Table Of ContentIDENTIFICATION AND QUANTIFICATION
OF DRUGS, METABOLITES,
DRUG METABOLIZING ENZYMES,
AND TRANSPORTERS
Concepts, Methods, and Translational Sciences
IDENTIFICATION AND
QUANTIFICATION
OF DRUGS,
METABOLITES, DRUG
METABOLIZING
ENZYMES, AND
TRANSPORTERS
Concepts, Methods, and
Translational Sciences
Edited by
S M
HUGUANG A
S K. C
WAPAN HOWDHURY
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Contributors
Farah Al Qaraghuli Department of Pharma- Liam Evans Hypha Discovery Ltd., Oxford-
ceutical Sciences, School of Pharmacy and shire,UnitedKingdom
Pharmaceutical Sciences, The State University RaymondEvers DepartmentofPharmacokinet-
of New York at Buffalo, Buffalo, NY, United ics,PharmacodynamicsandDrugMetabolism,
States Merck&CoInc.,Kenilworth,NJ,UnitedStates
Ravindra Varma Alluri Clinical Pharmacology Robert S. Foti Pharmacokinetics, Pharmaco-
andSafetySciences,R&DBioPharmaceuticals, dynamics and Drug Metabolism, Merck
AstraZeneca,Cambridge,UnitedKingdom Research Laboratories, Boston, MA, United
Sophie M.A. Argon Department of Pharma- States
ceutics, School of Pharmacy, University of Adrian J. Fretland DMPK, Research and Early
Washington,Seattle,WA,UnitedStates Development, Oncology R&D, AstraZeneca,
PiyushBajaj DrugSafetyResearchandEvalua- Waltham,MA,UnitedStates
tion,TakedaPharmaceuticalInternationalCo., Christopher Gemski Translational Research
Cambridge,MA,UnitedStates Bioassay and Immunogenicity Group, Drug
TashingaE.Bapiro DMPK,ResearchandEarly MetabolismandPharmacokineticDepartment,
Development, Oncology R&D, AstraZeneca, Takeda Pharmaceuticals International Co.,
Cambridge,UnitedKingdom Cambridge,MA,UnitedStates
Abdul Basit Department of Pharmaceutics, Anima Ghosal Independent Consultant,
University of Washington, Seattle; Depart- Piscataway,NJ,UnitedStates
ment of Pharmaceutical Sciences, Washington Jia Hao Drug Metabolism, Gilead Sciences
StateUniversity,Spokane,WA,UnitedStates Inc,FosterCity,CA,UnitedStates
Andreas Brink Roche Pharma Research Satyajeet Haridas Translational Research Bio-
and Early Development, Roche Innovation assay and Immunogenicity Group, Drug
CenterBasel,F.Hoffmann-LaRocheLtd,Basel, MetabolismandPharmacokineticDepartment,
Switzerland Takeda Pharmaceuticals International Co.,
Tingting Cai Laboratory Testing Division, Cambridge,MA,UnitedStates
WuXiAppTec,Nanjing,China SimonHauri RochePharmaResearchandEarly
Jose Castro-Perez Agios Pharmaceuticals, Inc., Development, Roche Innovation Center Basel,
Cambridge,MA,UnitedStates F.Hoffmann-LaRocheLtd,Basel,Switzerland
Jae H. Chang Preclinical Development, ORIC Nina Isoherranen Department of Pharma-
Pharmaceuticals, South San Francisco, CA, ceutics, University of Washington, Seattle,
UnitedStates WA,UnitedStates
Eugene Chia-Te Chen Department of Drug Wenying Jian DMPK, Janssen R&D, Spring
MetabolismandPharmacokinetics,Genentech, House,PA,UnitedStates
SouthSanFrancisco,CA,UnitedStates KevinJohnson DrugMetabolismandPharma-
Marie Croft Pharmaron ABS, Germantown, cokinetics, Genentech, South San Francisco,
MD,UnitedStates CA,UnitedStates
xi
xii
Contributors
BarryJones DMPK,Research andEarly Devel- Kaushik Mitra Department of Drug Metabo-
opment, Oncology R&D, AstraZeneca, Cam- lism and Pharmacokinetics, Janssen Research
bridge,UnitedKingdom and Development, Springhouse, PA, United
RobertS.Jones DrugMetabolismandPharma- States
cokinetics, Genentech, South San Francisco, Diana Montgomery Department of Pharma-
CA,UnitedStates cokinetics, Pharmacodynamics and Drug
JanFelixJoseph FreieUniversitaetBerlin,Insti- Metabolism,Merck&CoInc.,Kenilworth,NJ,
UnitedStates
tute of Pharmacy—Pharmaceutical Analysis;
Freie Universitaet Berlin, Department of AlexandraL.Orton DMPK,ResearchandEarly
Biology, Chemistry, Pharmacy, CoreFacility Development, Oncology R&D, AstraZeneca,
BioSupraMol,Berlin,Germany Cambridge,UnitedKingdom
S. Cyrus Khojasteh Drug Metabolism and KatieH.Owens DepartmentofPharmaceutics,
Pharmacokinetics, Genentech, South San School of Pharmacy, University of Washing-
Francisco,CA,UnitedStates ton,Seattle,WA,UnitedStates
Yurong Lai Drug Metabolism, Gilead Sciences Axel P€ahler Roche Pharma Research and
Inc,FosterCity,CA,UnitedStates Early Development, Roche Innovation Center
Hoa Le Drug Metabolism, Gilead Sciences, Basel, F. Hoffmann-La Roche Ltd, Basel,
Switzerland
FosterCity,CA,UnitedStates
Xiaomin Liang Drug Metabolism, Gilead Sci- Maria Kristina Parr Freie Universitaet Berlin,
InstituteofPharmacy—PharmaceuticalAnaly-
encesInc,FosterCity,CA,UnitedStates
sis,Berlin,Germany
Liming Liu Product Development, Curon
Shefali Patel DMPK, Janssen R&D, Spring
Biopharmaceutical Ltd, Shanghai, People’s
House,PA,UnitedStates
RepublicofChina
Filipe Lopes Roche Pharma Research and Ichiko D. Petrie Department of Pharma-
ceutics, School of Pharmacy, University of
Early Development, Roche Innovation Center
Washington,Seattle,WA,UnitedStates
Basel, F. Hoffmann-La Roche Ltd, Basel,
Switzerland RichardPhipps HyphaDiscoveryLtd.,Oxford-
JustinQ.Ly DrugMetabolismandPharmacok- shire,UnitedKingdom
inetics, Genentech, South San Francisco, CA, Chandra Prakash Agios Pharmaceuticals, Inc.,
UnitedStates Cambridge,MA,UnitedStates
Shuguang Ma Drug Metabolism and Pharma- Bhagwat Prasad DepartmentofPharmaceutics,
cokinetics,GenentechInc.,SouthSanFrancisco, UniversityofWashington,Seattle;Department
CA,UnitedStates of Pharmaceutical Sciences, Washington State
RoshiniMarkandu DMPK,ResearchandEarly University,Spokane,WA,UnitedStates
Development, Oncology R&D, AstraZeneca, Isabelle Ragueneau-Majlessi Department of
Cambridge,UnitedKingdom Pharmaceutics, School of Pharmacy, Univer-
Rosalinde Masereeuw Division of Pharmacol- sityofWashington,Seattle,WA,UnitedStates
ogy, Utrecht Institute for Pharmaceutical Venkatesh Pilla Reddy DMPK, Research and
Sciences,Utrecht,TheNetherlands Early Development, Oncology R&D; Depart-
J. Eric McDuffie Investigative & Mechanistic ments of Modeling and Simulation, Early
Oncology Drug Metabolism and Pharma-
Toxicology,JanssenResearch&Development,
cokinetics, R&D Oncology, AstraZeneca,
SanDiego,CA,UnitedStates
Cambridge,UnitedKingdom
xiii
Contributors
EllenRiddle DrugMetabolism,GileadSciences Caisheng Wu School of Pharmaceutical Sci-
Inc,FosterCity,CA,UnitedStates ences,XiamenUniversity,Xiamen,China
Qian Ruan Pharmaceutical Candidate Charac- Graeme C. Young GlaxoSmithKline Research
terization, Bristol-Myers Squibb, Princeton, and Development Ltd., David Jack Centre,
NJ,UnitedStates Ware,UnitedKingdom
Simone Schadt Roche Pharma Research and Jingjing Yu Department of Pharmaceutics,
Early Development, Roche Innovation Center School of Pharmacy, University of Washing-
Basel, F. Hoffmann-La Roche Ltd, Basel, ton,Seattle,WA,UnitedStates
Switzerland Lushan Yu Institute of Drug Metabolism and
DhavalK.Shah DepartmentofPharmaceutical PharmaceuticalAnalysis, ZhejiangUniversity,
Sciences, School of Pharmacy and Pharma- Hangzhou,People’sRepublicofChina
ceuticalSciences,TheStateUniversityofNew Su Zeng Institute of Drug Metabolism and
YorkatBuffalo,Buffalo,NY,UnitedStates PharmaceuticalAnalysis, ZhejiangUniversity,
Julia Shanu-Wilson Hypha Discovery Ltd., Hangzhou,People’sRepublicofChina
Oxfordshire,UnitedKingdom Haeyoung Zhang Department of Pharma-
Kelly MacLennan Staiger Drug Metabolism, ceutics, University of Washington, Seattle,
GileadSciencesInc,FosterCity,CA,UnitedStates WA,UnitedStates
JonathanSteele HyphaDiscoveryLtd.,Oxford- Wanying Zhang Department of Pharma-
shire,UnitedKingdom ceutical Sciences, School of Pharmacy and
Manthena V.S. Varma Medicine Design, Pharmaceutical Sciences, The State Univer-
sity of New York at Buffalo, Buffalo, NY,
Worldwide R&D, Pfizer Inc., Groton, CT,
UnitedStates
UnitedStates
Matthew P. Wagoner Drug Safety Research Andy Z.X. Zhu Department of Drug Metabo-
lism and Pharmacokinetics, Takeda Pharma-
and Evaluation, Takeda Pharmaceutical Inter-
nationalCo.,Cambridge,MA,UnitedStates ceuticals International Co., Cambridge, MA,
UnitedStates
Naidong Weng DMPK, Janssen R&D, Spring
Mingshe Zhu MassDefect Technologies, Prince-
House,PA,UnitedStates
ton,NJ,UnitedStates
Stephen Wrigley Hypha Discovery Ltd.,
Oxfordshire,UnitedKingdom
Foreword
It is my great pleasure to write the fore- identified many breakthroughs that have
word for this excellent book. The study of ultimately contributed to bringing drugs to
drugmetabolismanddispositionisamature patientswith anurgentneedforbettertrea-
science, but still essential in drug discovery tments.Itisverygratifyingthatthenumber
and development. Indeed, a quick search of ofdrugsapprovedbytheFDAisincreasing
PubMed, using “drug metabolism” as the steadily with 38 NMEs (new molecular en-
search term, came up with more than tity)and10BLAs(biologicallicenseapplica-
18,000 hits. One of the earlier articles is by tion) approved by the US Food and Drug
BernardBrodie,consideredtobethefounder Administration (FDA) in 2019 [2]. The level
of modern pharmacology and a major con- ofinnovationisbestillustratedbyabouthalf
tributor to the study of drug metabolism. of the approved drugs in 2019 having a
Hisarticle—publishedintheJournalofPhar- breakthroughdesignation.Drugmetabolism
macy and Pharmacology in 1956—is titled andpharmacokinetics(DMPK)scientistsare
“PathwaysofDrugMetabolism”anditisbased fullyembeddedinprojectteamsindrugdis-
onalectureattheUniversityofLondon[1]. coveryandworkhandinhandwithmedici-
It describes his work over the decade and nal chemists on the identification of drugs
some of it is based on collaborations with with superior properties. These scientists
other pioneers in the field such as Julius have become very good at dialing out the
Axelrod. One sentence still resonates very “knownunknowns”suchasmetabolismbycy-
muchanditactuallycoversmuchofthema- tochrome P450 enzymes. However, this has
terial described in this book: actuallyresultedinhavingtodealwithmuch
morecomplexdrugdispositionpathwaysin-
Finally,itisthoughtthatadetailedknowl- volvinglesswell-studieddrugmetabolizing
edge of enzymes involved in drug “detoxifica- enzymes and also more and more drug
tion” might help the medicinal chemist to transporters. This trend is also fueled by a
developcompoundsofeitherhighorlowstabil- shift toward more and more drugs having
ity in the body, whichever would be more de- beyond the “rule of five” molecular proper-
sirable ingaining a desired therapeutic result. ties [3]. Indeed, the average molecular
weight of drugs approved by the FDA in
Drugmetabolismscienceshaveadvanced 2016 and 2017 was more than 500 Da and
tremendouslysincethepublicationofthisar- the median clogP of drugs approved from
ticle and this progress was to a large extent 2008to2017isnow3.3,inpartdrivenbytry-
enabled by advances in the availability of ing, for example, to disrupt protein-protein
biochemicalreagentsandbioanalyticaltech- interactions. An additional consequence of
niques, in particular mass spectrometry. this shift in properties is that in vitro to
Bothacademicandindustrialscientistshave in vivo extrapolation of ADME properties
xv
xvi
Foreword
to predict the human pharmacokinetics has latter topic is covered in Chapter 5. The last
become more complex and the same can be chapter in the first section (Chapter 6)
said of predicting drug-drug interaction— focuses on accelerator mass spectrometry,
many of which now involve drug trans- a powerful tool to study ADME and the
porters. If anything, the involvement of absolute bioavailability inhumans.
DMPKscientistsisnowmoreimportantthan The second section addresses “Drug me-
everbecauseofthepursuitofnovelmolecular tabolizing enzymes, transporters and drug drug
modalities in academia and in the pharma- interactions.” Cytochrome P450-mediated
ceutical industry; a few that come to mind and non-cytochrome P450-mediated metab-
are antibody-drug conjugates, protein de- olism is discussed in Chapters 7 and 8.
graders, and macrocyclic peptides. DMPK Invitrotoinvivopredictionofdrug-drugin-
scientists can help make drugs out of these teraction is described in Chapter 9 while
hard-to-drugmodalities.Finally,theintegra- Chapter 10 specifically focuses on the role
tionofthisdiversearrayofinvitroandinvivo of transporters in drug disposition and
data is enabled by computational modeling drug-drug interaction, and Chapter 11 on
and simulation. Physiologically based phar- the clinical relevance of these drug-drug
macokinetic(PBPK)modelingisaverypow- interactions.MakingpredictionsusingPBPK
erfultooltopredicthumanpharmacokinetics modeling relies on accurate knowledge of
and study drug-drug interaction as well as physiologic constants and, most recently,
thepharmacokineticsinspecial populations mass spectrometry has been used to deter-
and infants. Pharmacokinetic/pharmaco- mine the abundance of drug metabolizing
dynamicmodelinganditsextension,quanti- enzymesandtransportersinvarioustissues.
tative systems pharmacology, can help This is addressed in Chapter 12. Finally,
translatepreclinicalfindingstotheclinic. Chapter 13 focuses on disease-drug interac-
Thisbookiscomprisedofmanyexcellent tionsmediatedbytherapeuticproteinssuch
chapters written by experts in the field that as interleukins.
address some of the challenges highlighted The third section focuses on “Strategy re-
in the previous paragraph. The first section lated to drug metabolism and safety.” Metabo-
focuses on “Techniques for identifying and lites in safety testing continues to be a
quantifying drugs and metabolites.” Chapters highlyrelevantareaofresearchindrugdis-
1–3 introduce the readers to the latest ad- coveryanddevelopmentanditisaddressed
vances in bioanalysis, in particular mass inChapter14.Acloselookatthe(patent)lit-
spectrometryfor theanalysisofdrugs,their eratureindicatesthatfindingdrugsthatbal-
metabolites,andendogenousbiomarkers.In ancepotencyandmetabolicstabilitycanstill
contrast to 20 years ago, high-resolution be problematic, and therefore many com-
mass spectrometry hasnow become routine panies incorporate deuterium to enhance
and it has had a great impact on the study metabolite stability and lower the dose—
of biotransformation. Of course, mass spec- seeChapter15fordetails.Thefocusonnovel
trometryisfrequentlynotsufficienttoiden- chemicalspaceandmoremoleculardiversity
tify the definitive structure of a metabolite, has introduced more chirality in molecules
and hence Chapter 4 focuses on strategies andtheimpactofthatonpharmacology,tox-
forgeneratingmetabolitesandcharacteriza- icology,anddrugmetabolismisdescribedin
tionviaNMR.Supercriticalfluidchromatog- Chapter 16. Drug-induced liver injury and
raphy has become easier to interface and it newpredictivemodelsforrenalinjuryarede-
can greatly facilitate chiral separation; the scribed in Chapters 17 and 18, respectively.
xvii
Foreword
Finally, Chapter 19 focuses on immuno- excellent set of chapters that describe the
genicity as a key component of antibody state of the art in ADME sciences as it
development. relates to drug discovery and development.
Thefourthandlastsectionisdedicatedto The efforts by all authors, and in particular
“Translational sciences.” The use of geneti- the editors Shuguang Ma and Swapan
cally modified rodents is explored further Chowdhury, are greatly appreciated. This
in Chapter 20, while Chapter 21 speaks to book will be used as a resource for many
theuseofCRISPRtoadvanceinvitroADME years to come.
models. In vitro to in vivo extrapolation of Cornelis E.C.A. Hop
hepatic and renal clearance is discussed in
Chapter 22. The breadth of our science is
References
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in translational sciences. Last, but by no Pharmacol.8(1)(1956)1–17.
means least, Chapter 24 elegantly defines [2] A. Mullard, 2019 FDA drug approvals, Nat. Rev.
DrugDiscov.19(2)(2020)79–84.
ways to predict the human efficacious dose
[3] M.D. Shultz, Two decades under the influence of
usingPK/PDmodeling.
theruleoffiveandthechangingpropertiesofap-
As is the case with many compilations, a proved oral drugs, J. Med. Chem. 62 (4) (2019)
lot of effort has gone into assembling an 1701–1714.
