Coagulation Factor IX (Recombinant) - BeneFIX; Antihemophilic factor B; nonacog alfa; rFIX
Status - approved; marketed
Organizations involved:
Pfizer – Manuf.; R&D; Tech. World. mark.; Parent
Wyeth –Former
Genetics Institute, Inc. – Former
Swedish Orphan Biovitrum AB (Sobi) - Former
Baxter Hyland Immuno – Former
Oxford University – Tech.
Washington Research Foundation – Tech.
BTG plc – Tech.
Columbia University – Tech.; Patent dispute
Cross ref.: See the Factor IX Products entry in the Blood Products section for further information about Factor IX.
Description: Coagulation Factor IX (Recombinant) or BeneFIX is a lyophilized (freeze-dried) formulation of nonacog alfa, a purified recombinant human Factor IX single-chain glycoprotein expressed by a transformed Chinese hamster ovary (CHO) cell line. The primary 415-amino acid sequence of nanacog alfa is identical to the Ala148 allelic form of human plasma-derived Factor IX. Although post-translation processing modifications and its final structure differ slightly from human Factor IX, nonacog alfa has structural and functional characteristics similar to those of human Factor IX. BeneFIX is a high purity, homogenous product, i.e., it is predominantly a single glycoprotein as shown by SDS-polyacrylamide gel electrophoresis, with a molecular weight of ~55 kDa.
The potency (in international units or I.U.) of BenFIX is determined using an in vitro one-stage clotting assay against the World Health Organization (WHO) International Standard for Factor IX concentrate. One Factor IX international unit is the amount of Factor IX activity present in 1 mL of pooled, normal, human plasma. The specific activity of BeneFIX is greater than or equal to 240 I.U. per milligram of protein.
BeneFIX is formulated as a sterile, nonpyrogenic, powder packaged in single-dose vials for intravenous administration of 250, 500, 1,000 I.U., or 2000 IU along with a diluent vial, a double-ended needle for reconstitution, a filter spike for withdrawal, an infusion set, and two alcohol swabs. The 500 and 1000 I.U. dosage strengths are isotonic (match electrolyte concentration of blood) after reconstitution; and the 250 I.U. dosage strength has half the tonicity of these dosages after reconstitution. Actual Factor IX activity in I.U. is stated on the label of each vial. Vials contain a slight excess volume to assure withdrawal of the specified dose. BeneFIX contains no preservatives or added animal or human source components. After reconstitution (dilution for injection), the concentrations of excipients in the 500 and 1000 I.U. dosage strengths are 10 mM L-histidine, 1% sucrose, 260 mM glycine, and 0.005% polysorbate 80 (Tween 80). The excipient concentrations after reconstitution of the 250 I.U. dosage strength are half these amounts. All dosage strengths yield a clear colorless solution upon reconstitution. Because it is produced in a Chinese hamster ovary (CHO) cell line, BeneFIX may contain trace amounts of hamster protein. BeneFIX is stored at 2 -8˚C (refrigerated), and may also be stored at room temperature (< 25˚C) for up to 6 months.
Nomenclature: Factor IX, rDNA/Pfizer [BIO]; BeneFIX [TR]; Coagulation Factor IX (Recombinant) [FDA]; nonacog alfa [USAN INN]; Antihemophilic Factor IX [SY]; Factor IX [SY]; Factor IX and Factor IXa [MESH]; rFIX [SY]
Biological.: BeneFIX is used for treatment of Factor IX deficiency (hemophilia B; Christmas disease) to provide Factor IX replacement in patients with genetic deficiency of functional Factor IX. See the Factor IX Products entry or background about Factor IX and Factor IX deficiency.
History: BeneFIX was the first recombinant Factor IX to receive FDA approval, and the first Factor IX product not derived from human blood, providing an alternative inherently free from the risk of transmission of human blood-borne pathogens. Unlike all prior Factor IX products, BeneFIX is not dependent on blood/plasma donations. BeneFIX was the first product to be marketed and sold directly by the Genetics Institute (later Wyeth). Clinical testing began in Feb. 1995. Worldwide market applications were filed in Sept. 1996.
Companies.: BeneFIX was developed and is manufactured by Genetics Institute, Inc., FDA CBER est. no. 1163, which became a subsidiary of Wyeth in 1997, and subsequently has been merged into Wyeth. Wyeth markets BeneFIX in the U.S. BeneFIX has been marketed exclusively by Baxter Healthcare Corp. in Europe (product launch on Jan. 28, 1999).
In July 2007, Wyeth assumed from Baxter all marketing and distribution rights in Europe. This marked the end of the original 10-year marketing deal Genetics Inst. had made with Baxter for European marketing.
In Aug. 2007, Wyeth granted Biovitrum AB co-marketing rights in the Nordic countries (Denmark, Finland, Iceland, Norway and Sweden). With the exception of these countries, Wyeth markets BeneFIX worldwide.
In Feb. 2012, Sobi return returned to Pfizer co-promotion rights for BeneFIX in the Nordic region.
Manufacture: Transformed Chinese hamster ovary (CHO) cells are cultured in a defined serum-free medium lacking any added protein components. The mammalian cells are grown in serum-free medium, with no bovine serum or animal proteins present, which avoids potential contamination by serum-associated viruses and prions. Nanacog alfa is purified using four sequential chromatographic steps, followed by a final viral retention filtration step; and is diafiltered into an albumin-free formulation and lyophilized. No human or animal proteins are added during manufacture or formulation.
Factor IX is secreted by the transformed CHO cells into the growth medium, which is then removed from the cells and purified through a series of chromatographic steps: Q-Sepharose-FF (a form of IEC and pseudo-affinity chromatography) and Cellufine sulfate (heparin-like) affinity chromatography, followed by ceramic hydroxyapatite, and chelate EMD copper chromatography to remove trace contaminants. Affinity chromatography with monoclonal antibody in not used to prevent potential contamination with extraneous animal proteins (e.g., mouse protein). After chromatography, the purified product is formulated in a buffer of polysorbate 80, histidine, glycine, and sucrose.
BeneFIX is expressed in an established CHO cell line that has been extensively characterized and shown to be free of infectious agents. A reference batch of recombinant Factor IX-producing CHO cells has been cultured and distributed into hundreds of small vials stored in the company’s cell bank to assure consistency in production lots. The stored cell banks are free of blood or plasma products. The cells are maintained in a frozen state at -135˚C in secure, monitored, temperature-controlled freezers for future use. This secured lot of production cells (Master Cell Bank) is expected to last for over 100 years, ensuring future consistent and sufficient production of recombinant Factor IX.
To start production of BeneFIX, a vial of stored, frozen, recombinant Factor IX production cells (from the cell bank) is brought into an inoculum room. The cells are placed in a glass spinner flask along with culture media containing vitamins, amino acids, minerals, and sugar, with no human or animal components. As the cells multiply, express and release recombinant Factor IX into the media, they are repeatedly transferred to bigger spinner flasks. After about 3 or 4 weeks, these inoculum cells are harvested and transferred to a computer-controlled, closed-system bioreactor for large-scale production of rFIX. After a growth period in the bioreactor, typically 3 days, the production cells are filtered off and discarded.
The rFIX secreted in the culture media is purified using a series of four chromatography columns. No monoclonal antibodies are used for the purification of recombinant Factor IX. First, recombinant Factor IX solution is pumped onto an affinity chromatography column under conditions which cause the recombinant Factor IX and other materials to adhere or bind to the column matrix. Then the column is washed with a special salt solution to release culture media components. Bound recombinant Factor IX is released and washed from the column using another solution. A membrane filtration step is performed at the end of the purification process to assure sterility. This filtration step retains all molecules with apparent molecular weights >70,000 Daltons (70 kDa; such as large proteins and viral particles). Recombinant Factor IX is then lyophilized (freeze-dried), and the powder is filled into vials which are labeled and packaged. No human or animal source components are added during production.
FDA class: Biologic BLA
Approvals: Date = 1997211; first approval (BLA); orphan designation (granted 1/3/1994); Indication = treatment of hemophilia B
Date = 20070326; BLA supplement; Indication = (four changes) a 2000 IU dosage strength vial; a needleless reconstitution device; a prefilled diluent syringe; and a reduced diluent volume [i.e, the new BeneFIX R2 Kit].
Indications: [full text of "Indications and Usage” section from recent product insert/labeling]:
BeneFIX Coagulation Factor IX (Recombinant) is indicated for the control and prevention of hemorrhagic episodes in patients with hemophilia B (congenital factor IX deficiency or Christmas disease), including control and prevention of bleeding in surgical settings. BeneFIX is not indicated for the treatment of other factor deficiencies (e.g., factors II, VII, and X), nor for the treatment of hemophilia A patients with inhibitors to factor VIII, nor for the reversal of coumarin-induced anticoagulation, nor for the treatment of bleeding due to low levels of liver-dependent coagulation factors.
Status: The BLA was submitted on Sept. 5, 1996, received priority review, and approved on Feb. 11, 1997, an approval time of ~5.4 months (~.49 year). With orphan status, no other substantively identical product will be approvable and marketable in the U.S. for seven years from the approval date (Feb. 11, 2004) unless the product shows significant improvements in safety and/or efficacy. BeneFIX was brought to market in slightly more than three years. This relatively speedy approval was based on its bioequivalence to blood fraction product. Presuming this included abbreviated trials based on comparison with Baxter’s blood-derived product, the approval of BeneFIX has aspects of a generic approval [biogeneric, biosimilar, follow-on protein (FOP), follow-on biologic (FOB), etc.].
Associated with BeneFIX approval by FDA, Genetics Inst./Wyeth pledged to conduct Phase IV clinical trials including following for two years 36 patients receiving BeneFIX who had previously received plasma-derived products. Since information regarding BeneFIX in previously untreated patients is limited, Genetics Inst./Wyeth is continuing a clinical trial involving up to 50 previously untreated patients for up to five years to evaluate this patient population for safety, efficacy, and inhibitor formation.
A MAA application for European Union (EU) approval was filed on Aug. 16, 1996 and approved on Aug. 27, 1997. This approval included recognition as an orphan product, allowing fee exemptions (although EU “orphan medicinal products” regulations were not finalized at the time).
The March 2007 sBLA included a new more convenient needleless injection preparation process, eliminating the risk of needlesticks during reconstitution. The final infusion volume for BeneFIX could be reduced by up to 75%. A reduced diluent amount may allow less volume to infuse for some patients, particularly useful for those who infuse multiple vials of BeneFIX. With the introduction of a new 5 mL prefilled diluent syringe for all dosage strengths, including a new 2000 IU dosage strength vial, BeneFIX has the lowest infusion volume of hemophilia B therapies (when similar IU amounts are infused). For patients needing over 2,000 I.U./treatent, the 2000 IU dosage strength vial for BeneFIX enables use of fewer vials. Previously, 1,000 I.U. was the largest vial size.
On Aug. 20, 2007, Wyeth received European Union approval for a 2000 IU dosage strength vial (the largest unit dosage vial available); a prefilled diluent syringe; a 5 mL diluent volume for all dosage strength; and a needleless reconstitution device, which eliminates the risk of needlesticks during reconstitution. These changes make preparation of BeneFIX much simpler.
Tech. transfer: Patents reported on the U.S. insert are 4,994,371; 5,171,569; 5,714,583; 6,372,716; 6,627,737. U.S. 5,171,569, assigned to Oxford Univ. (National Research Development Corp.), covering basic composition (sequence) and use) received an extension to Feb. 11, 2011.
U.S. 6,372,716, covering formulations, expires in 2019.
U .S. 6,627,737, covering purification methods, expires in 2015.
Dr. G. Brownlee, Oxford University, and collaborators isolated and cloned the human Factor IX gene in 1982. BTG plc (formerly British Technology Group and National Research Development Corp.), the largest commercial technology/patent transfer firm, filed patent applications on behalf of Oxford Univ. covering the gene and recombinant Factor IX. At about the same time, Dr. Davies, University of Washington, and collaborators were carrying out similar work and a U.S. patent application was filed on their behalf by the Washington Research Foundation (WRF). BTG negotiated a license agreement for exclusive rights to the University of Washington patents, enabling BTG to offer consolidated, uncontested worldwide patent licensing rights for recombinant Factor IX. Univ. of Washington patents include U.S. 4,868,112 covering sequences and recombinant Factor IX.
BTG subsequently exclusively licensed mammalian cell culture production of recombinant Factor IX to Genetics Inst. BTG plc has reported receiving an estimated $10 million or more annually in royalties from sales of BeneFIX. [BTG has also licensed to Pharmaceutical Proteins Ltd. (PPL) rights to develop Factor IX from transgenic sheep; rights for Factor IX for certain gene therapies to Genetic Therapy, Inc., Transkaryotic Therapies Inc. and GeneMedicine Inc.; and rights for adeno-associated virus vector gene therapies to Avigen, Inc.]
Genetics Inst./Wyeth has received Factor IX U.S. process-related patents including 5,714,583, “Factor IX purification methods,” issued Feb. 3, 1998, concerning the use of chromatography column elutents with varying conductivities to optimize purification and recovery; and 4,770,999, “High yield production of active Factor IX,” issued Sep. 13, 1988, concerning the addition of vitamin K to the CHO cell culture medium to increase the production and yield of recombinant Factor IX. U.S 6,372,716, assigned to Genetics Inst., concerns stable formulations of recombinant Factor IX.
Genetics Inst. (now Wyeth) was a licensee of Columbia University’s patents concerning cotransformation, a broadly-useful genetic engineering method allowing selection and isolation of transformed cells. The original patents and license expired in 2000, but Columbia received another patent in 2002 and was again seeking royalties, which Wyeth and other companies challenged in court. Recently, the University decided not to continue to press infringement suits and seek royalties, but the patent office is reexaming the relevant patent, and the university could against pursue infringement and royalties at a later date. See the “Tech. transfer” section of the Recombinant DNA Products entry (#100) for further information.
Trials: Clinical testing of BeneFIX began in February, 1995. Phase II/III clinical trials evaluated BeneFIX for treatment of spontaneous bleeding and prevention of bleeding during surgeries and postoperatively in patients with hemophilia B. International Phase III trials involved numerous centers in the U.S., Canada, United Kingdom, Belgium, France, and Germany. Due to the rarity of hemophilia B, clinical studies of BeneFIX have involved only a relatively small number of patients. Scant data are available regarding the even rarer serious side effects that may occur with BeneFIX.
In clinical studies of BeneFIX involving a total of 64 patients (44 previously treated patients [PTPs], 11 previously untreated patients [PUPs], and the 9 patients participating only in a surgical study), more than 7 million I.U. were administered over a period of up to 18 months. Forty-five patients were evaluated for efficacy, all of whom were treated successfully for bleeding episodes on an on-demand basis or for the prevention of bleeds. Bleeding episodes that were managed successfully include hemarthroses and bleeding in soft tissue and muscle. Factor IX antibodies (inhibitors) developed in only one of 44 patients who had previously received plasma-derived products. The patient was able to continue treatment with BeneFIX with no anamnestic rise in inhibitors or anaphylaxis. Safety and efficacy in pediatric populations have not been determined in clinical trials.
Medical: Administration of BeneFIX increases plasma levels of Factor IX and can temporarily correct coagulation defects in hemophilia B patients. BeneFIX is considered safe and effective for the treatment of hemophilia B, including treatment of spontaneous bleeds and during surgery. Some patients may require higher doses of BeneFIX, compared to blood plasma-derived Factor IX products. Trials have shown that BeneFIX has a safety profile comparable to other Factor IX products. Factor IX antibody or “inhibitor” formation is not a significant problem with BeneFIX, but patients using BeneFIX shou2ld be monitored for the development of Factor IX inhibitors.
Market: Total sales of BeneFIX were $248 million in 2003; and $219 million in 2002, and $140 million in 1999. By Jan. 1999, only two years after approval, the majority of hemophilia B patients had switched to BeneFIX. The author’s rough guess for 2005 sales is $275-290 million, and for 2004 sale is $260 million.
BeneFIX has been reported to have about 70% of the market (about 3,000 patients in the U.S.), and almost 100% of the HIV and pediatric markets, for Factor IX products due to safety concerns (with the human-derived products).
The 2007 Average Wholesale Price (AWP) is $1.00/IU for the 250, 500, and 1,000 IU vials, with a Direct Price (Manufacturer’s discount price) of 0.83/IU (Red Book, 2007). These prices are unchanged from 2005.
Medicare reimbursement is set at $1.12/IU for for inpatient and home care, and at $1.01/IU for outpatient care. Estimated Acquisition Costs (for hospitals, treatment centers) is $0.78-0.79/IU [from NHF].
In its March 22, 2006 price list, FFF Enterprises, a major biologics distributor, reported its price as $0.90/IU (no change from 2005 or 2004).
Wyeth claims (3/2007) that as the only recombinant Factor IX product (unstated), “BeneFIX is the only recombinant clotting factor product for hemophilia B and inherently is free from the risk of transmission of human blood-borne pathogens, such as HIV, hepatitis viruses and parvovirus.”
On June 1, 2005 Wyeth launched the Wyeth Factor Resource Program to provide free hemophilia factor to customers who experience a temporary lapse in insurance coverage. Qualified individuals receive product for up to one year or until a 200,000 IU maximum is reached. Participants must have insurance and have been using Wyeth products ReFacto (Factor VIII) or BeneFIX (Factor IX) for three consecutive months when they apply. Other key components of the Program include the Patient Assistance Program and the Reimbursement Information Line.
Competition: In Jan. 2006, Biovitrum AB, Sweden, and Syntonix Pharmaceuticals, Inc. (U.S.), entered into a co-development and commercialization agreement for Syntonix’ long-acting, recombinant Factor IX product, FIX:Fc. The companies will jointly develop and commercialize FIX:Fc, and equally share the costs and profits for development and commercialization. Syntonix is responsible for marketing in North America and Biovitrum is responsible for marketing in Europe, Russia and the Middle East. The extended half-life of FIX:Fc product could enable effective treatment for both prophylaxis and on-demand therapy with less frequent intravenous injections.
Inspiration Biopharmaceuticals, Inc. is developing IB1001, an intravenous (IV) recombinant Factor IX. Phase I trials began in Feb. 2009.
Companies involvement:
Full monograph
144 Factor IX, rDNA/Pfizer
Nomenclature:
Factor IX, rDNA/Pfizer [BIO]
BeneFIX [TR]
Coagulation Factor IX (Recombinant) [FDA]
nonacog alfa [USAN INN]
Antihemophilic Factor IX [SY]
Factor IX [SY]
rFIX [SY]
molecular weight (kDa) = 55
FDA Class: Biologic BLA
Year of approval (FDA) = 1997
Date of 1st FDA approval = 19970212
(in format YYYYMMDD)
Biosimilars/biobetters-related U.S. Patents: | 2011, based on extension of 5,171,569, basic composition and use patent
2015, based on 6627737 purification patent
2019, based on 6372716, a formulation patent |
U.S. Patent Expiration Year: | 2011 |
U.S. Biosimilars Data Exclusivity Expiration: | 2009 |
U.S. Biosimilars Orphan Exclusivity Expiration: | 2002 |
U.S. Biosimilars Launchability Year: | 2011 |
U.S. Biobetters Launchability Year: | 2011 |
Biosimilars/biobetters-related EU Patents: | 2016, based on EP 0832200 formulation and EP 1571208 purification patent
2015 (based on EP 0758248 formulation patent) |
EU Patent Expiration Year: | 2016 |
EU Biosimilars Data Exclusivity Expiration: | 2009 |
EU Biosimilars Orphan Exclusivity Expiration: | 2004 |
EU Biosimilars Launchability Year: | 2016 |
EU Biobetters Launchability Year: | 2016 |
Index Terms:
animal/human products-free
antihemophilic factors
biopharmaceutical products
blood products
exempt from CBER lot release requirements
hamster source materials
recombinant DNA
rodent source materials
bioreactors, spinner culture
Chinese hamster ovary (CHO) cells
mammalian cell culture
rodent cells <!-- rodentcells -->
adw, hepatitis B virus subtype
adw, hepatitis B virus subtype
adw, hepatitis B virus subtype
cells, human <!-- humancellculture -->
Challenge Virus Standard (CVS; rabies virus)
chondrocytes, human
glycine
heparin-induced thrombocytopenia (HIT)
histidine
hydroxyapatite
Plasma (Human)
polysorbate 80 (Tween 80)
PYinsl yeast cells
sucrose
vitamin K
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
orphan status
priority review status
EU200 Currently Approved in EU
SM001 Commerical Marketed Product
US200 Currently Approved in US
EM001 Marketed Product in EU
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