Essential IND Strategies:Fundamental Considerationson the Road to SuccessDarren Warren

Drug Discovery & DevelopmentTarget IdentificationHTS (High Throughput Screening)Actives to HitsHit to LeadLead Optimization (SAR)Preclinical CandidateIND Enabling & Registration Support StudiesAPI Synthesis, Analytical & Scale-upClinical CandidatePhase I - IIINDAMarket IntroductionPhase IVFocus of Today’s Discussion

Early Clinical Goals & Regulatory Strategy1) Define drug development goals: Identify target patient population,rationale for clinical need, & initial region for clinical development.Goal should not simply be to file an IND, rather develop a drug therapy2) Initial clinical trials designed to characterize:Human DMPK, safety, pharmacologic activity & proof of concept3) Understand regulatory guidelines and precedented approaches forsimilar drugs (same chemical class & indication or relevant)4) Chemical development strategies: Develop robust API synthetic routethat yields appropriate physical/chemical properties; clinically suitabledrug formulations, with well characterized impurity profiles.5) IND-enabling pharmacology, DMPK & toxicology studies: Need toalign with clinical route of administration, dose schedules, & duration oftreatment. Design to identify: PK/PD responses, target organs, doseresponse, exposure multiples & safety margins

Drug Development .Highly Integrated Process Log D, SolubilityCytotoxicityProtein bindingPermeabilityMetabolismCYP AssaysPreclinicalDevelopmentDMPKIn Vitro ADME Bioanalytical methoddevelopment andvalidation Bioanalytical sampleanalysis PK & metaboliteprofilingChemical Services cGMP synthesis(grams to kilos) Process chemistry Analytical chemistry Process development& engineering PharmacologyMetabolism studiesAnimal modelsPharmacokineticsToxicology(IND & beyond)Clinical & RegulatorySupportClinical SciencesPh 1 Pk/Safety &Ph 2 Efficacy designsRegulatory strategies& submissions

Pitfall #1: The Rush to First DoseManufacture of test article, development of analytical and bioanalytical methods,development of appropriate dosing formulations and as necessary preformulationwork all occur before first dose.

API Supply TargetsItemPre-ClinicalRange-FindingGLP ToxicologyPhase IAPI Needed50 - 200 grams0.20 - 2.0 kilograms2 - 10 kilogramsProcess StatusDiscovery routeUpgraded to scaleable andreliableUpgraded to scaleable andreliable5 - 12 liters50 liters200 - 400 liters 95%97%NLT 98%ID/purity - TANPurity - TANPurity - weight-basedRelated substances - TANRelated substances - TANPreparation ScaleAPI PurityAnalytical MethodsSolvents - totalResidual Solvents - ICHGuidelinesCounter ionProcess residues

Map out your API SupplyAPI Need ForAmount, gramsDose Range Finding StudiesSalt ScreenPolymorph ScreenGLP Toxicology StudiesFormulations researchReference StandardPhase IStability StudySTProgram Contingency @ 25%STMass Contingency @ 25%Program ,711

Map out your API SupplyTarget Requirement * (1 (MC/100))AmountPreparedgramsMass Contingency, %(MC)Discovery Route 10100 - 200Some Process Research 10050Process research/Scale-UpChanges 25025Process Status

Typical API Specification – Phase IProperty/AttributeAppearanceIdentityElemental AnalysisPurityImpurity ProfileFormMelting RangeResidual SolventsHeavy MetalsResidual PalladiumResidue on IgnitionMoistureMethodSpecificationVisualOff-white powderH NMRMatches reference standardC NMRMatches reference standardMass Spectrum Matches reference standardConsistent with structureHPLCNLT 98.0%(w)HPLCNo single impurity 0.5%Total impurities NMT 2.0%XRPDReport resultDSCReport ResultGCConforms to ICH LimitsTitrationNMT 20 ppmICPNMT 10 ppmCombustionNMT 0.2%Karl FischerNMT 0.5%

API Preparation StrategiesModel CDose Range-Finding Lot – Toxicology/Phase I LotsStarting MaterialRegulatory StartingMaterialToxicology LotPhase I Lot

API Preparation StrategiesModel CDose Range-Finding Lot – Toxicology/Phase I LotsAdvantages Utilizes dose range findingdata to set needs Can control impurity profiledifference betweenToxicology and Phase I lots Balances scale-up risk andpreparation timeDisadvantages May have slight delay inToxicology lot deliveryComplete process run morethan 2 times

API Supplies – Program ProgressionItemPre-ClinicalAPI Amount, kilogramsProcess Status0.1 - 3Discovery RouteAPI for Clinical Trial MaterialsPhase IPhase IIPhase IIICommercialProduction1 - 1050 - 200 200 1,000ProcessProcess IntendedProcessProcess Locked forScaleable andfor CommercialDeveloped andRoutine ProductionReliableUseValidatedPreparation Scale, liters 1 - 50 - Kilo-Lab50 - 400 Small 2000 - Pilot Plant 2,000 - Semi 8,000Pilot PlantWorksAnalytical MethodsTAN-based HPLCWeight-based purity, related substances residualMethods fullyPuritysolvents, and other methods are developed and validated developed, validated,in proportion to the clinical phraseand locked forproduction supportAPI CharacterizationRudimentaryAPI Purity, %SpecificationscGMP Controls95ID and purityNoneCharacterization becomes morecomplete as process knowledge andexperience increasesComprehensiveusing validatedmethods9898 98 Specifications refined and tightened as processRequired application rigor increases with processknowledge and experience and phaseRoutine QualityControl testing forproduction supportand API release98 LockedFull routinecompliance

API Preparation Planning – Capacity ModelCAPACITY MODELStep Step Step Custom RMS-1S-1C1C2BATCH SEQUENCING PLANYield Amount Required ProcessTotal%kg-moles kgsOutput 595.143.182.5036.4MWUsage Amount Requiredm/m kg-moles kgs222.001.00 0.01623.60248.901.00 0.01624.0379. Specifics Batches CycleSize Usage MOCTimeliters%days50405065.0 Glass53.1 Glass66.7 Glass22153.03.04.0RunTimedays6.

DMPK & Early DevelopmentIn Vitro MetabolismPlasma stabilityProtein bindingBlood compatibilityMicrosomal/Hepatocyte stability & metabolismSpecies comparison in microsomes & hepatocytesDefine metabolic pathway and major metabolites;metabolite structure elucidationPharmacokinetics (PK) & Toxicokinetics (TK)Lead & Formulation selection, pilot PKBioavailability & complete PK profilesIn vivo metabolite profiling & ADME studies (using cold or radiolabeled compound). Conducted preclinically or early Phase 1.Critical for Non-clinical Species Selection &Prediction of Human DMPK Responses

DMPK & Early DevelopmentDrug-Drug Interaction (DDI)CYP Assays: Consider patient population & co-therapiesInhibition (cocktail & individual assays, IC50 and Ki)Mechanism based inhibitor (MBI) determinationInductionIn vitro induction in hepatocytesToxicology/TK data can provide early indicators of inductionEx vivo induction in liver from treated animalsReporter gene assays for induction of CYP1A2 & CYP3A4UGT enzyme inhibitionCell TransportDrug TransportersPermeability (Caco2, uni- and bidirectional)MDR1-MDCK bidirectional permeability (P-gp)Uptake transporter assaysCritical for predicting Drug-Drug Interactions; interpretingPK & tox outcomes; & prediction of human PK profiles

Pharmacokinetics & MetabolismPharmacokineticsDefine Active Drug Concentration & PK profiles(major & relevant metabolites)AUC, Cmax/Cmin, Tmax, T1/2, Vd, & ClCharacterize over range of dosages, including expected clinicaland toxicology dosages (1x-10x efficacious dosages)Single & Repeat-dose PK (3-7 days)Defines saturation of absorption, metabolism, clearance/excretion,accumulation, gender and species differencesADMENot generally required for INDWill need to identify and characterize major/relevant metabolitesHelpful to understand primary routes of excretion &tissue distribution

Drug SafetyNon-GLP & GLP Toxicology & SafetyPharmacology StudiesGLP vs non-GLPAny study can be conducted in accordance with GLPGLP incurs increased cost and timelinesGLP (only) required for extrapolation to humansSpecies SelectionSelection based on in vitro metabolism and PK dataMajor metabolites must be expressed in tox speciesRodent (mice, rats)Non-Rodent (dogs, nonhuman primates)Gottingen mini-pigs, rabbits, etc. as justifiedRequirement for two species may be waived(ex. no pharmacology in rodent species for biologics)

Drug SafetyNon-GLP & GLP Toxicology & SafetyPharmacology StudiesDose Administration & ScheduleShould be the same as intended clinical route & scheduleDose schedule: daily (or multiple daily) vs. cycle dosingOral: gavage, nasogastric route, oral tablet/capsule or solutionParenteral: intravenous, continuous intravenous infusion,subcutaneous, intramuscular, intraperitonealTopical: dermal, ocularRegional treatment: intra-tendon, intra-articular and intra-vitrealCharacterize dose-response relationshipMinimum of 3 dosagesGood separation between dosages to avoid exposure overlapDose to toxic effect or maximum feasible limit

Drug SafetyToxicology StudiesPilot Toxicology StudiesInitial toxicity readouts (single and multiple dose)Required in each species, non-GLPTolerability - define the Maximum Tolerated Dose (MTD):single dose; morbidity/mortality, GI distress, severe CNS effects,respiratory distress, immune reactionsRepeat Dose Range-Finding Toxicity:repeat dose 5-14 days; identify dose & exposure responses, targetorgan toxicity; major organ system pathology; dose-limitingtoxicities; repeat-dose TKA go/no-go decision often follows:Toxicity profile? PK profile? Dose limitations? Off target tox?Pitfall: not considering your formulations carefullyPitfall: not conducting complete / robust pilot tox studies

Drug SafetyToxicology StudiesIND-enabling (pivotal) GLPTypically 14-28 day repeat dose to support SAD & MADPh I clinical studiesIntended as survey studies. Expected to include endpointsrelevant to molecular class, anticipated toxicity, PD identificationDose selection intended to elicit toxicityPrimary endpoints are clinical pathology & anatomical pathologyassessments with TK profile correlatesGoals: Identify target organ toxicity/pathology, translationalpredictive safety biomarkers, assess reversibility or progression,assess local tolerance, determine adverse effects with NOAEL &exposure ratiosBasis for selecting initial clinical doses & escalation.

Drug SafetyIND-Enabling ToxicologySpecific assessments as indicatedLocal effects (ex. injection or application site)Specific safety biomarkers as appropriate(clinical pathology or specialty assay)Immunogenicity as warranted (anti-drug antibody)Immune suppression or cytokine stormCommon concerns / issuesBlood volume limitations for large animalsTA consumption substantialTA preferred same batch as Ph IMaintain purity of purpose IND enabling.Avoid discovery investigations; pitfall forincluding unneeded endpoints

Drug SafetyGenetic ToxicologyHazard Identification for DNA damage in form of mutationsor chromosomal damagePre-IND requirement for 2 in vitro assays: AMES &Mammalian in vitro Chromosomal Aberration assayRegistration Requirement for in vivo Chrome Ab assay(Micronucleus Test)Prudence in conducting all 3 assays pre-INDAdditional clarification assays as needed (example toshow epigenetic or mechanism based effects)Does not address potential genotoxic impurities in API

Drug SafetySafety PharmacologyRequirementsICH Core Battery:CNS & Respiratory (generally rat) andCardiovascular (generally canine or non-human primate)In vitro cardiovascular ion channel assessmentGI, Renal, others as target organs dictatePurpose and DesignsDetermine potential for untoward pharmacologySingle dose pharmacology study, top dose near MTDSmall molecule – commonly stand alone studiesBiological – incorporate endpoints into non-rodent tox studyOncology (end stage) – waived

Drug SafetyCore Safety Pharmacology StudiesCardiovascular AssessmentsIn vitro hERG (minimum, other ion channel assay as indicated)In vivo telemetry cardiovascular functional evaluations:blood pressure, heart rate, and ECG waveform analysesRespiratory Functional AssessmentsIn vivo respiratory assessment in rodentsPlethysmography measuring respiratory rate,tidal volume, and minute volumeCNS Functional AssessmentsIn vivo central nervous system functionalassessment in rodentsFunctional observational batteryMotor & behavioral activity

Development Timelines and ResourcesElementsBallpark Price ( , 000)In Vitro metabolismVariousBioanalytical method validation(per analyte, two species)65 - 80PK (per study)10 – 20MTD / DRF (two species)100 – 130Genetic toxicity85 - 100Safety pharmacology135 - 16028 Day: RatDogMonkey200 – 300250 – 350400 – 600 Candidate API supply BA Internal standard synthesis Formulations development IND preparation / publication?

Development Timelines and cal Development (6 - 8 months)Synthetic process improvement & production of gram batchesChemical synthesis process development for 1-10 Kg batchAPI characterization and stability establishedInitial non-clinical & clinical formulations developedDrug product characterization supporting early clinical useDrug Safety and Metabolism (8 – 10 months)In vitro DMPK studiesPilot pharmacokinetic & toxicology studiesDrug safety IND-enabling toxicity studies(14 - 28 day rodent and non-rodent)Genotoxicity assaysSafety pharmacology profileThe above are required elements, there are no real short-cuts

Development Timelines and ResourcesCase Study – Concordia PharmaceuticalsProgram OutcomeIND filed 10 - 12 months after lead selection, barring any technicalor safety issues. Requires parallel activities including INDpreparation and clinical plan determination(15-18 months for biologics)Commercially-viable prototype API process developed anddemonstratedTotal project cost 2.5 – 4.0 million (biologics can be more)Clinical entry with a well characterized molecule