title: Development of a high throughput equilibrium dialysis method.
abstract: The identification of large numbers of biologically active chemical entities during high throughput screening (HTS) necessitates the incorporation of new strategies to identify compounds with drug-like properties early during the lead prioritization and development processes. One of the major steps in lead prioritization is an assessment of compound binding to plasma proteins, because it affects both the pharmacokinetics and pharmacodynamics of the compound in vivo. Equilibrium dialysis is the preferred method to determine the free drug fraction, because it is less susceptible to experimental artifacts. However, even low-volume standard equilibrium dialysis is currently not amenable to the HTS format. Those considerations dictate the development of a high throughput equilibrium dialysis device, without compromising the analytical quality of the data. The present paper demonstrates successful development of a 96-well format equilibrium dialysis plate. Plasma protein binding of three drugs, propranolol, paroxetine, and losartan, with low, intermediate, and high binding properties, respectively, were chosen for assay validation. The data indicate that the apparent free fraction obtained by this method correlates with the published values determined by the traditional equilibrium dialysis techniques.
authors: Leads Discovery Department, Dupont Pharmaceuticals Company, Rt. 141 & Henry Clay Road, Wilmington, Delaware 19880, USA. ilona.kariv@dupontpharma.com
authors: I
authors: I
authors: Kariv
authors: H
authors: H
authors: Cao
authors: K R
authors: KR
authors: Oldenburg
mesh_terms: D000928:Antidepressive Agents / Q000493:pharmacokinetics*; D000959:Antihypertensive Agents / Q000493:pharmacokinetics*; D001798:Blood Proteins / Q000378:metabolism*; D003956:Dialysis / Q000379:methods; D004353:Drug Evaluation, Preclinical; D006801:Humans; D019808:Losartan / Q000493:pharmacokinetics*; D008297:Male; D017374:Paroxetine / Q000493:pharmacokinetics*; D011433:Propranolol / Q000493:pharmacokinetics*; D011485:Protein Binding / Q000502:physiology; D014462:Ultrafiltration / Q000379:methods
chemical_list: D000928:Antidepressive Agents; D000959:Antihypertensive Agents; D001798:Blood Proteins; D017374:Paroxetine; D011433:Propranolol; D019808:Losartan