title: Mechanistic investigations into the species differences in pinometostat clearance: impact of binding to alpha-1-acid glycoprotein and permeability-limited hepatic uptake.
abstract: 1. The plasma clearance of the first-in-class DOT1L inhibitor, EPZ-5676 (pinometostat), was shown to be markedly lower in human compared to the preclinical species, mouse, rat and dog. 2. This led to vertical allometry where various interspecies scaling methods were applied to the data, with fold-errors between 4 and 13. We had previously reported the elimination and metabolic pathways of EPZ-5676 were similar across species. Therefore, the aim of this work was to explore the mechanistic basis for the species difference in clearance for EPZ-5676, focusing on other aspects of disposition. 3. The protein binding of EPZ-5676 in human plasma demonstrated a non-linear relationship suggesting saturable binding at physiologically relevant concentrations. Saturation of protein binding was not observed in plasma from preclinical species. Kinetic determinations using purified serum albumin and alpha-1-acid glycoprotein (AAG) confirmed that EPZ-5676 is a high affinity ligand for AAG with a dissociation constant (Kd) of 0.24 μM. 4. Permeability limited uptake was also considered since hepatocyte CLintwas much lower in human relative to preclinical species. Passive unbound CLintfor EPZ-5676 was estimated using a correlation analysis of logD and data previously reported on seven drugs in sandwich cultured human hepatocytes. 5. Incorporation of AAG binding and permeability limited hepatic uptake into the well-stirred liver model gave rise to a predicted clearance for EPZ-5676 within 2-fold of the observed value of 1.4 mL min-1kg-1. This analysis suggests that the marked species difference in EPZ-5676 clearance is driven by high affinity binding to human AAG as well as species-specific hepatic uptake invoking the role of transporters.
authors.affiliation: a Epizyme Inc , Cambridge , MA , USA and.
authors.forename: Sherri A
authors.initials: SA
authors.lastname: Smith
authors.affiliation: b Charles River Laboratories , Montreal , QC , Canada.
authors.forename: Sandra
authors.initials: S
authors.lastname: Gagnon
authors.affiliation: a Epizyme Inc , Cambridge , MA , USA and.
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Waters
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism*; D001562:Benzimidazoles / Q000378:metabolism*; D004285:Dogs; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D051379:Mice; D009961:Orosomucoid / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D013045:Species Specificity
chemical_list: D000970:Antineoplastic Agents; D001562:Benzimidazoles; C583893:EPZ-5676; D009961:Orosomucoid
keywords: DOT1L inhibitor; EPZ-5676; IVIVE; fraction unbound; non-linear kinetics