title: Intracellular concentrations determine the cytotoxicity of adefovir,  cidofovir and tenofovir.
abstract: Lack of in vitro to in vivo translation is a major challenge in safety prediction during early drug discovery.One of the most common in vitro assays to evaluate the probability of a compound to cause adverse effects is a cytotoxicity assay. Cytotoxicity of a compound is often measured by dose–response curves assuming the administered doses and intracellular exposures are equal at the time of measurement.However, this may not be true for compounds with low membrane permeability or those which are substrates for drug transporters as intracellular concentrations are determined both by passive permeability and active uptake through drug transporters. We show here that three antiviral drugs, adefovir, cidofovir and tenofovir exhibit significantly increased cytotoxicity in HEK293 cells transfected with organic anion transporter (OAT) 1 and 3 compared to a lack of cytotoxicity in HEK293 wildtype cells. A further look at the media and intracellular drug concentrations showed that 24 h after dosing, all three drugs had higher intracellular drug concentrations than that of media in the HEK-OAT1 cells whereas the intracellular drug concentrations in the wildtype cells were much lower than the administered doses. Comparing cytotoxicity IC(50) values of adefovir, cidofovir and tenofovir based on administered doses and measured intracellular concentrations in HEK-OAT1 cells revealed that intracellular drug concentrations have significant impact on calculated IC(50) values. Tenofovir showed much less intrinsic cytotoxicity than adefovir and cidofovir using intracellular concentrations rather than media concentration. Our data suggest that for low permeable drugs or drugs that are substrates for drug transporters, the choice of cellular model is critical for providing an accurate determination of cytotoxicity.
authors.affiliation: Compound Safety Prediction, Pfizer Inc., Groton, CT 06340, USA. xun.zhang126@gmail.com
authors.forename: Xun
authors.initials: X
authors.lastname: Zhang
authors.forename: Ruduan
authors.initials: R
authors.lastname: Wang
authors.forename: Mary
authors.initials: M
authors.lastname: Piotrowski
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Karen L
authors.initials: KL
authors.lastname: Leach
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000032:analysis / Q000633:toxicity; D000998:Antiviral Agents / Q000032:analysis / Q000633:toxicity*; D000077404:Cidofovir; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells / Q000737:chemistry / Q000187:drug effects; D006801:Humans; D027362:Organic Anion Transport Protein 1 / Q000096:biosynthesis / Q000378:metabolism; D063065:Organophosphonates / Q000032:analysis / Q000633:toxicity*; D000068698:Tenofovir; D018675:Toxicity Tests
chemical_list: D000998:Antiviral Agents; D027362:Organic Anion Transport Protein 1; D063065:Organophosphonates; C053001:adefovir; D003596:Cytosine; D000068698:Tenofovir; D000225:Adenine; D000077404:Cidofovir