Status: approved; marketed

Organizations involved:

CSL Bioplasma, Inc. - Manuf.; R&D; Tech.; USA mark.

CSL Bioplasma AG - Intl. mark.; Parent

CSL Ltd. - Parent

Aventis Behring LLC - Former

Armour Pharmaceuticals - R&D; Tech.; Former

Scripps Research Institute - R&D; Tech.

New York Blood Center - Tech.

Cross ref: See also Factor IX Products entry (#727).

Description: Coagulation Factor IX (Human) or Mononine is a sterile, stable, lyophilized (freeze-dried) formulation of Factor IX manufactured from pooled fractionated Plasma (Human) and purified of extraneous plasma-derived proteins, including Factors II, VII and C, by immunoaffinity chromatography using matrix-bound murine (mouse) monoclonal antibody with specificity for human Factor IX as an affinity ligand, with viral inactivation by incubation with sodium thiocyanate. The Factor IX is then dissociated from the monoclonal antibody column, recovered, treated with sodium thiocyanate for viral inactivation, and further purified, including two ultrafiltration steps for virus removal. The various purification steps substantially reduce virus titers. Mononine is highly purified Factor IX, showing predominantly a single component by sodium dodecylsulfate polyacrylamide electrophoresis (SDS PAGE), and has activity not less than 190 Factor IX units per mg total protein.

Mononine is packaged as powder in vials containing either 250, 500 or 1,000 I.U. Factor IX along with diluent for reconstitution (Sterile Water for Injection, USP), a double ended needle for reconstitution, a vented filter spike for withdrawal, an infusion set, and alcohol swabs. Each vial contains a labeled amount of Factor IX. The 250, 500 and 1,000 I.U. vials are reconstituted with 2.5, 5.0 and 10.0 mL, respectively, of Sterile Water for Injection, USP. The reconstituted product contains approximately 100 Factor IX I.U. per mL. This is equivalent to 100-times the normal Factor IX potency of an equal volume of Plasma from healthy individuals. One Factor IX unit is equivalent to the Factor IX activity in 1 mL of normal, pooled Plasma, as determined by comparison with the World Health Organization standard for Factor IX. When reconstituted for use, the product is isotonic (equal to 0.12-0.18 M NaCl) and neutral in pH (6.6-7.4). Mononine should be stored at 2-8ûC (refrigerated) and has a shelf life of 24 months. It may be stored for one month at room temperature (up to 30ûC). The date of manufacture is "defined as the date of the first sterile filtration of the bulk."

Mononine contains detectable traces of murine (mouse) protein, up to 50 ng/100 I.U., residual from the murine monoclonal antibody used for purification. The traces of murine protein have not been associated with anamnestic responses, induction of significant levels of human anti-mouse antibodies (HAMA), or clinical adverse effects. Non-detectable levels of Factor II, VII and X (< 0.0025 I.U/ I.U. Factor IX) with a specific activity of 150 IU/mg protein are present. The product also contains histidine (about 10 mM), sodium chloride (about 0.066 M) and mannitol (about 3%). Hydrochloric acid and/or sodium hydroxide may be used to adjust the pH of the murine monoclonal antibody used for purification.

Nomenclature: Factor IX, Mab purif./CSL [BIO]; Mononine [TR]; Coagulation Factor IX (Human) [FDA]; 9001-28-9 [CAS RN]; Mono-IX [SY]; Antihemophilic factor B [SY]; Factor IX [SY]; Christmas factor [SY]; Blood coagulation factor IX [SY]; NDC 0053-7668-04; 0053-7668-02; 0053-7668-01 [NDC]

Companies: Mononine is manufactured by CSL Bioplasma Inc., CBER/FDA est. no. 1639 (formerly Aventis Behring LLC, before that, Centeon L.L.C. and Armour Pharmaceutical Co.), at its Kankakee, IL facility. It is marketed in the U.S. by CSL Bioplasma Behring LLC, and internationally by its parent company, CSL Bioplasma AG, both subsidiaries of CSL Ltd.. The product was originally developed by Armour Pharmaceutical.

Manufacture: Mononine is produced from pooled human blood plasma, either recovered plasma units or source plasma. After removal of the cryoprecipitated fraction, the remaining supernatant fluid is fractionated by ion exchange chromatography using DEAE Sepharose resin. The eluate is further purified by immunoaffinity chromatography through a column containing matrix-bound murine Factor IX monoclonal antibody. The monoclonal antibody-bound Factor IX protein is eluted from the immunoaffinity matrix using sodium thiocyanate solution, and the eluate is incubated in 1.5 M sodium thiocyanate elution buffer for at least 2.5 hours for virus inactivation. The incubated filtrate is ultrafiltered to remove thiocyanate and filtered through two sequential membranes, each with an approximate molecular weight cutoff of 100,000 Dalton (100 kDa). Dual ultrafiltration allows passage of Factor IX, while retaining many viruses. The filtrate is then concentrated, and adsorbed onto and eluted from an additional ion exchange resin (aminohexyl-Sepharose gel; AH Sepharose) to remove trace residual murine monoclonal antibodies. The eluate is dialyzed or diafiltered to achieve the approximate excipient concentrations. The pH is adjusted with hydrochloric acid and/or sodium hydroxide, and the solution is clarified and sterilized through a series of filters. The final filter has a porosity of 0.2 µm for sterile filtration. The product is filled into vials and lyophilized (freeze-dried). [Note, U.S. patent 6,280,729 and its Example 1 apparently describe aspects of the actual manufacture of Mononine].

Studies of the overall manufacturing process using product samples spiked with virus have shown total reduction of 11.56 log (i.e., 10 to the 11.56 power) for HIV; 10.24 log for sindbis virus; 11.64 log for murine encephalomyocarditis virus (EMC); 14.23 log for vesicular stomatitis virus (VSV); and 10.90 log for vaccinia virus. The major manufacturing processes which contribute to reduction/removal of viruses are: 1) monoclonal antibody purification (no reduction in HIV; reduces Sindbis virus by 2.76 log, EMC by .89 log, vesicular stomatitis virus (VSV) by 7.18 log, and vaccinia virus by 3.60 log); 2) sodium thiocyanate inactivation (reduces HIV by 4.16 log, no substantive reduction in other test viruses); and 3) ultrafiltration (the double ultrafiltration reduces HIV by 7.4 log, Sindbis virus by 7.48 log, and EMC by 7.75 log; a single ultrafiltration step reduces VSV by 7.05 log and vaccinia virus by 7.30 log). Similar studies using porcine parvovirus (PPV) as a model for human parvovirus B19 show an overall reduction of 11.64 log (> 4.28 for combined Mab chromatography and sodium thiocyanate; 2.26 log for AH Sepharose chromatography; and 5.10 log for ultrafiltration).

Aventis Behring was the first company (May 2000) in North America to manufacture Plasma-derived products exclusively from Plasma released after proprietary investigational polymerase chain reaction (PCR; Nucleic Acid Testing; NAT) screening for HIV-1, hepatitis B virus, and hepatitis C virus. In 2000, Aventis Behring also added PCR screening for parvovirus B19 and hepatitis A viruses, making the company the first in the world to screen plasma using proprietary investigational PCR/NAT screening for five viruses (HIV-1, hepatitis B, hepatitis C, parvovirus B19, and hepatitis A).

FDA class: Biologic PLA BLA

Approvals: Date = 19840221; first approval, PLA, for Coagulation Factor IX (Human) from Armour Pharmaceutical; Indication = replacement treatment and prophylaxis of the hemorrhagic complications of hemophilia B

Date = 19920820, first approval; PLA ref. no. 90-0030 and 90-0576, for Mononine (with solvent detergent inactivation and monoclonal antibody purification); approval allowed export of unlabeled final container product; orphan designation (granted 06/27/1989; expired 8/20/1999)

Date = 19930401; PLA for original 1984 formulation revoked

Date = 20000511; BLA supplement; Indication = addition of statemen regarding use of Nucleic Acid Testing (NAT) using Polymerase Chain Reaction (PCR) for HIV-1, hepatitis B and C virus detection

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

Coagulation Factor IX (Human), Mononine is indicated for the prevention and control of bleeding in Factor IX deficiency, also known as Hemophilia B or Christmas disease. Mononine is not indicated in the treatment of prophylaxis of hemophilia A patients with inhibitors to Factor VIII.

Coagulation Factor IX (Human), Mononine contains non-detectable levels of Factors II, VII and X (<0.0025 IU per Factor IX unit, using standard coagulation assays) and is, therefore, not indicated for replacement therapy of these clotting factors. Mononine is also not indicated in the treatment or reversal of coumarin-induced anticoagulation in a hemorrhagic state caused by hepatitis-induced lack of production of liver dependent coagulation factors.

Status: Coagulation Factor IX (Human) from Armour Pharmaceutical Co. was approved in 1984. Coagulation Factor IX (Human) with monoclonal antibody purification and solvent/detergent viral inactivation, Mononine, from Armour was approved on August 20, 1992. This approval included manufacturing changes, amendment of the establishment license, and allowed export of unlabeled final container product. With the 2000 approval, the original PLA was converted to a BLA.

For purposes of orphan designation, the FDA explicitly concluded that Mononine was a different "drug" from Alpha-Nine (without immunoaffinity purification) by virtue of increased safety. FDA particularly concluded that the manufacture of Mono-nine is much less likely than AlphaNine (heat inactivated) to result in transmission of hepatitis C virus.

In Jan. 2004, Mononine was approved in the European Union (EU) for administration by continuous intravenous infusion (for which it is not approved in the U.S.).

Tech. transfer: The product insert/labeling cites U.S. patent 5,055,557, Zimmerman, T.S., "Ultrapurification of factor IX and other vitamin K-dependent proteins," October 8, 1991, assigned to Scripps Clinic & Research Foundation (La Jolla, CA), now The Scripps Research Institute (TSRI). This and related patent properties have been nonexclusively licensed by CSL Behring. Related Zimmerman/Scripps patents include U.S. 5,639,857, June 17, 1997, and 5,614,500, March 25, 1997.

U.S. 5,055,557 describes use of a murine monoclonal antibody isolated from mice injected with highly purified Factor IX and manufactured by the in vivo murine ascites method. Monoclonal antibody with high affinity for human Factor IX is bound to cross-linked agarose (Sepharose 4B from Phar-macia) to form the immunoaffinity chromatography matrix. Plasma is poured through the chromatography column, with a contact time in the range of 2 to 4 hours. The column is washed with tris-lithium chloride buffer (e.g., 0.1 M lysine base and 0.5 M lithium chloride, pH 8) to remove non-antibody-bound proteins. This is followed by a second washing with tris-hydrochloride buffer (e.g., 0.05 M tris-hydrochloride, pH 8) to maximize removal of lithium ions from the column. Elution of purified Factor IX is accomplished with buffer containing sodium thiocyanate as an eluant. The Factor IX pool may be further concentrated by standard procedures such as pressure ultrafiltration using an Amicon stirred cell with a YM-10 membrane under 50 psi of nitrogen pressure. The immunoadsorbent column can be regenerated by elution of the purified protein off of the column, and can be repeatedly used indefinitely. This overall method provides Factor IX with purification on the order of 1,500-fold.

U.S. 6,280,729, assigned to Aventis Behring, now CSL Bioplasma, describes methods for protecting blood-derived Factor IX from proteases during purification and storage using high concentrations of organic or inorganic salt(s), e.g., magnesium, potassium, sodium and lithium chloride or sodium sulfate, to stabilize Factor IX, with specific activity >194 and free of Factor IXa, degraded forms of Factor IX, and other Factors. Example 1 appears to describe aspects of the manufacture of Mononine.

Solvent detergent virus inactivation was developed by and nonexclusively licensed from the New York Blood Center. For example, see U.S. patent 4,820,805. See the entry for Pooled Plasma, Solvent Detergent Treated (SD Plasma) (#799) for further information about solvent detergent viral inactivation, used primarily for inactivation of enveloped viruses (e.g., HIV, hepatitis B and C viruses).

Market.: The 2007 Average Wholesale Price (AWP) is $1.18/IU (no change from 2004) (Red Book, 2007).

Medicare reimbursement for inpatient and home care at $1.12/IU and outpatient at $0.51/IU, and Estimated Acquisition Costs (for hospitals, treatment centers) is $0.80-$0.93/IU [from NHF].

In its March 22, 2006 price list, FFF Enterprises, a major biologics distributor, reported its price as $0.89/IU ($0.81 in 2005; $0.94/IU in 2004).