Status: approved in the EU in 2007; U.S. filing had been expected as early as 2008

Organizations involved:

Chiron Corp. – R&D; Tech.; Former

Novartis AG – Manuf.; R&D; Tech.; World mark.

MedImmune, Inc. – Tech

Astra Zeneca plc – Parent

Cross ref.: See also the entry for Influenza Vaccine, H5N1/Novartis, which is essentially Optaflu but containing a single H1N1 (swine flu) influenza strain.

Description: Flucelvax or Optaflu is a cell cultured trivalent detergent-disrupted (inactivated) seasonal split-virion (subunit, not whole virus) influenza virus vaccine cultured in Madin Darby Canine Kidney (MDCK) cells in phosphate buffered saline (PBS). The virus is inactivated with ß-propiolactone, disrupted by the detergent cetyltrimethylammonium bromide and purified through several process steps. Each of the 3 virus strains is produced and purified separately then pooled to formulate the trivalent vaccine.

This is the first cell cultured influenza vaccine approved in a major market. As with other seasonal trivalent influenza vaccines, Flucelvax/Optaflu contains 15 mg/doses of hemagglutinin (HA) antigen (45 mg total) from the influenza strains three priimary officially recommended influenza strains for the upcoming/current flu season. Optaflu is preservative-free, non-adjuvanted and presented in pre-filled syringes filled up to 0.5 mL with phosphate buffered saline (PBS) buffer. Flucelvax does not contain any preservatives, such as thimerosal, or antibiotics. Each dose of FLUCELVAX may contain residual amounts of MDCK cell protein (≤8.4 mcg), protein other than HA (≤ 120 mcg), MDCK cell DNA (≤ 10 ng), polysorbate 80 (≤ 1125 mcg), cetyltrimethlyammonium bromide (≤ 13.5 mcg), and β-propiolactone (≤ 0.5 mcg), which are used in the manufacturing process.

Flucelvax is packaged as a suspension for injection in 0.5-mL single-dose pre-filled syringes. FLUCELVAX is a sterile, slightly opalescent suspension in phosphate buffered saline. The tip caps of the pre-filled syringes may contain natural rubber latex. The syringe and syringe plunger stopper are manufactured without natural rubber latex. Flucelvax is stored refrigerated at 2°C to 8°C (36ºF to 46ºF).

Nomenclature: Influenza Vaccine, MDCK cultured/Novartis [BIO]; Optaflu [TR]; Flucelvax [TR US]; NDc 63851-611-01; NDC 63851-611-11 [NDC]

Companies.: Novartis, formerly Chiron Vaccines, developed and manufactures this vaccine at facilities in Marburg, Germany. Novartis is investing in this facility to expand capacity for cell culture influenza vaccines production. Novartis holds worldwide marketing rights.

In May 2006, the U.S. Department of Health and Human Services (DHHS) granted Novartis a $220 million contract (Contract No. HHSO100200900101C) to develop cell-based influenza vaccines, with some of the funds to go towards the cost of a new facility. In July 2006, Novartis announced it would build a $600 million plant in Holly Springs, NC, to manufacture cell-cultured influenza vaccines, both seasonal and pandemic. The plant was expected to begin production as early as 2011 and be ready for full production as early as 2012, with an annual output of 50 million doses of a trivalent vaccine (with the U.S. market generally >100 million doses/year). In the event of a pandemic, the facility would have the capacity to make up to 150 million monovalent (single strain) doses each year within 6 months (of a pandemic declaration).

Upon approval, the vaccine was manufactured at Novartis facilities in Marburg, Germany, with manufacture to be assumed, at least for the U.S. market, by the Holly Springs facility upon completion. Manufacture: Novartis’ MDCK cell line has been adapted to grow in suspension under serum- free, protein-free (PF) conditions. The cell line has been passaged over 500 times from its initial establishment in culture. Early stages of passage were conducted with calf serum and/or in the presence of other animal derived components. In later stages, the use of human and animal derived components has been eliminated at the primary level.

Monovalent bulk vaccine (drug substance) components from culture and clarification of three distinct influenza virus strains are blended and formulated in phosphate buffered saline (PBS) to produce a trivalent bulk harvest.

The MDCK cell line used was established from the kidneys of an apparently normal, adult, male cocker spaniel dog is 1958 by Drs. S.H. Madin and N.B. Darby, University of California (Berkeley, CA). This Chiron/Novartis cell line has been passages over 500-times in suspensin under serum- and protein-free conditions. Early passages were conducted in calf serum and/or in the presne of other animal-derived products. In later stages of passage, the use of human- and animal-derived components was eliminated at the primary level. Cells used for monobulk manufacture are stored under liquid nitrogen. Working cell banks (WCBs) derived from the MDCK master cell bank are used for manufacture. The WCB represents 5 passages with ~10 population doublings from the MCB.

The starting material for each monovalent bulk culture is a singe vial of MDCK WCB. Cells are cultured in chemically-defined media to optimize cell growth during production. The WCB are expanded byt sequential passage in spinner flasks, followed by scale-up in larger fermentation vessels Virus propagatin in the fermenter is performed over 2-4 days. Bulk culture is centrifuged and filtered, producing the "clarified virus harvest," the end product is the infection cycle and fermentation process. This clarified, filtered bulk may be stored at room temperature (16-25C) in stainless steel vessels for up to 24 hours.

Combined inactivation followed by detergent splitting of the purified virions and separation of subunit antigens has been shown to be very effective in inactivation/elimination of both enveloped and nonenveloped viruses.

The virus is purified by chromatography and/or ultra/diafiltration steps, viruses are inactivated (killed) by addition of beta-propriolactone and disrupted using cetyltrimethylammonium bromide (CTAB) to solubilize the viral HA and NA surface antigens. Drug substance manufacture concludes with filtration of the concentrated into the final bulk vessen to obtain monovalent bulk. Each monovalent bulk is individually release and may be stored at 2-8˚C (refrigerated) for up to 18 months.

Basic research on cell cuture-based production and the first manufacturing runs in small fermenter systems began in 1992. Formal process development began in 1995. Material for preclinica toxicology studies (in rabbits) and for Phase I and II trials was manufactured in 2002 using a 100 L pilot scale manufacturing process. Upscale to a 2500 L fermatation process was completed in 2003. Material for Phase III studies for the EU was manufactured under GMP in 2004 and 2005 at the 2500 L scale, which is used for commercial manufacture.

Regarding theoretical TSE exposure, MDCK cells are not considered likely TSE hazards, since dogs are not a significant TSE source. However, the cells have been passaged over 500 times, including exposure to human- and animal-derived products, but none of these media components have been used at the primary level, and none have been used since 1996. The only secondary-level (risk) component is bovine milk (casein) obtained from New Zealand, which is used to manufacture media component amino acids. EMEA/EU notes, "In view of this situation, a TSE risk reduction of 10.sup.31 has been determined based on the dilution of the cells in protein free media since the passaging started in 1996." TSE Certificates of Suitability have been obtained for a MDCK chemically-defined media (CDM) medium component, recombinant insulin...The risk of transmitting TSE by Optaflu is very remote."

Serial radial diffusion (SRD) is used to determine antigen concentrations. Sterile filtration is performed after transportation, at the filling site, prior to filling into syringes. This is integrated into the filling and packaging facilities. During transportation, temperature is maintained at 2-8˚C (refrigerated).

Companies.: Optaflu was developed and is manufactured by Novartis Vaccines and Diagnostics Ltd., CBER/FDA est. no. 1750, in Speke, U.K.

In July 2006, Novartis announced it would build a new vaccine manufacturing facility in Holly Springs, NC. Financing includes a $220 million grant from the US Department of Health and Human Services (HHS) in June 2006. The new 300,000ft plant is will employ ~350 to produce influenza vaccines. The facility is being constructed on a 167-acre site. The plant is expected to be validated by 2011–2012 and operational by 2013. The initial investment is estimated at $267 million, but could eventually reach $600 million with subsequent phases of expansion. The Town Council provided the land free, after purchasing for its owner, and granted an additional $1m spending for site clearance. Holly Springs also received an $800,000 grant from the Golden LEAF Foundation to help fund infrastructure improvements in the business park In total, Novartis received cash grants, land, tax credits and other incentives worth more than $41 million from state and local governments. The town property taxes that Novartis will pay on the first phase of construction are expected to enable Holly Springs to recoup its investment within ten years. The total investment of approximately $600 million by Novartis includes the $220 million grant from the federal government. This facility will be the first manufacturing plant in the U.S. to produce cell culture-derived influenza vaccines. Once completed and approved for commercial production, the facility is expected to annually produce up to 50 million doses of seasonal trivalent flu vaccines, which will be for use in the U.S. In the event of an influenza pandemic, the facility has planned to have a capacity of up to 150 million monovalent doses annually within six months of a pandemic declaration.

Novartis is also making investments in its site in Marburg, Germany, to expand capacity for influenza cell culture vaccines production in Europe.

Indications: [Full text of the :"INDICATIONS AND USAGE" section of the product insert/labeling]:

FLUCELVAX is an inactivated vaccine indicated for active immunization for the prevention of influenza disease caused by influenza virus subtypes A and type B contained in the vaccine.

FLUCELVAX is approved for use in persons 18 years of age and older.

Status: On April 27, 2007, the Committee for Medicinal Products for Human Use (CHMP), European Union (EU), concluded that there is “a favourable benefit to risk ratio” for Optaflu and recommended its approval for “prophylaxis of influenza for adults, especially in those who run an increased risk of associated complications.” EU-wide approval was granted on June 13, 2007. Iceland and Norway also have approved Optaflu

Novartis in making Optaflu available in Germany and Austria for the 2007/2008 influenza season. It will be available in other European Union countries for the 2008/2009 influenza season. Novartis anticipates applying for U.S. licensing in 2008 (for the 2008/9 flu season).

Cell culture technology enables flexible, faster start-up of vaccine manufacturing. Novartis expects to better meet the growing need for seasonal influenza vaccines and quickly respond to a pandemic influenza threat. Also, those allergic to eggs and egg products can receive this vaccine, since it it contains no egg proteins.

indications: The EU indication upon approval was "prophylaxis of influenza in adults, especially in those who run an increased risk of associated complilcations."

Tech . transfer: In Aug. 2007, Novartis licensed reverse genetics technology from MedImmune, as subsidary of Asta Zeneca plc. See the Influenza Vaccine Products entry (#470) for further information.

Trials: A total of 3,400 people received the vaccine in its trials (leading up to EU approval). Optaflu fulfilled all European Union immunogenicity and safety criteria and was comparable to conventional egg-based vaccines in stimulating an immune response and tolerability.

The basis for FDA approval included data from clinical trials that found Flucelvax to be well tolerated, with an efficacy of 83.8% against antigenically-matched strains compared to placebo. A multinational, randomized, observer-blinded, placebo-controlled trial was performed to assess clinical efficacy and safety of Flucelvax during the 2007- 2008 influenza season in adults aged 18 to 49 years in the US, Finland and Poland. A total of 11,404 subjects received Flucelvax (N=3828), seasonal influenza vaccine Agriflu (N=3676) or placebo (N=3900) in a 1:1:1 ratio. In a separate study in adults aged 65 years and older Flucelvax demonstrated comparable immunogenicity to Agriflu for all three strains post-vaccination. Adverse reactions wre similar to those observed with administration of other seasonal influenza vaccines. Overall, in clinical studies, the most common (≥10 %) solicited adverse reactions occurring in adults 18 to 64 years within seven days of vaccination with Flucelvax were pain at the injection site, erythema (redness) at the injection site, headache, fatigue, myalgia and malaise. The most common (≥10 %) solicited adverse reactions occurring in adults 65 years of age or older within 7 days of vaccination were erythema at the injection site, fatigue, headache and malaise.

Medical: The vaccine is administered by a single 0.5 mL intramuscular injection, just like other influenza vaccines.