Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

161
Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...
161
Drug Binding to Blood Components01:30

Drug Binding to Blood Components

139
When drugs enter systemic circulation, they interact with various components of the blood, including proteins such as human serum albumin (HSA), α1-acid glycoprotein (AAG), lipoproteins, globulins, and red blood cells (RBCs).
HSA is the most abundant plasma protein and is vital in drug binding. It contains distinct drug-binding sites, with different drugs exhibiting affinity for specific sites. There are three main drug-binding domains for HSA: sites I, II, and III. These domains are...
139
Affinity Chromatography01:03

Affinity Chromatography

603
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
603
Drug Distribution: Plasma Protein Binding01:29

Drug Distribution: Plasma Protein Binding

5.5K
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...
5.5K
Protein-Drug Binding: Mechanism and Kinetics01:16

Protein-Drug Binding: Mechanism and Kinetics

398
Protein-drug binding refers to the interaction between drugs and proteins within the body. This binding process can occur intracellularly, involving drug interactions with enzymes or receptors within cells, or extracellularly, involving plasma proteins in the blood.
Various forces drive these interactions, including hydrogen bonds, hydrophobic interactions, ionic bonds, electrostatic interactions, and van der Waals forces. These bonds enable drugs to bind to specific sites on proteins,...
398
Factors Affecting Protein-Drug Binding: Drug-Related Factors01:18

Factors Affecting Protein-Drug Binding: Drug-Related Factors

96
Drug binding to proteins is a complex phenomenon influenced by various drug-related factors, each playing a significant role in the interaction between drugs and proteins within the body.
One crucial factor in drug-protein binding is the drug's lipophilicity or its affinity for fat. More lipophilic drugs tend to have higher binding extents. For example, highly lipophilic drugs like cloxacillin exhibit substantial protein binding, with as much as 95% of the drug binding to proteins. In...
96

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Novel Xanthene Derivatives for Neuroprotection in Alzheimer's Disease-Synthesis and Biological Assessment.

ChemMedChem·2026
Same author

Tolcapone-loaded nanostructured lipid carriers for improved oral delivery.

International journal of pharmaceutics·2026
Same author

Chitosan Derivatives: Challenges and Opportunities in the Green and Sustainable Transition Era.

Molecules (Basel, Switzerland)·2026
Same author

The biophysical characterization of the transcription factor MntR from Mycobacterium tuberculosis and its selected mutants.

International journal of biological macromolecules·2026
Same author

Thioxanthone-Mediated Cytoprotection Against Cisplatin Toxicity: Exploring the Potential Involvement of P-Glycoprotein Through Computational and Experimental Approaches.

Journal of xenobiotics·2026
Same author

Incremental value of implantable loop recorders in arrhythmia detection and management in cardiomyopathies: Prospective study.

Revista portuguesa de cardiologia : orgao oficial da Sociedade Portuguesa de Cardiologia = Portuguese journal of cardiology : an official journal of the Portuguese Society of Cardiology·2026

Related Experiment Video

Updated: Jun 22, 2025

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening
08:34

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

Published on: October 16, 2015

10.0K

Binding studies of promethazine and its metabolites with human serum albumin by high-performance affinity

Maria Miguel Coelho1,2,3, Rita Lima1,2, Ana Sofia Almeida1,2,3

  • 1Laboratory of Organic and Pharmaceutical Chemistry, Department of Chemical Sciences, Faculty of Pharmacy of the University of Porto, 4050-313, Porto, Portugal.

Analytical and Bioanalytical Chemistry
|July 4, 2024
PubMed
Summary

Purple Drank, a mixture of promethazine (PMZ) and codeine (COD), poses risks due to plasma protein binding. This study reveals PMZ and its metabolites strongly bind to human serum albumin, while codeine binds less, impacting drug interactions and toxicity.

Keywords:
Binding affinityDockingHigh-performance affinity chromatographyHuman serum albuminPromethazine“Purple Drank”

More Related Videos

Quantitative Structure-Activity Relationship, Activity Prediction, and Molecular Dynamics of Non-nucleotide Reverse Transcriptase Inhibitors
10:29

Quantitative Structure-Activity Relationship, Activity Prediction, and Molecular Dynamics of Non-nucleotide Reverse Transcriptase Inhibitors

Published on: May 9, 2025

536
Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets
10:16

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

Published on: March 12, 2019

44.2K

Related Experiment Videos

Last Updated: Jun 22, 2025

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening
08:34

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

Published on: October 16, 2015

10.0K
Quantitative Structure-Activity Relationship, Activity Prediction, and Molecular Dynamics of Non-nucleotide Reverse Transcriptase Inhibitors
10:29

Quantitative Structure-Activity Relationship, Activity Prediction, and Molecular Dynamics of Non-nucleotide Reverse Transcriptase Inhibitors

Published on: May 9, 2025

536
Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets
10:16

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

Published on: March 12, 2019

44.2K

Area of Science:

  • Pharmacology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Purple Drank, a combination of promethazine (PMZ) and codeine (COD), is popular for its psychoactive effects but carries risks of fatal events.
  • Drug binding to plasma proteins like human serum albumin (HSA) can alter drug efficacy, increase toxicity, and lead to interactions.

Purpose of the Study:

  • To investigate the binding affinity of PMZ, its metabolites (DMPMZ, PMZSO), and COD to HSA.
  • To determine the binding sites and enantioselectivity of these compounds on HSA.

Main Methods:

  • High-performance affinity chromatography (HPAC) using a zonal approach to measure binding percentages (%b).
  • Displacement experiments using warfarin and (S)-ibuprofen as site probes.
  • In silico molecular docking studies.

Main Results:

  • PMZ and its metabolites showed high HSA binding (>80%), while COD exhibited lower binding (65%).
  • All compounds bound to both site I and site II of HSA, with competition mainly occurring at site II.
  • Binding was found to be non-enantioselective for the chiral compounds.

Conclusions:

  • The strong binding of PMZ and its metabolites to HSA suggests a significant role in their pharmacokinetic and pharmacodynamic profiles.
  • Understanding these binding interactions is crucial for assessing the risks associated with Purple Drank consumption and potential drug interactions.