DoD Explores New DNA Vaccine Delivery Method

Connie Schmaljohn, senior research scientist at the U.S. Army Medical Research Institute of Infectious Diseases, announced that DoD scientists are exploring a new DNA vaccine delivery method at the webcast “Armed with Science: Research and Applications for the Modern Military”.

“DNA offers a number of advantages over conventional vaccine approaches, especially with regard to biodefense vaccines. This is important when rapid vaccine development is needed for a newly emerging disease threat or possibly for a genetically engineered biological warfare pathogen.”

One of the newest DNA vaccine delivery methods relies on technology known as the “gene gun,” which is capable of delivering the vaccine directly into cells. The needle-free vaccination method is more cost-effective and less painful for the recipient.

“The DNA is first coated onto very, very tiny gold beads, and those gold beads with the DNA are then put inside of a plastic device that’s about the size of a small flashlight. Inside that device is also a little canister of compressed helium gas. When the trigger of the gene gun device is pushed, the gas is released and it propels the gold coated with the DNA out of the device into the skin of the vaccine recipient.”

USAMRIID is conducting a human study of DNA vaccines using this delivery method. Schmaljohn’s research team has isolated small amounts of DNA from the Hantaan and Puumala viruses - known health threats to U.S. troops stationed in Europe and Asia to develop the vaccines. Both vaccines are in Phase I clinical testing, the first step toward licensure by the FDA.

“Its main goal is actually to prove that the vaccine is safe in humans, but of course, we’re also interested in determining if it’s inducing an immune response. The hantaviruses, once they infect humans, can cause one of two serious human illnesses: hemorrhagic fever with renal syndrome, which occurs in Asia and Europe, or hantavirus pulmonary syndrome, which occurs in the Americas. Today there’s more than 100,000 cases of hemorrhagic fever with renal syndrome each year, with the highest number occurring in China, Russia … Scandinavia and other parts of Europe.”

USAMRIID is producing a DNA vaccine for the Asian and European hantaviruses that can cause hemorrhagic fever with renal syndrome. This disease first was recognized as a threat during the Korean War.

These viruses are found in many types of rodents, including rats, field mice and meadow voles.

“They’re transmitted to humans in the aerosols of rodent’s urine, feces and saliva. The rodents that carry these viruses are persistently infected, and they show no signs of illness.”

Over tens of thousands of years, these viruses and rodents have formed a mutually exclusive relationship in which both have adapted to one another. While the virus doesn’t appear to affect the rodents’ health, the virus does pose significant risk to humans.

DNA Vaccines May Protect Against Pandemic Influenza

Inovio Biomedical Corporation (NYSE Amex: INO), a leader in DNA vaccine discovery, development and delivery, announced preclinical data from two studies of influenza DNA vaccines designed using its SynCon(TM) technology. The data indicated that the consensus H1N1 and H5N1 influenza vaccines achieved significant increases in immune response and provided protection against influenza virus strains not genetically matched to the vaccine, which is a requirement for conventional vaccines to be effective and a limitation to their effectiveness, i.e. they can’t protect against newly emerging strains. Dr. Niranjan Sardesai, Senior VP, Research & Development of VGX Pharmaceuticals, which merged with Inovio Biomedical on June 1, 2009, presented this data at the 12th Annual Meeting of the American Society of Gene Therapy held in San Diego, CA, May 27 - 30, 2009, in a presentation entitled, “Improved Challenge Outcome Following Intradermic Vaccination by Electroporation of a Consensus H5N1 Influenza DNA Vaccine in Non-Human Primates.”

In the first study, the scientists immunized non-human primates with a consensus H5N1 avian influenza vaccine (VGX-3400). Delivered using the Company’s in vivo electroporation device, the vaccine achieved high levels of protective antibody titers against unmatched clade 1 and multiple unmatched clade 2 H5N1 viruses. The animals were then challenged with the unmatched A/Vietnam/1203/04 strain. There was a significant reduction in average viral load (amount of virus in the blood stream) and decreased symptoms of disease in all the vaccinated animals compared to unvaccinated controls. Animals vaccinated by the intradermal route had a greater than 10,000-fold reduction in average viral load compared to untreated controls, an unprecedented result in comparative studies with similar animal models. These results were recently published in the May issue of the Journal of Virology.

In the second study, the scientists immunized mice with the SynCon(TM) H1HA consensus DNA vaccine. They then challenged the mice with the unmatched H1N1 virus that caused the 1918 Spanish flu. This virus caused the most devastating pandemic of the last century, killing over 40 million people worldwide. While all the control mice died by day 8 after being challenged with a lethal dose of the virus, all the animals vaccinated with the consensus H1HA vaccine survived (100% protection) to the end of the experiment. In addition to measuring survival, the researchers observed significant protection from infection-associated morbidity in the H1HA immunized mice.

Dr. Sardesai stated, “Influenza viruses mutate and develop multiple divergent strains very rapidly, allowing them to potentially escape protection afforded by existing vaccines. On the other hand, developing influenza vaccines effective against these newly emerging, or genetically unmatched, strains has been challenging. These animal data are encouraging and suggest that we could effectively cross-protect against unmatched influenza strains, even against a pandemic virus such as the one that killed millions of people in 1918. We are now testing our SynCon(TM) H1N1-targeted vaccines against the newly emergent swine-origin influenza A (H1N1).”

Dr. J. Joseph Kim, now Inovio’s Chief Executive Officer, said, “The ability to universally protect people from influenza strains that are not exactly like the ones that make up the existing vaccines we have would be an important contribution to modern medicine. We are enthusiastic about the potential of our vaccine approach and results to date, and we have filed an IND for VGX-3400, a candidate vaccine targeting avian influenza H5N1. We look forward to moving Inovio’s first influenza program into the clinic.”

VGX Pharmaceuticals, which merged with Inovio Biomedical on June 1, 2009, and its collaborators at the University of Pennsylvania developed the Company’s novel SynCon(TM) technology to design DNA-based vaccines with the capability to provide protection against unmatched sub-types and strains of pathogens. Using the SynCon(TM) technology, the scientists created universal influenza vaccines targeting both seasonal and avian influenza strains. VGX Pharmaceuticals created a set of SynCon(TM) DNA vaccines based on influenza HA, NA, and NP proteins from strains H1N1, H2N2, H3N2, and H5N1. This designer approach can potentially allow Inovio to rapidly develop a universal influenza vaccine containing DNA targeting all these strains simultaneously by formulating the individual component plasmids together against these target strains which make up the majority of seasonal and pandemic influenza.

About Inovio Biomedical Corporation
Inovio Biomedical is engaged in the discovery, development, and delivery of a new generation of vaccines, called DNA vaccines, focused on cancers and infectious diseases. The company’s SynCon(TM) technology enables the design of DNA-based vaccines better-targeted to desired immune system mechanisms and capable of providing cross-protection against evolving, unmatched strains of pathogens such as influenza. Inovio’s electroporation DNA delivery technology uses brief, controlled electrical pulses to increase cellular DNA vaccine uptake. Initial human data has shown this method can safely and significantly increase gene expression and immune responses. Inovio’s clinical programs include HPV/cervical cancer (therapeutic) and HIV vaccines. An IND has been filed for an avian influenza vaccine. Partners and collaborators include Merck, Tripep, University of Southampton, University of Pennsylvania, and HIV Vaccines Trial Network. Inovio’s product candidates and technologies are protected by an extensive global intellectual property portfolio. More information is available at www.inovio.com.

Pneumococcal Vaccine Program for Developing Countries

The current price of the pneumococcal vaccine is about $70 in US and European countries. The Advanced Market Commitment (AMC) program will provide incentive for pharmaceutical companies to develop vaccines against diseases raging in poor countries by guarantee of fixed price purchase. G8 countries - among them UK, Canada, Italy, Norway and Russia will invest nearly $1.5 billion to the Advanced Market Commitment (AMC) program. Private philanthropic foundations, like the Gates Foundation, will also participate in the program. This program may help to bring down price of the pneumococcal vaccine to $3.5 for poor countries. About 1.6 million people die every year from pneumococcal diseases.

Vaccination is important for preventing pneumonia in both children and adults. Vaccinations against Haemophilus influenzae and Streptococcus pneumoniae in the first year of life have greatly reduced the role these bacteria play in causing pneumonia in children. Vaccinating children against Streptococcus pneumoniae has also led to a decreased incidence of these infections in adults because many adults acquire infections from children. Hib vaccine is now widely used around the globe. The childhood pneumococcal vaccine is still as of 2009 predominantly used in high-income countries, though this is changing. In 2009, Rwanda became the first low-income country to introduce pneumococcal conjugate vaccine into their national immunization program.

A vaccine against Streptococcus pneumoniae is also available for adults. In the U.S., it is currently recommended for all healthy individuals older than 65 and any adults with emphysema, congestive heart failure, diabetes mellitus, cirrhosis of the liver, alcoholism, cerebrospinal fluid leaks, or those who do not have a spleen. A repeat vaccination may also be required after five or ten years.

Influenza vaccines should be given yearly to the same individuals who receive vaccination against Streptococcus pneumoniae. In addition, health care workers, nursing home residents, and pregnant women should receive the vaccine. When an influenza outbreak is occurring, medications such as amantadine, rimantadine, zanamivir, and oseltamivir can help prevent influenza.

Lethality of H1N1 Influenza Virus Increasing

As the World Health Organization raised its pandemic alert to phase 6, the top of its six-point scale, a new analysis released yesterday (June 10) of the latest peptide genomic data for the H1N1 influenza virus shows that the current global outbreak of H1N1 is increasing in its capacity for lethality. The new sequence data on PubMed of the past two weeks through June 10, 2009 showed an increase in the Replikin Count* of the Replikin Lethality Gene in the pB1 genomic area from a mean of 2 in 2008 to a mean of 3.2 in 2009 (p<0.001). The Replikin Count of the Lethality Gene in 836 previous H1N1 influenza virus isolates has remained essentially unchanged (at 2) since 1933.

These analyses were conducted by the Boston-based biotech firm Replikins, Ltd. (www.replikins.com) using its FluForecast® software. A year ago (4/7/08), using the same software, the firm predicted the current H1N1 virus outbreak, and last month (5/23/09) an increase in the Replikin Count of the Replikin Infectivity Gene in the hemagglutinin area indicated a marked increase in infectivity of the evolving H1N1 virus.

"Last month the H1N1 genomic data indicated some bad and some good news. While it indicated an increase in the infectivity of the H1N1 virus, its lethality appeared to remain relatively low," noted Sam Bogoch MD PhD, chairman of Replikins Ltd. "However, the FluForecast® analysis of new data of the past few weeks , through June 10th, on 144 new specimens published on PubMed, indicate an increase in the current H1N1 outbreak's capacity for lethality. Since the software also permitted the automated analysis of all sequence data available on PubMed for all previous years, it was noted that this is the first such significant increase in the Replikin Count of the H1N1 Lethality Gene in 76 years. This is cause for concern and an accelerated vaccine effort."

For both the Infectivity Gene and the Lethality Gene, a significant increase in Replikin Count has invariably been followed by an increase in infectivity or lethality in influenza. Both the Replikin Infectivity Gene and the Replikin Lethality Gene have been found to act independently in all common influenza strains in human, swine, and bird hosts.

The company recently announced that it has made available for testing against H1N1 a Two-Punch™ PanFlu™ vaccine. The same vaccine system has been successfully tested against H5N1 in chickens.

About Replikins Ltd.
Replikins, LLC. (www.replikins.com), a Boston-based biotech company, and Replikins,Ltd., develop and market novel forecasting tools and synthetic vaccines to fight virulent rapidly replicating diseases including bird flu, malaria, and HIV. The company’s predictive products and vaccines in development are based upon the company’s discovery of Replikins, a new group of peptides related to the rapid replication function in viral and other diseases. The company has designed unique products to predict the emergence of virulent strains of particular diseases(FluForecast™) and is designing synthetic vaccines specifically tailored to combat a given strain and against shared properties of several strains (Syntope™ vaccines). The company is partnering with governments and the private sector in providing predictive tools and vaccines in furtherance of the public health initiative to prevent and combat epidemics.

Baxter Advances to Full-Scale Production of A/H1N1 Vaccine

Baxter International Inc. (NYSE: BAX) announced today that it has completed testing and evaluation of the A/H1N1 influenza virus and is now in full-scale production of a commercial A/H1N1 vaccine using its Vero cell culture technology. Baxter received an A/H1N1 strain from the U.S. Centers for Disease Control and Prevention [a World Health Organization (WHO) Collaborating Center] in early May and is diligently working to deliver a pandemic vaccine for use as early as July.

WHO raised the pandemic alert level to phase 6, indicating a global influenza pandemic involving the 2009 A/H1N1 strain. Baxter is in contact with WHO and other global public health authorities regarding the pandemic. A number of national public health authorities have existing pandemic agreements with Baxter that allow them to place orders for a vaccine now that a pandemic has been declared by WHO. These public health authorities will be evaluating their needs to determine their orders for vaccine supply. Despite the company’s existing obligations to supply vaccine under a pandemic phase 6 alert, Baxter is also committed to working with WHO to allocate a portion of the company’s commercial production to address global public health issues deemed most urgent.

Using its Vero cell technology, Baxter has received European Medicines Agency (EMEA) approval for a mock-up pandemic vaccine called CELVAPAN, the brand name for the company’s pandemic vaccine. The qualification, development and manufacturing processes used in gaining mock-up licensure for CELVAPAN apply as the company uses this new influenza A/H1N1 virus strain to produce a pandemic vaccine. The CELVAPAN EMEA licensure supports fast track approval of a pandemic vaccine containing the A/H1N1 virus strain. Baxter will submit the A/H1N1 vaccine for approval upon completion of initial manufacturing runs.

Baxter’s research and development, manufacturing capabilities and pandemic planning expertise allow the company to efficiently develop candidate vaccines against potentially emerging influenza viruses. Baxter believes that its Vero cell technology can be used to safely and reliably produce a vaccine in response to this global public health issue. It is possible that Baxter’s Vero cell technology may offer advantages, in that it may allow more rapid production and delivery of pandemic vaccines.

US Influenza Vaccine Antigen and Adjuvant Orders for 2009 (H1N1)

HHS has contracted with these manufacturers that currently hold U.S. licenses for flu vaccine as part of the National Strategy for Pandemic Influenza. In May 2009, HHS issued new orders on these contracts to produce a bulk supply of vaccine antigen and adjuvant and to produce pilot (also called investigational) lots of a 2009 H1N1 vaccine. Most will be stored in bulk, and a small amount will be prepared as vaccine for use in clinical studies to evaluate vaccine safety and the dosage required for a protective effect. This research will include studies with adjuvant to determine its safety and the effect it would have on the immune system’s response.

Influenza Vaccines for the new Virus Strain A (H1N1)

The first doses of influenza A (H1N1) vaccine could be available within five to six months after identification of the new virus. Prior to this date, small quantities of experimental vaccine will be made available to immunize volunteers in clinical trials to select the best vaccine formulations. Regulatory approval for the vaccines will be conducted by national authorities in parallel with the manufacturing process. National regulatory authorities have put into place expedited processes that do not compromise on the quality and safety of the vaccine. Delays in production could result from poor growth of the virus strain used to make the vaccine.

There are currently more than a dozen influenza vaccine manufacturers with licenses to produce vaccines. Upon request, the vaccine strain will be made available to each of them, as well as to other qualified vaccine manufacturers who are preparing to make influenza A (H1N1) vaccine, but do not yet have a licensed seasonal influenza vaccine.

Projections made for the production capacity of a vaccine for H5N1 cannot be automatically assumed to be the same for an H1N1 vaccine. H5N1 and H1N1 are distinct viruses, so the amount of antigen needed to make an effective H1N1 vaccine may differ from that for H5N1. Therefore, it is not possible to make a precise estimate. However, notwithstanding these considerations, and based on a global survey made by WHO on May 15, 2009, a maximum 4.9 billion could be produced in 12 months. This would only be the case if a number of conditions are met— notably that production yields for influenza A (H1N1) vaccine are equivalent to those usually obtained for seasonal vaccines, and that manufacturers use the vaccine formulation which has been shown for H5N1 influenza as being the most “dose-sparing” (i.e. those which use the smaller quantity of active principle). A more conservative estimate of global capacity is at least 1 to 2 billion doses per year. This does not translate automatically into numbers of persons vaccinated, as we do not yet know how many doses of the vaccine will be needed to achieve protection.

Close to 90% of the global capacity today is located in Europe and North America, with further significant manufacturing capacity in Australia and Japan. However, during the past five years, six manufacturers in developing countries have begun to acquire the technology to produce influenza vaccines and have received technical and financial support from WHO.

US Response to Pandemic Influenza Declaration

Vaccine Market Leading Companies and Government Bodies

Leading companies and government bodies in the biodefense vaccines and therapeutics community:

Advanced BioAdjuvants
Aldevron, LLC
AMAR International, Inc.
AVI BioPharma
Battelle
Bavarian Nordic A/S
Bavarian Nordic GmbH
CBMS-JVAP
Center for Biosecurity of UPMC and University of Pittsburgh
Center for Disease Prevention and Control
Chimerix, Inc.
CUBRC
Dalrymple & Associates, LLC
Defense Threat Reduction Agency
Department of Health and Human Services
Department of Homeland Security
DNAvaccine.com
Drinker, Biddle & Reath
DynPort Vaccine Company LLC, A CSC Company
Elusys Therapeutics, Inc.
Emergent BioSolutions, Inc.
Endacea, Inc.
Evolva SA
Evolva, Genetic Chemistry Inc.
Fabiani & Company
Food and Drug Administration
Fraunhofer USA Center for Molecular Biotechnology
Functional Genetics, Inc.
GenPhar, Inc.
GlaxoSmithKline
HDR CUH2A
Human Genome Sciences, Inc.
Humanetics Corporation
Implicit Bioscience
Infectious Diseases Society of America
Ingenuity Systems Inc.
Integrated Biotherapeutics Inc.
INTERPOL
IQ Therapeutics BV
John Paul II Stem Cell Research Institute/Cellular Engineering Technologies
JPEO-CBD
Kansas State University
Lovelace Respiratory Research Institute
McKenna Long & Aldridge LLP
MedImmune
Midwest Research Institute
National Defence
National Institute of Allergy and Infectious Diseases, NIH
Neugenesis
Orion Integrated Biosciences Inc.
PATH Malaria Vaccine Initiative
Patterson School
PharmAthene, Inc.
PPD
PRTM Management Consultants, LLC
Quintiles Transnational Corp.
Scripps Research Institute
SIGA Technologies
Tetraphase Pharmaceuticals, Inc.
Texas A&M University System and the Bush School of Government and Public Service
The Future, Inc.
The Texas A&M University System
United Therapeutics
University of Maryland, School of Medicine, Baltimore
VaxDesign Corporation
VaxInnate Corporation
Vital Probes, Inc.
Vivante GMP Solutions, Inc.

Influenza Vaccine Side Effects

Different side effects can be associated with the influenza shot and LAIV.

The influenza shot: The viruses in the influenza shot are killed (inactivated), so you cannot get the flu from a influenza shot. Some minor side effects:

• Aches
• Fever (low)
• Soreness
• Redness
• Local swelling where the shot was given

If these side effects occur, they begin soon after the shot and usually last 1 to 2 days. Almost all people who receive influenza vaccine have no serious problems from it. However, on rare occasions, flu vaccination can cause serious problems, such as severe allergic reactions.

LAIV (FluMist®): The viruses in the nasal-spray vaccine are weakened and do not cause severe symptoms often associated with influenza illness. (In clinical studies, transmission of vaccine viruses to close contacts has occurred only rarely.)

In adults, side effects from LAIV (FluMist®) can include

• sore throat
• runny nose
• cough
• headache

In children, side effects from LAIV (FluMist®) can include

• fever
• muscle aches
• vomiting
• headache
• wheezing
• runny nose