antihemophilic factor, von willebrand factor complex (human)
Dosage Form: injection
FULL PRESCRIBING INFORMATION
Indications and Usage for Alphanate
Hemophilia A
Alphanate®, Antihemophilic Factor/von Willebrand Factor Complex (Human), is indicated for the control and prevention of bleeding in patients with Factor VIII (FVIII) deficiency due to hemophilia A.1
von Willebrand Disease
Alphanate is indicated for surgical and/or invasive procedures in adult and pediatric patients with von Willebrand Disease (VWD) in whom desmopressin (DDAVP®) is either ineffective or contraindicated. It is not indicated for patients with severe VWD (Type 3) undergoing major surgery.
Alphanate Dosage and Administration
For Intravenous Use Only
Antihemophilic Factor (AHF) potency (FVIII:C activity) is expressed in International Units (IU) FVIII/vial on the product label. Additionally, Alphanate contains von Willebrand Factor:Ristocetin Cofactor (VWF:RCo), which is expressed in IU VWF:RCo/vial for the treatment of VWD.
Hemophilia A
- Treatment with Alphanate should be initiated under the supervision of a physician experienced in the treatment of hemophilia.
- Dosage and duration of treatment depend on the severity of the FVIII deficiency, the location and extent of bleeding, presence of inhibitors, and the patient’s clinical condition. Careful control of replacement therapy is especially important in cases of major surgery or life-threatening bleeding episodes.
- Dosing requirements and frequency of dosing is calculated on the basis of an expected initial response of 2% of normal FVIII:C increase per IU FVIII:C/kg body weight administered. 2,3
The expected in vivo peak increase in FVIII level expressed as IU/dL (or % normal) can be estimated using the following formulas:
Dosage (units) = body weight (kg) x desired FVIII rise (IU/dL or % normal) x 0.5 (IU/kg per IU/dL)
OR
IU/dL (or % normal) = Total Dose (IU)/body weight (kg) x 2
Thus, an administered AHF dose of 50 IU/kg will be expected to increase the circulating FVIII level to 100% of normal (100 IU/dL).
Doses administered should be titrated to the patient’s clinical response, including individualized needs, severity of the deficiency, severity of the hemorrhage, presence of inhibitors, and FVIII level desired. Patients may vary in their pharmacokinetic (e.g., half-life, in vivo recovery) and clinical responses to Alphanate. Although the dose can be estimated by the calculations above, it is highly recommended that, whenever possible, appropriate laboratory tests including serial FVIII activity assays be performed.
| Hemorrhagic event | Dosage (IU FVIII:C/kg Body Weight) |
Minor hemorrhage:
| FVIII:C levels should be brought to 30% of normal (15 IU FVIII/kg twice daily) until hemorrhage stops and healing has been achieved (1–2 days). |
Moderate hemorrhage:
| FVIII:C levels should be brought to 50% (25 IU FVIII/kg twice daily). Treatment should continue until healing has been achieved (2–7 days, on average). |
Major hemorrhage:
| FVIII:C levels should be brought to 80–100% for at least 3–5 days (40–50 IU FVIII/kg twice daily). Following this treatment period, FVIII levels should be maintained at 50% (25 IU FVIII/kg twice daily) until healing has been achieved. Major hemorrhages may require treatment for up to 10 days. Intracranial hemorrhages may require prophylaxis therapy for up to 6 months. |
| Surgery | Prior to surgery, the levels of FVIII:C should be brought to 80–100% of normal (40–50 IU FVIII/kg). For the next 7–10 days, or until healing has been achieved, the patient should be maintained at 60–100% FVIII levels (30–50 IU FVIII/kg twice daily). |
Dosing requirements and frequency of dosing is calculated on the basis of an expected initial response of 2% FVIII:C increase per IU FVIII:C/kg body weight (i.e., 2% per IU/kg) and an average half-life for FVIII:C of 12 hours. If dosing studies have determined that a particular patient exhibits a lower than expected response and shorter half-life, the dose and the frequency of dosing should be adjusted accordingly. Failure to achieve the expected plasma FVIII:C level or to control bleeding after an appropriately calculated dosage may be indicative of the development of an inhibitor (an antibody to FVIII:C). Its presence should be documented and the inhibitor level quantitated by appropriate laboratory procedures. Treatment with AHF in such cases must be individualized .4-6
Plasma FVIII levels should be monitored periodically to evaluate individual patient response to the dosage regime. Depending on the level of the inhibitor and/or clinical response, it may be appropriate to use an alternative ‘bypass’ therapeutic agent.
von Willebrand Disease
- Treatment with Alphanate should be initiated under the supervision of a physician experienced in the treatment of VWD.
- The ratio of VWF:RCo to FVIII in Alphanate varies by lot, so dosage should be re-evaluated whenever lot selection is changed.
- Dosage and duration of treatment depend on the severity of the VWF deficiency, the location and extent of bleeding, and the patient’s clinical condition. Careful control of replacement therapy is especially important in cases of major surgery or life-threatening bleeding episodes. Overdosage resulting in FVIII levels above 150% should be avoided.
The median incremental in vivo recoveries of VWF:RCo and FVIII:C were 3.12 (IU/dL)/(IU/kg) [mean, 3.29 ± 1.46 (IU/dL)/(IU/kg); range: 1.28 to 5.73 (IU/dL)/(IU/kg)] for VWF:RCo and 1.95 (IU/dL)/(IU/kg) [mean, 2.13 ± 0.58 (IU/dL)/(IU/kg); range: 1.33 to 3.32 (IU/dL)/(IU/kg)] for FVIII:C.
The following table provides dosing guidelines for pediatric and adult patients with von Willebrand Disease.7-10
| Minor Surgery/Bleeding | ||
| VWF:RCo | Target FVIII:C Activity Levels | |
| Pre-operative/pre- procedure dose: | Adults: 60 IU VWF:RCo/kg body weight.
Pediatrics: 75 IU VWF:RCo/kg body weight. | 40-50 IU/dL |
| Maintenance dose: | Adults: 40 to 60 IU VWF:RCo/kg body weight at 8 to 12 hour intervals as clinically needed for 1-3 days.
Pediatrics: 50 to 75 IU VWF:RCo/kg body weight at 8 to 12 hour intervals as clinically needed for 1-3 days. | 40-50 IU/dL |
| Safety Monitoring: | Peak and trough at least once daily | Peak and trough at least once daily |
| Therapeutic Goal (Trough)* : | >50 IU/dL | >50 IU/dL |
| Safety Parameter†: | Should not exceed 150 IU/dL | Should not exceed 150 IU/dL |
| ||
| Major Surgery/Bleeding | ||
| VWF:RCo | Target FVIII:C Activity Levels | |
| Pre-operative/pre- procedure dose: | Adults: 60 IU VWF:RCo/kg body weight. Pediatrics: 75 IU VWF:RCo/kg body weight. | 100 IU/dL |
| Maintenance dose: | Adults: 40 to 60 IU VWF:RCo/kg body weight at 8 to 12 hour intervals as clinically needed for atleast 3-7 days. Pediatrics: 50 to 75 IU VWF:RCo/kg body weight at 8 to 12 hour intervals as clinically needed for atleast 3-7 days. | 100 IU/dL |
| Safety Monitoring: | Peak and trough at least daily | Peak and trough at least daily |
| Therapeutic Goal (Trough)* : | >50 IU/dL | >50 IU/dL |
| Safety Parameter† : | Should not exceed 150 IU/dL | Should not exceed 150 IU/dL |
Reconstitution
Always Use Aseptic Technique
- Warm diluent (Sterile Water for Injection, USP) and concentrate (Alphanate) to at least room temperature (but not above 37 °C).
- Remove the plastic flip off cap from the diluent vial.
- Gently swab the exposed stopper surface with a cleansing agent such as alcohol trying to avoid leaving any excess cleansing agent on the stopper.
- Open the Mix2Vial® package by peeling away the lid (Figure 1). Leave the Mix2Vial in the clear outer packaging.
- Place the diluent vial upright on an even surface and hold the vial tight and pick up the Mix2Vial in its clear outer packaging. Holding the diluent vial securely, push the blue end of the Mix2Vial vertically down through the diluent vial stopper (Figure 2).
- While holding onto the diluent vial, carefully remove the clear outer packaging from the Mix2Vial set, ensuring the Mix2Vial remains attached to the diluent vial (Figure 3).
- Place the product vial upright on an even surface, invert the diluent vial with the Mix2Vial attached.
- While holding the product vial securely on a flat surface, push the clear end of the Mix2Vial set vertically down through the product vial stopper (Figure 4). The diluent will automatically transfer out of its vial into the product vial. (NOTE: If the Mix2Vial is connected at an angle, the vacuum may be released from the product vial and the diluent will not transfer into the product vial.)
- With the diluent and product vials still attached to the Mix2Vial, gently swirl the product vial to ensure the product is fully dissolved (Figure 5). Reconstitution requires less than 5 minutes. Do not shake the vial.
- Disconnect the Mix2Vial into two separate pieces (Figure 6) by holding each vial adapter and twisting counterclockwise. After separating, discard the diluent vial with the blue end of the Mix2Vial.
- Draw air into an empty, sterile syringe. Keeping the product vial upright with the clear end of the Mix2Vial attached, screw the disposable syringe onto the luer lock portion of the Mix2Vial device by pressing and twisting clockwise. Inject air into the product vial.
- While keeping the syringe plunger depressed, invert the system upside down and draw the reconstituted product into the syringe by pulling the plunger back slowly (Figure 7).
- When the reconstituted product has been transferred into the syringe, firmly hold the barrel of the syringe and the clear vial adapter (keeping the syringe plunger facing down) and unscrew the syringe from the Mix2Vial (Figure 8). Hold the syringe upright and push the plunger until no air is left in the syringe. Attach the syringe to a venipuncture set.
- NOTE: If the same patient is to receive more than one vial of concentrate, the contents of two vials may be drawn into the same syringe through a separate unused Mix2Vial set before attaching to the venipuncture set.
- Use the prepared drug as soon as possible within 3 hours after reconstitution.
- After reconstitution, parenteral drug products should be inspected visually for particulate matter and discoloration prior to administration, whenever solution and container permit. When reconstitution procedure is strictly followed, a few small particles may occasionally remain. The Mix2Vial set will remove particles and the labeled potency will not be reduced.
- Discard all administration equipment after use into the appropriate safety container. Do not reuse.
Administration
Instruction for Use:
Alphanate is for intravenous use only after reconstitution. Use plastic disposable syringes. Do not refrigerate after reconstitution. Reconstituted Alphanate may be stored at room temperature (not to exceed 30 °C) prior to administration, but administer intravenously within three hours.
Discard any unused contents into the appropriate safety container.
Do not administer Alphanate at a rate exceeding 10 mL/minute.
Dosage Forms and Strengths
Alphanate is a sterile, lyophilized powder for intravenous injection after reconstitution. It is available in the following potencies:
250 IU FVIII/5 mL single dose vial
500 IU FVIII/5 mL single dose vial
1000 IU FVIII/10 mL single dose vial
1500 IU FVIII/10 mL single dose vial
Contraindications
Alphanate is contraindicated in patients who have manifested life-threatening immediate hypersensitivity reactions, including anaphylaxis, to the product or its components.
Warnings and Precautions
Anaphylaxis and Severe Hypersensitivity Reactions
Anaphylaxis and severe hypersensitivity reactions are possible. Should symptoms occur, treatment with Alphanate should be discontinued, and emergency treatment should be administered.
Neutralizing Antibodies
Development of procoagulant activity-neutralizing antibodies (inhibitors) has been detected in patients receiving FVIII-containing products. Carefully monitor patients treated with AHF products for the development of FVIII inhibitors by appropriate clinical observations and laboratory tests. No studies have been conducted with Alphanate to evaluate inhibitor formation. Therefore, it is not known whether there are greater, lesser or the same risks of developing inhibitors due to the use of this product than there are with other FVIII preparations. If expected plasma FVIII activity levels are not attained, or if bleeding is not controlled with an appropriate dose, an assay that measures FVIII inhibitor concentration should be performed. Patients with these inhibitors may not respond to treatment with Antihemophilic Factor/von Willebrand Factor Complex (Human), or the response may be much less than would otherwise be expected; therefore, larger doses of Antihemophilic Factor/von Willebrand Factor Complex (Human) are often required. The management of bleeding in patients with inhibitors requires careful monitoring, especially if surgical procedures are indicated.4-6 Depending on the level of the inhibitor and/or clinical response, it may be appropriate to use an alternative ‘bypass’ therapeutic agent.
Reports in the literature suggest that patients with Type 3, severe von Willebrand Disease, may develop alloantibodies to von Willebrand factor (VWF) after replacement therapy.13 The risk of developing alloantibodies in patients with von Willebrand disease due to the use of this product is not known.
Thromboembolic Events
Thromboembolic events have been reported in von Willebrand Disease patients receiving AHF/VWF Complex (Human) replacement therapy, especially in the setting of known risk factors for thrombosis.14-16 In addition, endogenous high levels of FVIII have also been associated with thrombosis but no causal relationship has been established. In all VWD patients in situations of high thrombotic risk receiving coagulation factor replacement therapy, caution should be exercised and antithrombotic measures should be considered. See also ADVERSE REACTIONS (6).
Intravascular Hemolysis
Massive doses of AHF/VWF Complex (Human) have rarely resulted in acute hemolytic anemia, increased bleeding tendency or hyperfibrinogenemia.17 Alphanate contains blood group specific isoagglutinins and, when large and/or frequent doses are required in patients of blood groups A, B, or AB, the patient should be monitored for signs of intravascular hemolysis and falling hematocrit. Should this condition occur, thus leading to progressive hemolytic anemia, the administration of serologically compatible Type O red blood cells should be considered, the administration of Alphanate should be discontinued, and alternative therapy should be considered.
Vasomotor Reactions
Rapid administration of a FVIII concentrate may result in vasomotor reactions. Alphanate should not be administered at a rate exceeding 10 mL/minute.
Transmissible Infectious Agents
Because Alphanate is made from pooled human plasma, it may carry a risk of transmitting infectious agents, e.g., viruses, and theoretically, the Creutzfeldt-Jakob Disease (CJD) agent. Stringent procedures designed to reduce the risk of adventitious agent transmission have been employed in the manufacture of this product, from the screening of plasma donors and the collection and testing of plasma, through the application of viral elimination/reduction steps such as solvent detergent and heat treatment in the manufacturing process.18,19 Despite these measures, such products can still potentially transmit disease; therefore, the risk of infectious agents cannot be totally eliminated. See also DESCRIPTION (11).
Adverse Reactions
Serious adverse reactions observed in patients receiving Alphanate are anaphylaxis/hypersensitivity reactions. Thomboembolic events have also been observed in patients receiving Alphanate for VWD. See also WARNINGS AND PRECAUTIONS (5.3).
The most frequent adverse events reported with Alphanate in > 5% of patients are respiratory distress, pruritus, rash, urticaria, face edema, paresthesia, pain, fever, chills, joint pain and fatigue.
Clinical Trial Experience
Because clinical trials are conducted under widely varying conditions, adverse reaction rates observed in the clinical trials of a drug cannot be directly compared to rates in clinical trials of another drug and may not reflect the rates observed in clinical practice.
Hemophilia A
In a clinical study with Alphanate, regardless of causality, 14 of 23 (60.9%) patients experienced a total of 47 adverse events. Twenty-three (48.9%) of the AEs were mild, 19 (40.4%) were moderate and 5 (10.6%) were severe.
Two of 23 (8.7%) patients experienced 10 serious AEs during the study. None of the SAEs were considered to be related to study drug.
The most common AEs were pain (combined terms), with 14 episodes occurring in 7 patients, and headache, with 3 episodes occurring in 2 patients. None of the symptoms were judged to be related to treatment (see Table 3).
| Adverse Event | Number of Episodes | Number of Subjects with signs/symptoms | % |
Body as a whole | 23 | 11 | 47.8% |
| Pain (combined terms) | 14 | ||
| Headache | 3 | ||
| Accidental injury | 2 | ||
| Asthenia | 1 | ||
Cellulitis | 1 | ||
| Chest pain | 1 | ||
| Flu syndrome | 1 | ||
Digestive system | 6 | 4 | 17.4% |
| Dyspepsia | 1 | ||
| GI disorder | 1 | ||
| Hepatitis | 1 | ||
| Nausea | 1 | ||
| Tooth disorder | 1 | ||
| Vomiting | 1 | ||
Respiratory system | 5 | 4 | 17.4% |
| Cough increased | 1 | ||
| Lung disorder | 1 | ||
| Pharyngitis | 1 | ||
| Respiratory disorder | 1 | ||
| Rhinitis | 1 | ||
| Musculoskeletal system | 3 | 3 | 13.0% |
| Bone disorder | 2 | ||
| Bone necrosis | 1 | ||
Skin and appendages | 3 | 3 | 13.0% |
| Acne | 1 | ||
| Dry skin | 1 | ||
| Sweating | 1 | ||
Nervous system | 2 | 2 | 8.7% |
Insomnia | 1 | ||
| Somnolence | 1 | ||
Hemic and lymphatic system | 2 | 2 | 8.7% |
| Anemia | 1 | ||
| Ecchymosis | 1 | ||
Urogenital system | 2 | 2 | 8.7% |
| Abnormal ejaculation | 1 | ||
| Cutaneous moniliasis | 1 | ||
Special senses | 1 | 1 | 4.3% |
| Eye disorder | 1 |
VWD
In the prospective clinical study of Alphanate (A-SD/HT)** in patients with von Willebrand Disease, adverse events occurred in 18 of 36 (50.0%) subjects (irrespective of causality) and 53 of 204 (26.0%) infusions. Most of the AEs were unrelated to study drug, however, and the proportion of subjects experiencing an AE possibly, probably, or definitely related to study drug was 5 of 36 subjects (13.9%) treated with Alphanate.
The proportion of subjects with at least one serious AE, regardless of causality was 3 of 36 subjects (8.3%). There were no subjects who reported at least one serious AE possibly, probably, or definitely related to study drug.
Overall, AEs, regardless of causality, were observed in association with 53 of 204 (26.0%) infusions of Alphanate across all parts of these studies. Most AEs were unrelated to study drug, however, and the proportion of infusions associated with AEs possibly, probably, or definitely related to study drug was 14 of 204 infusions (6.9%).
The proportion of infusions associated with serious AEs, regardless of causality, was only 5 of 204 (2.5%) infusions of Alphanate. There were no observed serious AEs possibly, probably or definitely related to study drug.
**Alphanate, Solvent Detergent, non heat-treated (A-SD), is the previous formulation and Alphanate, Solvent Detergent/Heat Treated (A-SD/HT), is the current formulation. Both products are biochemically equivalent and demonstrate similar in vivo pharmacokinetic profiles. Both products are also similarly effective for the treatment of bleeding episodes and provide adequate hemostasis for surgical and invasive procedures, even the absence of bleeding time correction, in subjects with moderate and severe VWD.
The most common AEs regardless of causality are listed in Table 4.
| Adverse Event | Number of Episodes | Number of Subjects with signs/symptoms | % of Subjects |
Body as a whole | 42 | 14 | 33.3% |
| Pain (combined terms) | 31 | ||
| Headache | 4 | ||
| Edema face | 3 | ||
| Fever | 2 | ||
Digestive system | 16 | 10 | 27.8% |
| Nausea | 12 | ||
| Constipation | 2 | ||
| Vomiting | 2 | ||
Skin and appendages | 10 | 7 | 19.4% |
| Puritis | 6 | ||
| Rash | 4 | ||
Respiratory system | 4 | 2 | 5.6% |
Respiratory disorder | 4 | ||
Nervous system | 6 | 2 | 5.6% |
| Paresthesia | 3 | ||
| Anxiety | 3 | ||
Hemic and Lymphic system | 3 | 1 | 2.8% |
| Anemia | 3 |
One incident of pulmonary embolus was reported that was considered to have a possible relationship to the product. This subject received the dose of 60 IU VWF:RCo/kg body weight and the FVIII:C level achieved was 290%.
In the retrospective study conducted to determine the efficacy and safety of Alphanate (A-SD/HT) in a surgical or invasive procedure setting as peri-operative prophylaxis against excessive bleeding, see CLINICAL STUDIES (14), 3 out of 39 subjects (7.7%) experienced 6 adverse drug reactions. Four were considered mild and 2 were considered moderate. No subject discontinued their treatment due to an adverse reaction. The adverse drug reactions were pruritus, paresthesia (2 events) and hemorrhage (all considered mild), and one event each of moderate hematocrit decrease and orthostatic hypotension.
One adverse event (pain) related to the treatment with heat-treated Alphanate (A-SD/HT) was reported on the four pediatric patients with von Willebrand Disease during the course of the prospective study and none of the five pediatric subjects in the retrospective clinical study.
Post-Marketing Experience
The following adverse reactions have been identified during post-approval use of Alphanate (A-SD/HT). Because these reactions are reported voluntarily from a population of uncertain size, it is not always possible to reliably estimate their frequency or establish a causal relationship to drug exposure.
Among patients treated with Alphanate (A-SD/HT), cases of allergic/hypersensitivity reactions (including urticaria, rash, pruritus, chest tightness shortness of breath, wheezing, flushing, palpitations, nausea, and vomiting) have been reported.
The following represents the most frequently reported adverse reactions: fever, chills, headache, joint pain, and fatigue. In addition, one case was reported for swelling of the parotid gland, pulmonary embolus, femoral venous thrombosis, seizure, and brief cardiorespiratory arrest.
Drug Interactions
None known.
USE IN SPECIFIC POPULATIONS
Pregnancy
Pregnancy Category C. Animal reproduction studies have not been conducted with Alphanate. It is also not known whether Alphanate can cause fetal harm when administered to a pregnant woman or affect reproductive capacity. Alphanate should be given to a pregnant woman only if clearly needed.
Labor and Delivery
No human or animal data. Use only if clearly needed.
Nursing Mothers
No human or animal data. Use only if clearly needed.
Pediatric Use
8.4.1 Hemophilia A Indication
Clinical trials for safety and effectiveness in pediatric hemophilia A patients 16 years of age and younger have not been conducted.
8.4.2 VWD Indication
The hemostatic efficacy of Alphanate has been studied in 20 pediatric subjects with VWD 18 years of age and under. Based on the data from a subset of these subjects, age had no effect on the pharmacokinetics of VWF:RCo. There were no clinically important differences between pediatric patients and adults.
Geriatric Use
No human or animal data. Use only if clearly needed.
Alphanate Description
Alphanate is a sterile, lyophilized concentrate of FVIII (AHF) and von Willebrand Factor (VWF).
Alphanate is prepared from pooled human plasma by cryoprecipitation of FVIII, fractional solubilization, and further purification employing heparin-coupled, cross-linked agarose which has an affinity to the heparin binding domain of VWF/FVIII:C complex.20 The product is treated with a mixture of tri-n-butyl phosphate (TNBP) and polysorbate 80 to inactivate enveloped viruses. The product is also subjected to an 80 °C heat treatment step for 72 hours to inactivate enveloped and non-enveloped viruses. However, no procedure has been shown to be totally effective in removing viral infectivity from coagulation factor products.
Alphanate is labeled with the antihemophilic factor potency (FVIII:C activity) in International Units (IU) FVIII/vial and with VWF:RCo activity expressed in IU VWF:RCo/vial. The activities are referenced to their respective international standards established by the World Health Organization. One IU of FVIII or one IU of VWF:RCo is approximately equal to the amount of FVIII or VWF:RCo activity in 1 mL of freshly-pooled human plasma.
Alphanate contains Albumin (Human) as a stabilizer, resulting in a final container concentrate with a specific activity of at least 5 FVIII:C IU/mg total protein. Prior to the addition of the Albumin (Human) stabilizer, the specific activity is significantly higher. When reconstituted as directed, the composition of Alphanate is as follows:
| Component | Concentration |
| FVIII:C activity | 40 – 180 IU/mL |
| VWF:RCo activity | NLT 0.4 VWF:RCo IU per 1 IU of FVIII:C |
| Albumin (Human) | 0.3 – 0.9 g/100 mL |
| Calcium | NMT 5 mmol/L |
| Glycine | NMT 750 μg per FVIII:C IU |
| Heparin | NMT 1.0 U/mL |
| Histidine | 10 – 40 mmol/L |
| Imidazole | NMT 0.1 mg/mL |
| Arginine | 50 – 200 mmol/L |
| Polyethylene Glycol and Polysorbate 80 | NMT 1.0 μg per FVIII:C IU |
| Sodium | NMT 10 mEq/vial |
| Tri-n-butyl Phosphate (TNBP) | NMT 0.1 μg per FVIII:C IU |
NMT= not more than
NLT= not less than
Viral Reduction Capacity
The solvent detergent treatment process has been shown by Horowitz, et al., to provide a high level of viral inactivation without compromising protein structure and function.21 The susceptibility of human pathogenic viruses such as Human Immunodeficiency viruses (HIV), hepatitis viruses, as well as marker viruses such as Sindbis virus (SIN, a model for Hepatitis C virus) and Vesicular Stomatitis virus (VSV, a model for large, enveloped RNA virus), to inactivation by organic solvent detergent treatment has been discussed in the literature.22
In vitro inactivation studies to evaluate the solvent detergent treatment (0.3% Tri-n-butyl Phosphate and 1.0% Polysorbate 80) step in the manufacture of Alphanate demonstrated a log inactivation of ≥ 11.1 for HIV-1, ≥ 6.1 for HIV-2, ≥ 4.1 for VSV and ≥ 4.7 for SIN. No residual virus was detected after solvent detergent treatment in any of these studies.
Additional steps in the manufacturing process of Alphanate were evaluated for virus elimination capability. The dry heat cycle of 80 °C for 72 hours was shown to inactivate greater than 5.8 logs of Hepatitis A virus (HAV). Precipitation with 3.5% polyethylene glycol (PEG) and heparin-actigel-ALD chromatography are additional steps studied using Bovine Herpes virus (BHV, a model for Hepatitis B virus), Bovine Viral Diarrhea virus (BVD, a second model for Hepatitis C virus), human Poliovirus Sabin type 2 (POL, a model for Hepatitis A virus), Canine Parvovirus (CPV, a model for Parvovirus B19) and HIV-1.
Table 5 summarizes the reduction factors for each virus validation study performed for the manufacturing process of Alphanate.
Virus (Model Virus for) | 3.5% PEG Precipitation | Solvent–Detergent | Column Chromatography | Lyophilization | Dry Heat Cycle (80 °C, 72 h) | Total Log Removal |
| BHV (HBV) | < 1.0 | ≥ 8.0 | 7.6 | 1.3 | 2.1 | ≥ 19.0 |
| BVD (HCV) | < 1.0 | ≥ 4.5 | < 1.0 | < 1.0 | ≥ 4.9 | ≥ 9.4 |
| POL (HAV) | 3.3 | – | < 1.0 | 3.4 | ≥ 2.5 | ≥ 9.2 |
| CPV (B19) | 1.2 | – |
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