V accine clinical trial -Quality, include control of cell substrate

Vaccine clinical trial-Quality, include control of cell substrateYwan-Feng LiCenter for Drug Evaluation

4-7-2011IThe views expressed in this presentation are not necessary those of Center for Drug Evaluation-Taiwan

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The views offered here do not necessarily reflect official positions of TFDA

4-7-2011*Heterogenicity of the biological/biotechnological productsPeptides(20-30 a,a,)rDNA proteins(Mab, fusion protein)AllergensTraditional vaccinesBlood/plasma productsGene/cell therapy P.Tissue engineering P.rDNA-derived vaccines

4-7-2011*ScopeVaccineInformation from research stageClinical trial-INDCell substrate-Testing for adventitious agentsQuality, starts from phase 1 Quality, (almost) finalizes at phase 3 and continues through product life spanCollaboration from all parties makes a trial going

Type of vaccineVaccines represent the most diverse type of productsAttenuated or killed pathogens (bacteria, virus, parasites) (~traditional vaccine)Purified and recombinant protein Synthetic peptidesPolysaccharide (free or conjugate to carrier)DNA, viral vectors(Cell-based product)

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Preventive versus therapeutic vaccines 4-7-2011*In general:

CharacteristicPreventiveTherapeuticPopulationHealthy subjectUsually patientClinical outcomeDecrease microbial infection and/or transmissionCure or postpone disease progression(usually as a 2nd line strategy)RegimenLow dose, episodicUsually high dose, continual (more like a drug)Evaluations in early trialSafety, immunogenicitySafety, immunogenicity

Preventive versus therapeutic vaccines 4-7-2011*

CharacteristicPreventiveTherapeuticRegulatory evaluationEmphasis on safetyEfficacyBenefit/risk assessmentEfficacyPublic expectation Highly concern and sensitive to the potential risksLess concern regarding the potential risksHowever, quality and assessment of the vaccine (Ag and adjuvant) is the same for either type of vaccine

First licensed cancer therapeutic vaccine-Provenge (FDA)Autologous dendritic cells, activated by prostatic acid phosphatase (plus GM-CSF)2010, approved by FDA to treat asymptomatic or minimally symptomatic metastatic hormone-refractory prostate cancer2011, FDA approved Dendreon's request to increase production capacityFDA approved "36 additional workstations at the company's New Jersey facility, adding to the 12 already approved" 2011 (Mar.), US Medicare proposed to cover the cost of $93,000/patient prostate cancer vaccine

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Type of vaccine INDNew vaccinesInclude addition or change of adjuvant Modification of original productFormulation (e.g., lyophilized vs. liquid)StrengthRoute of administrationChange in indication, age group, schedule, etc.Concomitant administration with other vaccine

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4-7-2011*Vaccines in development/trial, Taiwan, 2011Ag typeVirus vaccine Polysaccharide conjugate vaccine rDNA protein vaccineIndicationInfectious diseaseCancer

ReferenceIn generalGuideline on the requirements for quality documentation concerning biological IMP in clinical trials, draft, EMA, 2010Guideline on strategies to identify and mitigate risks for FIH clinical trials with investigational medicinal products, EMA, 2007VaccinesWHO: Biologicals TRSJapan NIID: Minimum requirements for biological products Pharmacopeia: Ph. Eur, USP4-7-2011*

Quality of a product(EMA)NDA, to ensure a consistent, state-of-the art quality of a productIMP, quality attributes related to safety aspectsNature of product, clinical phase, patient population, nature/severity of illness, duration of trial.IMP documentation, M3 of CTDIMP should be produced in accordance with the principles and the detailed guidelines of GMP..*4-7-2011

4-7-2011*A clinical trialStarts with the quality/control of the test drugQuality of a biological/biotechnological productInclude safety issues, e.g., impurity, adventitious agent (e.g., bacteria/fungi, mycoplasma, virus)Test drug used in animal toxicity studies be representative of the material for human studySo as to support a phase 1 study with end points of safety and preliminary immunogenicity

Characterization -Ag vs. therapeutic drugFor Ag, immunogenicity is the desired effect, therefore, concept of certain characteristics is different (e.g., product-related impurity, which would otherwise cause undesired immunogenicity for protein drug)In general, extent of characterization is less for an Ag (e.g., product-related impurity) Thus, Process = quality is more likely to be the case for Ag. Therefore, the approach to establish a design space or platform technology is less likely to apply to vaccine product

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4-7-2011*ScopeVaccineInformation from research stageClinical trial-INDCollaboration from all parties makes a trial going

4-7-2011*Information from research stageScience A vast amount of information has generated from basic researchHowever, most information is yet to be interpreted and thus transferable to development Provide rationale based on disease pathogenesis, and identify Ag candidateControl of materials Raw materials, starting materials, solvents, reagents, catalysts, e.g., Source, history of the cell substrateHistory of construction of the expression plasmid

4-7-2011*Information from research stageSafety informationPlan to obtain and document relevant safety data from research studies even they are designed to assess biologic effects. This is an effective approach to lunch preclinical safety evaluationExtent and design of toxicity studies could depend on how much prior info existEspecially for vaccine product

4-7-2011*Control of materialsDocuments, starting from research stageOrigin, lineageHistory of passage, testingMedia component, e.g., FBS, trypsinAll of the documents be transferable to R& D stageEstablishment of a cell bank or virus bank~GMP Storage, inventory, identification, handling, GMPQualification of cell and virus bankContract labGLP/GMP status

4-7-2011*History of a virus strainExample (FDA, Review of Vero cell banks for Rotarix, 2008)The Serotype G1 HRV strain (genotype P[8]) which GSK used to make vaccine product is designated RIX4414. It was derived from strain 89-12, initially developed by Avant Therapeutics, Inc. ---------------. The virus was isolated in ------------- from a child in Cincinnati with a natural case of rotavirus with mild diarrhea. This original isolate was passaged 26 times in primary African Green monkey kidney cells (AGMK) by Avant for use as seed material. The P26 virus was ------- passaged by -- --------------- AVANT, -------, which passaged the seed virus an additional 7 passages to P33. This was the material that was clinically tested ---------------. The additional 7 passages were performed in an AGMK cell line that ------- characterized in --------.

4-7-2011*Raw material of animal sourceCOA of FBS (partially shown)

4-7-2011*Raw material of animal sourceCOA of porcine trypsin

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Biosafety control

Combination of testings (starting material, UPB, intermediates..) and demonstrating production process to remove a wide variety of potential infectious viruses

4-7-2011*CMCControl of materials (before phase 1)Raw materials, starting materials, solvents, reagents, catalystsBiologically-sourced materials, TSE concernSource, history, and generating of cell substrateExpression constructCell banking system, characterization, and testing Non-viral agentEndogenous and adventitious virusesTumorigenicity, case dependent

4-7-2011*A reference- Ancillary materials (AMs) for cell-based productReagent and materials that are NOT intended to be present in the final product, e.g., FBS, digestion enzymes, GF, cytokines, antibiotics, media Vendor qualification(cGMP), audit/inspection recordQuality control testing programDocumentation Grade, traceability, or country of origin/source ( animal-derived AMs)Batch analytical results Stability assessment during use

4-7-2011*Risk classification of AMsUSPRisk tier 1Low-risk, highly qualified materials with intended use as therapeutic drug or biologic, medical device, or implantable materialTherapeutic gradeE.g, HSA, insulin, IL-12, antibioticsCertificate of analysis (COA)Assess removal from final product

4-7-2011*Risk classification of AMs USPRisk tier 2Low-risk, well-characterized materials with intended use as AMs, produced in compliance with GMPsFor use in drug, biologic, or medical device manufacture, e.g., growth factor, proteolytic enzymes, density gradient media (Exclude most animal-derived materials)COAAssess removal from final productVendor audit

4-7-2011*Risk tier 3Moderate-risk materials not intended for use as AMsFor in vitro diagnostic use or reagent grade materials, e.g, growth factors, culture media, chemicalsCOAConfirm critical test result shown in COADevelop internal specifications, eventuallyAssess removal from final productVendor audit

Risk classification of AMs USP

4-7-2011*Risk classification of AMs USPRisk tier 4High-risk materialsToxin, most animal-derived materialsFeeder cells, ascites-derived Ab, cholera toxin, animal-derived additives (e.g., FBS)COAConfirm critical test result shown in COADevelop internal specifications, eventuallyAssess removal from final productVendor auditSource animal, country of origin, adventitious agent testing

4-7-2011*Recent guidance- Cell substrateGuidance for industry: Characterization and qualification of cell substrates and other biological starting materials used in the production of viral vaccines for the prevention and treatment of infectious disease, FDA, 2010Recommendations for the evaluation of animal cell cultures as substrates for the manufacture of biological medicinal products and for the characterization of cell banks, draft, WHO, 2010

4-7-2011*Ideal substrate to produce biological/biotechnological productsWHO Technical report series, No. 878, 1998Permanent/continuous cell lineMCB, WCBQuality controlledSerum-free and/or protein-free media

Nature Reviews, June 2010, vol.10, p.441-Cell line identificationIncidence of misidentification in 1977 was 16%, 1999 was 18%ATCC working group ASN-0002 (BOX), currently developing a standard for human cell line authentication

4-7-2011*Cell substrateFrom embryonic egg for Flu vaccineHarvest: allantoic fluid, manual or automated systemsInoculation: into allantoic cavity, manually or automated system

4-7-2011*Cell lines for the production of vaccinesLicensed vaccine

Cell substrateVaccineTypeOriginLive attenuatedInactivatedPrimary tissues or cellsCalf lymph, mouse brain, chicken egg, chicken embryo cellSmall pox, Influenza,Measles, MumpsJEV, Influenza, RabiesDiploid cellsHuman (MRC-5, WI-38)Rubella,Varicella/Zoster Poliovirus, HAV, RabiesContinuous cells(Non-tumorigenic)Monkey (Vero)Small pox, RotavirusPoliovirus, JEV, RabiesContinuous cells(Tumorigenic )Canine (MDCK)-InfluenzaNon-mammalian cellsYeast (S. cerevisiae)Insect (Hi-5)-HBV, HPV (rDNA product)

4-7-2011*Qualification of the cell bank-Biosafety testsNon-viral agentSterility, Mycoplasma, (Mycobacteria, Spiroplasma)Adventitious or endogenous virusesGeneral (in vitro and in vivo test, retrovirus)Specific (cell line dependent)Tumorigenicity, case dependent

Qualification of the cell bank-Virus tests* Specific tests for :Cell lines derived from human, NHP, or other cell lines as appropriate.Culture media using animal-derived components ( e.g., bovine or porcine) USP, Ph. Eur.

4-7-2011*TumorigenicityTumorigenicity (when not known, test on EPC)Cells form tumor in animal (nude mice), Hela as + control, medium/2n cells as control, 12 wks, 4 monthsOncogenicity (when T+ and for product of prophylactic use)Agents (e.g., virus, DNA) induce host cell to form tumor (newborn animal), negative control, 4 monthsCell substrate w/ or w/o tumorigenicity (in trial or licensed)

Traditional vaccinerDNA protein product(drug or vaccine)T- O-Yes (inactivated, attenuated)YesT+ O-Yes (Inactivated)YesT+ (O+)e.g., rodent cellsNo Yes

4-7-2011*PCVPorcine circovirus types 1 and 2 are both small sscDNA viruses and common in pigs. Neither PCV1 nor PCV2 are known to infect or cause illness in humans, however PCV2 may cause illness in pigs. Detecting PCV1 DNA in Rotarix and PCV1/PCV2 DNA in RotaTeq vaccine productsViruses derived from Vero MCB and carried through manufacture process to products PCV1 DNA is present in poliovirus harvests, but not in final bulk or container (due to inactivation step)

Source: FDA vaccine advisor committee meeting (May 7, 10)

4-7-2011*PCVRotaTeq (Merck)A live, oral pentavalent vaccine that contains 5 live reassortant rotaviruses, parent strains were isolated from human and bovine hostsPackage insert (Sep. 2010)

RotarixA live, oral vaccine derived from human 89-12 strain (G1P[8] type)Package insert (2010)

FDA actionsAdditional testings are requiredFDA (Dec., 2010) requested information regarding Plans that the manufacturers may have to implement additional adventitious agent testing methods as part of their manufacturing process as these methods become available including, but not limited to, screening for PCV and PCV DNA, andAny additional in-process testing for adventitious agents that they may have recently added, but not reported to FDA.4-7-2011*

4-7-2011*Validation of viral clearance stepsFor rDNA vaccine produced from mammalian/insect cell lines, why virus testing alone is not enough?Due to limitations of testing methodsSensitivity, susceptibility (indicator cell, animal model)Sampling of test materialReference: Validation of Biopharmaceutical purification processes for virus clearance evaluation, Allan Darling, Mol. Biotec. Vol. 21, 2002

4-7-2011*Allan Darling, Molecular Biotechnology, vol. 21, 2002Besides DL of test method, ability to detect low concentrations of virus is also limited by statistical samplingProbability that a sample v does not contain virus is p(0) = ((V-v)/V)nIf V>>v, above equation be simplified by Poisson distributionp(0) = e-cv, c = (In p)/-vIf v=1 mL, c=10-1000 virus particles/LProbability of 1 mL will not contain a virus particle c 10 100 1000 p(0) 0.99 0.90 0.37

4-7-2011*A clinical trial-IND dossierBiological/biotechnological productsPoorly characterized, e.g., virus vaccine, cell-based vaccineWell-characterized, e.g., rDNA protein vaccine, DNA vaccine Technical related documentClinical study proposalInvestigator brochure CMCPharmacology and toxicology (PK, when appropriate)Clinical

CMCA summary report with supporting documents, e.g., batch analysis, stability dataA valid description which reveals all necessary components to demonstrate the quality and control of the test drugPresent data in tabular form with brief narrative highlighting the main pointsCTD format is a valid reference to organize the dossier After phase 1, any change such as cell line, process, manufacture site, will require comparability

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4-7-2011*CTD M3 (partially shown)3.2.S DRUG SUBSTANCE ()3.2.S.1 General Information ()3.2.S.1.1 Nomenclature ()3.2.S.1.2 Structure ()3.2.S.1.3 General Properties ()3.2.S.2.Manufacture ()3.2.S.2.1 Manufacturer(s) ()3.2.S.2.2 Description of manufacturing process and process controls ()3.2.S.2.3 Control of materials ()3.2.S.2.4 Controls of critical steps and intermediates ()3.2.S.2.5 Process validation and/or evaluation (/)3.2.S.2.6 Manufacturing process development ()3.2.S.3 Characterization ()

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3.2.S.2.Manufacture

3.2.S.2.2 Description of manufacturing process and process controls

()3.2.S.2.3 Control of materials/Cell bankVirus seed lotidentityviabilitypurity`Unprocessed bulkexpression construct 3.2.S.2.4 Controls of critical steps and intermediatesDNA/(conjugation)3.2.S.2.5 Process validation and/or evaluation /

4-7-2011*CMC summary-Phase 1Manufacturer ad manufacturing process Flow diagram and description, batch sizeControls which relate to product safetyFor rDNA products derived from cell lines of human or animal origin, validation of the viral clearance procedureFor inactivated vaccines, a validation of the inactivation processFor live vaccines, a demonstration of the attenuating characteristics

4-7-2011*CMC summary-Phase 1Control of materialsRaw materials, starting materials, solvents, reagents, catalystsBiologically-sourced materials, TSE concernSource, history, and generating of cell substrate and viral/bacterial seedExpression constructCell/virus/bacteria banking system, characterization, and testing Non-viral agentAdventitious and endogenous virusesTumorigenicity, case dependent

CMC summary-Phase 1Analytical methodPharmacopeiaNon pharmacopeiaA brief descriptionQualification of safety related methodE.g., HCP, host cell DNA, residual reagent

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4-7-2011*CMC summary-Phase 1Drug substance (Ag, adjuvant, novel excipient)CharacterizationSpecification (preliminary), e.g., identity, strength, potency, and purity (& impurity)E.g., HCP, DNA, residual reagentsDrug productAdjuvant, excipients, diluentsDosage form, compositionPremix, on-site mix (adjuvant, dilution, reconstitution)Specification (preliminary)

4-7-2011*CMC summaryPhase 1StabilityAt least cover the duration of trialIn-use stability information, e.g., after mixing, dilution, reconstitution, multiple withdrawing Batch analytical result, DS and DPBatch for animal and clinical studiesSame batch/formulation is recommended for vaccine productsIf not, describe (to compare)CMCAnimal study might be useful

4-7-2011*Phase-in of validationValidation of manufacturing processes and analytical methods goes along with the clinical development In phase 1, safety related method requires certain extent of validation E.g., Host cell DNA, protein, viral test (e.g., PCR)Limit - specificity and LODQuantitation- more extensive validation

Potency assayCorrelation to in-vivo biological activity should be justifiedPotency should be in the stability study, even it is not proven to be stability-indicating in the early trial

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4-7-2011*Phase-in of validation Bioassay (potency)No need for LOD, LOQAt phase 1/2SpecificityPrecisionRepeatability, e.g., Samples from several independent preparations of the same stockSame sample, well to well, %CV

4-7-2011*Phase-in of validation Bioassay (potency)At phase 3 and NDAPrecisionIntermediate precision, e.g., plate to plate, day to day, analyst to analystReproducibility, inter-laboratoryRobustnessApply changes that probably will not happen (e.g., increase in Rx time, temperature change)Pushing the systemLinearity/range/accuracy

4-7-2011*USP Verification of compendial proceduresUsers of compendial analytical procedures are not required to validate procedure, but documented evidence of suitability should be established under actual conditions of use Not for microbiological proceduresSome of the analytical performance characteristics for validation study, may be used for verification processE.g., specificity is a key parameter, potential interference fromDrug substance from different suppliers may have different impurity profilesDrug product contains different excipients, additives

4-7-2011*Pre-clinical preparationAg (and adjuvant)Analytical methods, to characterize Ag/adjuvant, specification set up, and stability indicatingFormulation, delay optimization until beyond phase 1 and achieves proof of conceptTo maintain stability during trialEnable use in animal Tox and FIH studyBioanalytical assays, to monitor immune response in vivoVaccine-specific parametersIdentify infections

4-7-2011*After Phase 1Formulation for next clinical studiesDevelop/optimize manufacturing processesOptimize analytical test methodsUpdate CMC section of IND

4-7-2011*Phase 2Update clinical suppliesIdentify critical process parametersOptimize manufacturing processSelect doses for phase 3 studiesEstablish formulation and container/closureUpdate phase 2 package

4-7-2011*Phase 3Select commercial manufacturing and packaging sitesPrepare pharmaceutical development reportScale upProcess validationEstablish stability studiesPrepare CMC section of NDA

Lot release, in general4-7-2011*Marion Gruber, FDA-vaccine review , 2008

Tests after mixing, an example4-7-2011*Marie-Chantal Uwamwezi, GSK-malaria vaccine, 2010

4-7-2011*Product development-a reference to checkUS National Institute of Allergy and Infectious Diseases-VaccinesInstruction and advise to HIV vaccine researchersInformation regarding (HIV) vaccines in all aspectsPreclinical master contract (HIV vaccines)Manufacture GMP pilot lots of vaccine for testing in humans, or lots for testing in nonhuman primates Perform tests for safety, immunogenicity and other preclinical testing of vaccine candidates Preparation of FDA submissions leading up to human trials

4-7-2011*Overview of product development of a vaccineSource: US NIAID

Thank you!4-7-2011*

SPARE SLIDES4-7-2011*

4-7-2011*Phase 2 & 3- incremental requirementPC characterization, to more detailCharacter(s) affected by manufacture processBatch informationComparability due to changes such as process, scaleStability data, updateManufacturing and controls, updateSpecification, updatePhase-in validationProcess validation (phase 3 or NDA)Analytical method validation (phase 3 or NDA)

4-7-2011*Phase 1 3- incremental requirementPaul-Ehrlich-Institut

Different role and thus point of viewScientistContract manufacturerSponsorRegulatory agency

4-7-2011*Concentration of test drug

ScientistIf only the purified protein is potent, as expected. Concentration is not a major issue.Sponsor Concentration affects injection volume, either for animal study or clinical trial.Volume/dose for sc, id, im is less than iv, thus more concentrate formulation is required for route other than iv.Concentration affects the vaccine formulation, if adjuvant is to be used.Contract manufacturerConcentration be clarified before agreement. It depends on the capacity of the production.Process improvement or modificationTimeline, costRegulationIf only the stability of product is demonstrated, concentration is not a major issue.

4-7-2011*Lot and quantity of the test material

RegulationDifferent lots can be used for animal and clinical study, if only they are comparableSponsorSame or different lot for toxicity and clinical study Timeline, contract, costQuantity for pharmacology studies (may be research grade)Quantity for toxicity studiesOverage of 15-20%, plus consideration of formulation and vialing (e.g., 10 dose/vial, last 2 doses will use 1 vial) (Quantity for specification and stability testings, retention samples)Quantity for clinical trial

4-7-2011*On site mixing-Ag and adjuvant come from different manufacturer

RegulationIn-use stabilityAt least potency and sterility tests are required. Robustness of the method of on-site mixingDuring trial, if no change in the CMC (manufacture, specification..), tests are done on one representative lot. SponsorDesign the method of mixingWhat are the test items required at this conditionWho/where would be able to perform the in-use stability tests

4-7-2011*Estimate cost of the test materialsExamples

ManufacturerIf there is no DMF already in place, additional cost is needed to prepare documentAny test/study by contract lab (e.g., viral clearance, cell bank qualification) is at additional cost Sponsor~10% overage for each filling/mixing step or each component.Usually 30-40% of the total quantity are used for clinical trial, the rest of the products are used in animal studies (e.g., immunogenicity, toxicity) and tests (e.g, in-use stability)Final quantity ordered need to take all experiments, tests, and reserve samples into consideration

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