title: In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis B virus isolates.
abstract: Hepatitis B virus (HBV) infection causes major public health problems worldwide. The clinical limitation of current antiviral drugs for HBV, such as lamivudine, is the emergence of drug-resistant viral strains during prolonged antiviral therapy. Cepharanthine hydrochloride (CH), a natural alkaloid-derived compound, has been reported to possess potent activity against various viruses. The present study was performed to evaluate the in vitro activity of CH against clinical wild-type and lamivudine-resistant HBV isolates in transiently transfected cells. HBV DNA was extracted from serum samples collected both before lamivudine therapy and at the time of viral breakthrough and was amplified by polymerase chain reaction (PCR). The amplicons were cloned into a novel expression vector, pHY106, which can initiate the intracellular HBV replication cycle after cell transfection. Following transfection of the cloned amplicon into HepG2 cells, a drug susceptibility assay was performed. The level of viral antigen, HBeAg, was determined by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time PCR (Q-PCR) was used for determining the amount of intracellular HBV DNA. Heat stress cognate 70 (Hsc70), a host protein required for HBV replication, was also analyzed by reverse transcription PCR (RT-PCR) to explore the possible antiviral mechanism of CH. The results showed that CH inhibited replication and HBeAg production by either wild-type or lamivudine-resistant HBV clinical isolates in a dose-dependent manner. The Hsc70 mRNA was also downregulated significantly. In conclusion, CH is active against both wild-type and lamivudine-resistant HBV clinical isolates, and its activity may be associated with its inhibition of host Hsc70.
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver Diseases, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, China.
authors.forename: Yu-Bing
authors.initials: YB
authors.lastname: Zhou
authors.forename: Ya-Feng
authors.initials: YF
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Li-Yun
authors.initials: LY
authors.lastname: Zheng
authors.forename: Xiao-Ang
authors.initials: XA
authors.lastname: Yang
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Jin-Hua
authors.initials: JH
authors.lastname: Jiang
authors.forename: Fang
authors.initials: F
authors.lastname: Ma
authors.forename: De-Tao
authors.initials: DT
authors.lastname: Yin
authors.forename: Chang-Yu
authors.initials: CY
authors.lastname: Sun
authors.forename: Qing-Duan
authors.initials: QD
authors.lastname: Wang
mesh_terms: D000328:Adult; D000998:Antiviral Agents / Q000009:adverse effects / Q000494:pharmacology*; D044182:Benzylisoquinolines / Q000009:adverse effects / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002681:China; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D024882:Drug Resistance, Viral; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006509:Hepatitis B / Q000097:blood / Q000188:drug therapy* / Q000821:virology; D006513:Hepatitis B e Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D006801:Humans; D020128:Inhibitory Concentration 50; D019259:Lamivudine / Q000627:therapeutic use*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*; D052117:Benzodioxoles
chemical_list: D000998:Antiviral Agents; D044182:Benzylisoquinolines; D004279:DNA, Viral; D050883:HSC70 Heat-Shock Proteins; D006513:Hepatitis B e Antigens; D019259:Lamivudine; D012333:RNA, Messenger; D011994:Recombinant Proteins; C494371:HSPA8 protein, human; C006947:cepharanthine; D052117:Benzodioxoles