Vaccine clinical trial -Quality, include control of cell substrate

1
Vaccine clinical trial-Quality, include control
of cell substrate

• Ywan-Feng Li
• Center for Drug Evaluation
• 4-7-2011
• ????????????(I)

The views expressed in this presentation are not
necessary those of Center for Drug
Evaluation-Taiwan
2
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The views offered here do not necessarily reflect
official positions of TFDA 
3
Heterogenicity of the biological/biotechnological
products
Peptides (20-30 a,a,)
Tissue engineering P.
rDNA proteins (Mab, fusion protein)
Gene/cell therapy P.
Blood/plasma products
rDNA-derived vaccines
Allergens
Traditional vaccines
Process defines product
4
Scope

• Vaccine
• Information from research stage
• Clinical trial-IND
• Cell substrate-Testing for adventitious agents
• Quality, starts from phase 1
• Quality, (almost) finalizes at phase 3 and
  continues through product life span
• Collaboration from all parties makes a trial going

5
Type of vaccine

• Vaccines represent the most diverse type of
  products
• Attenuated or killed pathogens (bacteria, virus,
  parasites) (traditional vaccine)
• Purified and recombinant protein
• Synthetic peptides
• Polysaccharide (free or conjugate to carrier)
• DNA, viral vectors
• (Cell-based product)

6
Preventive versus therapeutic vaccines
In general
Characteristic Preventive Therapeutic
Population Healthy subject Usually patient
Clinical outcome Decrease microbial infection and/or transmission Cure or postpone disease progression (usually as a 2nd line strategy)
Regimen Low dose, episodic Usually high dose, continual (more like a drug)
Evaluations in early trial Safety, immunogenicity Safety, immunogenicity
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Preventive versus therapeutic vaccines
Characteristic Preventive Therapeutic
Regulatory evaluation Emphasis on safety Efficacy Benefit/risk assessment Efficacy
Public expectation Highly concern and sensitive to the potential risks Less concern regarding the potential risks
However, quality and assessment of the vaccine (Ag and adjuvant) is the same for either type of vaccine However, quality and assessment of the vaccine (Ag and adjuvant) is the same for either type of vaccine However, quality and assessment of the vaccine (Ag and adjuvant) is the same for either type of vaccine
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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 cancer
• 2011, FDA approved Dendreon's request to increase
  production capacity
• FDA 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

9
Type of vaccine IND

• New vaccines
• Include addition or change of adjuvant
• Modification of original product
• Formulation (e.g., lyophilized vs. liquid)
• Strength
• Route of administration
• Change in indication, age group, schedule, etc.
• Concomitant administration with other vaccine

10
Vaccines in development/trial, Taiwan, 2011

• Ag type
• Virus vaccine
• Polysaccharide conjugate vaccine
• rDNA protein vaccine
• Indication
• Infectious disease
• Cancer

11
Reference

• In general
• Guideline on the requirements for quality
  documentation concerning biological IMP in
  clinical trials, draft, EMA, 2010
• Guideline on strategies to identify and mitigate
  risks for FIH clinical trials with
  investigational medicinal products, EMA, 2007
• Vaccines
• WHO Biologicals TRS
• Japan NIID Minimum requirements for biological
  products
• Pharmacopeia Ph. Eur, USP

12
Quality of a product(EMA)

• NDA, to ensure a consistent, state-of-the art
  quality of a product
• IMP, quality attributes related to safety aspects
• Nature of product, clinical phase, patient
  population, nature/severity of illness, duration
  of trial.
• IMP documentation, M3 of CTD
• IMP should be produced in accordance with the
  principles and the detailed guidelines of GMP..

13
A clinical trialStarts with the quality/control
of the test drug

• Quality of a biological/biotechnological product
• Include 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 study
• So as to support a phase 1 study with end points
  of safety and preliminary immunogenicity

14
Characterization -Ag vs. therapeutic drug

• For 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

15
Scope

• Vaccine
• Information from research stage
• Clinical trial-IND
• Collaboration from all parties makes a trial going

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Information from research stage

• Science
• A vast amount of information has generated from
  basic research
• However, most information is yet to be
  interpreted and thus transferable to development
• Provide rationale based on disease pathogenesis,
  and identify Ag candidate
• Control of materials
• Raw materials, starting materials, solvents,
  reagents, catalysts, e.g.,
• Source, history of the cell substrate
• History of construction of the expression plasmid

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Information from research stage

• Safety information
• Plan 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 evaluation
• Extent and design of toxicity studies could
  depend on how much prior info exist
• Especially for vaccine product

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Control of materials

• Documents, starting from research stage
• Origin, lineage
• History of passage, testing
• Media component, e.g., FBS, trypsin
• All of the documents be transferable to R D
  stage
• Establishment of a cell bank or virus bank
• GMP
• Storage, inventory, identification, handling, GMP
• Qualification of cell and virus bank
• Contract lab
• GLP/GMP status

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History of a virus strain

• Example (FDA, Review of Vero cell banks for
  Rotarix, 2008)
• The Serotype G1 HRV strain (genotype P8) 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 --------.

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Raw material of animal sourceCOA of FBS
(partially shown)
21
Raw material of animal sourceCOA of porcine
trypsin
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Scope

• Vaccine
• Information from research stage
• Clinical trial-IND
• Cell substrate-Testing for adventitious agents
• Quality, starts from phase 1
• Quality, (almost) finalizes at phase 3 and
  continues through product life span
• Collaboration from all parties makes a trial going

23
Biosafety control Combination of testings
(starting material, UPB, intermediates..) and
demonstrating production process to remove a wide
variety of potential infectious viruses
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CMC

• Control of materials (before phase 1)
• Raw materials, starting materials, solvents,
  reagents, catalysts
• Biologically-sourced materials, TSE concern
• Source, history, and generating of cell substrate
• Expression construct
• Cell banking system, characterization, and
  testing
• Non-viral agent
• Endogenous and adventitious viruses
• Tumorigenicity, case dependent

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A reference- Ancillary materials (AMs) for
cell-based product

• Reagent 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 record
• Quality control testing program
• Documentation
• Grade, traceability, or country of origin/source
  ( animal-derived AMs)
• Batch analytical results
• Stability assessment during use

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Risk classification of AMsUSPlt1043gt

• Risk tier 1
• Low-risk, highly qualified materials with
  intended use as therapeutic drug or biologic,
  medical device, or implantable material
• Therapeutic grade
• E.g, HSA, insulin, IL-12, antibiotics
• Certificate of analysis (COA)
• Assess removal from final product

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Risk classification of AMs USPlt1043gt

• Risk tier 2
• Low-risk, well-characterized materials with
  intended use as AMs, produced in compliance with
  GMPs
• For use in drug, biologic, or medical device
  manufacture, e.g., growth factor, proteolytic
  enzymes, density gradient media (Exclude most
  animal-derived materials)
• COA
• Assess removal from final product
• Vendor audit

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Risk classification of AMs USPlt1043gt

• Risk tier 3
• Moderate-risk materials not intended for use as
  AMs
• For in vitro diagnostic use or reagent grade
  materials, e.g, growth factors, culture media,
  chemicals
• COA
• Confirm critical test result shown in COA
• Develop internal specifications, eventually
• Assess removal from final product
• Vendor audit

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Risk classification of AMs USPlt1043gt

• Risk tier 4
• High-risk materials
• Toxin, most animal-derived materials
• Feeder cells, ascites-derived Ab, cholera toxin,
  animal-derived additives (e.g., FBS)
• COA
• Confirm critical test result shown in COA
• Develop internal specifications, eventually
• Assess removal from final product
• Vendor audit
• Source animal, country of origin, adventitious
  agent testing

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Recent guidance- Cell substrate

• Guidance 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, 2010
• Recommendations 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

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Ideal substrate to produce biological/biotechnolog
ical products

• WHO Technical report series, No. 878, 1998
• Permanent/continuous cell line
• MCB, WCB
• Quality controlled
• Serum-free and/or protein-free media
• Nature Reviews, June 2010, vol.10, p.441-
• Cell line identification
• Incidence of misidentification in 1977 was 16,
  1999 was 18
• ATCC working group ASN-0002 (BOX), currently
  developing a standard for human cell line
  authentication
  

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Cell substrateFrom embryonic egg for Flu vaccine
Harvest allantoic fluid, manual or automated
systems
Inoculation into allantoic cavity, manually or
automated system
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Cell lines for the production of vaccines

• Licensed vaccine

Cell substrate Cell substrate Vaccine Vaccine
Type Origin Live attenuated Inactivated
Primary tissues or cells Calf lymph, mouse brain, chicken egg, chicken embryo cell Small pox, Influenza, Measles, Mumps JEV, Influenza, Rabies
Diploid cells Human (MRC-5, WI-38) Rubella, Varicella/Zoster Poliovirus, HAV, Rabies
Continuous cells (Non-tumorigenic) Monkey (Vero) Small pox, Rotavirus Poliovirus, JEV, Rabies
Continuous cells (Tumorigenic ) Canine (MDCK) - Influenza
Non-mammalian cells Yeast (S. cerevisiae) Insect (Hi-5) - HBV, HPV (rDNA product)
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Qualification of the cell bank-Biosafety tests

• Non-viral agent
• Sterility, Mycoplasma, (Mycobacteria,
  Spiroplasma)
• Adventitious or endogenous viruses
• General (in vitro and in vivo test, retrovirus)
• Specific (cell line dependent)
• Tumorigenicity, case dependent

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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.
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Tumorigenicity

• Tumorigenicity (when not known, test on EPC)
• Cells form tumor in animal (nude mice), Hela as
  control, medium/2n cells as control, 12 wks, ?4
  months
• Oncogenicity (when T and for product of
  prophylactic use)
• Agents (e.g., virus, DNA) induce host cell to
  form tumor (newborn animal), negative control,
  ?4 months
• Cell substrate w/ or w/o tumorigenicity (in trial
  or licensed)

Traditional vaccine rDNA protein product (drug or vaccine)
T- O- Yes (inactivated, attenuated) Yes
T O- Yes (Inactivated) Yes
T (O) e.g., rodent cells No Yes
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PCV

• Porcine 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 products
• Viruses 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)
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PCV

• RotaTeq (Merck)
• A live, oral pentavalent vaccine that contains 5
  live reassortant rotaviruses, parent strains were
  isolated from human and bovine hosts
• Package insert (Sep. 2010)
• Rotarix
• A live, oral vaccine derived from human 89-12
  strain (G1P8 type)
• Package insert (2010)

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FDA actionsAdditional testings are required

• FDA (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, and
• Any additional in-process testing for
  adventitious agents that they may have recently
  added, but not reported to FDA.

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Validation of viral clearance steps

• For rDNA vaccine produced from mammalian/insect
  cell lines, why virus testing alone is not
  enough?
• Due to limitations of testing methods
• Sensitivity, susceptibility (indicator cell,
  animal model)
• Sampling of test material
• Reference Validation of Biopharmaceutical
  purification processes for virus clearance
  evaluation, Allan Darling, Mol. Biotec. Vol. 21,
  2002

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Allan Darling, Molecular Biotechnology, vol. 21,
2002

• Besides DL of test method, ability to detect low
  concentrations of virus is also limited by
  statistical sampling
• Probability that a sample v does not contain
  virus is
• p(0) ((V-v)/V)n
• If Vgtgtv, above equation be simplified by Poisson
  distribution
• p(0) e-cv, c (In p)/-v
• If v1 mL, c10-1000 virus particles/L
• Probability of 1 mL will not contain a virus
  particle
• c 10 100 1000
• p(0) 0.99 0.90 0.37

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Scope

• Vaccine
• Information from research stage
• Clinical trial-IND
• Cell substrate-Testing for adventitious agents
• Quality, starts from phase 1
• Quality, (almost) finalizes at phase 3 and
  continues through product life span
• Collaboration from all parties makes a trial going

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A clinical trial-IND dossier

• Biological/biotechnological products
• Poorly characterized, e.g., virus vaccine,
  cell-based vaccine
• Well-characterized, e.g., rDNA protein vaccine,
  DNA vaccine
• Technical related document
• Clinical study proposal
• Investigator brochure
• CMC
• Pharmacology and toxicology
• (PK, when appropriate)
• Clinical

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CMC

• A summary report with supporting documents, e.g.,
  batch analysis, stability data
• A valid description which reveals all necessary
  components to demonstrate the quality and control
  of the test drug
• Present data in tabular form with brief narrative
  highlighting the main points
• CTD 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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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 bank)?????(Virus seed lot)?????????? ???????????identity?viability?purity????????????? ?????? Unprocessed bulk?????? ????????????,??expression construct ?????
3.2.S.2.4 Controls of critical steps and intermediates ???????????????,??DNA/?????? ????????????(conjugation)???????
3.2.S.2.5 Process validation and/or evaluation ??????????????/??,?????????????????????????
?? (?????)
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CMC summary-Phase 1

• Manufacturer ad manufacturing process
• Flow diagram and description, batch size
• Controls which relate to product safety
• For rDNA products derived from cell lines of
  human or animal origin, validation of the viral
  clearance procedure
• For inactivated vaccines, a validation of the
  inactivation process
• For live vaccines, a demonstration of the
  attenuating characteristics

48
CMC summary-Phase 1

• Control of materials
• Raw materials, starting materials, solvents,
  reagents, catalysts
• Biologically-sourced materials, TSE concern
• Source, history, and generating of cell substrate
  and viral/bacterial seed
• Expression construct
• Cell/virus/bacteria banking system,
  characterization, and testing
• Non-viral agent
• Adventitious and endogenous viruses
• Tumorigenicity, case dependent

49
CMC summary-Phase 1

• Analytical method
• Pharmacopeia
• Non pharmacopeia
• A brief description
• Qualification of safety related method
• E.g., HCP, host cell DNA, residual reagent

50
CMC summary-Phase 1

• Drug substance (Ag, adjuvant, novel excipient)
• Characterization
• Specification (preliminary), e.g., identity,
  strength, potency, and purity ( impurity)
• E.g., HCP, DNA, residual reagents
• Drug product
• Adjuvant, excipients, diluents
• Dosage form, composition
• Premix, on-site mix (adjuvant, dilution,
  reconstitution)
• Specification (preliminary)

51
CMC summaryPhase 1

• Stability
• At least cover the duration of trial
• In-use stability information, e.g., after mixing,
  dilution, reconstitution, multiple withdrawing
• Batch analytical result, DS and DP
• Batch for animal and clinical studies
• Same batch/formulation is recommended for vaccine
  products
• If not, describe (to compare)
• CMC
• Animal study might be useful

52
Phase-in of validation

• Validation 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 LOD
• Quantitation- more extensive validation

53
Potency assay

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

54
Phase-in of validation Bioassay (potency)

• No need for LOD, LOQ
• At phase 1/2
• Specificity
• Precision
• Repeatability, e.g.,
• Samples from several independent preparations of
  the same stock
• Same sample, well to well, CVlt15-20
• (Linearity/range/accuracy)

55
Phase-in of validation Bioassay (potency)

• At phase 3 and NDA
• Precision
• Intermediate precision, e.g.,
• plate to plate, day to day, analyst to analyst
• Reproducibility, inter-laboratory
• Robustness
• Apply changes that probably will not happen
  (e.g., increase in Rx time, temperature change)
• Pushing the system
• Linearity/range/accuracy

56
USPlt1226gt Verification of compendial procedures

• Users 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 procedures
• Some of the analytical performance
  characteristics for validation study, may be
  used for verification process
• E.g., specificity is a key parameter, potential
  interference from
• Drug substance from different suppliers may have
  different impurity profiles
• Drug product contains different excipients,
  additives

57
Pre-clinical preparation

• Ag (and adjuvant)
• Analytical methods, to characterize Ag/adjuvant,
  specification set up, and stability indicating
• Formulation, delay optimization until beyond
  phase 1 and achieves proof of concept
• To maintain stability during trial
• Enable use in animal Tox and FIH study
• Bioanalytical assays, to monitor immune response
  in vivo
• Vaccine-specific parameters
• Identify infections

58
After Phase 1

• Formulation for next clinical studies
• Develop/optimize manufacturing processes
• Optimize analytical test methods
• Update CMC section of IND

59
Phase 2

• Update clinical supplies
• Identify critical process parameters
• Optimize manufacturing process
• Select doses for phase 3 studies
• Establish formulation and container/closure
• Update phase 2 package

60
Phase 3

• Select commercial manufacturing and packaging
  sites
• Prepare pharmaceutical development report
• Scale up
• Process validation
• Establish stability studies
• Prepare CMC section of NDA

61
Lot release, in general
Marion Gruber, FDA-vaccine review , 2008
62
Tests after mixing, an example
Marie-Chantal Uwamwezi, GSK-malaria vaccine, 2010
63
Product development-a reference to check

• US National Institute of Allergy and Infectious
  Diseases-Vaccines
• Instruction and advise to HIV vaccine researchers
• Information regarding (HIV) vaccines in all
  aspects
• Preclinical 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

64
Overview of product development of a vaccine
Source US NIAID
65

• Thank you!

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SPARE SLIDES
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Phase 2 3- incremental requirement

• PC characterization, to more detail
• Character(s) affected by manufacture process
• Batch information
• Comparability due to changes such as process,
  scale
• Stability data, update
• Manufacturing and controls, update
• Specification, update
• Phase-in validation
• Process validation (phase 3 or NDA)
• Analytical method validation (phase 3 or NDA)

68
Phase 1 3- incremental requirement
Paul-Ehrlich-Institut
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Phase 1 3- incremental requirement
Paul-Ehrlich-Institut
70
Phase 1 3- incremental requirement
Paul-Ehrlich-Institut
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Different role and thus point of view
Contract manufacturer
Scientist
Regulatory agency
Sponsor
72
Concentration of test drug
Scientist If 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 manufacturer Concentration be clarified before agreement. It depends on the capacity of the production. Process improvement or modification Timeline, cost
Regulation If only the stability of product is demonstrated, concentration is not a major issue.
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Lot and quantity of the test material
Regulation Different lots can be used for animal and clinical study, if only they are comparable
Sponsor Same or different lot for toxicity and clinical study Timeline, contract, cost Quantity for pharmacology studies (may be research grade) Quantity for toxicity studies Overage 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
74
On site mixing-Ag and adjuvant come from
different manufacturer
Regulation In-use stability At least potency and sterility tests are required. Robustness of the method of on-site mixing During trial, if no change in the CMC (manufacture, specification..), tests are done on one representative lot.
Sponsor Design the method of mixing What are the test items required at this condition Who/where would be able to perform the in-use stability tests
75
Estimate cost of the test materialsExamples
Manufacturer If there is no DMF already in place, additional cost is needed to prepare document Any 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
76
Thank you