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Related Concept Videos

The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

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Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
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Physiological models with protein binding in pharmacokinetics offer a sophisticated approach to understanding drug disposition. These models consider drug-protein interactions, enabling them to effectively predict drug concentrations in different organs and tissues. This precision aids in accurate drug dosing, providing a significant advantage over conventional models. A key process within these models is equilibration, which ensures that drug concentrations achieve a steady state within the...
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Drug concentration is the quantity of a drug present in a biological sample. Measuring drug amounts in biological samples allows the clinician to understand how a drug is absorbed, distributed, metabolized, and excreted. Samples can be obtained through invasive or non-invasive methods. Invasive techniques involve surgical or parenteral interventions to gather blood, cerebrospinal fluid, or tissue biopsy. Conversely, non-invasive approaches provide samples like urine, feces, and saliva.
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Drugs predominantly attach to plasma proteins, with only a small percentage remaining unbound. The unbound portion can be calculated as one minus the bound fraction. Acidic drugs form large, inactive complexes by reversibly binding to plasma albumin, which prevents them from diffusing across biological barriers. These drug-protein complexes act as reservoirs for the drugs. As the concentration of unbound drugs decreases, these complexes quickly dissociate to release the free drug, maintaining...
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Deriving protein binding-corrected chemical concentrations for in vitro testing.

Aditya R Kolli1

  • 1PMI R&D, Philip Morris Products S.A., Neuchâtel, Switzerland.

Clinical and Translational Science
|August 22, 2023
PubMed
Summary

Accurate in vitro testing requires accounting for protein binding. This study developed models to calculate corrected chemical concentrations, ensuring physiologically relevant results for drug development and toxicology.

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Area of Science:

  • Pharmacology
  • Toxicology
  • Biochemistry

Background:

  • In vitro testing traditionally uses cell culture media with 5%-10% fetal bovine serum (FBS).
  • Variable unbound extracellular chemical concentrations can occur due to physicochemical properties and testing conditions, complicating the assessment of chemical potency and concentration-response relationships.
  • Accurate interpretation of in vitro data necessitates understanding the unbound fraction of chemicals.

Purpose of the Study:

  • To develop and apply protein binding models for calculating corrected chemical concentrations in vitro.
  • To determine the significance of protein binding corrections for chemicals with varying binding affinities in FBS-containing media.
  • To provide a tool for researchers to easily calculate these adjusted concentrations.

Main Methods:

  • Utilized one- and two-protein binding models to estimate protein binding-corrected chemical concentrations.
  • Evaluated the protein binding of ceftizoxime, moxifloxacin, and nicotine, noting their low binding affinity in 5%-10% FBS.
  • Highlighted the necessity of corrections for moderate and highly protein-bound chemicals.

Main Results:

  • Low protein binding (<5%) was observed for ceftizoxime, moxifloxacin, and nicotine in 5%-10% FBS, considered negligible.
  • Significant variations in unbound concentrations were found for moderate and highly protein-bound chemicals in standard cell culture media.
  • Protein binding corrections are crucial for accurate in vitro concentration-response assessments.

Conclusions:

  • In vitro pharmacological and toxicological assessments require protein binding-adjusted concentrations for physiological relevance.
  • Standard in vitro testing conditions with 5%-10% FBS can lead to inaccurate unbound concentrations for many chemicals.
  • An Excel tool is provided to facilitate the calculation of protein binding-corrected concentrations for in vitro studies.