed the strongest antiviral effect on LucUbiNeoET replicons at nontoxic concentrations. Tested at 100 µg/mL, KgF25 had a high inhibitory effect on HCV replication, comparable to that of 0.01 µM daclatasvir or 1 µM telaprevir. This fraction also inhibited HCVcc infection by mostly targeting the entry step. KgF25 inhibited HCV entry in a pan-genotypic manner by directly inactivating free viral particles. Its antiviral effects were mediated by the transcriptional upregulation of the haem oxygenase-1 gene and interferon antiviral response. Three constituents, namely, benzene, 1,1'-(oxydiethylidene)bis (1), carbamic acid, (4-methylphenyl)-, 1-phenyl (2), and 6-phenyl, 4-(1'-oxyethylphenyl) hexene (3), were identified from the active fraction KgF25 by GC-MS. Khaya grandifoliola contains ingredients capable of acting on different steps of the HCV life cycle.
authors: Laboratory of Pharmacology and Toxicology, Department of Biochemistry, Faculty of Science, University of Yaounde I, PO Box 812, Yaounde, Cameroon.|Laboratory of Applied Biochemistry, Department of Biological Sciences, Faculty of Science, University of Ngaoundéré, Ngaoundéré, Cameroon.
authors: Borris Rosnay Tietcheu
authors: BR
authors: Galani
authors: CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019, UMR 8204, CIIL, Centre d'Infection et d'Immunité de Lille, Univ. Lille, 59000, Lille, France.
authors: Marie-Emmanuelle
authors: ME
authors: Sahuc
authors: Institute of Experimental Immunology and Hepatology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Gabriele
authors: G
authors: Sass
authors: Laboratory of Pharmacology and Toxicology, Department of Biochemistry, Faculty of Science, University of Yaounde I, PO Box 812, Yaounde, Cameroon.
authors: Frédéric Nico
authors: FN
authors: Njayou
authors: Institute of Experimental Immunology and Hepatology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Christine
authors: C
authors: Loscher
authors: Department of Organic Chemistry, Faculty of science, University of Yaounde I, Yaounde, Cameroon.
authors: Pierre
authors: P
authors: Mkounga
authors: CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019, UMR 8204, CIIL, Centre d'Infection et d'Immunité de Lille, Univ. Lille, 59000, Lille, France.
authors: Gaspard
authors: G
authors: Deloison
authors: CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019, UMR 8204, CIIL, Centre d'Infection et d'Immunité de Lille, Univ. Lille, 59000, Lille, France.
authors: Priscille
authors: P
authors: Brodin
authors: CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019, UMR 8204, CIIL, Centre d'Infection et d'Immunité de Lille, Univ. Lille, 59000, Lille, France.
authors: Yves
authors: Y
authors: Rouillé
authors: Institute of Experimental Immunology and Hepatology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Gisa
authors: G
authors: Tiegs
authors: CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019, UMR 8204, CIIL, Centre d'Infection et d'Immunité de Lille, Univ. Lille, 59000, Lille, France.
authors: Karin
authors: K
authors: Séron
authors: Laboratory of Pharmacology and Toxicology, Department of Biochemistry, Faculty of Science, University of Yaounde I, PO Box 812, Yaounde, Cameroon. pmoundipa@hotmail.com.
authors: Paul Fewou
authors: PF
authors: Moundipa
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D045744:Cell Line, Tumor; D002850:Chromatography, Gel / Q000379:methods; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D016174:Hepacivirus / Q000187:drug effects*; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D029582:Meliaceae / Q000737:chemistry; D024301:Plant Bark / Q000737:chemistry; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D010936:Plant Extracts