Using capillary electrophoresis/frontal analysis to screen drugs interacting with human serum proteins.
We have used capillary electrophoresis in the frontal analysis mode (CE/FA) to determine the binding capacity of beta-adrenoceptor blocking drugs to individual serum proteins, serum protein mixtures and human serum. The free drug concentration was directly measured from the height of the frontal peak and used to calculate the bound drug concentration. From the bound drug concentration, the percentage of drug bound to the serum proteins alpha1-acid glycoprotein (AGP) and human serum albumin (HSA) was then determined. In addition to determining the percent of a drug bound to a protein, the drug-protein association constant (Ka) was determined for AGP binding to beta-blockers. The data-estimated association constants were consistent with literature values. The CE/FA studies on the beta-adrenoceptor blocking drugs and the serum proteins indicated that HSA, AGP, high density lipoprotein (HDL), and low density lipoprotein (LDL) were the main contributors to serum binding for this series of compounds. The serum-drug binding data sorted the beta-adrenoceptor blocking drugs into high and low binding categories. The protein mixture (AGP + HSA + HDL + LDL) resulted in dividing the beta-blockers into the same high/low rankings. The protein mixture (AGP + HSA + HDL + LDL) was amenable to automation, did not autoaggregate, and had constant concentrations for the proteins.
McDonnell
P A
PA
The R. W. Johnson Pharmaceutical Research Institute, Spring House, PA 19477, USA. pmcdonne@prius.jnj.com
Caldwell
G W
GW
Masucci
J A
JA
D000319:Adrenergic beta-Antagonists / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism*; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000097:blood / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000097:blood / Q000378:metabolism; D009961:Orosomucoid / Q000378:metabolism; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism
D000319:Adrenergic beta-Antagonists; D001798:Blood Proteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D009961:Orosomucoid; D012709:Serum Albumin