Status: approved; marketed

Organizations involved:

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

Instituto Grifols, S.A. –Intl. mark.

Probitas S.A. – Parent

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

Green Cross Corp. – Former

Yoshitomi Pharmaceutical Industries, Ltd. – Former

Mitsubishi Chemical Corp. – Former

Abbott Laboratories – R&D; Tech.; Former

New York Blood Center, Inc. – Tech.

Cross ref: See the Factor VIII Products entry (#715). See also other similar plasma-derived Factor VIII:vWF entries, e.g., Koate-DVI (#724) and Humate-P (#726).

Description: Antihemophilic Factor (Human) Solvent Detergent/Heat Treated or Alphanate is a lyophilized (freeze-dried) formulation of a noncovalently bound complex of Antihemophilic Factor (Human) or Factor VIII with von Willebrand’s factor (vWF), i.e., Factor VIII:vWF, manufactured from pooled Plasma, with manufacture including viral inactivation using the solvent detergent process (tri-n-butyl phosphate and polysorbate 80) and dry heat pasteurization (80˚C) of the product in final container vials. The Factor VIII:wWF complex is the major form in which Factor VIII exists in vivo, with the von Willebrand’s factor acting to stabilize Factor VIII against degradation and proteolysis.

Alphanate was the first Antihemophilic factor (AHF) product in the U.S. to incorporate both solvent detergent and heat treatment methods of viral inactivation. Alphanate is doubly viral inactivated (as is Koate-DVI).

Alphanate is packaged in single dose vials along with a double-ended transfer needle and microaggregate filter for intravenous administration in the treatment of hemophilia A or acquired Factor VIII deficiency. Each Alphanate vial is labeled with its Antihemophilic Factor content (Factor VIII activity) expressed in International Units (IU) per vial (referenced to the WHO International Standard). Alphanate contains Albumin (Human) as a stabilizer. The final product has specific activity in the range of 150 units/mg protein. If the Factor VIII were cleaved from the bound von Willebrand’s Factor molecules, the specific activity of the Factor VIII would be in the range of 1,000 units/mg protein. When reconstituted with the appropriate volume of Sterile Water for Injection, USP, Alphanate contains 0.3-0.9 g Albumin (Human)/100 mL; not more than (NMT) 5 mmol calcium/L; NMT 750 µg glycine/IU Factor VIII; NMT 1.0 U heparin/mL; 10-40 mmol histidine/L; NMT 0.1 mg imidazole/mL; 50-200 mmol arginine/L; NMT 1.0 µg polyethylene glycol (PEG)/IU Factor VIII; NMT 1.0 µg polysorbate 80/IU Factor VIII; NMT 10 mEq sodium/vial; and NMT 0.1 µg tri-n-butyl phosphate/IU Factor VIII. Alphanate should be stored at 2-8˚C (refrigerated). It may be stored at room temperature not to exceed 30˚C for up to 2 months.

Nomenclature: Factor VIII:vWF/Grifols [BIO]; Alphanate [TR with 1994 approval]; Profilate SD [TR prior to 1994 approval]; Antihemophilic Factor (Human) Solvent Detergent/Heat Treated [FDA]; Antihemophilic Factor [USAN]; Antihemophilic Factor, Human [former USAN]; Antihemophilic Factor USP [USP]; Blood-coagulation factor VIII, complex [CAS]; 9001-27-8 [CAS RN]; Factor VIII:von Willebrand’s factor complex [SY]; Antihemophilic Factor VIII [SY]; Factor VIII:C [SY]

Companies.: Alphanate 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.. later a subsidiary of 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. Alpha-nate is marketed in the U.S. by Grifols Biological Inc., and inter-nationally 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. and manufactured at Abbott facilities in South Pasadena, CA. This facility and its products were acquired by (licensed transferred to) Alpha Therapeutic on Aug. 15, 1978.

In 2007, Grifols is in the process of validating a new state-of- the-art bulk processing and aseptic filling facility at its manufacturing complex in Los Angeles, CA. The facility uses proprietary equipment and process flow designs pioneered by Grifols Engineering S.A., a wholly owned subsidiary of Grifols S.A. When fully validated and FDA approved, the new facility will be used for final container filling of Alphanate and other Grifols products.

Manufacture: Source units of Plasma, negative for HIV, hepatitis B virus, and other contamination, may include recovered Plasma, Source Plasma Salvaged, and Source Plasma (bulk cryoprecipitate). After Cohn-Oncley cold ethanol fractionation of pooled plasma, the cryoprecipitate is dissolved and fractionated (precipitated) further with polyethylene glycol (PEG). The supernatant from PEG fractionation is treated with the solvent detergent viral inactivation process involving mixture with 0.30% ± 0.2% v/w tri(n-butyl) phosphate (TNBP) solvent and 1.00% ± 0.05% w/w polysorbate 80 (Tween 80) detergent for at least six hours at 27˚C ± 3˚C. Treated material is further purified by heparin affinity chromatography, i.e., fed into a chromatography column containing heparin ligand coupled to a cross-linked agarose matrix with affinity for the heparin binding domain of the Factor VIII:vWF complex. The heparin ligands also bind vWF molecules and a small amount of extraneous proteins. The column is eluted with calcium chloride which releases the bound proteins. The eluate is ultrafiltered.

Another precipitation step (“Selective Salt Precipitation”) is performed. This separates Factor VIII:vWF complexes from extraneous proteins. Sodium chloride and glycine are added to the eluate and the mixture is incubated for some time at 15-20˚C. The sodium chloride causes extraneous proteins bound to Factor VIII:vWF complexes to separate from the heavier complexes, yielding a highly enriched precipitate of Factor VIII:vWF complex. The precipitate is centrifuged, resulting in bulk product with specific activity in the range of 150 units/mg protein. Albumin (Human) at a concentration of 0.05%-1% is added as a stabilizer, glycine is added, and albumin concentration is adjusted to 0.02%. The product is sterile filtered, filled in vials, lyophilized (freeze-dried), followed by heat treatment for viral inactivation at 80˚C for 72 hours (particularly, to inactivate non-lipid enveloped viruses, e.g., hepatitis A virus, not affected by solvent detergent inactivation), and sealed. Final containers are negative for microbial contamination, including HIV and hepatitis B virus.

The various manufacturing steps each contribute to viral inactivation and/or removal – PEG precipitation; solvent detergent inactivation; heparin column chromatography; lyophilization; and dry heat. Overall virus removal/reduction is ≥ 13.2 log for HIV-1; ≥ 6.2 log for HIV-2; ≥ 4.1 log for vesicular stomatitis virus (VSV); ≥ 4.7 log for sindbis virus; > 19.0 log for bovine herpes virus (BHV); > 9.2 log for poliovirus; ≥ 7.9 log for hepatitis A virus (HAV); ≥ 9.4 log for bovine viral diarrheal virus (BVDV); and 5.3 log for canine papillomavirus (CPV). In vitro inactivation studies sponsored by Alpha to evaluate the solvent detergent treatment step used an assay with a sensitivity of 2 logs for the marker viruses, vesicular stomatitis virus (VSV) and sindbis virus. The studies showed a kill of 6.8 log for VSV and 6.4 log for sindbis virus. Greater than 10 logs for HIV-1 and greater than 6 logs of HIV-2 were inactivated.

FDA class: Biologic PLA

CBER class: Blood And Blood Derivatives

Approvals: Date = 19710430; first approval, PLA, granted to Abbott Labs.

Date = 19780815; product license withdrawn from Abbott Labs. and granted (reissued; PLA/ELA ref. no. 78-262 and 78-268) to Alpha Therapeutic Corp. (new owner)

Date = 19840210; PLA supplement (no. 83-280); Indication = implementation of heat treatment step during manufacture for viral inactivation (apparently wet heat, in solution, pasteurization) (product then known as Profilate)

Date = 19850904; PLA supplement (ref. no. 85-090); Indication = change in viral inactivation processing to dry heating (pasteurization) of final container vials

Date = 19890719; PLA supplement (ref. no. 88-170); Indication = addition of solvent detergent viral inactivation process during manufacture (product then Profilate SD)

Date = 19940719; PLA supplemental (ref. no. 90-0218; Indication = addition of a heparin agarose chromatographic purification step and a precipitation step to the manufacturing process; approval also provided for export of frozen paste, frozen bulk solution and unlabeled final container product

Date = 20070131; BLA supplement; Indications: = treatment of congenital von Willebrand’s Disease in connection with surgery or other invasive procedures in whom desmopressin (DDAVP ) is either ineffectiveor contraindicated

Date = 20070201; BLA supplement; Indication = for the treatment of congenital von Willebrand Disease (VWD) in connection with surgery or other invasive procedures

Date = 20070427; BLA supplement; Indication = for adult and pediatric patients with von Willebrand disease for the prevention of excessive bleeding pre and post surgery

Indications: [full text of the "INDICATIONS AND USAGE” section from product insert/labeling, 5/14/07]:

1.1 Hemophilia A or Acquired Factor VIII Deficiency: Antihemophilic Factorlvon Willebrand Factor Complex (Human), Alphanate, is indicated for the prevention and control of bleeding in patients with Factor VIII deficiency due to hemophilia A or acquired Factor VIII deficiency.

1.2 von Willebrand Disease: Antihemophilic Factorhon Willebrand Factor Complex (Human), Alphanate, is also indicated for surgical and/or invasive procedures in patients with von Willebrand Disease (VWD) in whom desmopressin (DDAVP)i s either ineffective or contraindicated. It is not indicated for patients with severe VWD (Type 3) undergoing major surgery.

Status: With its Jan. 2007 approval, Alphanate became the first and only dual inactivated (solvent detergent and heat treatment) and affinity chromatography purified antihemophilic factor/von Willebrand factor complex that is FDA approved for the treatment of von Willebrand’s disease. In connection with this approval, Grifolts must conduct a postmarketing study to further evaluate safety and efficacy of the product in type 3 von Willebrand’s disease patients undergoing major surgeries.

Tech. transfer: Solvent detergent viral inactivation technology 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).

Trials: In clinical studies with Alphanate to support its 2007 approval for use in von Willebrand’s patients undergoing surgery, 120 major, moderate and minor surgical procedures were performed in 76 patients. Based on predefined criteria for efficacy, more than 90% of patients had favorable outcomes. In these studies, 15.8 percent of subjects and 5.7 percent of infusions were accompanied by adverse reactions, most commonly itching, pharyngitis, paresthesia (a sensation of numbness and tingling on the skin) and headache, swelling of the face, and rash and chills.

Medical: Successful management of surgery or invasive procedures in mildly, moderately and severely affected von Willebrand’s patients routinely requires correction of the bleeding defect. In the absence of correction of the bleeding defect, patients may suffer from prolonged bleeding and delayed wound healing.

Market: The 2007 Average Wholesale Price (AWP) is $1.00/IU for all presentations (unchanged from 2004) (Red Book, 2007).

Medicare inpatient and home care reimbursement is set at $0.90/IU, and at $0.51/IU for outpatient care. Estimated Acquisition Costs (for hospitals, treatment centers) is $.61-.66/IU [data from NHF].

In its March 22, 2006 price list, FFF Enterprises, a major biologics distributor, reported $0.59/IU for all sizes of Alphanate (same in 2005 and 2004).