Rachael S. Felberbaum, Ph.D. Daniel D. Adams, J.D.

Protein Sciences Corporation

White Paper A Revolutionary Approach to Influenza Vaccines

Recombinant technology allows perfect genetic match to circulating virus without the use of live influenza virus Cell-based manufacture enables fast, inexpensive production with high yields FluBlok : the first recombinant influenza vaccine made with speed and reliability PanBlok : the pandemic version of FluBlok the solution for pandemic influenza

Influenza is a continually evolving challenge Influenza is a leading cause of illness and death around the world, resulting on average in more than 1 200,000 hospitalizations and ranging from 3,000 to 49,000 deaths annually in the U.S. alone. New strains of influenza are continually emerging and lead to epidemics every winter. Every so often, new influenza strains achieve pandemic potential that in the past have been associated with mortality rates as staggering as >70%. Vaccination against influenza is an effective measure to prevent disease. However, because of the mutability of the virus, annual adjustment of influenza vaccines is required to provide effective protection against circulating strains. An independent market study found that the worldwide influenza vaccine market is growing rapidly and is predicted to more than double in size to over $4.4 2 billion by 2016.

Current vaccines for influenza are limited Influenza vaccines are traditionally composed of partially purified hemagglutinin (HA) proteins derived from the most prevalent circulating strains of influenza virus. The vaccines marketed today in the U.S. are made by infecting embryonated chicken eggs with live influenza virus in order to propagate the virus and generate ingredients for the vaccine. Once sufficient virus has been produced, it is harvested, inactivated and the HA proteins isolated and partially purified. Although in use for more than half a century, this manufacturing technology is fraught with limitations. Production cycles are long, taking on average 6 months to generate vaccine from the time a new strain is identified. In a field where new strains continually emerge and have the potential to reach pandemic proportions in a matter of months, this timeframe is unacceptable. In addition, chicken egg technology suffers from susceptibility to avianderived influenza strains, requires adaptation of the virus to achieve high yields in chicken eggs (and, therefore, results in deviation from the circulating virus), necessitates biocontainment facilities and produces a vaccine that is not suitable for people with egg-related allergies.

Recombinant technology offers the solution The greatest potential for substantially shortening the time and increasing the reliability of influenza vaccine production lies in the use of recombinant DNA technologies. - U.S. Presidents Council of 3 Advisors on Science and Technology At Protein Sciences, we have developed a proprietary Baculovirus Expression Vector System (BEVS) platform technology that enables us to solve these problems and produce high quality recombinant
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Statistics according to the Centers for Disease Control and Prevention. Datamonitor, 2007. 3 Presidents Council of Advisors on Science and Technology. Report to the President on Reengineering the Influenza Vaccine Production Enterprise to Meet the Challenges of Pandemic Influenza. Executive Office of the President: Washington, DC, August 2010; www.whitehouse.gov/sites/default/files/microsites/ostp/PCAST-Influenza-Vaccinology-Report.pdf.

Rachael S. Felberbaum, Ph.D. Daniel D. Adams, J.D. Protein Sciences Corporation vaccine quickly, reliably and at cost. This platform leverages the natural infection process of insect cells by baculoviruses. We genetically re-engineer the baculovirus using the genetic code for a desired protein, which reprograms the insect cells it infects to generate large quantities of that protein. In the case of influenza, we program insect cells to produce recombinant hemagglutinin (rHA) that we formulate into either FluBlok, our seasonal trivalent vaccine, or PanBlok, our monovalent pandemic version. In both cases, the end result is highly purified influenza vaccine whose production requires no biocontainment in the U.S. (no live influenza viruses are used at any stage of production) or adaptation for growth in eggs and can be used in people with egg-related allergies. Furthermore, production time is fast vaccine can be produced in just weeks from the time of strain identification.

Figure 1. The BEVS platform can be used to rapidly generate vaccine against any strain of influenza. Baculoviruses are genetically engineered to express recombinant hemagglutinin (rHA) that matches the circulating virus strain(s). The baculoviruses are then added to a large-scale culture of expresSF+ insect cells that can grow to high densities in our proprietary serum-free media. Following harvest, rHA is purified and formulated with PBS into vaccine. This entire process can be completed within 2 months or less.

FluBlok for seasonal influenza FluBlok, our recombinant protein-based seasonal influenza vaccine, has shown excellent performance in the clinic and a Biologics License Application (BLA) for approval in adults 18 years and older is under review at the FDA. FluBlok contains three full-length rHA proteins that correspond to the current seasons circulating influenza strains. Each of the three rHAs is produced individually and the purified monovalent bulks are then blended and filled into single-dose vials that contain a total of 135 g of rHA (45 g of each). This is three times the amount of active ingredient than is contained in the leading marketed influenza vaccines. This formulation is economically possible to manufacture because of the high yields of protein generated by the BEVS platform. In clinical trials, this higher antigen content has resulted in substantially improved serum hemagglutination inhibition (HI) antibody responses, a well-accepted surrogate marker of influenza vaccine efficacy. FluBlok safety has been assessed in over 3,000 adults and children and the data demonstrate an excellent safety profile. When approved, FluBlok will be the 2

Rachael S. Felberbaum, Ph.D. Daniel D. Adams, J.D. Protein Sciences Corporation first recombinant licensed influenza vaccine, the first cell-derived influenza vaccine licensed in the U.S. and the first influenza vaccine that can be safely used for vaccination of people with egg allergies. FluBlok Advantages Higher antigen content Potentially superior immunogenicity, especially in the elderly and immunocompromised No live influenza viruses used in production No antibiotics, adjuvants or preservatives (e.g., thimerosal) No egg-related byproducts

PanBlok for pandemic influenza PanBlok is the pandemic version of FluBlok. The vaccine contains a single rHA that corresponds to a circulating influenza virus that has the potential to cause a pandemic. Phase I clinical trials with PanBlok have been completed in the U.S. and Australia, and Phase II clinical development is underway in Japan through a partnership with UMN Pharma, a development-stage biotechnology company that is partnered with Astellas Pharma. The data suggest that PanBlok is safe and that administration of PanBlok can effectively prime vaccine recipients for enhanced immune responses upon subsequent vaccination. Pandemic viruses rarely simply appear but rather can be identified as they are emerging. The Centers for Disease Control and Prevention (CDC) and World Health Organization (WHO) perform continual monitoring to detect new viruses before they become a realized threat. When a new virus is identified, the CDC/WHO publishes its genetic sequence. We are able to clone and express the relevant rHA protein from this genetic information using our BEVS platform and generate a master baculovirus bank. We have more than 200 such banks in storage and, once FluBlok receives regulatory approval, we will vastly increase this number. This preparation will enable us to significantly increase our ability to respond quickly to a pandemic, which already takes only a matter of weeks; should a particular vaccine be needed, we can begin its commercial manufacture within two weeks of notification and have the first doses ready to be released within 30 days thereafter. PanBlok Advantages Manufacturing technology provides ability for a specific and rapid response to a pandemic strain emergence Priming response has been demonstrated Adjuvanted formulation results in significant dose sparing No live influenza viruses used in production No antibiotics or preservatives (e.g., thimerosal) No egg-related byproducts

Meeting the Needs of the Developing World The 2009 H1N1 swine flu pandemic highlighted the lack of global preparedness that exists to combat new pandemic outbreaks of influenza. The outdated chicken egg technology failed to generate sufficient supplies of vaccine to populations in the developed counties in which the vaccine was manufactured, let alone the developing countries that have no such capability. Obviously, the developing world cannot afford to construct billion dollar pandemic flu vaccine facilities that will stand idle for most of their existence, not to mention that the workforce could hardly be financed and be ready to turn on production at a moments notice. The beauty of BEVS technology is that it can be successfully used to manufacture vaccines in any monoclonal antibody facility. Worldwide there exists over 2.5 million liters of such cell culture capacity all of which could be used to produce PanBlok in an emergency. Technology transfer from us could occur within three weeks and, thereafter, such facilities could produce an estimated 6 billion doses of pandemic vaccine in a matter of months. Alternatively, the cost of a BEVS production facility with capacity to produce sufficient pandemic vaccines to protect most countries is about $35 million (compared to $1 billion). These facilities could also be used when pandemic influenza vaccine is not 3

Rachael S. Felberbaum, Ph.D. Daniel D. Adams, J.D. Protein Sciences Corporation needed to produce recombinant vaccines or monoclonal antibodies for a multitude of other diseases that are rampant in the developing world, such as Marburg and Ebola, thus providing additional rationale for their construction. With BEVS technology, there can be enough for all.