Enzyme–Inhibitor Interactions and a Simple, Rapid Method for Determining Inhibition Modality

@article{Buker2019EnzymeInhibitorIA,
  title={Enzyme–Inhibitor Interactions and a Simple, Rapid Method for Determining Inhibition Modality},
  author={Shane M. Buker and Paula Ann Boriack-Sjodin and Robert A Copeland},
  journal={SLAS Discovery},
  year={2019},
  volume={24},
  pages={515 - 522}
}
Contemporary chemical biology and drug discovery are increasingly focused on the discovery of inhibitory molecules that interact with enzyme targets in specific ways, such as allosteric or orthosteric binding. Hence, there is increasing interest in evaluating hit compounds from high-throughput diversity screening to determine their mode of interaction with the target. In this work, the common inhibition modalities are reviewed and clarified. The impact of substrate concentration, relative to… 

Figures from this paper

High-Throughput Mechanism of Inhibition

This work revisits the common inhibition modalities and illustrates the impact of substrate concentration relative to Km upon the pattern of changes in IC50 that are expected for increasing substrate concentration.

Optimizing model comparison for enzymatic mechanism analysis

This work examines the use of the modular equation permutations, that can be produced through binding curve summation, to fit and evaluate the interactions of abietic acid with protein tyrosine phosphatase nonreceptor type 11.

Response to the Article “Enzyme–Inhibitor Interactions and a Simple, Rapid Method for Determining Inhibition Modality”

  • Ryan Walsh
  • Biology
    SLAS discovery : advancing life sciences R & D
  • 2019
This article was quite disappointed in its one-sided biased endorsement of classical inhibition models and the omission of this point greatly reduces the overall usefulness of a review.

On-flow enzymatic inhibitor screening: The emerging success of liquid chromatography-based assays

This review covers articles from the last decade that combine the use of varied immobilization methods on different solid supports and several equipment setups in on-flow systems, emphasizing the performance and capacity of recognizing and identifying biologically active compounds in various matrices.

Quantitative Measurements of Pharmacological and Toxicological Activity of Molecules

Toxicity and pharmacological activity scales of molecules, in particular toxicants, xenobiotics, drugs, nutraceuticals, etc., are described by multiples indicators, and the most popular is the median

Resurrecting the phoenix: When an assay fails.

This review is a step-by-step attempt at reintroducing fundamental biochemical concepts within the context of an enzyme assay, delineating probable causes for failure and potential approaches to get an assay back up and running.

Explicit Treatment of Non‐Michaelis‐Menten and Atypical Kinetics in Early Drug Discovery **

This review strives to present an overview of enzyme kinetic mechanisms that are atypical and, oftentimes, do not conform to the classical MM kinetics to enable effective screening and characterisation of small‐molecule inhibitors with desirable physiological outcomes.

Allosteric Site on SHIP2 Identified Through Fluorescent Ligand Screening and Crystallography: A Potential New Target for Intervention.

It is posit that binding of ligands to this site restrains L4 loop motions that are key to interdomain communications that accompany high catalytic activity with phosphoinositide substrate, and may, therefore, be a future druggable target for medicinal chemistry.

Antimalarial Flavonoid-Glycoside from Acacia pennata with Inhibitory Potential Against PfDHFR-TS: An In-silico Study

FG17 was computed as a non-toxic, bioavailable, synthetically accessible compound and a better enzyme inhibitor than RJ1 to conclude that FG17 may be used as a lead scaffold to design antimalarial agents against PfDHFR-TS in the future.

References

SHOWING 1-10 OF 18 REFERENCES

Non‐Competitive Inhibition by Active Site Binders

  • Y. Blat
  • Biology, Chemistry
    Chemical biology & drug design
  • 2010
Tools like alternative substrates, testing the enzyme reaction in the reverse direction and monitoring inhibition time dependence can be applied to enable distinction between ‘badly behaving’ active site binders and true exosite inhibitors.

High-Throughput Determination of Mode of Inhibition in Lead Identification and Optimization

A high-throughput MOI determination method for evaluating thousands of compounds using an existing screening infrastructure by measuring the ratio of IC50 or percent inhibition at 2 carefully chosen substrate or ligand concentrations to define an inhibitor as competitive, uncompetitive, or noncompetitive.

A simple kinetic method for rapid mechanistic analysis of reversible enzyme inhibitors.

  • C. LaiJoe C. Wu
  • Biology, Chemistry
    Assay and drug development technologies
  • 2003
Among 76 Csp3 inhibitors analyzed, 70 were found to be non-mutually exclusive inhibitors, suggesting the existence of an allosteric site(s) in CSP3 for effective inhibition, and the implication of this observation is discussed.

Matrix-Based Activity Pattern Classification as a Novel Method for the Characterization of Enzyme Inhibitors Derived from High-Throughput Screening

A novel method for determining the MOI of a compound without the need for curve fitting of the enzyme inhibition data is described and in good agreement with the known MOI and compares favorably with the previously described IC50-shift method.

A selective inhibitor of PRMT5 with in vivo and in vitro potency in MCL models.

EPZ015666 is an orally available inhibitor of PRMT5 enzymatic activity in biochemical assays with a half-maximal inhibitory concentration (IC50) of 22 nM and broad selectivity against a panel of other histone methyltransferases.

Evaluation of enzyme inhibitors in drug discovery. A guide for medicinal chemists and pharmacologists.

  • R. Copeland
  • Biology, Chemistry
    Methods of biochemical analysis
  • 2005
This work has shown that knowing Inhibitor Modality is important for Structure-Based Lead Organization and Associating Cellular Effects with Target Enzyme Inhibition should Require a Certain Affinity for the target Enzyme.

ATP-competitive inhibitors of the mitotic kinesin KSP that function via an allosteric mechanism.

A new class of selective KSP inhibitors that are active against ispinesib-resistant forms of KSP are described, suggesting that they compete with ATP binding via a novel allosteric mechanism.

Exosite Interactions Determine the Affinity of Factor X for the Extrinsic Xase Complex*

Substrate interactions at exosites, sites removed from the active site of VIIa within the enzyme complex, determine affinity and binding specificity in the productive recognition of factor X by the VIIa-TF complex.

Exosites determine macromolecular substrate recognition by prothrombinase.

Limits to the common approach of inferring the basis of factor Xa specificity with active site mutants or the targeting the active site offactor Xa with reversible inhibitors for therapeutic purposes are suggested.