Rapid Manufacture and Release of a GMP Batch of Zaire Ebolavirus Glycoprotein Vaccine Made Using Recombinant Baculovirus-Sf9 Insect Cell Culture Technology

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Tim Hahn SVP, Global Manufacturing Operations Novavax, Inc. 10 March 2015

1 Presentation Overview

• Ebola Background • Novavax Ebola/Mak Glyoprotein Vaccine • Novavax saponin-based Matrix-M™ Adjuvant • Rapid Manufacturing Timeline • Pre-clinical Data

2 2 Ebola Background

• A new virus was discovered in 1976 ‒ Caused hemorrhagic fever ‒ Later named genus Ebolavirus ‒ Five Ebola virus species including Zaire Ebola virus (EBOV) • From 1976 to 2013 ‒ Several outbreaks of Ebola Virus Disease (EBV) ‒ Average EVD case fatality rate is about 50% ‒ Most outbreaks in central and west Africa ‒ Largest number of cases previously reported was 425 in 2000 outbreak ‒ Estimate of total EVD deaths from 1976 to 2013 is 1590. • On-going 2014 (to 2015) outbreak as of March 4, 2015 ‒ 23,969 cases ‒ 9807 deaths • 6-times the total from 1976 to 2013

3 3 Span of Endemic

• Most cases in Guinea, Liberia, and Sierra Leone

• Infections also reported in: • Nigeria • Democratic Republic of Congo • Senegal • United States • Spain •Mali • United Kingdom

4 Presentation Overview

• Ebola Background • Novavax Ebola/Mak Glyoprotein Vaccine • Novavax saponin-based Matrix-M™ Adjuvant • Rapid Manufacturing Timeline • Pre-clinical Data

5 5 Guinea Ebola (EBOV) Gene Sequence for Cluster 3 Virus Science September 12, 2014

• In Sierra Leone May 2014 at funeral of a Guinea EVD case, attendees were infected with two distinct Ebola cluster 1,2 viruses leading to cluster 3 sustained human-to-human transmission. • “Because many of the mutations alter protein sequences and other biologically meaningful targets, they should be monitored for impact on diagnostics, “In memoriam: Tragically, five co-authors, who contributed greatly to public health and research vaccines, and therapies efforts in Sierra Leone, contracted EVD and lost their critical to outbreak battle with the disease before this manuscript could be response.” published”

6 6 Novavax EBOV/Mak GP recombinant nanoparticle vaccine

Recombinant EBOV/Makona Glycoprotein (GP) • GenBank #AIG96283 • EBOV [H.sapiens -wt/SLE/2014/Makona-G3798] • Full length, unmodified GP gene • Synthetic, codon optimized • Cloned into a baculovirus vector rBV-GP

CLONING EXPRESSION PURIFICATION Baculovirus vector rBV-GP infected EBOV GP rBV-GP Sf9 cells nanoparticles EBOV GP 30-40nm GPrBV-GP Sf9 cells

7 Novavax EBOV/Makona Glycoprotein (GP) Vaccine

1Nonsynonymous AA: 2014 EBOV/Makona GP and 1976 EBOV/Mayinga GP

20 AA changes

mAb 13F6

1Gire, et al, Science 12 Sept 2014

8 Anti-Ebola ELISA EC50 of mAb 13C6, 6D8 and KZ52

Ebola mAb ELISA 4 KZ52, 13C6, 6D8

13F6 1

0 1e-5 1e-4 0.001 0.01 0.1 1 10 100 1000 10000 conccentration (ug/ml) 4-PmAb Fit: y = (A - D)/( 1 + (x/C)^B ) + D: A B ECC (g/mL)D R^2 c13c6 (Group01: Dilution vs Values) 0.0539 1.14 0.062350 3.46 0.998 H13F6 (Group02: Dilution vs Values) 0.0485 1.26 1.18e+08 1.53e+05 0.999 c6D8 (Group03:13C6 Dilution vs Values) 0.0243 1.02 0.0882 0.06233.67 0.999 KZ52 (Group04: Dilution vs Values) 0.0645 1.21 0.34 3.64 0.999 ______13F6 * - Weighting: Fixed 6D8 0.0882 KZ52 0.34

2 aa change in 13F6 epitope on Ebola GP nanoparticle results in loss of recognition

9 Structure of Ebolavirus Glycoprotein

• The crystal structure of Ebolavirus GP reveals a three- lobed chalice-like structure.

• The three GP1 subunits (colored blue and green), mediate attachment to new host cells and are tethered together by the three GP2 subunits (white).

• GP2 forms the protein machinery that drives fusion of the viral membrane with the host cell.

• The human antibody KZ52 (yellow) binds the GP at the base of the chalice, where it bridges GP1 to GP2, before fusion of the membranes

Research conducted by J.E. Lee, M.L. Fusco, W.B. Oswald, A.J. Hessell, D.R. Burton, and E.O. Saphire (The Scripps Research Institute). Research Funding: U.S. National Institutes of Health, the Burroughs Wellcome Fund, and the Canadian Institutes of Health Research. Operation of the ALS is supported by the U.S. Department of Energy, Office of Basic Energy Sciences. Publication about this research: J.E. Lee, M.L. Fusco, W.B. Oswald, A.J. Hessell, D.R. Burton, and E.O. Saphire, "Structure of the Ebola virus glycoprotein bound to an antibody from a human survivor," Nature 454, 177 (2008).

10 Comparison of crystal structure and vaccine nanoparticle

GP1

GP2

2D TEM – Manual coloration

11 Presentation Overview

• Ebola Background • Novavax Ebola/Mak Glyoprotein Vaccine • Novavax saponin-based Matrix-M™ Adjuvant • Rapid Manufacturing Timeline • Pre-clinical Data

1212 Matrix-M Adjuvant Background - Saponins

• Quillaja saponaria, the soap bark tree, is an evergreen tree native to Chile

• Saponins (glycosides of terpenes and steroids) are extracted and purified from the bark of the tree

General uses of saponins:

 Food – foaming agent in beverages (produce a stable foam in beer)

 Cosmetics – creams, facial cleansers, shampoos

 Pharmaceuticals – Immune booster (adjuvant)

1313 Matrix-M Adjuvant Background - Nanoparticle

1414 Presentation Overview

• Ebola Background • Novavax Ebola/Mak Glyoprotein Vaccine • Novavax saponin-based Matrix-M™ Adjuvant • Rapid Manufacturing Timeline • Pre-clinical Data

1515 Phase I: Manufacture 200L scale EBOV/Mak GP

Master rBV 50L Wave Working rBV EBOV-GP EBOV-GP

Sf9 WCB Culture flasks 50L Wave 200L Bioreactor

Control Sf9 cells Harvest Cell Count

16 Purification EBOV/Mak GP

200LCell Bioreactorharvest

CellCentrifugation harvest Cell lysis: GP trimers Filtration Anion exchange

EBOV GP Affinity Cation exchange 0.2 µm filtration 30 – 40nm

17 EBOV/Mak GP Vaccine Manufacturing Timeline

18 EBOV/Mak GP Vaccine Manufacturing Timeline

Trigger Event

19 EBOV/Mak GP Vaccine Manufacturing Timeline

Project Start Date

20 EBOV/Mak GP Vaccine Manufacturing Timeline

Passage 1 recombinant baculovirus obtained

21 EBOV/Mak GP Vaccine Manufacturing Timeline

Master Virus Seed (passage 2) available

22 EBOV/Mak GP Vaccine Manufacturing Timeline

Drug substance manufacturing initiated to coordinate with MVS availability

23 EBOV/Mak GP Vaccine Manufacturing Timeline

The purification process needed to be defined prior to bioreactor harvest (and use available buffers)

24 EBOV/Mak GP Vaccine Manufacturing Timeline

Purification process defined in time to update SOPs and batch records

25 EBOV/Mak GP Vaccine Manufacturing Timeline

Drug product formulated and filled days after DS was manufactured. Formulation was based on protein assay

26 EBOV/Mak GP Vaccine Manufacturing Timeline

A critical item was the development of a “potency” assay for release and stability studies. This was never on the primary critical path

27 EBOV/Mak GP Vaccine Manufacturing Timeline

MVS, DS, and DP release testing occurred in parallel

28 EBOV/Mak GP Vaccine Manufacturing Timeline

Batch and QC record review occurred in parallel with processing and testing

29 EBOV/Mak GP Vaccine Manufacturing Timeline

A GMP batch for a Phase I clinical study was released 3 months from project start

30 Matrix-M Manufacturing was Matrix-M Manufacturing Timeline completed in 6 weeks and was never on the critical path

31 Presentation Overview

• Ebola Background • Novavax Ebola/Mak Glyoprotein Vaccine • Novavax saponin-based Matrix-M™ Adjuvant • Rapid Manufacturing Timeline • Pre-clinical Data

3232 EBOV/Makona GP vaccine protected mice against lethal challenge with Ebola virus

Mice were challenged on day 42 with an intraperitoneal injection of 1,000 pfu mouse adapted EBOV/Mayinga 1976 strain

1One animal with an unrelated abdominal abscess and was dropped from the study.

33 Vaccine Response Story to be continued…

Wednesday, March 11, 2:15-3:00

34 Perspectives on Recombinant Baculovirus-Sf9 Platform Development and Manufacturing Process

H7N9 Influenza VLP Vaccine

3535 Comparable Upstream Processes for VLP and Nanoparticle Vaccines

Advantages: • One master cell bank • Rapid generation of recombinant baculovirus • Platform cell expansion process • Known purification options

3636 3-month Response Time from Gene Sequence to GMP Batch Release

Keys to success: • One master cell bank • Rapid generation of recombinant baculovirus • Platform cell expansion process • Known purification options • Rapid process confirmation at small scale • Rapid assay development • Forward processing of intermediates • Project management of timeline • Collaboration with suppliers and contract service providers • Utilization of single-use technology

3737 Recombinant Baculovirus-Sf9 Technology Enables a Rapid Response as a Countermeasure to Infectious Agent Threats

Viral Threats Response

Process Development & GMP Production

• Vaccine candidate development • Process & assay development • Preclinical study expertise • 1000L GMP production Clinical study expertise • Regulatory expertise • QC and QA for testing and release

38 Ebola GP Vaccine - Author Recognition

39 Ebola GP Vaccine – Employee Acknowledgements