The Hummel–Dreyer method: impact in pharmacology

Abstract

Reviews devoted to drug-protein binding present mostly a variety of analytical methods applied to studying the unique process when the protein reversibly binds a low-molecular-weight substance-the drug. This report, however, reviews the great majority of papers presenting the application of one particular method introduced by Hummel and Dreyer in 1962: Measurement of protein-binding phenomena by gel filtration. The main criterion in reviewing published papers was the application of the Hummel-Dreyer method when the counterpart substances were the drug and the human plasma protein.

Ligand substitutions between ruthenium–cymene compounds can control protein versus DNA targeting and anticancer activity

“…Samples containing 20 mM K-cacodylate (pH 6.0), 5 μM NCP and RAED-C:NCP ratios of 1:1, 2:1, 4:1 and 10:1 were incubated at room temperature for up to 48 h. Kinetic experiments of samples containing a 2:1 RAED-C:NCP ratio were carried out to determine the rate constant of the binding reaction (by monitoring the NCP–RAED-C adduct peak area) and the time required for an equilibrium to be established. The rate constant k was determined assuming pseudo-first-order kinetics and the binding constant K by using the equation r =( nKc )/(1+ Kc ) ( r , bound drug:NCP molar ratio; n , number of binding sites; c , concentration of unbound drug)35. The equation was solved for different RAED-C:NCP ratios by nonlinear regression analysis using Polymath 6.1.…”

A Ruthenium Antimetastasis Agent Forms Specific Histone Protein Adducts in the Nucleosome Core

“…The rate constant k was determined assuming pseudo-first-order kinetics and the binding constant K by adaptation of the Hummel-Dreyer method using the equation r = (nKc)/A C H T U N G T R E N N U N G (1+Kc) (r = bound drug:NCP molar ratio; n = number of binding sites; c = concentration of unbound drug). [40] The equation was solved for different RAPTA-C:NCP ratios by nonlinear regression analysis using Polymath 6.1. To assess the distribution of drug between the histone and DNA components, RAPTA-C-treated NCP was first denatured with 6 m guanidinium-HCl buffer prior to SEC fractionation.…”

Chromatographic analysis of drug interactions in the serum proteome

“…77 This method was modified for work with HPLC in 1978 and was validated by examining the binding that takes place between HSA and warfarin. 78 The HPLC version of the Hummel-Dreyer method has since been used for the analysis of many drug-protein systems. Examples of these systems have included the interactions of HSA with buspirone, ceftrixone, diazepam, isradipine, phenobarbitol, phenytoin, propranolol, warfarin, carvedilol, furosemide, phenylbutazone and pirprofen; the binding of AGP with isradipine, propranolol, propanfenone, and carvedilol; and the binding of HDL or LDL with amlodipine isradipine, or propranolol.…”

“…7 In addition, this approach has been modified for use in CE. 7,71 The Hummel-Dreyer method has been utilized in both binding studies 78 and competition studies, as has been shown in work with ISRP columns. 7,79 In addition, the Hummel-Dreyer technique has been employed with both serum proteins and other proteins or binding agents (e.g., enzymes, polysaccharides, and cyclodextrins).…”