MDCK as a substrate for flu antibody creation.

MDCK as a substrate for flu immunization creation Solvay Pharmaceuticals Presentation to VRBPAC November 16, 2005

Agenda Background Solvay and flu Background Solvay’s MDCK venture Safety investigation of MDCK-based antibody Conclusions

Solvay Pharmaceuticals Division of Solvay Group Global – main 40 pharma 12,800 workers 2004 deals: $2.8 billion Major R&D destinations Marietta, GA Dijon and Paris, France Hanover, Germany Weesp, The Netherlands

Solvay Pharmaceuticals & Influenza Egg-based antibody First flu immunization in Europe (1950) Uninterrupted supply for a long time Total > 250 million measurements Fourth supplier around the world; more than 50 nations

Solvay Pharmaceuticals & Influenza immunization generation offices – Weesp, The Netherlands US Headquarters Marietta, GA Countries supplied with Solvay’s egg-based flu immunization

Why a cell-based antibody venture? Eggs are open creation framework inclined to tainting Availability of eggs is defenseless against avian sicknesses

Why MDCK? Expansive vulnerability to flu infections Substantial involvement in flu research and observation High infection yields Economically possible Favorable proportion of infection to polluting influences

Solvay Pharmaceuticals’ MDCK-based flu antibody extend More than 10 years experience Microcarrier, sans serum Preclinical and clinical advancement system License conceded in The Netherlands Commercial scale office License to be redesigned to business scale item, trailed by overall permitting, including US

Solvay Pharmaceuticals’ MDCK-based immunization office Solvay Pharmaceuticals, Weesp, The Netherlands

MDCK security evaluation Identify and measure potential dangers of cell line Quantify end of these potential dangers by downstream handling (DSP) Quantify potential dangers of cell substrate staying for the immunization beneficiary CBER’s Defined-Risks Approach Lewis AM Jr, Krause P, Peden K. Dev Biol. 2001, vol 106, pp 513-535

MDCK wellbeing evaluation Cell line portrayal Downstream handling Final item security

MDCK security appraisal Cell line portrayal Passage history Adventitious operators Tumorigenicity

1a. Cell portrayal - History 1958: Isolation from pooch kidney 1964: Deposition at American Type Culture Collection (ATCC) No presentation of Bovine Spongiform Encephalopathy-like operators 1991: Preparation of ATCC working stock 1992: Preparation of Master Cell Bank and Working Cell Bank

1a. Cell portrayal - Passage history 0: Isolation from puppy kidney 49: Deposition at American Type Culture Collection (ATCC) 52: Preparation of ATCC working stock 56: Preparation of Master Cell Bank 57: Preparation of Working Cell Bank entries utilized for immunization produce 97: Preparation of Extended Cell Bank sections utilized for security evaluation

1b. Cell portrayal - Adventitious specialists Mycoplasmas Bacterial and contagious sterility In vitro co-development in locator cell lines In vivo testing in different species Electron microscopy Retrovirus testing No unusual operators found in Solvay’s MDCK

1b. Cell portrayal - Adventitious specialists Specific testing for normally happening canine infections Specific testing for infections to which MDCK is helpless No extrinsic operators found in Solvay’s MDCK

1c. Cell portrayal – Tumorigenicity Phenotypic trademark; tumor development in creature models Concern of introduction of antibody beneficiary to Intact cells Cellular segments Residual cell DNA Unlikely in immunization beneficiary because of allograft dismissal

1c. Cell portrayal – Tumorigenicity Tumorigenic capability of in place cells 4 week and 6 month study in grown-up insusceptible lacking naked mice Tumorigenic capability of cell lysates 6 month study in grown-up and infant bare mice, infant rats and infant hamsters Oncogenic capability of DNA 6 month study in grown-up and infant naked mice, infant rats and infant hamsters

Incidence Nodule size (mm 2 ) Complete relapse 40 days 6 mos Negative control 0/26 - 10 1 MDCK, p98 0/26 - 10 3 MDCK, p98 0/26 - 10 5 MDCK, p98 18/26 2 6 5 10 7 MDCK, p98 30/30 28 27 4 10 7 MDCK, p56 (ATCC) 23/26 2 0 23 Positive control (10 7 HeLa) 25/26 109 sac 1c. Cell portrayal – Tumorigenicity Intact cells : 6 month study in 4 week bare mice

1c. Cell portrayal – Tumorigenicity Intact cells : 6 month study in 4 week naked mice * Non-MDCK

1c. Cell portrayal – Tumorigenicity Lysate of 10 7 cells : 6 month oncogenicity study

1c. Cell portrayal – Tumorigenicity 0.1mg MDCK-DNA : 6 month oncogenicity study

1c. Cell portrayal – Tumorigenicity Characterization of tumors in DNA study Histopathology One mouse: histiocytic tumor in liver One mouse: lymphoma in thoracic pit Spontaneous tumors not unforeseen in safe inadequate bare mice

1c. Cell portrayal – Tumorigenicity Follow-up studies with MDCK cell DNA Larger study in bare mice Fetal and neonatal wellbeing study in rats

1c. Cell portrayal – Tumorigenicity rundown Moderate tumorigenic potential in invulnerable insufficient creatures appeared at measurements levels ≥ 10 5 Majority of knobs incompletely relapse; complete relapse in 5/25 at 10 5 and in 4/28 at 10 7 Increases with section level and/or adjustment to sans serum development Histopathology at entry 98 is in concurrence with the writing for MDCK cells

1c. Cell portrayal – Tumorigenicity synopsis Tumorigenic potential not saw in safe equipped creatures Lysates of 10 7 MDCK cells not tumorigenic 0.1 mg MDCK-DNA not thought to be oncogenic; confirmative studies started

MDCK security appraisal 1. Cell line portrayal Downstream handling (DSP) Elimination of in place cells Elimination of cell DNA

MDCK-based immunization generation

2a. DSP - Elimination of in place cells

2a. DSP - Elimination of in place cells

2b. DSP - Elimination of DNA Benzonase

2b. DSP - Elimination of DNA content freedom element detail: < 10 nanograms for every dosage

2. DSP - Summary Adequate purging and testing warrants antibody wellbeing Solvay is resolved to take after the most recent logical bits of knowledge and administrative direction

MDCK security appraisal Cell line portrayal Downstream handling Final item security Pre-clinical experience Clinical experience

MDCK-based immunization – Pre-clinical experience

Local resilience No neighborhood bothering Systemic lethality No unfavorable impacts Pyrogenicity No particular increment in body temp Mutagenicity No increment in number of micronuclei Anaphylaxis No dynamic hypersensitivity Passive hypersensitivity great to egg-based MDCK-based antibody – Pre-clinical experience

MDCK-based antibody – Clinical experience 14 randomized, twofold visually impaired studies 1,023 subjects on MDCK-based subunit immunization Population: 18-60, more than 60, patients at-danger for complexities for flu, atopic patients and subjects with an egg-sensitivity Major destinations: Show immunogenic non-mediocrity and tantamount wellbeing to existing egg-based subunit flu immunization

MDCK-based antibody – Clinical experience Local and systemic reactogenicity profile is practically identical to egg-based antibody Reactions are minor and brief No startling wellbeing discoveries Non-substandard in immunogenicity MDCK-based antibody has similar wellbeing and immunogenicity profile as egg-based immunization

Conclusions Solvay is certain MDCK is a protected substrate for inactivated flu antibodies The utilization of MDCK will enhance unwavering quality of flu antibody supply, and improve pandemic readiness Solvay will seek after authorizing around the world, including US

Iris de Bruijn, Ph.D. Worldwide Clinical Director, Influenza Vaccines Ed Geuns, Pharm.D. Chief, Regulatory Affairs Michael Hare, B.S. Supervisor, Regulatory Affairs Alex Kersten, D.V.M. Senior Scientist, Pre-Clinical Development Jeroen Medema, MSc. Senior Scientist, Vaccines Peter Finn, MRCVS., FRCPath. Expert Michael Williams, B.S. VP, Viral Products The Biologics Consulting Group Ruth Wolff, Ph.D. Chief, Therapeutics The Biologics Consulting Group Participants

MDCK as a substrate for flu antibody generation Solvay Pharmaceutic