V accine 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 新醫藥品法規人才培訓課程（I） The views expressed in this presentation are not necessary those of Center for Drug Evaluation-Taiwan

2. 本次內容僅代表查驗中心之觀點及經驗分享凡涉及政策方向及法規解釋適用，應依衛生主管機關之指示為準本次內容僅代表查驗中心之觀點及經驗分享凡涉及政策方向及法規解釋適用，應依衛生主管機關之指示為準 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:

7. Preventive versus therapeutic vaccines

8. 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

16. 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

17. 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

18. 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

19. History of a virus strain Example (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 waspassaged 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 --------.

20. Raw material of animal sourceCOA of FBS (partially shown)

21. Raw material of animal sourceCOA of porcine trypsin

22. 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. Research stage Cell/virus seed & raw material Cell/virus bank and unprocessed bulk Contract lab Viral clearance steps Contract lab Purified bulk (Calculation of estimated particles/dose) Biosafety control Combination of testings (starting material, UPB, intermediates..) and demonstrating production process to remove a wide variety of potential infectious viruses

24. 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

25. 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

26. Risk classification of AMsUSP<1043> 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

27. Risk classification of AMs USP<1043> 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

28. Risk classification of AMs USP<1043> 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

29. Risk classification of AMs USP<1043> 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

30. Recent guidance- Cell substrate • Guidance for industry: Characterization and qualification of cell substrates andother 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

31. Ideal substrate to produce biological/biotechnological 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

32. Cell substrateFrom embryonic egg for Flu vaccine Inoculation: into allantoic cavity, manually or automated system Harvest: allantoic fluid, manual or automated systems

33. Cell lines for the production of vaccines • Licensed vaccine

34. 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

35. 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.

36. 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)

37. 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)

38. 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 (G1P[8] type) • Package insert (2010)

39. 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.

40. 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

41. 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 V>>v, above equation be simplified by Poisson distribution • p(0) = e-cv, c = (In p)/-v • If v=1 mL, c=10-1000 virus particles/L • Probabilityof 1 mL will not contain a virus particle c 10 100 1000 p(0) 0.99 0.90 0.37

42. 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

43. 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

44. 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

45. CTD M3 (partially shown) • 3.2.S DRUG SUBSTANCE (原料藥) • 3.2.S.1 General Information (一般資料) • 3.2.S.1.1 Nomenclature (命名) • 3.2.S.1.2 Structure (結構式) • 3.2.S.1.3 General Properties (一般性質) • 3.2.S.2.Manufacture (製造) • 3.2.S.2.1 Manufacturer(s) (製造者) • 3.2.S.2.2 Description of manufacturing process and process controls (製程敍述及製程管制) • 3.2.S.2.3 Control of materials (物質管制) • 3.2.S.2.4 Controls of critical steps and intermediates (關鍵步驟及中間體管制) • 3.2.S.2.5 Process validation and/or evaluation (製程確效及/或評估) • 3.2.S.2.6 Manufacturing process development (製造程序的發展) • 3.2.S.3 Characterization (特性)

46. 舉例 (原料藥部分)

47. 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)