SolvingTheWorld’sHardestProblems

February23st,2017

2

TheRoleofToxicologyinDrugDevelopment

soladosis facit venenum

3

Toxicology• Toxicologyisthestudyoftheadverse

effectsofchemical,physical,orbiologicalagentsonpeople,animals,andtheenvironment.

• Toxicologistsarescientiststrainedtoinvestigate,interpretandcommunicatethenatureofthoseeffects.

• Factorsthatinfluencechemicaltoxicityincludethedosage(andwhetheritisacuteorchronic);therouteofexposure,thespecies,age,sexandenvironment.

• Therelationshipbetweendoseanditseffectsontheexposedorganismisofhighsignificanceintoxicology.

“Solelythedosedeterminesthatathingisnotapoison.”Paracelsus(German-Swissphysician),1493-1541.

Water,inexcess,istoxic.

4

• Needtoprovenewdrugsaresafe:– Beforefirstadministrationtohumans– Beforelaterclinicaltrials

• Is“safe”arealisticgoal?• WhatdoesNonclinicaltoxicologyreallydo?– Hazardidentification– Riskassessment

WhyDoToxicologyTesting?

5/10/17

5

HazardIdentificationvsRiskAssessment

HAZARD RISK

6

IfToxicologyTestingisneededtoprovedrugs“safe”forhumanuse,wheredoesToxicologyfitinDrugDevelopment?

5/10/17

7

Mile-HighViewofDrugDevelopment

Discovery• TargetValidation

• LeadDiscovery&Optimization

Pre-ClinicalEvaluations• Safety• Efficacy

ClinicalTrials• PhaseI• PhaseII

GettingtotheMarket• PhaseIII

Pre-clinical Clinical

ToxicologyineveryPhase=Nonclinical

8

ToxicologySupportofDrugDevelopment

Discovery

• LimitedToxicologysupport;primarilyintheformofintellectualinput(consultation)andlimitedscreening

LeadDiscovery

• Non-GLPinvitroscreens;limitedinvivoscreening

Pre-clinicalEval

• GLPGeneralToxicology,GeneticToxicologyandSafetyPharmacology

Clinical

• GLPReproductiveToxicology,subchronic GeneralToxicology;Pre-Oncogenicity dose-ranging;InvestigativeToxicology

Market

• GLPChronicGeneralToxicology,ReproductiveToxicology,Immunotoxicology,Oncogenicity

PostMarketing

• InvestigativeToxicology;Pharmacovigilance (consultation)

95/10/17

NonclinicalDeliverablesDiscoveryPhase

LeadCompound

Intellectualinput:Earlyleadsandtargetliabilityassessment

CandidateSelection-18months -12months -6months

“PHARMACY”Prelimform.ForSAandFTIHstudies

Assistancewithformulationsforanimalmodels

BIOANALYSISAssaydevelopment,preliminarymetabolismstudiessupportforSAandPharmacy

TASUPPLY Synth~300g,doserangingstudies

Supplyof~50gcompoundfromMedChem

Startsynth28daytoxandFTIHsupplies

SAFETYASSESSMENTGenotox(insilico SAR)

Adhocscreens (teratogenicity,cytotoxicity,mutagenicity,HERG,ratCVetc)orbiomarkersBasedonpriorknowledge

Formulation,speciesselectionforGLP

Bacterialmutagenicityscreen

Non-GLPRatToxscreens

Mammaliangenotoxicity(invitro)

10

• Thecrossoverpointbetweendrugdiscoveryanddrugdevelopment.– Provideinformationonmechanism(s)ofactionofadrug– Providesanearlyindicationofthepotentialforsomekindsoftoxiceffects,allowingadecisiontoterminateadevelopmentprogrambeforespendingtoomuchmoney.

• Invitromethodsarewidelyusedfor:– Screeningandrankingchemicals– Studyingcell,tissue,ortargetspecificeffects– Improvesubsequentstudydesign

InVitroToxicology

11

• Invitromethodsareusually– Lessexpensivetorunthaninvivostudies– Fasterthaninvivostudies– Compliantwiththe3Rsinitiative

• Replacement• Reduction• Refinement

– Somewhatlesspredictiveoftoxicityinintactorganisms

InVitroToxicology

12

TargetValidation

LeadDiscovery

LeadOptimization

Pre-ClinicalEvaluation

PhaseI

PhaseII

PhaseIII

Registration&LaunchPhaseIV

WhereDoInVitroModelsFitinDrugDevelopment?

Disease/EfficacyModels

• Highbiologicalrelevance

• Understandingofanimal– versushuman–relevantModeofAction

•Medium/Lowthroughputacceptable

Predictivetests:earlyprioritizationofdrugcandidates

• Fastturnaroundtime• High/mediumthroughput

• Lowcompoundrequirement

• Qualitativeassessmenttoguidemedicinalchemistry

InvestigativePlatforms

• Highbiologicalrelevance

• UnderstandingofhumanModeofAction

• Lowthroughputacceptable

Traditionally,thisiswhere90%ofinvitrotestingtookplace.

13

PredictiveToxicologyCompoundRelated

Toxicity

•Insilico flags

•Similaritytopreviouscompounds

•Predictivetox signaturese.g.BioMAP*,connectivitymapping

• Selectivity(offtargetevaluation)

TargetRelatedToxicity

EarlySafetyPrediction(ESP)

TailoredSafetyEvaluation

Supplement invivoefficacystudies(ifapplicable)withtoxicologyendpoints

Includenon-standardelementsinearlytoxicologystudies

Includeadditionalinvitro/invivoassaysinscreeningcascadee.g.Mitoactivity,myocyte,podocytes

• SecondaryPharmacology

• Genotoxicity

• Cardiotoxicity

• Hepatotoxicity

• Physicochemicalproperties

CoreStrategies

14

• CrossPharmareviewofrationale,strategiesandmethodologiesforinvitropharmacologicalprofilingat4majorPharmacompanies(AZ,GSK,Novartis&Pfizer)– NineteenmoleculartargetswereidenticalinallfourPharmacompanies– Another25targetsweretestedin3outof4ofthecompanies– Recommendedminimalpaneloftargets(44targets)

SecondaryPharmacologyTargets

GPCRsAdenosine(A2a)

Adrenergic(a1a,a2a,b1,b2)Cannabanoid (CB1,CB2)

Peptidergic (CCK-A,ET-A,V1a)Dopamine(D1,D2)Histamine(H1,H2)Opioid (d,k,µ)

Muscarinic (M1,M2,M3)Serotonin(5HT1a,5HT1b,5HT2a,5HT2b)

Ionchannels

Voltagegated(Cav1.2,hERG,KCNQ1/KCNE1,Nav1.5)

Ligand gated(nACHR a1ora4,GABA-a,NR1,5HT3)

Enzymes

AchECOX1COX2MAOaPDE3APDE4DLCK

TransportersDAT,NET,SERT

NuclearHormoneReceptorsAR,GR

Bowesetal.NatureReviewsDrugDiscovery11,909(2012)

15

• Thegoalistoincreasebetterdrugqualityselectionthroughtheuseofphysiologicallyrelevantinvitroculturesfortarget/diseaseinvestigation,drugdispositionandmetabolism,andtoxicityprofilingBEFORE yougointoanyanimal

16

• Resultsfromnonclinicaltoxicologystudiesshould,ataminimum:– Establishasafestartingdoseforclinicalstudies– Provideinformationonadrug-treatmentregimenthatwouldproducetheleasttoxicity

– Assesstargetorgantoxicityanditsreversibility– Provideinsightintobiomarkersforclinicalmonitoring

InVivoToxicology- Purpose

17

NonclinicalDeliverablesCandidatetoFirstTimeinHumans

TOXICOLOGY

Output:SummariesforIND/CIB

FinalReportsforIND

-3months -6monthsCandidateSelection FTIH

GLPGeneticToxAmesTestMouseLymphomaMicronucleus

Doserangingnon-rodent

Doseranginginrats(ifrequired) RepeatdoseTox

28Dayrodent28Daynon-rodent

SafetyPharmacologyRatIrwinstudyRatRespiratoryNon-rodentCVinvitroHerg assay

Deliver28DayToxdrug

Definelongtermrouteandfinalisolation

Startprepof6monthtoxsupplies

InitialOccupationalHazardEval.forAPI

Toxicokinetics &MajorMetaboliteID

5/10/17

18

NonclinicalDeliverablesFTIHto‘ProofofConcept’(PoC)

5/10/17

+6months EndPhase(+9months)

InitiateProgramofInvestigativestudiesasrequired.

SubChronic Toxicity3/6monthrepeatdosetox studies(rodent&non-rodent)possibleenabler

Pre-Onco Studies[4yrs.before NDA/MAA]

SupplyDS3/6monthtox &preOnco

Preparedrugsupply9/12monthTox.&Carc studies

ReproductiveToxicology

RodentEFDStudy

Rabbitdoserange

Rodentfemalefertilitystudy

RabbitEFD

Supplydrugsubstanceforreprotox

BioA supportforhumanbioavailabilitystudy

AdditionalOcc.&Enviro.Safetystudies(withCHESGroup)

JapanDevelopment:Ratacute,clinicalroute

FTIH PoC (~18M)

Teratology

19

NonclinicalDeliverablesPoC toMarket

5/10/17

Phase2B MarketCommitPhase3 Filing(L-12)

Rat&MouseOncostudies(3yrspre-file)

Non-rodent9/12Month

Rat6month(ifnotdoneprev.)

Ratmalefertility

RatPre/PostnatalTox

CompilationofNDA/MAA

RespondtoReg.Questions

Othersupportingstudies - bridging,combo,newroutes,regionalrequirements,investigativetox.JapanDevelopment - ‘recovery’animaldataforJNDA

+continuingactivitiesinChemistry/PharmaceuticaldepartmentsandBioAnalysis.

ImmunoToxstudiesifreq.

JuvenileTox studies(timingdependentonclinicalplan)

20

NonclinicalActivitiesandDeliverablesPost-MarkettoEndofTALifeCycle

5/10/17

BIOANALYSIS

TOXICOLOGY

CHEMISTRY

ALLDEPARTMENTS

LineExtensionsIntellectualPropertydefence

RegulatoryUpdatesNewsubmissionsEmergingissuesOTCproducts

21

• The numberandtypesofstudiesrequireddependonthetherapeuticindication.

• Drugsforlife-threateningillnessesrequirefewerstudiestoreachtheclinic.

• Ingeneral,animalstudiesareconductedintwospecies,onerodent(e.g.,rat,mouse)andonenon-rodent(e.g.,dog,nonhumanprimate).Biologicsmayrequireonlyonespecies.– Why2species?

• Otherspecies(e.g.,rabbits,ferrets,hamsters,mini-pigs)maybeusedforspecialstudies(e.g.,vaccinestudies).

CommentsonNonclinicalSafetyStudies

22

• KeyAssumptions– Otherorganismscanserveasaccuratepredictivemodelsoftoxicityinhumans.

– Selectionofanappropriatemodeltouseiskeytoaccuratepredictioninhumans.

– Understandingthestrengthsandweaknessesofanyparticularmodelisessentialtounderstandingtherelevanceofspecificfindingstohumans.

• Caveat– Drugsshowingsafetyandefficacyinpreclinicalanimalmodelsmayshowverydifferentpharmacologicalproperties whenadministeredtohumans

AnimalModelsinToxicologyTesting

23

• Developmentofproperpreclinicalmodelswhichcanefficientlypredictdrugbehaviorinhumansisessentialpriortotestingadruginahumansubject.

• TheFDAandotherregulatoryagenciesaremoreandmorerequiringSponsorstoprovidedatatosupportselectionofthespecificspecies(andevenstrains)usedtosupporttestingofnewdrugs.

AnimalModelsinToxicologyTesting

24

WhatisthefutureofToxicologyinDrugDevelopment?

5/10/17

25

CurrentSituation• Currently,drugdevelopmentisalong

andcostlyprocessinlargepartduetothefailureofdrugcandidatesidentifiedininitialinvitroscreenstoperformasintendedinhumans.

• Betweentheinitialtargetidentification/validationandhumanclinicaltrials,liesalargegulfthatisbridgedthroughtheuseofinvivoanimalmodelswhichareusedtopredictthepotentialefficacyandsafetyinhumans.

• Thistraditionalpathofdrugdiscoveryisinefficient atbest,withonly~9%ofcompoundsachievingMarketApproval.

26

• Naturereviewsdrugdiscovery– Toxicityisleadingcauseofattritionatallstagesofdrugdevelopmentprocess– Majorityofsafety-relatedattritionoccursinpreclinicalstudies– Earlypredictionpreclinicalsafetyliabilities:design/selectionofbetterdrug

candidates– Betterdrugcandidatespossessincreasedprobabilityofmarketing/approval

Safety-RelatedAttrition:GeneralConsensus

NatureReviewsDrugDiscovery6,636-649(2007);NatureReviewsDrugDiscovery3,711-715(2004)

27

Safety-RelatedAttrition:OneCompany

7.9

21

2.1 2.6

22

5

0.23

21

18

0.00

5.00

10.00

15.00

20.00

25.00

Term

inationfore

achcrite

rion(%

)

Majorreasonsforprojectterminationswithin1Company(2004-2013)

28

Safety-RelatedAttrition:OneCompany

0

5

10

15

20

25

30

Cardiovascular

Liver

CNS

Gastrointestinal

2007-2012

29

• Twomaincontributorstothisinefficiencylieintheinvitroandinvivomodelsusedtocharacterizethecandidatemolecules.

• Thetranslatabilityofin-vitrotoin-vivotoclinicaloutcomeischallenging.– Akeyproblemisthatleadcompoundsaretypicallyvalidatedandoptimizedin

oversimplifiedculturemodelswhichlackintegrativephysiology.– Pre-clinicalinvivocharacterizationiscurrentlydependentonanimalmodels

whenthefinalcustomer(thepatient)ishuman.– However,canageneticallystablebreedofrodentsinacontrolled

environmentrepresentageneticallydiversepopulationofhumanpatientslivingindiverseenvironments?• Forexample,humansoftenmetabolizechemicalcompoundsverydifferentlyfromthe

testanimalspecies.

• Analternatepathfordrugdiscoverymaybenefitfromtheuseofmorehumanrelevant,complexinvitromodelsthatintegratetheinvivophysiology.Efficacy,ADME,drugdispositionintissueandToxicityarecriticalbreakpointsindrugdevelopmentandcouldgivethegreatestreturnonutilizationoftheseimprovedmodelsystems.

RootCauseofthisInefficiency

30

PrimaryScreeningandAnimalandHumanTrials

32

BACK-UPSLIDES

33

• Regulationstoensuretheintegrityofdatafromnonclinicalstudies.

• IntheUSA,theGLPsareadministeredbytheFDA,andarelaidoutin21CFRPart58

• Otherregulatoryagencies(OECD,EPA)havetheirownsetsofGLPregulationsthataresimilartobutnotidenticaltothoseoftheFDA.

• DefinitivepreclinicalstudiesmustbeGLPcompliant

GoodLaboratoryPractices

34

SalientPointsaddressedbyinvitroModelsforDrugSafety

RequirementsforaninvitromodelinthisSituation OrganRelevantPhysiology– Low

ThroughputSpecificeffect– HighThroughput§ Predictivetests:earlyprioritization

ofdrugcandidates- Fastturnaroundtime- High/mediumthroughput- Lowcompoundrequirement- Qualitativeassessmentto

guidemedicinalchemistry

§ Highlystandardized,thoroughlycharacterizedinvitromodelwithlowtechnical&biologicalvariability

§ Easeofuse,simplehandling§ Amenabletoautomation§ Readoutallowsquickevaluation

anddecisionmaking

§ Mechanistictests:de–riskingofinvivofindingsindevelopmentstages- Highbiologicalrelevance- Understandingofanimal–

versushuman–relevantModeofAction

- Quantitativeassessmenttomake«GO/NOGO»decision

§ ModeofActionunknown:Ascloseaspossiblerecapitulationofinvivosituation– invivophenotypedisplayedininvitrosystem

§ Humanrelevantphysiology§ Abilitytocompareanimalversus

humanataninvitrolevel§ Quantitativemeasurements

possible

OrganRelevantP

hysio

logy–

LowThrou

ghpu

t

Specificeffect–High

Throughput

35

PotentialUsesofComplexInVitroCulturesinDrugDevelopment

36

Adipose

Heart

FemaleReproduction

Liver

BloodVessel

Muscle

Kidney

Gut

Lung

Brain

Skin

NIHFundedandIndustrySupportedSingleand

Multi-OrganChipProjects

ParticipatingInstitutionsCharlesStartDraperLabGladstoneInstituteClevelandClinicClevelandClinic;FlocellUniversityofCalifornia,SanFranciscoUniversityofIllinoisCharlesStarkDraperLabUniversityofWashingtonUniversityofCalifornia,SanDiegoYaleUniversityVanderbiltUniversityWyssInstitute,HarvardMedicalSchoolUniversityofCalifornia,BerkeleyJohnsHopkinsUniversityColumbiaUniversityMorgridge InstituteforResearch,Inc.MassachusettsInstituteofTechnologyBaylorCollegeofMedicineWashingtonUniversityGlaxoSmithKline

37

BloodBrainBarrier

• MIMETAS• Blood:Brain BarrierConsortium

– TheaimoftheprojectistodevelopamorepredictiveinvitroBBBmodelbytheintegrationofphysiologicallyrelevantfactorsintheOrganoPlateTM.

– Basedoncriteriasetbytheconsortium,theprojectwillstartwithanon-competitivephasefocusingentirelyonthedevelopmentoftheBBBmodel.

– Themodelwillbevalidatedwithaselectedcompoundlibraryandcomparedwithexistingdatatosetthebasisforinvitrotoinvivoextrapolation.

https://www.youtube.com/watch?v=L_VEJAZ5J6U

38

SynovialJointModel

A)Schematicrepresentationofthesynovialjoint,adaptedfromStrandV.etal.NatureReviewsDrugDiscovery6,75-92(January2007).B)SynovialmodelintheOrganoPlateTM

39

LiverOrganoid Model

40

WhatIsTheBestModelofEfficacy?

ClinicalTrial(<80% attrition?)

Relevantcellandtissue

MolecularFingerprintsBiomarkers

Genetics

Emergingpath

Traditionalpath

MolecularTarget

RecombinantCellularAssay

Animalmodels

ClinicalTrial(95%attrition)