title: In silico analysis and experimental validation of azelastine hydrochloride (N4) targeting sodium taurocholate co-transporting polypeptide (NTCP) in HBV therapy.
abstract: OBJECTIVE
  The aim of this study was to explore sodium taurocholate co-transporting polypeptide (NTCP) exerting its function with hepatitis B virus (HBV) and its targeted candidate compounds, in HBV therapy.
  
  
  METHODS
  Identification of NTCP as a novel HBV target for screening candidate small molecules, was used by phylogenetic analysis, network construction, molecular modelling, molecular docking and molecular dynamics (MD) simulation. In vitro virological examination, q-PCR, western blotting and cytotoxicity studies were used for validating efficacy of the candidate compound.
  
  
  RESULTS
  We used the phylogenetic analysis of NTCP and constructed its protein-protein network. Also, we screened compounds from Drugbank and ZINC, among which five were validated for their authentication in HepG 2.2.15 cells. Then, we selected compound N4 (azelastine hydrochloride) as the most potent of them. This showed good inhibitory activity against HBsAg (IC50 = 7.5 μm) and HBeAg (IC50 = 3.7 μm), as well as high SI value (SI = 4.68). Further MD simulation results supported good interaction between compound N4 and NTCP.
  
  
  CONCLUSIONS
  In silico analysis and experimental validation together demonstrated that compound N4 can target NTCP in HepG2.2.15 cells, which may shed light on exploring it as a potential anti-HBV drug.
authors: College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Sichuan University, Chengdu, 610064, China.
authors: L-L
authors: LL
authors: Fu
authors: J
authors: J
authors: Liu
authors: Y
authors: Y
authors: Chen
authors: F-T
authors: FT
authors: Wang
authors: X
authors: X
authors: Wen
authors: H-Q
authors: HQ
authors: Liu
authors: M-Y
authors: MY
authors: Wang
authors: L
authors: L
authors: Ouyang
authors: J
authors: J
authors: Huang
authors: J-K
authors: JK
authors: Bao
authors: Y-Q
authors: YQ
authors: Wei
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D003198:Computer Simulation; D015195:Drug Design; D056945:Hep G2 Cells; D006509:Hepatitis B / Q000188:drug therapy*; D006515:Hepatitis B virus / Q000187:drug effects*; D006801:Humans; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D029363:Organic Anion Transporters, Sodium-Dependent / Q000235:genetics / Q000378:metabolism*; D010793:Phthalazines / Q000737:chemistry / Q000494:pharmacology*; D010802:Phylogeny; D027981:Symporters / Q000235:genetics / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D029363:Organic Anion Transporters, Sodium-Dependent; D010793:Phthalazines; D027981:Symporters; C071908:sodium-bile acid cotransporter; C020976:azelastine