Status: approved, but not marketed in U.S. (discontinued)

Organizations involved:

Grifols Biological Inc. – Manuf.; Tech.; USA mark.

Istituto Grifols S.A. – Intl. mark

Probitas S.A. – Parent

Alpha Therapeutic Corp. – R&D; Tech.; Former

Green Cross Corp. – Former inv.

Yoshitomi Pharmaceutical Industries, Ltd. – Former

Mitsubishi Chemical Corp. – Former

New York Blood Center – Tech.

Abbott Laboratories – R&D; Tech.; Former.

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

Description: Coagulation Factor IX (Human) Solvent Detergent Treated/Virus Filtered or AlphaNine SD is a lyophilized (freeze-dried) formulation of purified Factor IX prepared from cold ethanol fractionated Plasma (Human). Manufacturing includes ion exchange chromatography (DEAE cellulose), dual affinity chromatography, virus inactivation using the solvent detergent [tri-n-butyl phosphate (TNBP) and polysorbate 80] and wet heat (pasteurization) methods, and virus removal by nano-filtration. The final reconstituted product contains >150 IU Factor IX per mg of protein (generally 230 IU/mg).

AlphaNine SD is supplied as a sterile lyophilized powder in single use vials along with 10 mL Sterile Water for Injection, USP (10 ml) for reconstitution prior to intravenous injection, along with a double-ended needle and microaggregate filter for use in administration. Each vial of AlphaNine SD is labeled with the number of plasma equivalents of Factor IX/vial, expressed in International Units (IU) referenced to the WHO International Standard. One plasma equivalent is defined as the activity contained in one mL of normal plasma. AlphaNine SD also contains not more than 0.04 unit heparin, 1 mg dextrose, 2.5 µg polysorbate 80 (Tween 80) and 0.25 µg tri(n-butyl)phosphate (TNBP) per IU of Factor IX. AlphaNine SD should be stored at 2-8˚C (refrigerated) and has a shelf life of 12 months. The date of manufacture is defined as the date of the first sterile filtration of the bulk. Levels of Factor VII (proconvertin), Factor II prothrombin) and Factor X (Stuart-Prower Factor) in the product are below the limit of detection (less than 0.04 Factor VII unit, less than 0.05 Factor II unit, and less than 0.05 Factor X unit per IU Factor IX).

First generation AlphaNine manufacturing included a wet heptane/heat treatment step for viral inactivation and ion exchange chromatography, and provided product with a specific activity of 80-120 units/mg of protein. Factor IX has a theoretical maximum specific activity of 250 units/mg of protein. Second generation AlphaNine introduced a further dual affinity chromatography step, resulting in a product with a specific activity of 200 units/mg protein. The current purification process, including dual affinity chromatography and nanofiltration, achieves a specific activity of 230 units/mg of protein.

Solvent detergent viral inactivation eliminates over 12 log of HIV-1; 6 log of HIV-2; over 4.9 log of vesicular stomatitis virus (VSV); and over 5.3 log of Sindbis virus. The ion exchange and dual affinity chromatography steps, respectively, eliminate an additional 1.4 and 4.7 logs of Sindbis virus (a total of 11.4 log). Nano-fil-tration removes additional viruses, including parvovirus, hepatitis A virus (HAV), some nonenveloped viruses, and particulates larger than 15 nm in diameter.

Companies.: Alphanine SD is manufactured by Grifols Biological Inc., CBER/FDA est. no. 1694, previously Alpha Therapeutic Corp. (originally independent, later a subsidiary of Green Cross Corp., a subsidiary of Yoshitomi Pharmaceutical Industries, Ltd., acquired by Mitsubishi Chemical), CBER/FDA lic. no. 0744. Alpha was acquired by Probitas Pharma S.A. in July 2003 and merged into its Instituto Grifols subsidiary. Alphanine SD is marketed in the U.S. by Grifols Biological Inc., and internationally by Istituto Grifols S.A. and affiliates (previously Alpha Therapeutic Corp. and affiliates, including Green Cross and Yoshitomi). The product was originally developed by Abbott Labs., which divested its plasma derivatives business to Alpha in the late 1970s.

Nomenclature: Factor IX SD [BIO]; AlphaNine SD [TR]; Coagulation Factor IX (Human) Solvent Detergent Treated/Virus Filtered [FDA]; 9001-28-9 [CAS RN]; Antihemophilic factor B [SY]; Factor IX [SY]; Christmas factor [SY]; Blood coagulation factor IX [SY EINECS]; NDC 49669-3600-2 [NDC]

Manufacture: AlphaNine SD is manufactured from pooled human blood plasma [Plasma (Human)] from which cryoprecipitate (Factor VIII) has been removed. This cryoprecipitate-poor plasma is fractionated (Cohn cold ethanol process), and Factor IX is isolated using diethylaminoethyl (DEAE) cellulose ion-exchange chromatography (probably DEAE Sepha-rose from Amersham Pharmacia Biotech or similar matrix). The eluate is treated using the solvent detergent process, involving addition of low levels of tri(n-butyl)phosphate (TNBP) and poly-sorbate 80 (Tween 80), for viral inactivation. The product is incubated with the solvent detergent mixture for six hours at 27 ± 3˚C. The Factor IX protein is then precipitated with barium citrate and further purified by two sequential affinity chromatography steps utilizing an unspecified polysaccharide as the ligand. This further reduces residual amounts of Factor X and increases purity. The product is filtered (nano-filtered) through a 15 nanometer filtration membrane to remove viruses (both enveloped and nonenveloped viruses, including parvovirus B19 and hepatitis A virus). Vials are filled and the product is lyophilized (freeze-dried), sealed, labeled, and packaged. As a result of adding the nanofiltration step to the dual chromatography and other partitioning steps, the final product has a specific activity averaging 229 (± l23) units of factor IX per mg of protein (based on a theoretical maximum of 250 units/mg).

Studies involving production lots spiked with virus prior to the solvent detergent process have shown that this and subsequent manufacturing steps reduce HIV-1 by at least 12.2 log (i.e., 1012.2); HIV-2 by at least 6.00 log; Sindbis virus (a model for hepatitis C virus) by at least 5.30 log; and vesicular stomatitis virus (VSV) by at least 4.90 log. The ion exchange (DEAE Sepharose) chromatography step has been shown to reduce the concentration of Sindbis virus by 1.41 log; and the two sequential affinity chromatography steps reduce viral concentration by 4.70 log. Altogether, independent of the solvent detergent, heat treatment, and nanofiltration processes, the purification steps used are capable of reducing virus concentrations by about 6 log. Note, viral reduction (inactivation) of AlphaNine (the 1st generation product) was achieved by a wet solvent heating method, involving maintaining the product in n-heptane suspension at 60˚C for 20 hours.

FDA class: Biologic PLA

CBER class: Blood And Blood Derivatives

Approvals: Date = unknown; approval date for Coagulation Factor IX (Human) manufactured by the Abbott Labs. [product no longer in current CBER/FDA databases/listings]

Date = 19780815; Licenses for the Abbott Labs., S. Pasadena, CA, manufacturing facilities and products transferred to a new owner, Alpha Therapeutic

Date = 19841029; PLA supplement; added wet heat treatment/solvent viral inactivation [61˚C, 20 hr. in heptane]

Date = 19901231, PLA (later converted to BLA STN 103249/0) for heptane heat treated Factor IX; first approval for the solvent detergent inactivated product (AlphaNine SD); orphan designation (granted 07/05/1990; expired 12/31/1997); Note, this is the first approval reported in the CBER/FDA database and other FDA sources for Coagulation Factor IX (Human) from Alpha Therapeutic Corp.

Date = 19920826; PLA supplement no. 91-0350; Indication = amendment regarding solvent/detergent processing; inclussion of solvent detergent treatment and removal of heat treatment with n-heptane

(solvent detergent treated and virus filtered) – inclussion of Nanofiltration for virus removal Date = 19960607; PLA suplement; Indication = (solvent detergent treated and virus filtered, i.e., the current, product) inclusion of nanofiltration for virus removal

Indications: [full text of the "INDICATIONS AND USAGE” section of product insert/labeling, 6/2012]:

AlphaNine SD is indicated for the prevention and control of bleeding in patients with Factor IX deficiency due to hemophilia B. AlphaNine SD contains low, non-therapeutic levels of Factors II, VII, and X, and, therefore, is not indicated for the treatment of Factor II, VII or X deficiencies. This product is also not indicated for the reversal of coumarin anticoagulant-induced hemorrhage, nor in the treatment of hemophilia A patients with inhibitors to Factor VIII.

Status: See the Regulatory section of the entry for Factor IX VH (Bebu-lin VH) (#734) for information about an FDA ruling involving a later, comparable product that was eventually ruled superior to AlphaNine (at the time), granted Orphan designation, and allowed to enter the market (despite AlphaNine SD having previously received orphan designation).

As of June 2012, AlphaNine SD was approved in 16 countries, including the U.S. and several European countries, including Germany and the U.K.

Tech. transfer: Solvent detergent viral 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 (unchanged from 2004) (Red Book, 2007).

Medicare reimbursement is set at $1.12/IU for inpatient and home care, and at $0.51/IU for outpatient care. Estimated Acquisition Costs (for hospitals, treatment centers) is $0.65/IU [from NHF].

In its March 22, 2006 price list, FFF Enterprises, a major biologics distributor, reported $0.72/IU for all sizes (same in 2005 and 2004).

Alpha Therapeutic (now Grifols) reported that in clinical trials in patients undergoing surgery, the amount used ranged from 3,295-52,200 IU, costing $3,460 to $54,810 (at rate of $1.05/IU). The average total units used per subject (mean) was 28,512 IU costing $29,938. For knee surgery, the mean used was 29,983 IU, for hernia repair was 30,720 IU, and for dental surgery was 5,703 IU.