Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics

ABSTRACT The invention of a therapeutic begins by characterizing features that differentiate healthy versus diseased states; this often presents as changes in the concentration of an analyte. Examples include elevated blood glucose in diabetes, high cholesterol in heart disease, and protein aggregat...

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Main Authors: Rebecca A. Kohnz, Dan Zhou, Bin Lou, Huifang Yao, David McKenney, Dhiraj Dokwal, Ruth Villanueva, Heidi Kocalis, Jeanine E. Ballard, Jennifer Piesvaux, Stephen F. Previs
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:Pharmacology Research & Perspectives
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Online Access:https://doi.org/10.1002/prp2.70099
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author Rebecca A. Kohnz
Dan Zhou
Bin Lou
Huifang Yao
David McKenney
Dhiraj Dokwal
Ruth Villanueva
Heidi Kocalis
Jeanine E. Ballard
Jennifer Piesvaux
Stephen F. Previs
author_facet Rebecca A. Kohnz
Dan Zhou
Bin Lou
Huifang Yao
David McKenney
Dhiraj Dokwal
Ruth Villanueva
Heidi Kocalis
Jeanine E. Ballard
Jennifer Piesvaux
Stephen F. Previs
author_sort Rebecca A. Kohnz
collection DOAJ
description ABSTRACT The invention of a therapeutic begins by characterizing features that differentiate healthy versus diseased states; this often presents as changes in the concentration of an analyte. Examples include elevated blood glucose in diabetes, high cholesterol in heart disease, and protein aggregation in neurodegeneration. Analyte concentrations reflect the (im)balance of synthetic and degradation rates; as such, aberrant biochemical kinetics drive the changes in endpoint concentration that define disease biology. Therapeutics aim to reset the concentration of a disease marker via modulation of biochemical kinetics. This is easy to understand for drugs directly targeting an enzyme in a pathway but, although less obvious, this can also be at the core of protein: protein interactions. For instance, stimulation of the insulin receptor changes the flux of several biochemical substrates (across multiple tissues); similarly, modulation of proprotein convertase subtilisin/kexin type 9‐low density lipoprotein (PCSK9‐LDL) receptor interactions alters cholesterol trafficking. These classic examples underscore the importance of studying biochemical kinetics at a clinical level. Here, we discuss how kinetic studies link disease biology with mechanism of action elucidation and screening. This has an immediate impact on (i) enabling in vitro‐in vivo correlations in early discovery, (ii) enhancing exposure‐response models aiding in human dose prediction, and (iii) providing support for biomarker plans, including clinical diagnostics. Mechanism of action studies can also influence modality selection; e.g., knowledge regarding target kinetics is needed when making decisions surrounding the development of a reversible inhibitor vs. an irreversible covalent modifier, or an intervention that affects target levels such as those which enhance protein degradation or reduce protein synthesis.
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spelling doaj-art-fed2bc076b7b48259b9371a5d84771ea2025-08-20T02:07:24ZengWileyPharmacology Research & Perspectives2052-17072025-06-01133n/an/a10.1002/prp2.70099Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical KineticsRebecca A. Kohnz0Dan Zhou1Bin Lou2Huifang Yao3David McKenney4Dhiraj Dokwal5Ruth Villanueva6Heidi Kocalis7Jeanine E. Ballard8Jennifer Piesvaux9Stephen F. Previs10Merck & co., Inc. South San Francisco California USAMerck & co., Inc. West Point Pennsylvania USAMerck & co., Inc. South San Francisco California USAMerck & co., Inc. Rahway New Jersey USAMerck & co., Inc. South San Francisco California USAMerck & co., Inc. South San Francisco California USAMerck & co., Inc. South San Francisco California USAMerck & co., Inc. South San Francisco California USAMerck & co., Inc. West Point Pennsylvania USAMerck & co., Inc. Boston Massachusetts USAMerck & co., Inc. Rahway New Jersey USAABSTRACT The invention of a therapeutic begins by characterizing features that differentiate healthy versus diseased states; this often presents as changes in the concentration of an analyte. Examples include elevated blood glucose in diabetes, high cholesterol in heart disease, and protein aggregation in neurodegeneration. Analyte concentrations reflect the (im)balance of synthetic and degradation rates; as such, aberrant biochemical kinetics drive the changes in endpoint concentration that define disease biology. Therapeutics aim to reset the concentration of a disease marker via modulation of biochemical kinetics. This is easy to understand for drugs directly targeting an enzyme in a pathway but, although less obvious, this can also be at the core of protein: protein interactions. For instance, stimulation of the insulin receptor changes the flux of several biochemical substrates (across multiple tissues); similarly, modulation of proprotein convertase subtilisin/kexin type 9‐low density lipoprotein (PCSK9‐LDL) receptor interactions alters cholesterol trafficking. These classic examples underscore the importance of studying biochemical kinetics at a clinical level. Here, we discuss how kinetic studies link disease biology with mechanism of action elucidation and screening. This has an immediate impact on (i) enabling in vitro‐in vivo correlations in early discovery, (ii) enhancing exposure‐response models aiding in human dose prediction, and (iii) providing support for biomarker plans, including clinical diagnostics. Mechanism of action studies can also influence modality selection; e.g., knowledge regarding target kinetics is needed when making decisions surrounding the development of a reversible inhibitor vs. an irreversible covalent modifier, or an intervention that affects target levels such as those which enhance protein degradation or reduce protein synthesis.https://doi.org/10.1002/prp2.70099biochemical pathwaydrug discoveryisotope tracersmass spectrometrymetabolic activityorgan crosstalk
spellingShingle Rebecca A. Kohnz
Dan Zhou
Bin Lou
Huifang Yao
David McKenney
Dhiraj Dokwal
Ruth Villanueva
Heidi Kocalis
Jeanine E. Ballard
Jennifer Piesvaux
Stephen F. Previs
Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
Pharmacology Research & Perspectives
biochemical pathway
drug discovery
isotope tracers
mass spectrometry
metabolic activity
organ crosstalk
title Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
title_full Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
title_fullStr Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
title_full_unstemmed Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
title_short Elucidation of Mechanism of Action in Drug Invention: Using Stable Isotope Tracers to Unravel Biochemical Kinetics
title_sort elucidation of mechanism of action in drug invention using stable isotope tracers to unravel biochemical kinetics
topic biochemical pathway
drug discovery
isotope tracers
mass spectrometry
metabolic activity
organ crosstalk
url https://doi.org/10.1002/prp2.70099
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