HDM-PAMPA to predict gastrointestinal absorption, binding percentage, equilibrium and kinetics constants with human serum albumin and using 2 end-point measurements.
The parallel artificial membrane permeability assay (PAMPA) is a high-throughput screening (HTS) technique developed to predict passive permeability through numerous different biological membranes, such as the gastrointestinal tract (GIT), the blood brain barrier (BBB), and the dermal layer. PAMPA is based on an artificial membrane, such as hexadecane (HDM), which separates two compartments (i.e., a donor and an acceptor compartment). In the present study, an HDM-PAMPA method was developed with human serum albumin (HSA) under iso-pH and gradient-pH conditions to predict the percentage of binding, dissociation/association constants (Kdand Ka, respectively) and dissociation/association kinetic rates (koffand kon, respectively) between a given drug and HSA. Thanks to the kinetic properties of PAMPA, a two end-point assay was implemented to obtain all three properties. The assay was used to measure basic, acidic, and amphoteric compounds. The protein was free in solution, allowing a direct comparison between this assay and equilibrium dialysis (ED). The developed PAMPA enabled screening of up to 96 compounds in a single run, generating valuable information on absorption and distribution in a high-throughput and high-repeatable manner.
School of Pharmaceutical Sciences, University of Geneva-University of Lausanne CMU, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland.
Alban
A
Bujard
School of Pharmaceutical Sciences, University of Geneva-University of Lausanne CMU, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland.
Charlotte
C
Petit
School of Pharmaceutical Sciences, University of Geneva-University of Lausanne CMU, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland.
Pierre-Alain
PA
Carrupt
School of Pharmaceutical Sciences, University of Geneva-University of Lausanne CMU, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland.
Serge
S
Rudaz
School of Pharmaceutical Sciences, University of Geneva-University of Lausanne CMU, 1 rue Michel-Servet, 1211 Geneva 4, Switzerland. Electronic address: julie.schappler@unige.ch.
Julie
J
Schappler
D000473:Alkanes / Q000378:metabolism / Q000493:pharmacokinetics*; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D023381:Endpoint Determination / Q000379:methods* / Q000592:standards; D005544:Forecasting; D065569:Gastrointestinal Absorption / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D008567:Membranes, Artificial; D011485:Protein Binding / Q000502:physiology; D012709:Serum Albumin / Q000378:metabolism*
D000473:Alkanes; D008567:Membranes, Artificial; D012709:Serum Albumin; C007932:n-hexadecane
Binding percentage; Dissociation/association constants; Dissociation/association microconstants; Gastrointestinal tract absorption; Human serum albumin; PAMPA; Passive permeability