# Patent

## Title
Treatment of coagulation disease by administration of recombinant VWF

## Abstract

The present invention provides methods of treating coagulation disease, including hemophilia and von Willebrand disease by administering recombinant von Willebrand Factor alone or in combination with Factor VIII.

## Background
CROSS-REFERENCES TO RELATED APPLICATIONS
The present application claims the benefit of U.S. Patent Application No. 61/495,884, filed Jun. 10, 2011, U.S. Patent Application No. 61/511,901, filed Jul. 26, 2011, and U.S. Patent Application No. 61/523,790, filed Aug. 15, 2011, the disclosures of which are expressly incorporated herein by reference in their entireties for all purposes.


BACKGROUND OF THE INVENTION
Coagulation diseases, such as von Willebrand Disease (VWD) and Hemophilia, generally result from a deficiency in the coagulation cascade. “von Willebrand Disease” refers to the group of diseases caused by a deficiency of von Willebrand factor. Von Willebrand factor helps blood platelets clump together and stick to the blood vessel wall, which is necessary for normal blood clotting. Hemophilia A refers to a deficiency of Factor VIII activity, whereas Hemophilia B refers to a Factor IX deficiency. Current treatment for these coagulopathies includes a replacement therapy using pharmaceutical preparations comprising the normal coagulation factor.
Replacement therapy in VWD and Hemophilia A patients involves the repeated administration of preparations containing normal coagulation factors via intravenous infusion, which can constitute a heavy load on the life of these patients, particularly when venous access is difficult to achieve. It would be advantageous if the frequency of infusions could be reduced. One potentially viable therapy is to stabilize Factor VIII through its association with a second molecule, such as von Willebrand Factor (VWF), with the result that plasma half-life of Factor VIII is increased.
VWF is a glycoprotein circulating in plasma as a series of multimers ranging in size from about 500 to 20,000 kD. The full length of cDNA of VWF has been cloned; the propolypeptide corresponds to amino acid residues 23 to 764 of the full length prepro-VWF (Eikenboom et al (1995) Haemophilia 1, 77 90). Multimeric forms of VWF are composed of 250 kD polypeptide subunits linked together by disulfide bonds. VWF mediates the initial platelet adhesion to the sub-endothelium of the damaged vessel wall, with the larger multimers exhibiting enhanced hemostatic activity. Multimerized VWF binds to the platelet surface glycoprotein Gp1bα, through an interaction in the A1 domain of VWF, facilitating platelet adhesion. Other sites on VWF mediate binding to the blood vessel wall. Thus, VWF forms a bridge between the platelet and the vessel wall that is essential to platelet adhesion and primary hemostasis under conditions of high shear stress. Normally, endothelial cells secrete large polymeric forms of VWF and those forms of VWF that have a lower molecular weight arise from proteolytic cleavage. The multimers of exceptionally large molecular masses are stored in the Weibel-Pallade bodies of the endothelial cells and liberated upon stimulation by agonists such as thrombin and histamine.
That FVIII pharmacokinetics are a function of VWF levels is supported by several previous observations. Reduction of FVIII binding activity in von Willebrand Disease (VWD), due to either reduced VWF protein levels or lowered FVIII binding affinity, results in reduced steady-state levels of endogenous FVIII (summarized in Castaman et al., Disorders of Hemostasis 88(1):94-108 (2003), and improving survival of VWF has been proposed as a viable strategy for improving FVIII stability (Denis et al., Thromb Haemost. 2008 February; 99(2):271-8; Turecek et al., Blood, 2006, 108(11): Abstract 1002). Among severe Hemophilia A patients, a correlation between pre-infusion VWF levels and the half-life of infused FVIII has been demonstrated by Fijnvandraat and colleagues (Fijnvandraat, et al., Br J Haematol. 1995 October; 91(2):474-6). In that study, patients with 200-300% of average VWF levels were seen to have a FVIII half-life of 15-29 hours compared to a mean of 12.5 hours in patients with normal VWF levels. In another study, patients with blood group O were demonstrated to have significantly lower VWF levels and shorter FVIII half-lives (15.3 hours) compared with those with blood group A (19.7 hours) (Vlot, et al. Thromb Haemost. 2000 January; 83(1):65-9). Chemically modified VWF has been shown to prolong survival of rFVIII (Turecek et al., J. Thromb. Haemost. 2007 Jul. 9; 5(2) abstract available at: http/www.blackwellpublishing.com/isth2007/abstract.asp?id=64898). As such, co-administration of rVWF and rFVIII is a viable strategy for the treatment of coagulation diseases such as von Willebrand Disease and Hemophilia A.
BRIEF SUMMARY OF THE INVENTION
Accordingly, the present invention provides methods and combinations for treating coagulation disease by administering recombinant von Willebrand Factor (rVWF) alone or in combination with recombinant Factor VIII (rFVIII) to a subject in need thereof, with the result that the in-vivo half-life of Factor VIII is increased.
In one aspect, the present invention provides a method for treating Von Willebrand Disease or Hemophilia A in a subject in need thereof, the method comprising: administering to the subject recombinant Von Willebrand Factor (rVWF) such that Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein the rVWF is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, and wherein the rVWF has a higher specific activity than plasma derived Von Willebrand Factor.
In further embodiments and in accordance with the above, methods of the invention include co-administering to the subject recombinant Von Willebrand Factor (rVWF) and recombinant Factor VIII (rFVIII).
In further embodiments and in accordance with any of the above, the rVWF and rFVIII are administered together in a single composition.
In further embodiments and in accordance with any of the above, the subject is administered between 1.0 IU/kg VWF:RCo and 150 IU/kg VWF:RCo per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 2 IU/kg VWF:RCo and 50 IU/kg VWF:RCo per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 5 IU/kg VWF:RCo and 40 IU/kg VWF:RCo per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 10 IU/kg VWF:RCo and 20 IU/kg VWF:RCo per dose.
In further embodiments and in accordance with any of the above, the rVWF used in methods of the invention is matured in vitro by treatment with Furin.
In further embodiments and in accordance with any of the above, the rVWF is produced through expression in a Chinese Hamster Ovary (CHO cell culture).
In further embodiments and in accordance with any of the above, the rFVIII and rVWF are produced through expression in the same cell culture.
In further embodiments, and in accordance with any of the above, the subject is administered rVWF no more than once every other day.
In further embodiments and in accordance with any of the above, the subject is administered rVWF no more than twice a week.
In further aspects and in accordance with any of the above, the high molecular weight VWF multimer composition maintains the at least 20% VWF decamers or higher order multimers for at least 3 hours post-administration.
In further embodiments and in accordance with any of the above, the Factor VIII half-life is extended by about 5 hours.
In further embodiments and in accordance with any of the above, the Factor VIII half-life is extended for at least 12 hours.
In further embodiments and in accordance with any of the above, the Factor VIII half-life is extended for at least 24 hours.
In further embodiments and in accordance with any of the above, the Factor VIII half-life is extended for at least 36 hours.
In further embodiments and in accordance with any of the above, the Factor VIII half-life is extended for at least 48 hours.
In further embodiments and in accordance with any of the above, wherein the Factor VIII half-life is extended for at least 72 hours.
In further embodiments and in accordance with any of the above, the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 2:1 and 1:4.
In further embodiments and in accordance with any of the above, the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 3:2 and 1:3.
In further embodiments and in accordance with any of the above, the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 1:1 and 1:2.
In further embodiments and in accordance with any of the above, 23 the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is about 3:4.
In further embodiments and in accordance with any of the above, the rVWF has a specific activity of about 20-150 mU/μg.
In further embodiments and in accordance with any of the above, the high molecular weight VWF multimer composition comprises at least 30% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the high molecular weight VWF multimer composition comprises at least 40% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the high molecular weight VWF multimer composition comprises at least 50% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the high molecular weight VWF multimer composition comprises at least 60% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the high molecular weight VWF multimer composition comprises at least 70% VWF decamers or higher order multimers.
In further aspects and in accordance with any of the above, the present invention provides a method for treating Hemophilia A or Von Willebrand Disease in a subject in need thereof, the method comprising: administering to the subject recombinant Von Willebrand Factor (rVWF) such that Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein: (a) the rVWF has a higher specific activity than plasma derived Von Willebrand Factor, wherein the specific activity of rVWF is about 20-150 mU/μg; and (b) the FVIII half-life is at least 1.5 times higher as compared to FVIII half-life in a subject administered plasma derived Von Willebrand Factor.
In further aspects and in accordance with any of the above, the present invention provides a method for treating Hemophilia A or Von Willebrand Disease in a subject in need thereof, the method comprising: administering to the subject recombinant Von Willebrand Factor (rVWF) such that Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein: (a) the rVWF is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, (b) the rVWF has a higher specific activity than plasma derived Von Willebrand Factor, wherein the specific activity of rVWF is at least about 20-150 mU/μg; and (c) the FVIII half-life is at least 1.5 times higher as compared to FVIII half-life in a subject administered plasma derived Von Willebrand Factor.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24 hours post-administration of the rVWF is at least 90% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24 hours post-administration is at least 100% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 36 hours post-administration is at least 80% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 48 hours post-administration is at least 50% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 6 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 12 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 18 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 24 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 36 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 48 hours post-administration.
In further embodiments and in accordance with any of the above, the higher order rVWF multimers are stable for at least 72 hours post-administration.
In further aspects and in accordance with any of the above, the present invention provides a method for treating Hemophilia A or Von Willebrand Disease in a subject in need thereof, the method comprising: administering to the subject recombinant Von Willebrand Factor (rVWF).
In further embodiments and in accordance with any of the above, the method comprises co-administering to the subject recombinant Factor VIII (rFVIII) and recombinant Von Willebrand Factor (rVWF).
In further embodiments and in accordance with any of the above, the rFVIII and rVWF are administered together in a single composition.
In further embodiments and in accordance with any of the above, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 2:1 and 1:4.
In further embodiments and in accordance with any of the above, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 3:2 and 1:3.
In further embodiments and in accordance with any of the above, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is between 1:1 and 1:2.
In further embodiments and in accordance with any of the above, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) administered to the subject is about 3:4.
In further embodiments and in accordance with any of the above, the rVWF is matured in vitro by treatment with Furin.
In further embodiments and in accordance with any of the above, the rVWF is expressed in mammalian cell culture.
In further embodiments and in accordance with any of the above, the rFVIII is expressed in mammalian cell culture.
In further embodiments and in accordance with any of the above, the mammalian culture comprises CHO cells.
In further embodiments and in accordance with any of the above, the rFVIII and rVWF are co-expressed in the same culture.
In further embodiments and in accordance with any of the above, the rFVIII and rVWF are co-purified.
In further embodiments and in accordance with any of the above, the rFVIII and rVWF are purified separately.
In further embodiments and in accordance with any of the above, a rFVIII/rVWF complex is reconstituted prior to administration.
In further embodiments and in accordance with any of the above, rVWF is treated with Furin prior to reconstituting the rFVIII/rVWF complex.
In further embodiments and in accordance with any of the above, the reconstituted rFVIII/rVWF complex is treated with Furin.
In further embodiments and in accordance with any of the above, the Furin is recombinant Furin.
In further embodiments and in accordance with any of the above, the subject is administered rVWF no more than once daily.
In further embodiments and in accordance with any of the above, the subject is administered rVWF no more than once every other day.
In further embodiments and in accordance with any of the above, the subject is co-administered rVWF no more than once every third day.
In further embodiments and in accordance with any of the above, the subject is administered rVWF no more than once every fourth day.
In further embodiments and in accordance with any of the above, the subject is administered between 1.5 IU/kg FVIII:C and 150 IU/kg FVIII:C per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 10 IU/kg FVIII:C and 100 IU/kg FVIII:C per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 25 IU/kg FVIII:C and 75 IU/kg FVIII:C per dose.
In further embodiments and in accordance with any of the above, the subject is administered between 40 IU/kg FVIII:C and 60 IU/kg FVIII:C per dose.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24 hours post-administration is at least 90% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24 hours post-administration is at least 100% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 36 hours post-administration is at least 80% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 48 hours post-administration is at least 50% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
In further embodiments and in accordance with any of the above, the rVWF administered to the subject has a HMW VWF multimer composition comprising at least 10% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the rVWF administered to the subject has a HMW VWF multimer composition comprising at least 20% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the rVWF administered to the subject has a HMW VWF multimer composition comprising at least 30% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the rVWF administered to the subject has a HMW VWF multimer composition comprising at least 40% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, the rVWF administered to the subject has a HMW VWF multimer composition comprising at least 50% VWF decamers or higher order multimers.
In further embodiments and in accordance with any of the above, higher order rVWF multimers are stable in vitro for at least 3 hours post-administration.
In further embodiments and in accordance with any of the above, higher order rVWF multimers are stable in vitro for at least 6 hours post-administration.
In further embodiments and in accordance with any of the above, wherein higher order rVWF multimers are stable in vitro for at least 12 hours post-administration.
In further embodiments and in accordance with any of the above, wherein higher order rVWF multimers are stable in vitro for at least 18 hours post-administration.
In further embodiments and in accordance with any of the above, higher order rVWF multimers are stable in vitro for at least 24 hours post-administration.
In further embodiments and in accordance with any of the above, higher order rVWF multimers are stable in vitro for at least 36 hours post-administration.
In further embodiments and in accordance with any of the above, wherein higher order rVWF multimers are stable in vitro for at least 48 hours post-administration.
In further embodiments and in accordance with any of the above, higher order rVWF multimers are stable in vitro for at least 72 hours post-administration.
In further embodiments and in accordance with any of the above, endogenous FVIII activity is stabilized for at least 12 hours.
In further embodiments and in accordance with any of the above, endogenous FVIII activity is stabilized for at least 24 hours.
In further embodiments and in accordance with any of the above, endogenous FVIII activity is stabilized for at least 36 hours.
In further embodiments and in accordance with any of the above, endogenous FVIII activity is stabilized for at least 48 hours.
In further embodiments and in accordance with any of the above, endogenous FVIII activity is stabilized for at least 72 hours.
In further embodiments and in accordance with any of the above, wherein co-administered rFVIII activity is stabilized for at least 12 hours.
In further embodiments and in accordance with any of the above, co-administered rFVIII activity is stabilized for at least 24 hours.
In further embodiments and in accordance with any of the above, co-administered rFVIII activity is stabilized for at least 36 hours.
In further embodiments and in accordance with any of the above, co-administered rFVIII activity is stabilized for at least 48 hours.
In further embodiments and in accordance with any of the above, co-administered rFVIII activity is stabilized for at least 72 hours.
In further embodiments and in accordance with any of the above, co-administered rFVIII activity is stabilized by extending the half-life of the rFVIII in vivo.
In further embodiments and in accordance with any of the above, FVIII half-life is extended by about five hours as compared to a patient administered pdFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 36 hours in a patient administered said rFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 48 hours in a patient administered said rFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 72 hours in a patient administered said rFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 90 hours in a patient administered said rFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 120 hours in a patient administered said rFVIII.
In further embodiments and in accordance with any of the above, at least 1% of the co-administered rFVIII activity is maintained for at least 168 hours in a patient administered said rFVIII.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1. A schematic of the study design assessing tolerability and safety after single doses of rVWF:rFVIII.
FIG. 2. Pharmacokinetic data. (A) provides PK data for rVWF/rFVIII and pdVWF/pdFVIII. (B) provides data on progressive loss of high molecular weight rVWF upon exposure to ADAMTS13.
FIG. 3. Pharmacokinetic data showing FVIII PK for rVWF/rFVIII and pdVWF/pdFVIII.
FIG. 4. Table summarizing patient demographics for the study.
FIG. 5. Pharmacokinetic data of rVWF/rFVIII and pdVWF/pdFVIII treatment of Cohort 4A.
FIG. 6. SDS-PAGE data on VWF multimer cleavage by ADAMTS13.
FIG. 7. Pharmacokinetic data of rVWF/rFVIII and pdVWF/pdFVIII treatment of Cohort 4A.
FIG. 8. Summary of adverse events from the study.
FIG. 9. rVWF PK parameters data from the study.
FIG. 10. rVWF PK data from Patient 1.
FIG. 11. rVWF PK data from Patient 2.
FIG. 12. rVWF PK data from Patient 3.
FIG. 13. Comparison of rVWF PK in Patients 1, 2 and 3—comparison of the VWF parameters and FVIII activity.
FIG. 14. FVIII activity across all cohorts in the study.
FIG. 15. rVWF PK in patients of Cohort 2—comparison of VWF parameters and FVIII activity.
FIG. 16. Summary of pharmacokinetic parameters for Factor VIII procoagulant activity (FVIII:C) for patients in Cohort 2.
FIG. 17. rVWF PK in all patients of Cohort 3—comparison of plasma parameters for VWF and FVIII.
FIG. 18. Summary of pharmacokinetic parameters for Factor VIII procoagulant activity (FVIII:C) for patients in Cohort 3.
FIG. 19. Data from patients in Cohort 4.
FIG. 20. Summary of pharmacokinetic parameters for Factor VIII procoagulant activity (FVIII:C) for patients in Cohort 4A.
FIG. 21. Pharmacokinetic data of FVIII:C comparing co-administered rVWF and rFVIII to Advate Pivotal.
FIG. 22. Antibody summary for a subject receiving co-administered pdVWF/pdFVIII or rVWF/rFVIII.



DETAILED DESCRIPTION OF THE INVENTION
Overview
The present invention provides compositions and methods for treating coagulation disease in a subject by administering recombinant von Willebrand Factor (rVWF) alone or in combination with Factor VIII (which can be recombinant or plasma derived). In some aspects, the compositions and methods of the present invention are used for treating coagulation diseases such as von Willebrand Disease (VWD) or Hemophilia A.
In one aspect, rVWF administered to the subject provides increased stability for in vivo Factor VIII (FVIII) activity as compared to FVIII stability due to administered plasma derived VWF, allowing for lower doses and/or frequency of treatment than in traditional treatments for coagulation diseases. Increased stability of FVIII activity and levels of FVIII can be measured using methods known in the art and described herein, including standard assays such as one-stage clotting assays, chromogenic assays, and immunoassays (see for example Lippi et al., Blood Coagulation & Fibrinolysis, 2009, 20(1): 1-3 and Chandler et al., Am J. Clin. Pathol., 2003, 120:34-39, each of which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to assays of FVIII level and activity). As shown in FIG. 20, average FVIII half-life was increased by rVWF by 5.2 hours over the half-life for patients receiving pdVWF. FIGS. 2 and 7 also show increases in FVIII activity in patients administered rVWF as compared to those administered pdVWF: FIG. 2 shows that average FVIII half-life was increased by 4.7 hours for patients administered rVWF as compared to patients administered pdVWF, and the data from Cohort 4A in FIG. 7 shows an increase in FVIII half-life of 5.3 hours for patients administered rVWF as compared to patients administered pdVWF. In addition, as shown in FIG. 21, the half-life of plasma FVIII is increased by 12.27 hours in VWD patients receiving rVWF in combination with rFVIII as compared to patients receiving FVIII (i.e., Advate) alone.
In a further aspect, the administration of rVWF stabilizes endogenous and/or co-administered FVIII activity, with the result that the in vivo half-life and/or activity of FVIII is increased. In embodiments in which rVWF and FVIII are co-administered, the rVWF and FVIII can be administered to the subject together in a single composition. In further embodiments, neither rVWF nor FVIII are modified with a water soluble polymer. In other embodiments, either the rVWF or FVIII or both are modified with a water soluble polymer. As will be appreciated, in embodiments in which rVWF is co-administered with FVIII, the FVIII may be recombinant or plasma derived.
In further aspects and in accordance with any of the above, the rVWF administered to the subject is a high molecular weight multimer composition comprising decamers or higher order multimers of rVWF. As discussed above, the use of rVWF compositions of the invention provide therapeutic flexibility to dose (or re-dose) with or without FVIII (recombinant or plasma derived). In further embodiments, the rVWF administered to the subject is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers. In specific embodiments, the rVWF administered to subjects is not modified with a water soluble polymer.
In still further aspects, the rVWF administered to the subject has a higher specific activity than pdVWF.
In a still further aspect, the rVWF alone or in combination with pdFVIII or rFVIII is administered to the subject no more than twice a week.
In a yet further aspect, the rVWF is processed with Furin prior to administration to the subject. In certain embodiments, the rVWF is processed with recombinant Furin.
In further aspects, the rVWF of use in the present invention is produced in accordance with methods known in the art and described for example in US 2012/0035110, filed Jul. 8, 2011 and U.S. Pat. No. 8,173,597, issued May 8, 2012, each of which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to rVWF compositions and methods for producing those compositions.
In accordance with any of the above, rVWF alone or in combination with FVIII is used to treat patients with coagulation diseases, such as VWD and Hemophilia A. Patients with VWD have some level of FVIII, but the stability of that FVIII is generally compromised because these patients lack VWF. Treatment of VWD patients may in some embodiments involve an initial treatment with both rVWF and rFVIII followed by repeated administrations of rVWF alone. In other embodiments, the initial treatment may be with rVWF alone while subsequent repeated administrations are with both rVWF and rFVIII, or the initial and subsequent repeat administrations may all include a co-administration of both rVWF and rFVIII. Similarly, Hemophilia A patients (who lack FVIII) may receive an initial treatment of both rVWF and rFVIII, and subsequent repeat treatments may comprise the administration of rFVIII alone or rVWF alone. In other embodiments, the initial treatment may be rFVIII alone while the subsequent repeat treatments involve co-administration of rVWF and rFVIII.
DEFINITIONS
As used herein. “rVWF” refers to recombinant VWF.
As used herein, “rFVIII” refers to recombinant FVIII.
The term “recombinant” when used with reference, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.
As used herein, “recombinant VWF” includes VWF obtained via recombinant DNA technology. In certain embodiments, VWF proteins of the invention can comprise a construct, for example, prepared as in WO 1986/06096 published on Oct. 23, 1986 and U.S. patent application Ser. No. 07/559,509, filed on Jul. 23, 1990, in the name of Ginsburg et al., which is incorporated herein by reference with respect to the methods of producing recombinant VWF. The VWF in the present invention can include all potential forms, including the monomeric and multimeric forms. It should also be understood that the present invention encompasses different forms of VWF to be used in combination. For example, the VWF of the present invention may include different multimers, different derivatives and both biologically active derivatives and derivatives not biologically active.
In the context of the present invention, the recombinant VWF embraces any member of the VWF family from, for example, a mammal such as a primate, human, monkey, rabbit, pig, rodent, mouse, rat, hamster, gerbil, canine, feline, and biologically active derivatives thereof. Mutant and variant VWF proteins having activity are also embraced, as are functional fragments and fusion proteins of the VWF proteins. Furthermore, the VWF of the invention may further comprise tags that facilitate purification, detection, or both. The VWF described herein may further be modified with a therapeutic moiety or a moiety suitable imaging in vitro or in vivo.
As used herein, “plasma-derived VWF (pdVWF)” includes all forms of the protein found in blood including the mature VWF obtained from a mammal having the property of in vivo-stabilizing, e.g. binding, of at least one FVIII molecule.
The term “highly multimeric VWF” or “high molecular weight VWF” refers to VWF comprising at least 10 subunits, or 12, 14, or 16 subunits, to about 20, 22, 24 or 26 subunits or more. The term “subunit” refers to a monomer of VWF. As is known in the art, it is generally dimers of VWF that polymerize to form the larger order multimers. (see Turecek et al., Semin. Thromb. Hemost. 2010, 36(5): 510-521 which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings regarding multimer analysis of VWF).
As used herein, the term “factor VIII” or “FVIII” refers to any form of factor VIII molecule with the typical characteristics of blood coagulation factor VIII, whether endogenous to a patient, derived from blood plasma, or produced through the use of recombinant DNA techniques, and including all modified forms of factor VIII. Factor VIII (FVIII) exists naturally and in therapeutic preparations as a heterogeneous distribution of polypeptides arising from a single gene product (see, e.g., Andersson et al., Proc. Natl. Acad. Sci. USA, 83:2979-2983 (1986)). Commercially available examples of therapeutic preparations containing Factor VIII include those sold under the trade names of HEMOFIL M, ADVATE, and RECOMBINATE (available from Baxter Healthcare Corporation, Deerfield, Ill., U.S.A.).
As used herein, “plasma FVIII activity” and “in vivo FVIII activity” are used interchangeably. The in vivo FVIII activity measured using standard assays may be endogenous FVIII activity, the activity of a therapeutically administered FVIII (recombinant or plasma derived), or both endogenous and administered FVIII activity. Similarly, “plasma FVIII” refers to endogenous FVIII or administered recombinant or plasma derived FVIII.
As used herein “von Willebrand Disease” refers to the group of diseases caused by a deficiency of von Willebrand factor. Von Willebrand factor helps blood platelets clump together and stick to the blood vessel wall, which is necessary for normal blood clotting. As described in further detail herein, there are several types of Von Willebrand disease.
As used herein, the terms “hemophilia” or “haemophilia” refer to a group of disease states broadly characterized by reduced blood clotting or coagulation. Hemophilia may refer to Type A, Type B, or Type C hemophilia, or to the composite of all three diseases types. Type A hemophilia (hemophilia A) is caused by a reduction or loss of factor VIII (FVIII) activity and is the most prominent of the hemophilia subtypes. Type B hemophilia (hemophilia B) results from the loss or reduction of factor IX (FIX) clotting function. Type C hemophilia (hemophilia C) is a consequence of the loss or reduction in factor XI (FXI) clotting activity. Hemophilia A and B are X-linked diseases, while hemophilia C is autosomal. Common treatments for hemophilia include both prophylactic and on-demand administration of clotting factors, such as FVIII, FIX, including Bebulin®-VH, and FXI, as well as FEIBA-VH, desmopressin, and plasma infusions.
The terms “isolated,” “purified,” or “biologically pure” refer to material that is substantially or essentially free from components that normally accompany it as found in its native state. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. VWF is the predominant species present in a preparation is substantially purified. The term “purified” in some embodiments denotes that a nucleic acid or protein gives rise to essentially one band in an electrophoretic gel. In other embodiments, it means that the nucleic acid or protein is at least 50% pure, more preferably at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more pure. “Purify” or “purification” in other embodiments means removing at least one contaminant from the composition to be purified. In this sense, purification does not require that the purified compound be homogenous, e.g., 100% pure.
As used herein, “administering” (and all grammatical equivalents) includes intravenous administration, intramuscular administration, subcutaneous administration, oral administration, administration as a suppository, topical contact, intraperitoneal, intralesional, or intranasal administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route including parenteral, and transmucosal (e.g., oral, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.
The terms “therapeutically effective amount or dose” or “therapeutically sufficient amount or dose” or “effective or sufficient amount or dose” refer to a dose that produces therapeutic effects for which it is administered. For example, a therapeutically effective amount of a drug useful for treating hemophilia can be the amount that is capable of preventing or relieving one or more symptoms associated with hemophilia. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).
As used herein, the terms “patient” and “subject” are used interchangeably and refer to a mammal (preferably human) that has a disease or has the potential of contracting a disease.
As used herein, the term “about” denotes an approximate range of plus or minus 10% from a specified value. For instance, the language “about 20%” encompasses a range of 18-22%.
As used herein, the term “half-life” refers to the period of time it takes for the amount of a substance undergoing decay (or clearance from a sample or from a patient) to decrease by half.
Compositions of the Invention
The present invention utilizes compositions comprising recombinant von Willebrand Factor (rVWF) for treatment of coagulation disease, such as VWD and Hemophilia A. In some embodiments, the present invention utilizes rVWF in combination with Factor VIII (FVIII). The co-administered FVIII may be recombinant (rFVIII) or plasma derived (pdFVIII). In preferred aspects, the compositions of the present invention stabilize in vivo Factor VIII activity (also referred to herein as plasma Factor VIII activity) such that the in vivo half-life of Factor VIII is extended as compared to that in subjects that have not been administered rVWF or that have been administered pdVWF. Measuring the extent to which rVWF stabilizes FVIII activity (including extension of FVIII half-life) can be carried out using methods known in the art. The level of FVIII activity can be measured by, for instance, one-stage clotting assays, chromogenic assays, and immunoassays (see for example Lippi et al., Blood Coagulation & Fibrinolysis, 2009, 20(1): 1-3, European Pharmacopoeia (Ph. Eur., 3.sup.rd Ed. 1997: 2.7.4), and Chandler et al., Am J. Clin. Pathol., 2003, 120:34-39, each of which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to assays of FVIII level and activity).
In certain embodiments, VWF proteins of the invention may comprise a construct, for example, prepared as in WO 1986/06096 published on Oct. 23, 1986 and U.S. patent application Ser. No. 07/559,509, filed on Jul. 23, 1990, in the name of Ginsburg et al., which is incorporated herein by reference with respect to the methods of producing recombinant VWF. The VWF useful for the present invention includes all potential forms, including the monomeric and multimeric forms. One particularly useful form of VWF are homo-multimers of at least two VWFs. The VWF proteins may be either a biologically active derivative, or when to be used solely as a stabilizer for FVIII the VWF may be of a form not biologically active. It should also be understood that the present invention encompasses different forms of VWF to be used in combination. For example, a composition useful for the present invention may include different multimers, different derivatives and both biologically active derivatives and derivatives not biologically active.
In primary hemostasis VWF serves as a bridge between platelets and specific components of the extracellular matrix, such as collagen. The biological activity of VWF in this process can be measured by different in vitro assays (Turecek et al., Semin. Thromb. Hemost. 28: 149-160, 2002). The ristocetin cofactor assay is based on the agglutination of fresh or formalin-fixed platelets induced by the antibiotic ristocetin in the presence of VWF. The degree of platelet agglutination depends on the VWF concentration and can be measured by the turbidimetric method, e.g. by use of an aggregometer (Weiss et al., J. Clin. Invest. 52: 2708-2716, 1973; Macfarlane et al., Thromb. Diath. Haemorrh. 34: 306-308, 1975). The second method is the collagen binding assay, which is based on ELISA technology (Brown et Bosak, Thromb. Res. 43: 303-311, 1986; Favaloro, Thromb. Haemost. 83: 127-135, 2000). A microtiter plate is coated with type I or III collagen. Then the VWF is bound to the collagen surface and subsequently detected with an enzyme-labeled polyclonal antibody. The last step is the substrate reaction, which can be photometrically monitored with an ELISA reader. As provided herein, the specific Ristocetin Cofactor activity of the VWF (VWF:RCo) of the present invention is generally described in terms of mU/μg of VWF, as measured using in vitro assays.
An advantage of the rVWF compositions of the present invention over pdVWF is that rVWF exhibits a higher specific activity than pdVWF. In some embodiments, the rVWF of the invention has a specific activity of at least about 20, 22.5, 25, 27.5, 30, 32.5, 35, 37.5, 40, 42.5, 45, 47.5, 50, 52.5, 55, 57.5, 60, 62.5, 65, 67.5, 70, 72.5, 75, 77.5, 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150 or more mU/μg. In one embodiment, rVWF used in the methods described herein has a specific activity of from 20 mU/μg to 150 mU/μg. In another embodiment, the VWF has a specific activity of from 30 mU/μg to 120 mU/μg. In another embodiment, the rVWF has a specific activity from 40 mU/μg to 90 mU/μg. In yet another embodiment, the rVWF has a specific activity selected from variations 1 to 133 found in Table 1.







TABLE 1







Exemplary embodiments for the specific activity of rVWF found


in the compositions and used in the methods provided herein.








(mU/μg)












20
Var. 1


22.5
Var. 2


25
Var. 3


27.5
Var. 4


30
Var. 5


32.5
Var. 6


35
Var. 7


37.5
Var. 8


40
Var. 9


42.5
Var. 10


45
Var. 11


47.5
Var. 12


50
Var. 13


52.5
Var. 14


55
Var. 15


57.5
Var. 16


60
Var. 17


62.5
Var. 18


65
Var. 19


67.5
Var. 20


70
Var. 21


72.5
Var. 22


75
Var. 23


77.5
Var. 24


80
Var. 25


82.5
Var. 26


85
Var. 27


87.5
Var. 28


90
Var. 29


92.5
Var. 30


95
Var. 31


97.5
Var. 32


100
Var. 33


105
Var. 34


110
Var. 35


115
Var. 36


120
Var. 37


125
Var. 38


130
Var. 39


135
Var. 40


140
Var. 41


145
Var. 42


150
Var. 43


 20-150
Var. 44


 20-140
Var. 45


 20-130
Var. 46


 20-120
Var. 47


 20-110
Var. 48


 20-100
Var. 49


20-90
Var. 50


20-80
Var. 51


20-70
Var. 52


20-60
Var. 53


20-50
Var. 54


20-40
Var. 55


 30-150
Var. 56


 30-140
Var. 57


 30-130
Var. 58


 30-120
Var. 59


 30-110
Var. 60


 30-100
Var. 61


30-90
Var. 62


30-80
Var. 63


30-70
Var. 64


30-60
Var. 65


30-50
Var. 66


30-40
Var. 67


 40-150
Var. 68


 40-140
Var. 69


 40-130
Var. 70


 40-120
Var. 71


 40-110
Var. 72


 40-100
Var. 73


40-90
Var. 74


40-80
Var. 75


40-70
Var. 76


40-60
Var. 77


40-50
Var. 78


 50-150
Var. 79


 50-140
Var. 80


 50-130
Var. 81


 50-120
Var. 82


 50-110
Var. 83


 50-100
Var. 84


50-90
Var. 85


50-80
Var. 86


50-70
Var. 87


50-60
Var. 88


 60-150
Var. 89


 60-140
Var. 90


 60-130
Var. 91


 60-120
Var. 92


 60-110
Var. 93


 60-100
Var. 94


60-90
Var. 95


60-80
Var. 96


60-70
Var. 97


 70-150
Var. 98


 70-140
Var. 99


 70-130
Var. 100


 70-120
Var. 101


 70-110
Var. 102


 70-100
Var. 103


70-90
Var. 104


70-80
Var. 105


 80-150
Var. 106


 80-140
Var. 107


 80-130
Var. 108


 80-120
Var. 109


 80-110
Var. 110


 80-100
Var. 111


80-90
Var. 112


 90-150
Var. 113


 90-140
Var. 114


 90-130
Var. 115


 90-120
Var. 116


 90-110
Var. 117


 90-100
Var. 118


100-150
Var. 119


100-140
Var. 120


100-130
Var. 121


100-120
Var. 122


100-110
Var. 123


110-150
Var. 124


110-140
Var. 125


110-130
Var. 126


110-120
Var. 127


120-150
Var. 128


120-140
Var. 129


120-130
Var. 130


130-150
Var. 131


130-140
Var. 132


140-150
Var. 133





Var. = Variation






The rVWF of the present invention is highly multimeric comprising about 10 to about 40 subunits. In further embodiments, the multimeric rVWF produced using methods of the present invention comprise about 10-30, 12-28, 14-26, 16-24, 18-22, 20-21 subunits. In further embodiments, the rVWF is present in multimers varying in size from dimers to multimers of over 40 subunits (>10 million Daltons). The largest multimers provide multiple binding sites that can interact with both platelet receptors and subendothelial matrix sites of injury, and are the most hemostatically active form of VWF. As shown in the multimer analysis in FIG. 2 (bottom panel), application of ADAMTS13 will cleave the ultra-large rVWF multimers over time, but during production (generally through expression in cell culture), rVWF compositions of the present invention are generally not exposed to ADAMTS13 and retain their highly multimeric structure.
In one embodiment, a rVWF composition used in the methods described herein has a distribution of rVWF oligomers characterized in that 95% of the oligomers have between 6 subunits and 20 subunits. In other embodiments, the a rVWF composition has a distribution of rVWF oligomers characterized in that 95% of the oligomers have a range of subunits selected from variations 458 to 641 found in Table 2.







TABLE 2







Exemplary embodiments for the distribution of rVWF oligomers


found in the compositions and used in the methods provided herein.








Subunits












 2-40
Var. 458


 2-38
Var. 459


 2-36
Var. 460


 2-34
Var. 461


 2-32
Var. 462


 2-30
Var. 463


 2-28
Var. 464


 2-26
Var. 465


 2-24
Var. 466


 2-22
Var. 467


 2-20
Var. 468


 2-18
Var. 469


 2-16
Var. 470


 2-14
Var. 471


 2-12
Var. 472


 2-10
Var. 473


2-8
Var. 474


 4-40
Var. 475


 4-38
Var. 476


 4-36
Var. 477


 4-34
Var. 478


 4-32
Var. 479


 4-30
Var. 480


 4-28
Var. 481


 4-26
Var. 482


 4-24
Var. 483


 4-22
Var. 484


 4-20
Var. 485


 4-18
Var. 486


 4-16
Var. 487


 4-14
Var. 488


 4-12
Var. 489


 4-10
Var. 490


4-8
Var. 491


 6-40
Var. 492


 6-38
Var. 493


 6-36
Var. 494


 6-34
Var. 495


 6-32
Var. 496


 6-30
Var. 497


 6-28
Var. 498


 6-26
Var. 499


 6-24
Var. 500


 6-22
Var. 501


 6-20
Var. 502


 6-18
Var. 503


 6-16
Var. 504


 6-14
Var. 505


 6-12
Var. 506


 6-10
Var. 507


6-8
Var. 508


 8-40
Var. 509


 8-38
Var. 510


 8-36
Var. 511


 8-34
Var. 512


 8-32
Var. 513


 8-30
Var. 514


 8-28
Var. 515


 8-26
Var. 516


 8-24
Var. 517


 8-22
Var. 518


 8-20
Var. 519


 8-18
Var. 520


 8-16
Var. 521


 8-14
Var. 522


 8-12
Var. 523


 8-10
Var. 524


10-40
Var. 525


10-38
Var. 526


10-36
Var. 527


10-34
Var. 528


10-32
Var. 529


10-30
Var. 530


10-28
Var. 531


10-26
Var. 532


10-24
Var. 533


10-22
Var. 534


10-20
Var. 535


10-18
Var. 536


10-16
Var. 537


10-14
Var. 538


10-12
Var. 539


12-40
Var. 540


12-38
Var. 541


12-36
Var. 542


12-34
Var. 543


12-32
Var. 544


12-30
Var. 545


12-28
Var. 546


12-26
Var. 547


12-24
Var. 548


12-22
Var. 549


12-20
Var. 550


12-18
Var. 551


12-16
Var. 552


12-14
Var. 553


14-40
Var. 554


14-38
Var. 555


14-36
Var. 556


14-34
Var. 557


14-32
Var. 558


14-30
Var. 559


14-28
Var. 560


14-26
Var. 561


14-24
Var. 562


14-22
Var. 563


14-20
Var. 564


14-18
Var. 565


14-16
Var. 566


16-40
Var. 567


16-38
Var. 568


16-36
Var. 569


16-34
Var. 570


16-32
Var. 571


16-30
Var. 572


16-28
Var. 573


16-26
Var. 574


16-24
Var. 575


16-22
Var. 576


16-20
Var. 577


16-18
Var. 578


18-40
Var. 579


18-38
Var. 580


18-36
Var. 581


18-34
Var. 582


18-32
Var. 583


18-30
Var. 584


18-28
Var. 585


18-26
Var. 586


18-24
Var. 587


18-22
Var. 588


18-20
Var. 589


20-40
Var. 590


20-38
Var. 591


20-36
Var. 592


20-34
Var. 593


20-32
Var. 594


20-30
Var. 595


20-28
Var. 596


20-26
Var. 597


20-24
Var. 598


20-22
Var. 599


22-40
Var. 600


22-38
Var. 601


22-36
Var. 602


22-34
Var. 603


22-32
Var. 604


22-30
Var. 605


22-28
Var. 606


22-26
Var. 607


22-24
Var. 608


24-40
Var. 609


24-38
Var. 610


24-36
Var. 611


24-34
Var. 612


24-32
Var. 613


24-30
Var. 614


24-28
Var. 615


24-26
Var. 616


26-40
Var. 617


26-38
Var. 618


26-36
Var. 619


26-34
Var. 620


26-32
Var. 621


26-30
Var. 622


26-28
Var. 623


28-40
Var. 624


28-38
Var. 625


28-36
Var. 626


28-34
Var. 627


28-32
Var. 628


28-30
Var. 629


30-40
Var. 630


30-38
Var. 631


30-36
Var. 632


30-34
Var. 633


30-32
Var. 634


32-40
Var. 635


32-38
Var. 636


32-36
Var. 637


32-34
Var. 638


34-40
Var. 639


36-38
Var. 640


38-40
Var. 641





Var. = Variation






In one embodiment, a rVWF composition can be characterized according to the percentage of rVWF molecules that are present in a particular higher order rVWF multimer or larger multimer. For example, in one embodiment, at least 20% of rVWF molecules in a rVWF composition used in the methods described herein are present in an oligomeric complex of at least 10 subunits. In another embodiment, at least 20% of rVWF molecules in a rVWF composition used in the methods described herein are present in an oligomeric complex of at least 12 subunits. In yet other embodiments, a rVWF composition used in the methods provided herein has a minimal percentage (e.g., has at least X %) of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer (e.g., a multimer of at least Y subunits) according to any one of variations 134 to 457 found in Table 3 to Table 5.







TABLE 3







Exemplary embodiments for the percentage of rVWF molecules that are present in a


particular higher order rVWF multimer or larger multimer found in the compositions


and used in the methods provided herein.









Minimal Number of Subunits in rVWF Multimer














6
8
10
12
14
16

















Minimal Percentage
10%
Var. 134
Var. 152
Var. 170
Var. 188
Var. 206
Var. 224


of rVWF Molecules
15%
Var. 135
Var. 153
Var. 171
Var. 189
Var. 207
Var. 225



20%
Var. 136
Var. 154
Var. 172
Var. 190
Var. 208
Var. 226



25%
Var. 137
Var. 155
Var. 173
Var. 191
Var. 209
Var. 227



30%
Var. 138
Var. 156
Var. 174
Var. 192
Var. 210
Var. 228



35%
Var. 139
Var. 157
Var. 175
Var. 193
Var. 211
Var. 229



40%
Var. 140
Var. 158
Var. 176
Var. 194
Var. 212
Var. 230



45%
Var. 141
Var. 159
Var. 177
Var. 195
Var. 213
Var. 231



50%
Var. 142
Var. 160
Var. 178
Var. 196
Var. 214
Var. 232



55%
Var. 143
Var. 161
Var. 179
Var. 197
Var. 215
Var. 233



60%
Var. 144
Var. 162
Var. 180
Var. 198
Var. 216
Var. 234



65%
Var. 145
Var. 163
Var. 181
Var. 199
Var. 217
Var. 235



70%
Var. 146
Var. 164
Var. 182
Var. 200
Var. 218
Var. 236



75%
Var. 147
Var. 165
Var. 183
Var. 201
Var. 219
Var. 237



80%
Var. 148
Var. 166
Var. 184
Var. 202
Var. 220
Var. 238



85%
Var. 149
Var. 167
Var. 185
Var. 203
Var. 221
Var. 239



90%
Var. 150
Var. 168
Var. 186
Var. 204
Var. 222
Var. 240



95%
Var. 151
Var. 169
Var. 187
Var. 205
Var. 223
Var. 241





Var. = Variation













TABLE 4







Exemplary embodiments for the percentage of rVWF molecules that are present in a


particular higher order rVWF multimer or larger multimer found in the compositions


and used in the methods provided herein.









Minimal Number of Subunits in rVWF Multimer














18
20
22
24
26
28

















Minimal Percentage
10%
Var. 242
Var. 260
Var. 278
Var. 296
Var. 314
Var. 332


of rVWF Molecules
15%
Var. 243
Var. 261
Var. 279
Var. 297
Var. 315
Var. 333



20%
Var. 244
Var. 262
Var. 280
Var. 298
Var. 316
Var. 333



25%
Var. 245
Var. 263
Var. 281
Var. 299
Var. 317
Var. 335



30%
Var. 246
Var. 264
Var. 282
Var. 300
Var. 318
Var. 336



35%
Var. 247
Var. 265
Var. 283
Var. 301
Var. 319
Var. 337



40%
Var. 248
Var. 266
Var. 284
Var. 302
Var. 320
Var. 338



45%
Var. 249
Var. 267
Var. 285
Var. 303
Var. 321
Var. 339



50%
Var. 250
Var. 268
Var. 286
Var. 304
Var. 322
Var. 340



55%
Var. 251
Var. 269
Var. 287
Var. 305
Var. 323
Var. 341



60%
Var. 252
Var. 270
Var. 288
Var. 306
Var. 324
Var. 342



65%
Var. 253
Var. 271
Var. 289
Var. 307
Var. 325
Var. 343



70%
Var. 254
Var. 272
Var. 290
Var. 308
Var. 326
Var. 344



75%
Var. 255
Var. 273
Var. 291
Var. 309
Var. 327
Var. 345



80%
Var. 256
Var. 274
Var. 292
Var. 310
Var. 328
Var. 346



85%
Var. 257
Var. 275
Var. 293
Var. 311
Var. 329
Var. 347



90%
Var. 258
Var. 276
Var. 294
Var. 312
Var. 330
Var. 348



95%
Var. 259
Var. 277
Var. 295
Var. 313
Var. 331
Var. 349





Var. = Variation













TABLE 5







Exemplary embodiments for the percentage of rVWF molecules that are present in a


particular higher order rVWF multimer or larger multimer found in the compositions


and used in the methods provided herein.









Minimal Number of Subunits in rVWF Multimer














30
32
34
36
38
40

















Minimal Percentage
10%
Var. 350
Var. 368
Var. 386
Var. 404
Var. 422
Var. 440


of rVWF Molecules
15%
Var. 351
Var. 369
Var. 387
Var. 405
Var. 423
Var. 441



20%
Var. 352
Var. 370
Var. 388
Var. 406
Var. 424
Var. 442



25%
Var. 353
Var. 371
Var. 389
Var. 407
Var. 425
Var. 443



30%
Var. 354
Var. 372
Var. 390
Var. 408
Var. 426
Var. 444



35%
Var. 355
Var. 373
Var. 391
Var. 409
Var. 427
Var. 445



40%
Var. 356
Var. 374
Var. 392
Var. 410
Var. 428
Var. 446



45%
Var. 357
Var. 375
Var. 393
Var. 411
Var. 429
Var. 447



50%
Var. 358
Var. 376
Var. 394
Var. 412
Var. 430
Var. 448



55%
Var. 359
Var. 377
Var. 395
Var. 413
Var. 431
Var. 449



60%
Var. 360
Var. 378
Var. 396
Var. 414
Var. 432
Var. 450



65%
Var. 361
Var. 379
Var. 397
Var. 415
Var. 433
Var. 451



70%
Var. 362
Var. 380
Var. 398
Var. 416
Var. 434
Var. 452



75%
Var. 363
Var. 381
Var. 399
Var. 417
Var. 435
Var. 453



80%
Var. 364
Var. 382
Var. 400
Var. 418
Var. 436
Var. 454



85%
Var. 365
Var. 383
Var. 401
Var. 419
Var. 437
Var. 455



90%
Var. 366
Var. 384
Var. 402
Var. 420
Var. 438
Var. 456



95%
Var. 367
Var. 385
Var. 403
Var. 421
Var. 439
Var. 457





Var. = Variation






In accordance with the above, the rVWF composition administered to the subject (with or without FVIII) generally comprises a significant percentage of high molecular weight (HMW) rVWF multimers. In further embodiments, the HMW rVWF multimer composition comprises at least 10%-80% rVWF decamers or higher order multimers. In further embodiments, the composition comprises about 10-95%, 20-90%, 30-85%, 40-80%, 50-75%, 60-70% decamers or higher order multimers. In further embodiments, the HMW rVWF multimer composition comprises at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% decamers or higher order multimers.
Assessment of the number and percentage of rVWF multimers can be conducted using methods known in the art, including without limitation methods using electrophoresis and size exclusion chromatography methods to separate VWF multimers by size, for example as discussed by Cumming et al, (J Clin Pathol. 1993 May; 46(5): 470-473, which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to assessment of VWF multimers). Such techniques may further include immunoblotting techniques (such as Western Blot), in which the gel is immunoblotted with a radiolabelled antibody against VWF followed by chemiluminescent detection (see for example Wen et al., (1993), J. Clin. Lab. Anal., 7: 317-323, which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to assessment of VWF multimers). Further assays for VWF include VWF:Antigen (VWF:Ag), VWF:Ristocetin Cofactor (VWF:RCof), and VWF:Collagen Binding Activity assay (VWF:CBA), which are often used for diagnosis and classification of Von Willebrand Disease. (see for example Favaloro et al., Pathology, 1997, 29(4): 341-456, which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to assays for VWF).
In further embodiments, higher order rVWF multimers of the invention are stable for about 1 to about 90 hours post-administration. In still further embodiments, the higher order rVWF multimers are stable for about 5-80, 10-70, 15-60, 20-50, 25-40, 30-35 hours post-administration. In yet further embodiments, the higher order rVWF multimers are stable for at least 3, 6, 12, 18, 24, 36, 48, 72 hours post-administration. In certain embodiments the stability of the rVWF multimers is assessed in vitro.
In one embodiment, higher order rVWF multimers used in the compositions and methods provided herein have a half life of at least 12 hour post administration. In another embodiment, the higher order rVWF multimers have a half life of at least 24 hour post administration. In yet other embodiments, the higher order rVWF multimers have a half life selected from variations 642 to 1045 found in Table 6.







TABLE 6







Exemplary embodiments for the half-life of higher order rVWF multimers


found in the compositions and used in the methods provided herein.








Hours












at least 1
Var. 642


at least 2
Var. 643


at least 3
Var. 644


at least 4
Var. 645


at least 5
Var. 646


at least 6
Var. 647


at least 7
Var. 648


at least 8
Var. 649


at least 9
Var. 650


at least 10
Var. 651


at least 11
Var. 652


at least 12
Var. 653


at least 14
Var. 654


at least 16
Var. 655


at least 18
Var. 656


at least 20
Var. 657


at least 22
Var. 658


at least 24
Var. 659


at least 27
Var. 660


at least 30
Var. 661


at least 33
Var. 662


at least 36
Var. 663


at least 39
Var. 664


at least 42
Var. 665


at least 45
Var. 666


at least 48
Var. 667


at least 54
Var. 668


at least 60
Var. 669


at least 66
Var. 670


at least 72
Var. 671


at least 78
Var. 672


at least 84
Var. 673


at least 90
Var. 674


 2-90
Var. 675


 2-84
Var. 676


 2-78
Var. 677


 2-72
Var. 678


 2-66
Var. 679


 2-60
Var. 680


 2-54
Var. 681


 2-48
Var. 682


 2-45
Var. 683


 2-42
Var. 684


 2-39
Var. 685


 2-36
Var. 686


 2-33
Var. 687


 2-30
Var. 688


 2-27
Var. 689


 2-24
Var. 690


 2-22
Var. 691


 2-20
Var. 692


 2-18
Var. 693


 2-16
Var. 694


 2-14
Var. 695


 2-12
Var. 696


 2-10
Var. 697


2-8
Var. 698


2-6
Var. 699


2-4
Var. 700


 3-90
Var. 701


 3-84
Var. 702


 3-78
Var. 703


 3-72
Var. 704


 3-66
Var. 705


 3-60
Var. 706


 3-54
Var. 707


 3-48
Var. 708


 3-45
Var. 709


 3-42
Var. 710


 3-39
Var. 711


 3-36
Var. 712


 3-33
Var. 713


 3-30
Var. 714


 3-27
Var. 715


 3-24
Var. 716


 3-22
Var. 717


 3-20
Var. 718


 3-18
Var. 719


 3-16
Var. 720


 3-14
Var. 721


 3-12
Var. 722


 3-10
Var. 723


3-8
Var. 724


3-6
Var. 725


3-4
Var. 726


 4-90
Var. 727


 4-84
Var. 728


 4-78
Var. 729


 4-72
Var. 730


 4-66
Var. 731


 4-60
Var. 732


 4-54
Var. 733


 4-48
Var. 734


 4-45
Var. 735


 4-42
Var. 736


 4-39
Var. 737


 4-36
Var. 738


 4-33
Var. 739


 4-30
Var. 740


 4-27
Var. 741


 4-24
Var. 742


 4-22
Var. 743


 4-20
Var. 744


 4-18
Var. 745


 4-16
Var. 746


 4-14
Var. 747


 4-12
Var. 748


 4-10
Var. 749


4-8
Var. 750


4-6
Var. 751


 6-90
Var. 752


 6-84
Var. 753


 6-78
Var. 754


 6-72
Var. 755


 6-66
Var. 756


 6-60
Var. 757


 6-54
Var. 758


 6-48
Var. 759


 6-45
Var. 760


 6-42
Var. 761


 6-39
Var. 762


 6-36
Var. 763


 6-33
Var. 764


 6-30
Var. 765


 6-27
Var. 766


 6-24
Var. 767


 6-22
Var. 768


 6-20
Var. 769


 6-18
Var. 770


 6-16
Var. 771


 6-14
Var. 772


 6-12
Var. 773


 6-10
Var. 774


6-8
Var. 775


 8-90
Var. 776


 8-84
Var. 777


 8-78
Var. 778


 8-72
Var. 779


 8-66
Var. 780


 8-60
Var. 781


 8-54
Var. 782


 8-48
Var. 783


 8-45
Var. 784


 8-42
Var. 785


 8-39
Var. 786


 8-36
Var. 787


 8-33
Var. 788


 8-30
Var. 789


 8-27
Var. 790


 8-24
Var. 791


 8-22
Var. 792


 8-20
Var. 793


 8-18
Var. 794


 8-16
Var. 795


 8-14
Var. 796


 8-12
Var. 797


 8-10
Var. 798


10-90
Var. 799


10-84
Var. 800


10-78
Var. 801


10-72
Var. 802


10-66
Var. 803


10-60
Var. 804


10-54
Var. 805


10-48
Var. 806


10-45
Var. 807


10-42
Var. 808


10-39
Var. 809


10-36
Var. 810


10-33
Var. 811


10-30
Var. 812


10-27
Var. 813


10-24
Var. 814


10-22
Var. 815


10-20
Var. 816


10-18
Var. 817


10-16
Var. 818


10-14
Var. 819


10-12
Var. 820


12-90
Var. 821


12-84
Var. 822


12-78
Var. 823


12-72
Var. 824


12-66
Var. 825


12-60
Var. 826


12-54
Var. 827


12-48
Var. 828


12-45
Var. 829


12-42
Var. 830


12-39
Var. 831


12-36
Var. 832


12-33
Var. 833


12-30
Var. 834


12-27
Var. 835


12-24
Var. 836


12-22
Var. 837


12-20
Var. 838


12-18
Var. 839


12-16
Var. 840


12-14
Var. 841


14-90
Var. 842


14-84
Var. 843


14-78
Var. 844


14-72
Var. 845


14-66
Var. 846


14-60
Var. 847


14-54
Var. 848


14-48
Var. 849


14-45
Var. 850


14-42
Var. 851


14-39
Var. 852


14-36
Var. 853


14-33
Var. 854


14-30
Var. 855


14-27
Var. 856


14-24
Var. 857


14-22
Var. 858


14-20
Var. 859


14-18
Var. 860


14-16
Var. 861


16-90
Var. 862


16-84
Var. 863


16-78
Var. 864


16-72
Var. 865


16-66
Var. 866


16-60
Var. 867


16-54
Var. 868


16-48
Var. 869


16-45
Var. 870


16-42
Var. 871


16-39
Var. 872


16-36
Var. 873


16-33
Var. 874


16-30
Var. 875


16-27
Var. 876


16-24
Var. 877


16-22
Var. 878


16-20
Var. 879


16-18
Var. 880


18-90
Var. 881


18-84
Var. 882


18-78
Var. 883


18-72
Var. 884


18-66
Var. 885


18-60
Var. 886


18-54
Var. 887


18-48
Var. 888


18-45
Var. 889


18-42
Var. 890


18-39
Var. 891


18-36
Var. 892


18-33
Var. 893


18-30
Var. 894


18-27
Var. 895


18-24
Var. 896


18-22
Var. 897


18-20
Var. 898


20-90
Var. 899


20-84
Var. 900


20-78
Var. 901


20-72
Var. 902


20-66
Var. 903


20-60
Var. 904


20-54
Var. 905


20-48
Var. 906


20-45
Var. 907


20-42
Var. 908


20-39
Var. 909


20-36
Var. 910


20-33
Var. 911


20-30
Var. 912


20-27
Var. 913


20-24
Var. 914


20-22
Var. 915


22-90
Var. 916


22-84
Var. 917


22-78
Var. 918


22-72
Var. 919


22-66
Var. 920


22-60
Var. 921


22-54
Var. 922


22-48
Var. 923


22-45
Var. 924


22-42
Var. 925


22-39
Var. 926


22-36
Var. 927


22-33
Var. 928


22-30
Var. 929


22-27
Var. 930


22-24
Var. 931


24-90
Var. 932


24-84
Var. 933


24-78
Var. 934


24-72
Var. 935


24-66
Var. 936


24-60
Var. 937


24-54
Var. 938


24-48
Var. 939


24-45
Var. 940


24-42
Var. 941


24-39
Var. 942


24-36
Var. 943


24-33
Var. 944


24-30
Var. 945


24-27
Var. 946


27-90
Var. 947


27-84
Var. 948


27-78
Var. 949


27-72
Var. 950


27-66
Var. 951


27-60
Var. 952


27-54
Var. 953


27-48
Var. 954


30-90
Var. 955


30-84
Var. 956


30-78
Var. 957


30-72
Var. 958


30-66
Var. 959


30-60
Var. 960


30-54
Var. 961


30-48
Var. 962


30-45
Var. 963


30-42
Var. 964


30-39
Var. 965


30-36
Var. 966


30-33
Var. 967


33-90
Var. 968


33-84
Var. 969


33-78
Var. 970


33-72
Var. 971


33-66
Var. 972


33-60
Var. 973


33-54
Var. 974


33-48
Var. 975


33-45
Var. 976


33-42
Var. 977


33-29
Var. 978


33-36
Var. 979


36-90
Var. 980


36-84
Var. 981


36-78
Var. 982


36-72
Var. 983


36-66
Var. 984


36-60
Var. 985


36-54
Var. 986


36-48
Var. 987


36-45
Var. 988


36-42
Var. 989


36-39
Var. 990


39-90
Var. 991


39-84
Var. 992


39-78
Var. 993


39-72
Var. 994


39-66
Var. 995


39-60
Var. 996


39-54
Var. 997


39-48
Var. 998


39-45
Var. 999


39-42
Var. 1000


42-90
Var. 1001


42-84
Var. 1002


42-78
Var. 1003


42-72
Var. 1004


42-66
Var. 1005


42-60
Var. 1006


42-54
Var. 1007


42-48
Var. 1008


42-45
Var. 1009


45-90
Var. 1010


45-84
Var. 1011


45-78
Var. 1012


45-72
Var. 1013


45-66
Var. 1014


45-60
Var. 1015


45-54
Var. 1016


45-48
Var. 1017


48-90
Var. 1018


48-84
Var. 1019


48-78
Var. 1020


48-72
Var. 1021


48-66
Var. 1022


48-60
Var. 1023


48-54
Var. 1024


54-90
Var. 1025


54-84
Var. 1026


54-78
Var. 1027


54-72
Var. 1028


54-66
Var. 1029


54-60
Var. 1030


60-90
Var. 1031


60-84
Var. 1032


60-78
Var. 1033


60-72
Var. 1034


60-66
Var. 1035


66-90
Var. 1036


66-84
Var. 1037


66-78
Var. 1038


66-72
Var. 1039


72-90
Var. 1040


72-84
Var. 1041


72-78
Var. 1042


78-90
Var. 1043


78-84
Var. 1044


84-90
Var. 1045





Var. = Variation






In further aspects, rVWF of use in the present invention increases stability of plasma FVIII, which, as will be appreciated, may include pdFVII or rFVIII which has been administered to the patient or it may include FVIII endogenous to the patient, or any combination thereof. For example, as shown in FIG. 22, the half-life of plasma FVIII is increased in VWD patients receiving rVWF in combination with rFVIII as compared to patients receiving FVIII (i.e., Advate) alone. In further embodiments, rVWF increases half-life of FVIII by about 1.5-5-fold as compared to the half-life seen with patients receiving FVIII alone. In still further embodiments, rVWF increases half-life of FVIII by about 1.0-4.5, 1.5-4.0, 2.0-3.5, 2.5-3.0 fold. In one embodiment, administration of rVWF increases the stability of plasma FVIII, as compared to the administration of plasma-derived VWF, by an amount selected from variations 1046 to 1089 found in Table 7. In a specific embodiment, administration of a rVWF/FVIII complex increases the stability of plasma FVIII, as compared to the administration of plasma-derived VWF, by an amount selected from variations 1046 to 1089 found in Table 7.







TABLE 7







Exemplary embodiments for the increase in half-life experienced by


plasma FVIII after administration of rVWF and rVWF/FVIII complexes,


as compared to after administration of plasma-derived VWF and


plasma-derived VWF/FVIII complexes.










Increase in




plasma FVIII




half-life






at least 1.5-fold
Var.




1046



at least 2.0-fold
Var.




1047



at least 2.5-fold
Var.




1048



at least 3.0-fold
Var.




1049



at least 3.5-fold
Var.




1050



at least 4.0-fold
Var.




1051



at least 4.5 fold
Var.




1052



at least 5.0-fold
Var.




1053



1.5-fold
Var.




1054



2.0-fold
Var.




1055



2.5-fold
Var.




1056



3.0-fold
Var.




1057



3.5-fold
Var.




1058



4.0-fold
Var.




1059



4.5-fold
Var.




1060



5.0-fold
Var.




1061



1.5-5.0 fold
Var.




1062



1.5-4.5 fold
Var.




1063



1.5-4.0 fold
Var.




1064



1.5-3.5 fold
Var.




1065



1.5-3.0 fold
Var.




1066



1.5-2.5 fold
Var.




1067



1.5-2.0 fold
Var.




1068



2-5.0 fold
Var.




1069



2-4.5 fold
Var.




1070



2-4.0 fold
Var.




1071



2-3.5 fold
Var.




1072



2-3.0 fold
Var.




1073



2-2.5 fold
Var.




1074



2.5-5.0 fold
Var.




1075



2.5-4.5 fold
Var.




1076



2.5-4.0 fold
Var.




1077



2.5-3.5 fold
Var.




1078



2.5-3.0 fold
Var.




1079



3-5.0 fold
Var.




1080



3-4.5 fold
Var.




1081



3-4.0 fold
Var.




1082



3-3.5 fold
Var.




1083



3.5-5.0 fold
Var.




1084



3.5-4.5 fold
Var.




1085



3.5-4.0 fold
Var.




1086



4-5.0 fold
Var.




1087



4-4.5 fold
Var.




1088



4.5-5.0 fold
Var.




1089





Var. = Variation






In still further embodiments, the increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In yet further embodiments, the increase in FVIII half-life is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF.
In one embodiment, the increase in the mean residence time (MRT) plasma FVIII is maintained for a time selected from variations 1090 to 1299 found in Table 8, after administration of rVWF. In a specific embodiment, the increase in the half life of plasma FVIII is maintained for a time selected from variations 1090 to 1299 found in Table 8, after administration of a rVWF/FVIII complex.







TABLE 8







Exemplary embodiments for the time plasma FVIII stability is


maintained after administration of rVWF and rVWF/FVIII complexes.








Hours












at least 6
Var. 1090


at least 12
Var. 1091


at least 18
Var. 1092


at least 24
Var. 1093


at least 30
Var. 1094


at least 36
Var. 1095


at least 42
Var. 1096


at least 48
Var. 1097


at least 54
Var. 1098


at least 60
Var. 1099


at least 66
Var. 1100


at least 72
Var. 1101


at least 78
Var. 1102


at least 84
Var. 1103


at least 90
Var. 1104


at least 96
Var. 1105


at least 102
Var. 1106


at least 108
Var. 1107


at least 114
Var. 1108


at least 120
Var. 1109


 6-120
Var. 1110


 6-114
Var. 1111


 6-108
Var. 1112


 6-102
Var. 1113


 6-96
Var. 1114


 6-90
Var. 1115


 6-84
Var. 1116


 6-78
Var. 1117


 6-72
Var. 1118


 6-66
Var. 1119


 6-60
Var. 1120


 6-54
Var. 1121


 6-48
Var. 1122


 6-42
Var. 1123


 6-36
Var. 1124


 6-30
Var. 1125


 6-24
Var. 1126


 6-18
Var. 1127


 6-12
Var. 1128


 12-120
Var. 1129


 12-114
Var. 1130


 12-108
Var. 1131


 12-102
Var. 1132


12-96
Var. 1133


12-90
Var. 1134


12-84
Var. 1135


12-78
Var. 1136


12-72
Var. 1137


12-66
Var. 1138


12-60
Var. 1139


12-54
Var. 1140


12-48
Var. 1141


12-42
Var. 1142


12-36
Var. 1143


12-30
Var. 1144


12-24
Var. 1145


12-18
Var. 1146


 18-120
Var. 1147


 18-114
Var. 1148


 18-108
Var. 1149


 18-102
Var. 1150


18-96
Var. 1151


18-90
Var. 1152


18-84
Var. 1153


18-78
Var. 1154


18-72
Var. 1155


18-66
Var. 1156


18-60
Var. 1157


18-54
Var. 1158


18-48
Var. 1159


18-42
Var. 1160


18-36
Var. 1161


18-30
Var. 1162


18-24
Var. 1163


 24-120
Var. 1164


 24-114
Var. 1165


 24-108
Var. 1166


 24-102
Var. 1167


24-96
Var. 1168


24-90
Var. 1169


24-84
Var. 1170


24-78
Var. 1171


24-72
Var. 1172


24-66
Var. 1173


24-60
Var. 1174


24-54
Var. 1175


24-48
Var. 1176


24-42
Var. 1177


24-36
Var. 1178


24-30
Var. 1179


 30-120
Var. 1180


 30-114
Var. 1181


 30-108
Var. 1182


 30-102
Var. 1183


30-96
Var. 1184


30-90
Var. 1185


30-84
Var. 1186


30-78
Var. 1187


30-72
Var. 1188


30-66
Var. 1189


30-60
Var. 1190


30-54
Var. 1191


30-48
Var. 1192


30-42
Var. 1193


30-36
Var. 1194


 36-120
Var. 1195


 36-114
Var. 1196


 36-108
Var. 1197


 36-102
Var. 1198


36-96
Var. 1199


36-90
Var. 1200


36-84
Var. 1201


36-78
Var. 1202


36-72
Var. 1203


36-66
Var. 1204


36-60
Var. 1205


36-54
Var. 1206


36-48
Var. 1207


36-42
Var. 1208


 42-120
Var. 1209


 42-114
Var. 1210


 42-108
Var. 1211


 42-102
Var. 1212


42-96
Var. 1213


42-90
Var. 1214


42-84
Var. 1215


42-78
Var. 1216


42-72
Var. 1217


42-66
Var. 1218


42-60
Var. 1219


42-54
Var. 1220


42-48
Var. 1221


 48-120
Var. 1222


 48-114
Var. 1223


 48-108
Var. 1224


 48-102
Var. 1225


48-96
Var. 1226


48-90
Var. 1227


48-84
Var. 1228


48-78
Var. 1229


48-72
Var. 1230


48-66
Var. 1231


48-60
Var. 1232


48-54
Var. 1233


 54-120
Var. 1234


 54-114
Var. 1235


 54-108
Var. 1236


 54-102
Var. 1237


54-96
Var. 1238


54-90
Var. 1239


54-84
Var. 1240


54-78
Var. 1241


54-72
Var. 1242


54-66
Var. 1243


54-60
Var. 1244


 60-120
Var. 1245


 60-114
Var. 1246


 60-108
Var. 1247


 60-102
Var. 1248


60-96
Var. 1249


60-90
Var. 1250


60-84
Var. 1251


60-78
Var. 1252


60-72
Var. 1253


60-66
Var. 1254


 66-120
Var. 1255


 66-114
Var. 1256


 66-108
Var. 1257


 66-102
Var. 1258


66-96
Var. 1259


66-90
Var. 1260


66-84
Var. 1261


66-78
Var. 1262


66-72
Var. 1263


 72-120
Var. 1264


 72-114
Var. 1265


 72-108
Var. 1266


 72-102
Var. 1267


72-96
Var. 1268


72-90
Var. 1269


72-84
Var. 1270


72-78
Var. 1271


 78-120
Var. 1272


 78-114
Var. 1273


 78-108
Var. 1274


 78-102
Var. 1275


78-96
Var. 1276


78-90
Var. 1277


78-84
Var. 1278


 84-120
Var. 1279


 84-114
Var. 1280


 84-108
Var. 1281


 84-102
Var. 1282


84-96
Var. 1283


84-90
Var. 1284


 90-120
Var. 1285


 90-114
Var. 1286


 90-108
Var. 1287


 90-102
Var. 1288


90-96
Var. 1289


 96-120
Var. 1290


 96-114
Var. 1291


 96-108
Var. 1292


 96-102
Var. 1293


102-120
Var. 1294


102-114
Var. 1295


102-108
Var. 1296


108-120
Var. 1297


108-114
Var. 1298


114-120
Var. 1299





Var. = Variation






In further aspects, the rVWF of the invention shows an increased effect on the stability of FVIII as compared to the effect of pdVWF. For example, as shown in FIG. 20, average FVIII half-life was increased by rVWF by 5.2 hours over the half-life for patients receiving pdVWF. In further embodiments, rVWF increases average FVIII half-life by about 1-15, 2-14, 3-13, 4-12, 5-11, 6-10, 5-9, 6-8 hours. In still further embodiments, rVWF increases FVIII half-life by about 10% to about 75% as compared to pdVWF. In yet further embodiments, rVWF increases FVIII half-life by about 10-80%, 15-65%, 20-60%, 25-55%, 30-50%, 35-45% as compared to pdVWF. In certain embodiments, the administration of rVWF increases the half life of FVIII by an amount selected from variations 1300 to 1643 found in Table 9, as compared to administration of plasma-derived VWF. In yet further embodiments, the average or percentage increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In yet further embodiments, the increase in FVIII half-life is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF.







TABLE 9







Exemplary embodiments for the increase in half-life experienced by


plasma FVIII after administration of rVWF and rVWF/FVIII complexes.








Increase in



half life












at least 1
hr
Var. 1300


at least 2
hr
Var. 1301


at least 3
hr
Var. 1302


at least 4
hr
Var. 1303


at least 5
hr
Var. 1304


at least 6
hr
Var. 1305


at least 7
hr
Var. 1306


at least 8
hr
Var. 1307


at least 9
hr
Var. 1308


at least 10
hr
Var. 1309


at least 11
hr
Var. 1310


at least 12
hr
Var. 1311


at least 13
hr
Var. 1312


at least 14
hr
Var. 1313


at least 15
hr
Var. 1314


1
hr
Var. 1315


2
hr
Var. 1316


3
hr
Var. 1317


4
hr
Var. 1318


5
hr
Var. 1319


6
hr
Var. 1320


7
hr
Var. 1321


8
hr
Var. 1322


9
hr
Var. 1323


10
hr
Var. 1324


11
hr
Var. 1325


12
hr
Var. 1326


13
hr
Var. 1327


14
hr
Var. 1328


15
hr
Var. 1329


1-15
hr
Var. 1330


1-14
hr
Var. 1331


1-13
hr
Var. 1332


1-12
hr
Var. 1333


1-11
hr
Var. 1334


1-10
hr
Var. 1335


1-9
hr
Var. 1336


1-8
hr
Var. 1337


1-7
hr
Var. 1338


1-6
hr
Var. 1339


1-5
hr
Var. 1340


1-4
hr
Var. 1341


1-3
hr
Var. 1342


1-2
hr
Var. 1343


2-15
hr
Var. 1344


2-14
hr
Var. 1345


2-13
hr
Var. 1346


2-12
hr
Var. 1347


2-11
hr
Var. 1348


2-10
hr
Var. 1349


2-9
hr
Var. 1350


2-8
hr
Var. 1351


2-7
hr
Var. 1352


2-6
hr
Var. 1353


2-5
hr
Var. 1354


2-4
hr
Var. 1355


2-3
hr
Var. 1356


3-15
hr
Var. 1357


3-14
hr
Var. 1358


3-13
hr
Var. 1359


3-12
hr
Var. 1360


3-11
hr
Var. 1361


3-10
hr
Var. 1362


3-9
hr
Var. 1363


3-8
hr
Var. 1364


3-7
hr
Var. 1365


3-6
hr
Var. 1366


3-5
hr
Var. 1367


3-4
hr
Var. 1368


4-15
hr
Var. 1369


4-14
hr
Var. 1370


4-13
hr
Var. 1371


4-12
hr
Var. 1372


4-11
hr
Var. 1373


4-10
hr
Var. 1374


4-9
hr
Var. 1375


4-8
hr
Var. 1376


4-7
hr
Var. 1377


4-6
hr
Var. 1378


4-5
hr
Var. 1379


5-15
hr
Var. 1380


5-14
hr
Var. 1381


5-13
hr
Var. 1382


5-12
hr
Var. 1383


5-11
hr
Var. 1384


5-10
hr
Var. 1385


5-9
hr
Var. 1386


5-8
hr
Var. 1387


5-7
hr
Var. 1388


5-6
hr
Var. 1389


6-15
hr
Var. 1390


6-14
hr
Var. 1391


6-13
hr
Var. 1392


6-12
hr
Var. 1393


6-11
hr
Var. 1394


6-10
hr
Var. 1395


6-9
hr
Var. 1396


6-8
hr
Var. 1397


6-7
hr
Var. 1398


7-15
hr
Var. 1399


7-14
hr
Var. 1400


7-13
hr
Var. 1401


7-12
hr
Var. 1402


7-11
hr
Var. 1403


7-10
hr
Var. 1404


7-9
hr
Var. 1405


7-8
hr
Var. 1406


8-15
hr
Var. 1407


8-14
hr
Var. 1408


8-13
hr
Var. 1409


8-12
hr
Var. 1410


8-11
hr
Var. 1411


8-10
hr
Var. 1412


8-9
hr
Var. 1413


9-15
hr
Var. 1414


9-14
hr
Var. 1415


9-13
hr
Var. 1416


9-12
hr
Var. 1417


9-11
hr
Var. 1418


9-10
hr
Var. 1419


10-15
hr
Var. 1420


10-14
hr
Var. 1421


10-13
hr
Var. 1422


10-12
hr
Var. 1423


10-11
hr
Var. 1424


11-15
hr
Var. 1425


11-14
hr
Var. 1426


11-13
hr
Var. 1427


11-12
hr
Var. 1428


12-15
hr
Var. 1429


12-14
hr
Var. 1430


12-13
hr
Var. 1431


13-15
hr
Var. 1432


13-14
hr
Var. 1433


14-15
hr
Var. 1434








at least 10%
Var. 1435


at least 15%
Var. 1436


at least 20%
Var. 1437


at least 25%
Var. 1438


at least 30%
Var. 1439


at least 35%
Var. 1440


at least 40%
Var. 1441


at least 45%
Var. 1442


at least 50%
Var. 1443


at least 55%
Var. 1444


at least 60%
Var. 1445


at least 65%
Var. 1446


at least 70%
Var. 1447


at least 75%
Var. 1448


at least 80%
Var. 1449


at least 85%
Var. 1450


at least 90%
Var. 1451


at least 95%
Var. 1452


at least 100%
Var. 1453


10%
Var. 1454


15%
Var. 1455


20%
Var. 1456


25%
Var. 1457


30%
Var. 1458


35%
Var. 1459


40%
Var. 1460


45%
Var. 1461


50%
Var. 1462


55%
Var. 1463


60%
Var. 1464


65%
Var. 1465


70%
Var. 1466


75%
Var. 1467


80%
Var. 1468


85%
Var. 1469


90%
Var. 1470


95%
Var. 1471


100% 
Var. 1472


 10-100%
Var. 1473


10-95%
Var. 1474


10-90%
Var. 1475


10-85%
Var. 1476


10-80%
Var. 1477


10-75%
Var. 1478


10-70%
Var. 1479


10-65%
Var. 1480


10-60%
Var. 1481


10-55%
Var. 1482


10-50%
Var. 1483


10-45%
Var. 1484


10-40%
Var. 1485


10-35%
Var. 1486


10-30%
Var. 1487


10-25%
Var. 1488


10-20%
Var. 1489


10-15%
Var. 1490


 15-100%
Var. 1491


15-95%
Var. 1492


15-90%
Var. 1493


15-85%
Var. 1494


15-80%
Var. 1495


15-75%
Var. 1496


15-70%
Var. 1497


15-65%
Var. 1498


15-60%
Var. 1499


15-55%
Var. 1500


15-50%
Var. 1501


15-45%
Var. 1502


15-40%
Var. 1503


15-35%
Var. 1504


15-30%
Var. 1505


15-25%
Var. 1506


15-20%
Var. 1507


 20-100%
Var. 1508


20-95%
Var. 1509


20-90%
Var. 1510


20-85%
Var. 1511


20-80%
Var. 1512


20-75%
Var. 1513


20-70%
Var. 1514


20-65%
Var. 1515


20-60%
Var. 1516


20-55%
Var. 1517


20-50%
Var. 1518


20-45%
Var. 1519


20-40%
Var. 1520


20-35%
Var. 1521


20-30%
Var. 1522


20-25%
Var. 1523


 25-100%
Var. 1524


25-95%
Var. 1525


25-90%
Var. 1526


25-85%
Var. 1527


25-80%
Var. 1528


25-75%
Var. 1529


25-70%
Var. 1530


25-65%
Var. 1531


25-60%
Var. 1532


25-55%
Var. 1533


25-50%
Var. 1534


25-45%
Var. 1535


25-40%
Var. 1536


25-35%
Var. 1537


25-30%
Var. 1538


 30-100%
Var. 1539


30-95%
Var. 1540


30-90%
Var. 1541


30-85%
Var. 1542


30-80%
Var. 1543


30-75%
Var. 1544


30-70%
Var. 1545


30-65%
Var. 1546


30-60%
Var. 1547


30-55%
Var. 1548


30-50%
Var. 1549


30-45%
Var. 1550


30-40%
Var. 1551


30-35%
Var. 1552


 35-100%
Var. 1553


35-95%
Var. 1554


35-90%
Var. 1555


35-85%
Var. 1556


35-80%
Var. 1557


35-75%
Var. 1558


35-70%
Var. 1559


35-65%
Var. 1560


35-60%
Var. 1561


35-55%
Var. 1562


35-50%
Var. 1563


35-45%
Var. 1564


35-40%
Var. 1565


 40-100%
Var. 1566


40-95%
Var. 1567


40-90%
Var. 1568


40-85%
Var. 1569


40-80%
Var. 1570


40-75%
Var. 1571


40-70%
Var. 1572


40-65%
Var. 1573


40-60%
Var. 1574


40-55%
Var. 1575


40-50%
Var. 1576


40-45%
Var. 1577


 45-100%
Var. 1578


45-95%
Var. 1579


45-90%
Var. 1580


45-85%
Var. 1581


45-80%
Var. 1582


45-75%
Var. 1583


45-70%
Var. 1584


45-65%
Var. 1585


45-60%
Var. 1586


45-55%
Var. 1587


45-50%
Var. 1588


 50-100%
Var. 1589


50-95%
Var. 1590


50-90%
Var. 1591


50-85%
Var. 1592


50-80%
Var. 1593


50-75%
Var. 1594


50-70%
Var. 1595


50-65%
Var. 1596


50-60%
Var. 1597


50-55%
Var. 1598


 55-100%
Var. 1599


55-95%
Var. 1600


55-90%
Var. 1601


55-85%
Var. 1602


55-80%
Var. 1603


55-75%
Var. 1604


55-70%
Var. 1605


55-65%
Var. 1606


55-60%
Var. 1607


 60-100%
Var. 1608


60-95%
Var. 1609


60-90%
Var. 1610


60-85%
Var. 1611


60-80%
Var. 1612


60-75%
Var. 1613


60-70%
Var. 1614


60-65%
Var. 1615


 65-100%
Var. 1616


65-95%
Var. 1617


65-90%
Var. 1618


65-85%
Var. 1619


65-80%
Var. 1620


65-75%
Var. 1621


65-70%
Var. 1622


 70-100%
Var. 1623


70-95%
Var. 1624


70-90%
Var. 1625


70-85%
Var. 1626


70-80%
Var. 1627


70-75%
Var. 1628


 75-100%
Var. 1629


75-95%
Var. 1630


75-90%
Var. 1631


75-85%
Var. 1632


75-80%
Var. 1633


 80-100%
Var. 1634


80-95%
Var. 1635


80-90%
Var. 1636


80-85%
Var. 1637


 85-100%
Var. 1638


85-95%
Var. 1639


85-90%
Var. 1640


 90-100%
Var. 1641


90-95%
Var. 1642


95%-100% 
Var. 1643





Var. = Variation






In still further aspects and in accordance with any of the above, the stabilization of FVIII activity by rVWF as compared to pdVWF can be measured by metrics in addition to FVIII half-life, including mean residence time (MRT) and area under curve (AUC). In exemplary embodiments, rVWF increases MRT by about 1-15 hours as compared to pdVWF. In further embodiments, rVWF increases MRT by about 1-25, 2-20, 3-15, 4-10, 5-9, 6-8 hours as compared to pdVWF. In still further embodiments, rVWF increases FVIII MRT by about 10-80%, 15-65%, 20-60%, 25-55%, 30-50%, 35-45% as compared to pdVWF. In yet further embodiments, the average or percentage increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In certain embodiments, the administration of rVWF increases the MRT of FVIII by an amount selected from variations 1644 to 1987 found in Table 10, as compared to administration of plasma-derived VWF.
In yet further embodiments, the increase in MRT is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF. In certain embodiments, the increase in MRT of FVIII caused by administration of rVWF is maintained for a time selected from variations 1090 to 1299 found in Table 8.







TABLE 10







Exemplary embodiments for the increase in FVIII mean residence


time after administration of rVWF and rVWF/FVIII complexes,


as compared to after administration of plasma-derived VWF and


plasma-derived VWF/FVIII complexes.








Increase in



half life












at least 1
hr
Var. 1644


at least 2
hr
Var. 1645


at least 3
hr
Var. 1646


at least 4
hr
Var. 1647


at least 5
hr
Var. 1648


at least 6
hr
Var. 1649


at least 7
hr
Var. 1650


at least 8
hr
Var. 1651


at least 9
hr
Var. 1652


at least 10
hr
Var. 1653


at least 11
hr
Var. 1654


at least 12
hr
Var. 1655


at least 13
hr
Var. 1656


at least 14
hr
Var. 1657


at least 15
hr
Var. 1658


1
hr
Var. 1659


2
hr
Var. 1660


3
hr
Var. 1661


4
hr
Var. 1662


5
hr
Var. 1663


6
hr
Var. 1664


7
hr
Var. 1665


8
hr
Var. 1666


9
hr
Var. 1667


10
hr
Var. 1668


11
hr
Var. 1669


12
hr
Var. 1670


13
hr
Var. 1671


14
hr
Var. 1672


15
hr
Var. 1673


1-15
hr
Var. 1674


1-14
hr
Var. 1675


1-13
hr
Var. 1676


1-12
hr
Var. 1677


1-11
hr
Var. 1678


1-10
hr
Var. 1679


1-9
hr
Var. 1680


1-8
hr
Var. 1681


1-7
hr
Var. 1682


1-6
hr
Var. 1683


1-5
hr
Var. 1684


1-4
hr
Var. 1685


1-3
hr
Var. 1686


1-2
hr
Var. 1687


2-15
hr
Var. 1688


2-14
hr
Var. 1689


2-13
hr
Var. 1690


2-12
hr
Var. 1691


2-11
hr
Var. 1692


2-10
hr
Var. 1693


2-9
hr
Var. 1694


2-8
hr
Var. 1695


2-7
hr
Var. 1696


2-6
hr
Var. 1697


2-5
hr
Var. 1698


2-4
hr
Var. 1699


2-3
hr
Var. 1700


3-15
hr
Var. 1701


3-14
hr
Var. 1702


3-13
hr
Var. 1703


3-12
hr
Var. 1704


3-11
hr
Var. 1705


3-10
hr
Var. 1706


3-9
hr
Var. 1707


3-8
hr
Var. 1708


3-7
hr
Var. 1709


3-6
hr
Var. 1710


3-5
hr
Var. 1711


3-4
hr
Var. 1712


4-15
hr
Var. 1713


4-14
hr
Var. 1714


4-13
hr
Var. 1715


4-12
hr
Var. 1716


4-11
hr
Var. 1717


4-10
hr
Var. 1718


4-9
hr
Var. 1719


4-8
hr
Var. 1720


4-7
hr
Var. 1721


4-6
hr
Var. 1722


4-5
hr
Var. 1723


5-15
hr
Var. 1724


5-14
hr
Var. 1725


5-13
hr
Var. 1726


5-12
hr
Var. 1727


5-11
hr
Var. 1728


5-10
hr
Var. 1729


5-9
hr
Var. 1730


5-8
hr
Var. 1731


5-7
hr
Var. 1732


5-6
hr
Var. 1733


6-15
hr
Var. 1734


6-14
hr
Var. 1735


6-13
hr
Var. 1736


6-12
hr
Var. 1737


6-11
hr
Var. 1738


6-10
hr
Var. 1739


6-9
hr
Var. 1740


6-8
hr
Var. 1741


6-7
hr
Var. 1742


7-15
hr
Var. 1743


7-14
hr
Var. 1744


7-13
hr
Var. 1745


7-12
hr
Var. 1746


7-11
hr
Var. 1747


7-10
hr
Var. 1748


7-9
hr
Var. 1749


7-8
hr
Var. 1750


8-15
hr
Var. 1751


8-14
hr
Var. 1752


8-13
hr
Var. 1753


8-12
hr
Var. 1754


8-11
hr
Var. 1755


8-10
hr
Var. 1756


8-9
hr
Var. 1757


9-15
hr
Var. 1758


9-14
hr
Var. 1759


9-13
hr
Var. 1760


9-12
hr
Var. 1761


9-11
hr
Var. 1762


9-10
hr
Var. 1763


10-15
hr
Var. 1764


10-14
hr
Var. 1765


10-13
hr
Var. 1766


10-12
hr
Var. 1767


10-11
hr
Var. 1768


11-15
hr
Var. 1769


11-14
hr
Var. 1770


11-13
hr
Var. 1771


11-12
hr
Var. 1772


12-15
hr
Var. 1773


12-14
hr
Var. 1774


12-13
hr
Var. 1775


13-15
hr
Var. 1776


13-14
hr
Var. 1777


14-15
hr
Var. 1778








at least 10%
Var. 1779


at least 15%
Var. 1780


at least 20%
Var. 1781


at least 25%
Var. 1782


at least 30%
Var. 1783


at least 35%
Var. 1784


at least 40%
Var. 1785


at least 45%
Var. 1786


at least 50%
Var. 1787


at least 55%
Var. 1788


at least 60%
Var. 1789


at least 65%
Var. 1790


at least 70%
Var. 1791


at least 75%
Var. 1792


at least 80%
Var. 1793


at least 85%
Var. 1794


at least 90%
Var. 1795


at least 95%
Var. 1796


at least 100%
Var. 1797


10%
Var. 1798


15%
Var. 1799


20%
Var. 1800


25%
Var. 1801


30%
Var. 1802


35%
Var. 1803


40%
Var. 1804


45%
Var. 1805


50%
Var. 1806


55%
Var. 1807


60%
Var. 1808


65%
Var. 1809


70%
Var. 1810


75%
Var. 1811


80%
Var. 1812


85%
Var. 1813


90%
Var. 1814


95%
Var. 1815


100% 
Var. 1816


 10-100%
Var. 1817


10-95%
Var. 1818


10-90%
Var. 1819


10-85%
Var. 1820


10-80%
Var. 1821


10-75%
Var. 1822


10-70%
Var. 1823


10-65%
Var. 1824


10-60%
Var. 1825


10-55%
Var. 1826


10-50%
Var. 1827


10-45%
Var. 1828


10-40%
Var. 1829


10-35%
Var. 1830


10-30%
Var. 1831


10-25%
Var. 1832


10-20%
Var. 1833


10-15%
Var. 1834


 15-100%
Var. 1835


15-95%
Var. 1836


15-90%
Var. 1837


15-85%
Var. 1838


15-80%
Var. 1839


15-75%
Var. 1840


15-70%
Var. 1841


15-65%
Var. 1842


15-60%
Var. 1843


15-55%
Var. 1844


15-50%
Var. 1845


15-45%
Var. 1846


15-40%
Var. 1847


15-35%
Var. 1848


15-30%
Var. 1849


15-25%
Var. 1850


15-20%
Var. 1851


 20-100%
Var. 1852


20-95%
Var. 1853


20-90%
Var. 1854


20-85%
Var. 1855


20-80%
Var. 1856


20-75%
Var. 1857


20-70%
Var. 1858


20-65%
Var. 1859


20-60%
Var. 1860


20-55%
Var. 1861


20-50%
Var. 1862


20-45%
Var. 1863


20-40%
Var. 1864


20-35%
Var. 1865


20-30%
Var. 1866


20-25%
Var. 1867


 25-100%
Var. 1868


25-95%
Var. 1869


25-90%
Var. 1870


25-85%
Var. 1871


25-80%
Var. 1872


25-75%
Var. 1873


25-70%
Var. 1874


25-65%
Var. 1875


25-60%
Var. 1876


25-55%
Var. 1877


25-50%
Var. 1878


25-45%
Var. 1879


25-40%
Var. 1880


25-35%
Var. 1881


25-30%
Var. 1882


 30-100%
Var. 1883


30-95%
Var. 1884


30-90%
Var. 1885


30-85%
Var. 1886


30-80%
Var. 1887


30-75%
Var. 1888


30-70%
Var. 1889


30-65%
Var. 1890


30-60%
Var. 1891


30-55%
Var. 1892


30-50%
Var. 1893


30-45%
Var. 1894


30-40%
Var. 1895


30-35%
Var. 1896


 35-100%
Var. 1897


35-95%
Var. 1898


35-90%
Var. 1899


35-85%
Var. 1900


35-80%
Var. 1901


35-75%
Var. 1902


35-70%
Var. 1903


35-65%
Var. 1904


35-60%
Var. 1905


35-55%
Var. 1906


35-50%
Var. 1907


35-45%
Var. 1908


35-40%
Var. 1909


 40-100%
Var. 1910


40-95%
Var. 1911


40-90%
Var. 1912


40-85%
Var. 1913


40-80%
Var. 1914


40-75%
Var. 1915


40-70%
Var. 1916


40-65%
Var. 1917


40-60%
Var. 1918


40-55%
Var. 1919


40-50%
Var. 1920


40-45%
Var. 1921


 45-100%
Var. 1922


45-95%
Var. 1923


45-90%
Var. 1924


45-85%
Var. 1925


45-80%
Var. 1926


45-75%
Var. 1927


45-70%
Var. 1928


45-65%
Var. 1929


45-60%
Var. 1930


45-55%
Var. 1931


45-50%
Var. 1932


 50-100%
Var. 1933


50-95%
Var. 1934


50-90%
Var. 1935


50-85%
Var. 1936


50-80%
Var. 1937


50-75%
Var. 1938


50-70%
Var. 1939


50-65%
Var. 1940


50-60%
Var. 1941


50-55%
Var. 1942


 55-100%
Var. 1943


55-95%
Var. 1944


55-90%
Var. 1945


55-85%
Var. 1946


55-80%
Var. 1947


55-75%
Var. 1948


55-70%
Var. 1949


55-65%
Var. 1950


55-60%
Var. 1951


 60-100%
Var. 1952


60-95%
Var. 1953


60-90%
Var. 1954


60-85%
Var. 1955


60-80%
Var. 1956


60-75%
Var. 1957


60-70%
Var. 1958


60-65%
Var. 1959


 65-100%
Var. 1960


65-95%
Var. 1961


65-90%
Var. 1962


65-85%
Var. 1963


65-80%
Var. 1964


65-75%
Var. 1965


65-70%
Var. 1966


 70-100%
Var. 1967


70-95%
Var. 1968


70-90%
Var. 1969


70-85%
Var. 1970


70-80%
Var. 1971


70-75%
Var. 1972


 75-100%
Var. 1973


75-95%
Var. 1974


75-90%
Var. 1975


75-85%
Var. 1976


75-80%
Var. 1977


 80-100%
Var. 1978


80-95%
Var. 1979


80-90%
Var. 1980


80-85%
Var. 1981


 85-100%
Var. 1982


85-95%
Var. 1983


85-90%
Var. 1984


 90-100%
Var. 1985


90-95%
Var. 1986


95%-100% 
Var. 1987





Var. = Variation






Further exemplary differences between pdVWF and rVWF are provided in the following table:











pdVWF
rVWF







Synthesized in endothelial cells and
Expressed in CHO cells


megakaryocytes



Post-translational modification of
Propeptide removal mediated


propeptide removal occurs intracellularly:
in vitro through exposure of


during passage of the protein to the Golgi
the pro-VWF to recombinant


and post-Golgi compartments
Furin


Glycosylation/ABO blood group glycans
Fully glycosylated/ABO blood


present
group gylcans absent


Consists of VWF subunits that have been
No exposure to ADAMTS13


exposed to plasma ADAMTS13
→ Intact VWF subunits


→ Ultra-large VWF multimers absent
→ Ultra-large VWF multimers


→ Subunits cleaved at TYR1605-MET1606
present



→ Subunit cleavage occurs 



upon ADAMTS13 exposure


Plasma-derived VWF concentrates contain
Higher specific activity than


other proteins incl. ADAMTS13,
pdVWF


hemagglutinins









In some embodiments, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) is between 3:1 and 1:5. In further embodiments, the ratio is between 2:1 and 1:4. In still further embodiments, the ratio is between 5:2 and 1:4. In further embodiments, the ratio is between 3:2 and 1:3. In still further embodiments, the ratio is about 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 2:3, 2:4, 2:5, 3:1, 3:2, 3:4, or 3:5. In further embodiments, the ratio is between 1:1 and 1:2. In yet further embodiments, the ratio is 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, or 2:1. In certain embodiments, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in a composition useful for a method described herein is selected from variations 1988 to 2140 found in Table 11.







TABLE 11







Exemplary embodiments for the ratio of rFVIII procoagulant activity


(IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo)


in compositions and used in methods provided herein.








(IU rFVIII:C) to



(IU rVWF:RCo)





4:1
Var.



1988


3:1
Var.



1989


2:1
Var.



1990


3:2
Var.



1991


4:3
Var.



1992


1:1
Var.



1993


5:6
Var.



1994


4:5
Var.



1995


3:4
Var.



1996


2:3
Var.



1997


3:5
Var.



1998


1:2
Var.



1999


2:5
Var.



2000


1:3
Var.



2001


1:4
Var.



2002


1:5
Var.



2003


1:6
Var.



2004


4:1-1:6
Var.



2005


4:1-1:5
Var.



2006


4:1-1:4
Var.



2007


4:1-1:3
Var.



2008


4:1-2:5
Var.



2009


4:1-1:2
Var.



2010


4:1-3:5
Var.



2011


4:1-2:3
Var.



2012


4:1-3:4
Var.



2013


4:1-4:5
Var.



2014


4:1-5:6
Var.



2015


4:1-1:1
Var.



2016


4:1-4:3
Var.



2017


4:1-3:2
Var.



2018


4:1-2:1
Var.



2019


4:1-3:1
Var.



2020


3:1-1:6
Var.



2021


3:1-1:5
Var.



2022


3:1-1:4
Var.



2023


3:1-1:3
Var.



2024


3:1-2:5
Var.



2025


3:1-1:2
Var.



2026


3:1-3:5
Var.



2027


3:1-2:3
Var.



2028


3:1-3:4
Var.



2029


3:1-4:5
Var.



2030


3:1-5:6
Var.



2031


3:1-1:1
Var.



2032


3:1-4:3
Var.



2033


3:1-3:2
Var.



2034


3:1-2:1
Var.



2035


2:1-1:6
Var.



2036


2:1-1:5
Var.



2037


2:1-1:4
Var.



2038


2:1-1:3
Var.



2039


2:1-2:5
Var.



2040


2:1-1:2
Var.



2041


2:1-3:5
Var.



2042


2:1-2:3
Var.



2043


2:1-3:4
Var.



2044


2:1-4:5
Var.



2045


2:1-5:6
Var.



2046


2:1-1:1
Var.



2047


2:1-4:3
Var.



2048


2:1-3:2
Var.



2049


3:2-1:6
Var.



2050


3:2-1:5
Var.



2051


3:2-1:4
Var.



2052


3:2-1:3
Var.



2053


3:2-2:5
Var.



2054


3:2-1:2
Var.



2055


3:2-3:5
Var.



2056


3:2-2:3
Var.



2057


3:2-3:4
Var.



2058


3:2-4:5
Var.



2059


3:2-5:6
Var.



2060


3:2-1:1
Var.



2061


3:2-4:3
Var.



2062


4:3-1:6
Var.



2063


4:3-1:5
Var.



2064


4:3-1:4
Var.



2065


4:3-1:3
Var.



2066


4:3-2:5
Var.



2067


4:3-1:2
Var.



2068


4:3-3:5
Var.



2069


4:3-2:3
Var.



2070


4:3-3:4
Var.



2071


4:3-4:5
Var.



2072


4:3-5:6
Var.



2073


4:3-1:1
Var.



2074


1:1-1:6
Var.



2075


1:1-1:5
Var.



2076


1:1-1:4
Var.



2077


1:1-1:3
Var.



2078


1:1-2:5
Var.



2079


1:1-1:2
Var.



2080


1:1-3:5
Var.



2081


1:1-2:3
Var.



2082


1:1-3:4
Var.



2083


1:1-4:5
Var.



2084


1:1-5:6
Var.



2085


5:6-1:6
Var.



2086


5:6-1:5
Var.



2087


5:6-1:4
Var.



2088


5:6-1:3
Var.



2089


5:6-2:5
Var.



2090


5:6-1:2
Var.



2091


5:6-3:5
Var.



2092


5:6-2:3
Var.



2093


5:6-3:4
Var.



2094


5:6-4:5
Var.



2095


4:5-1:6
Var.



2096


4:5-1:5
Var.



2097


4:5-1:4
Var.



2098


4:5-1:3
Var.



2099


4:5-2:5
Var.



2100


4:5-1:2
Var.



2101


4:5-3:5
Var.



2102


4:5-2:3
Var.



2103


4:5-3:4
Var.



2104


3:4-1:6
Var.



2105


3:4-1:5
Var.



2106


3:4-1:4
Var.



2107


3:4-1:3
Var.



2108


3:4-2:5
Var.



2109


3:4-1:2
Var.



2110


3:4-3:5
Var.



2111


3:4-2:3
Var.



2112


2:3-1:6
Var.



2113


2:3-1:5
Var.



2114


2:3-1:4
Var.



2115


2:3-1:3
Var.



2116


2:3-2:5
Var.



2117


2:3-1:2
Var.



2118


2:3-3:5
Var.



2119


3:5-1:6
Var.



2120


3:5-1:5
Var.



2121


3:5-1:4
Var.



2122


3:5-1:3
Var.



2123


3:5-2:5
Var.



2124


3:5-1:2
Var.



2125


1:2-1:6
Var.



2126


1:2-1:5
Var.



2127


1:2-1:4
Var.



2128


1:2-1:3
Var.



2129


1:2-2:5
Var.



2130


2:5-1:6
Var.



2131


2:5-1:5
Var.



2132


2:5-1:4
Var.



2133


2:5-1:3
Var.



2134


1:3-1:6
Var.



2135


1:3-1:5
Var.



2136


1:3-1:4
Var.



2137


1:4-1:6
Var.



2138


1:4-1:5
Var.



2139


1:5-1:6
Var.



2140





Var. = Variation






In specific aspects, the rVWF and/or the FVIII (recombinant or plasma derived) used in accordance with the present invention are not modified with any conjugation, post-translation or covalent modifications. In particular embodiments, the rVWF and/or FVIII of the present invention are not modified with a water soluble polymer, including without limitation, a polyethylene glycol (PEG), a polypropylene glycol, a polyoxyalkylene, a polysialic acid, hydroxyl ethyl starch, a poly-carbohydrate moiety, and the like.
In other aspects, the rVWF and/or the FVIII (recombinant or plasma derived) used in accordance with the present invention are modified through conjugation, post-translation modification, or covalent modification, including modifications of the N- or C-terminal residues as well as modifications of selected side chains, for example, at free sulfhydryl-groups, primary amines, and hydroxyl-groups. In one embodiment, a water soluble polymer is linked to the protein (directly or via a linker) by a lysine group or other primary amine. In one embodiment, the rVWF and/or FVIII proteins of the present invention may be modified by conjugation of a water soluble polymer, including without limitation, a polyethylene glycol (PEG), a polypropylene glycol, a polyoxyalkylene, a polysialic acid, hydroxyl ethyl starch, a poly-carbohydrate moiety, and the like.
Water soluble polymers that may be used to modify the rVWF and/or FVIII include linear and branched structures. The conjugated polymers may be attached directly to the coagulation proteins of the invention, or alternatively may be attached through a linking moiety. Non-limiting examples of protein conjugation with water soluble polymers can be found in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192, and 4,179,337, as well as in Abuchowski and Davis “Enzymes as Drugs,” Holcenberg and Roberts, Eds., pp. 367 383, John Wiley and Sons, New York (1981), and Hermanson G., Bioconjugate Techniques 2nd Ed., Academic Press, Inc. 2008.
Protein conjugation may be performed by a number of well known techniques in the art, for example, see Hermanson G., Bioconjugate Techniques 2nd Ed., Academic Press, Inc. 2008. Examples include linkage through the peptide bond between a carboxyl group on one of either the coagulation protein or water-soluble polymer moiety and an amine group of the other, or an ester linkage between a carboxyl group of one and a hydroxyl group of the other. Another linkage by which a coagulation protein of the invention could be conjugated to a water-soluble polymer compound is via a Schiff base, between a free amino group on the polymer moiety being reacted with an aldehyde group formed at the non-reducing end of the polymer by periodate oxidation (Jennings and Lugowski, J. Immunol. 1981; 127:1011-8; Femandes and Gregonradis, Biochim Biophys Acta. 1997; 1341; 26-34). The generated Schiff Base can be stabilized by specific reduction with NaCNBH3 to form a secondary amine. An alternative approach is the generation of terminal free amino groups on the polymer by reductive amination with NH4Cl after prior oxidation. Bifunctional reagents can be used for linking two amino or two hydroxyl groups. For example a polymer containing an amino group can be coupled to an amino group of the coagulation protein with reagents like BS3 (Bis(sulfosuccinimidyl)suberate/Pierce, Rockford, Ill.). In addition heterobifunctional cross linking reagents like Sulfo-EMCS (N-ε-Maleimidocaproyloxy) sulfosuccinimide ester/Pierce) can be used for instance to link amine and thiol groups. In other embodiments, an aldehyde reactive group, such as PEG alkoxide plus diethyl acetal of bromoacetaldehyde; PEG plus DMSO and acetic anhydride, and PEG chloride plus the phenoxide of 4-hydroxybenzaldehyde, succinimidyl active esters, activated dithiocarbonate PEG, 2,4,5-trichlorophenylcloroformate and P-nitrophenylcloroformate activated PEG, may be used in the conjugation of a coagulation protein.
In some aspects, the rVWF used in methods of the present invention has been matured in vitro with Furin. In further embodiments, the Furin is recombinant Furin.
In further aspects, the rVWF and/or rFVIII used in the methods of the present invention are produced by expression in a mammalian cell culture using methods known in the art. In particular embodiments, the mammalian culture comprises CHO cells. In further embodiments, the rVWF and the rFVIII are co-expressed in the same culture. In such embodiments, the rVWF and the rFVIII are purified together (co-purified) or separately using methods known in the art. In other embodiments, the rVWF and the rFVIII are expressed in different cultures.
In an exemplary embodiment, the rVWF of the invention comprises rVWF protein isolated from a CHO cell expression system. In a further embodiment, the propeptide removal is mediated in vitro through exposure of the pro-VWF to Furin—in a still further embodiment, the Furin used for propeptide removal is recombinant Furin. In as yet further embodiment, fully glycosylated/ABO blood group glycans are absent.
In yet further embodiments, the rVWF used in methods and compositions of the present invention by expression in a suitable eukaryotic host system. Examples of eukaryotic cells include, without limitation, mammalian cells, such as CHO, COS, HEK 293, BHK, SK-Hep, and HepG2; insect cells, e.g., SF9 cells, SF21 cells, S2 cells, and High Five cells; and yeast cells, e.g., Saccharomyces or Schizosaccharomyces cells. In one embodiment, the VWF can be expressed in yeast cells, insect cells, avian cells, mammalian cells, and the like. For example, in a human cell line, a hamster cell line, or a murine cell line. In one particular embodiment, the cell line is a CHO, BHK, or HEK cell line. Typically, mammalian cells, e.g., CHO cell from a continuous cell line, can be used to express the VWF of the present invention.
In certain embodiments, the nucleic acid sequence comprising a sequence coding for VWF can be a vector. The vector can be delivered by a virus or can be a plasmid. The nucleic acid sequence coding for the protein can be a specific gene or a biologically functional part thereof. In one embodiment, the protein is at least a biologically active part of VWF.
A wide variety of vectors can be used for the expression of the VWF and can be selected from eukaryotic expression vectors. Examples of vectors for eukaryotic expression include: (i) for expression in yeast, vectors such as pAO, pPIC, pYES, pMET, using promoters such as AOX1, GAP, GAL1, AUG1, etc; (ii) for expression in insect cells, vectors such as pMT, pAc5, pIB, pMIB, pBAC, etc., using promoters such as PH, p10, MT, Ac5, OpIE2, gp64, polh, etc., and (iii) for expression in mammalian cells, vectors such as pSVL, pCMV, pRc/RSV, pcDNA3, pBPV, etc., and vectors derived from viral systems such as vaccinia virus, adeno-associated viruses, herpes viruses, retroviruses, etc., using promoters such as CMV, SV40, EF-1, UbC, RSV, ADV, BPV, and β-actin.
In some embodiments of the present invention, the nucleic acid sequence further comprises other sequences suitable for a controlled expression of a protein such as promoter sequences, enhancers, TATA boxes, transcription initiation sites, polylinkers, restriction sites, poly-A-sequences, protein processing sequences, selection markers, and the like which are generally known to a person of ordinary skill in the art.
In certain embodiments, the cell-culture methods of the invention may comprise the use of a microcarrier. In some embodiments, the cell-cultures of the embodiments can be performed in large bioreactors under conditions suitable for providing high volume-specific culture surface areas to achieve high cell densities and protein expression. One means for providing such growth conditions is to use microcarriers for cell-culture in stirred tank bioreactors. The concept of cell-growth on microcarriers was first described by van Wezel (van Wezel, A. L., Nature 216:64-5 (1967)) and allows for cell attachment on the surface of small solid particles suspended in the growth medium. These methods provide for high surface-to-volume ratios and thus allow for efficient nutrient utilization. Furthermore, for expression of secreted proteins in eukaryotic cell lines, the increased surface-to-volume ratio allows for higher levels of secretion and thus higher protein yields in the supernatant of the culture. Finally, these methods allow for the easy scale-up of eukaryotic expression cultures.
The cells expressing VWF can be bound to a spherical or a porous microcarrier during cell culture growth. The microcarrier can be a microcarrier selected from the group of microcarriers based on dextran, collagen, plastic, gelatine and cellulose and others as described in Butler (1988. In: Spier & Griffiths, Animal Cell Biotechnology 3:283-303). It is also possible to grow the cells to a biomass on spherical microcarriers and subculture the cells when they have reached final fermenter biomass and prior to production of the expressed protein on a porous microcarrier or vice versa. Suitable spherical microcarriers can include smooth surface microcarriers, such as CytodexTM 1, CytodexTM 2, and CytodexTM 3 (GE Healthcare) and macroporous microcarriers such as CytoporeTM 1, CytoporeTM 2, CytolineTM 1, and CytolineTM 2 (GE Healthcare).
In certain embodiments, rVWF is expressed in cells cultured in cell culture media that produces high molecular weight rVWF. The terms “cell culture solution,” “cell culture medium or media,” and “cell culture supernatant” refer to aspects of cell culture processes generally well known in the art. In the context of the present invention, a cell culture solution can include cell culture media and cell culture supernatant. The cell culture media are externally added to the cell culture solution, optionally together with supplements, to provide nutrients and other components for culturing the cells expressing VWF. The cell culture supernatant refers to a cell culture solution comprising the nutrients and other components from the cell culture medium as well as products released, metabolized, and/or excreted from the cells during culture. In further embodiments, the media can be animal protein-free and chemically defined. Methods of preparing animal protein-free and chemically defined culture media are known in the art, for example in US 2008/0009040 and US 2007/0212770, which are both incorporated herein for all purposes and in particular for all teachings related to cell culture media. “Protein free” and related terms refers to protein that is from a source exogenous to or other than the cells in the culture, which naturally shed proteins during growth. In another embodiment, the culture medium is polypeptide free. In another embodiment, the culture medium is serum free. In another embodiment the culture medium is animal protein free. In another embodiment the culture medium is animal component free. In another embodiment, the culture medium contains protein, e.g., animal protein from serum such as fetal calf serum. In another embodiment, the culture has recombinant proteins exogenously added. In another embodiment, the proteins are from a certified pathogen free animal. The term “chemically defined” as used herein shall mean, that the medium does not comprise any undefined supplements, such as, for example, extracts of animal components, organs, glands, plants, or yeast. Accordingly, each component of a chemically defined medium is accurately defined. In a preferred embodiment, the media are animal-component free and protein free.
In certain embodiments, the culture of cells expressing VWF can be maintained for at least about 7 days, or at least about 14 days, 21 days, 28 days, or at least about 5 weeks, 6 weeks, 7 weeks, or at least about 2 months, or 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 months or longer. The cell density at which a cell-culture is maintained at for production of a recombinant VWF protein will depend upon the culture-conditions and medium used for protein expression. One of skill in the art will readily be able to determine the optimal cell density for a cell-culture producing an VWF. In one embodiment, the culture is maintained at a cell density of between about 0.5×106 and 4×107 cells/ml for an extended period of time. In other embodiments, the cell density is maintained at a concentration of between about 1.0×106 and about 1.0×107 cells/ml for an extended period of time. In other embodiments, the cell density is maintained at a concentration of between about 1.0×106 and about 4.0×106 cells/ml for an extended period of time. In other embodiments, the cell density is maintained at a concentration of between about 1.0×106 and about 4.0×106 cells/ml for an extended period of time. In yet other embodiments, the cell density may be maintained at a concentration between about 2.0×106 and about 4.0×106, or between about 1.0×106 and about 2.5×106, or between about 1.5×106 and about 3.5×106, or any other similar range, for an extended period of time. After an appropriate time in cell culture, the rVWF can be isolated from the expression system using methods known in the art.
In a specific embodiment, the cell density of the continuous cell culture for production of rVWF is maintained at a concentration of no more than 2.5×106 cells/mL for an extended period. In other specific embodiments, the cell density is maintained at no more than 2.0×106 cells/mL, 1.5×106 cells/mL, 1.0×106 cells/mL, 0.5×106 cells/mL, or less. In one embodiment, the cell density is maintained at between 1.5×106 cells/mL and 2.5×106 cells/mL.
In one specific embodiment of the cell cultures described above, the cell culture solution comprises a medium supplement comprising copper. Such cell culture solutions are described for example in US 2012/0035110, filed Jul. 8, 2011, which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to cell culture methods and compositions for producing recombinant VWF.
In further embodiments, subsequent to purification (separately or together) from a mammalian cell culture, the rFVIII/rVWF complex is reconstituted prior to administration. In still further embodiments, the rVWF is treated with Furin prior to or subsequent to reconstitution of the rFVIII/rVWF complex. In further embodiments, the Furin is recombinant Furin.
In still further embodiments, the rVWF of the invention is not exposed to ADAMTS13, with the result that ultra large (i.e., comprising 10 or more subunits) are present in rVWF compositions of the invention.
In specific aspects, the rVWF and/or the rFVIII used in methods of the present invention are contained in a formulation containing a buffer, a sugar and/or a sugar alcohol (including without limitation trehalose and mannitol), a stabilizer (such as glycine), and a surfactant (such as Polysorbate 80). In further embodiments, for formulations containing rFVIII, the formulation may further include sodium, histidine, calcium, and glutathione.
In one aspect, the formulations comprising rVWF and/or rFVIII are lyophilized prior to administration. Lyophilization is carried out using techniques common in the art and should be optimized for the composition being developed [Tang et al., Pharm Res. 21:191-200. (2004) and Chang et al., Pharm Res. 13:243-9 (1996)].
Methods of preparing pharmaceutical formulations can include one or more of the following steps: adding a stabilizing agent as described herein to said mixture prior to lyophilizing, adding at least one agent selected from a bulking agent, an osmolarity regulating agent, and a surfactant, each of which as described herein, to said mixture prior to lyophilization. A lyophilized formulation is, in one aspect, at least comprised of one or more of a buffer, a bulking agent, and a stabilizer. In this aspect, the utility of a surfactant is evaluated and selected in cases where aggregation during the lyophilization step or during reconstitution becomes an issue. An appropriate buffering agent is included to maintain the formulation within stable zones of pH during lyophilization.
The standard reconstitution practice for lyophilized material is to add back a volume of pure water or sterile water for injection (WFI) (typically equivalent to the volume removed during lyophilization), although dilute solutions of antibacterial agents are sometimes used in the production of pharmaceuticals for parenteral administration [Chen, Drug Development and Industrial Pharmacy, 18:1311-1354 (1992)]. Accordingly, methods are provided for preparation of reconstituted recombinant VWF (with or without recombinant Factor VIII) compositions comprising the step of adding a diluent to a lyophilized recombinant VWF composition of the invention.
The lyophilized material may be reconstituted as an aqueous solution. A variety of aqueous carriers, e.g., sterile water for injection, water with preservatives for multi dose use, or water with appropriate amounts of surfactants (for example, an aqueous suspension that contains the active compound in admixture with excipients suitable for the manufacture of aqueous suspensions). In various aspects, such excipients are suspending agents, for example and without limitation, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents are a naturally-occurring phosphatide, for example and without limitation, lecithin, or condensation products of an alkylene oxide with fatty acids, for example and without limitation, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example and without limitation, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example and without limitation, polyethylene sorbitan monooleate. In various aspects, the aqueous suspensions also contain one or more preservatives, for example and without limitation, ethyl, or n-propyl, p-hydroxybenzoate.
In certain embodiments, compositions of the present invention are liquid formulations for administration with the use of a syringe or other storage vessel. In further embodiments, these liquid formulations are produced from lyophilized material described herein reconstituted as an aqueous solution.
In a further aspect, the compositions of the invention further comprise one or more pharmaceutically acceptable carriers. The phrases “pharmaceutically” or “pharmacologically” acceptable refer to molecular entities and compositions that are stable, inhibit protein degradation such as aggregation and cleavage products, and in addition do not produce allergic, or other adverse reactions when administered using routes well-known in the art, as described below. “Pharmaceutically acceptable carriers” include any and all clinically useful solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like, including those agents disclosed above.
Administration of Compositions of the Invention for Methods of Treating Disease
One of the advantages of administering rVWF to subjects to treat coagulation disease is that the higher specific activity of rVWF as compared to pdVWF allows flexibility in the amount of rVWF administered and the number of times the subject is re-dosed with rVWF (with or without co-administered FVIII). In addition, rVWF compositions provide the further flexibility of re-dosing with rVWF alone after an initial co-administration of rVWF and FVIII, without need for additional dosing with FVIII. As will be appreciated and as is discussed in further detail herein, the co-administered FVIII may be recombinant or plasma derived.
In one aspect, the administration of rVWF in accordance with the invention results in higher plasma FVIII levels and/or activity in the subject than is seen with a subject administered pdVWF. As discussed above, increases in FVIII levels and activity can be measured using methods standard in the art, thus allowing for determination of appropriate dosages for rVWF with or without FVIII.
Single or multiple administrations of rVWF (with or without FVIII) are carried out with the dose levels and pattern being selected by the treating physician. For the prevention or treatment of disease, the appropriate dosage depends on the type of disease to be treated (e.g., von Willebrand disease), the severity and course of the disease, whether drug is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the drug, and the discretion of the attending physician.
In further embodiments and in accordance with any of the above, treatment of coagulation diseases such as VWD or Hemophilia A may involve an initial treatment of rVWF alone or in combination with FVIII, followed by one or more repeat doses of rVWF alone, rVWF plus FVIII together, or FVIII alone. The nature of the initial and then the subsequent repeat administrations will depend in part on the disease being treated. For example, patients with VWD have some level of FVIII, but the stability of that FVIII is generally compromised because these patients lack VWF. Treatment of VWD patients may thus in some embodiments involve an initial treatment with both rVWF and rFVIII followed by repeated administrations of rVWF alone. In other embodiments, the initial treatment may be with rVWF alone while subsequent repeated administrations are with both rVWF and rFVIII. In still other embodiments, the initial and subsequent repeat administrations may all include a co-administration of both rVWF and rFVIII. Similarly, Hemophilia A patients (who lack FVIII) may receive an initial treatment of both rVWF and rFVIII, and subsequent repeat treatments may comprise the administration of rFVIII alone or rVWF alone. In other embodiments, the initial treatment may be rFVIII alone while the subsequent repeat treatments involve co-administration of rVWF and rFVIII.
In further aspects, rVWF is administered to a subject in doses ranging from 0.5 IU/kg-200 IU/kg. In some embodiments, rVWF is administered in doses ranging from 1-190, 5-180, 10-170, 15-160, 20-150, 25-140, 30-130, 35-120, 40-110, 45-100, 50-90, 55-80, or 60-70 IU/kg. In further embodiments and in accordance with any of the above, rVWF (with or without FVIII) is administered to a subject at doses of between about 1 IU/kg to about 150 IU/kg rVWF. In still further embodiments, the rVWF and rFVIII is administered at doses of between 1.5 IU/kg to 150 IU/kg, 2 IU/kg to 50 IU/kg, 5 IU/kg to 40 IU/kg, 10 IU/kg to 20 IU/kg, 10 IU/kg to 100 IU/kg, 25 IU/kg to 75 IU/kg, and 40 IU/kg to 75 IU/kg. In still further embodiments, rVWF is administered at 2, 5, 7.5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 IU/kg. As will be appreciated and as is discussed further herein, appropriate dosages of rVWF (or rVWF together with FVIII) may be ascertained through use of established assays for determining blood level dosages in conjunction with appropriate dose-response data. In one embodiment, rVWF is administered to a subject in a dose selected from variations 2141 to 2338 in Table 12.







TABLE 12







Exemplary embodiments for the dosage of rVWF administered


to a subject according to the methods provided herein.








IU/kg












About 0.5
Var. 2141


About 1
Var. 2142


About 2
Var. 2143


About 3
Var. 2144


About 4
Var. 2145


About 5
Var. 2146


About 10
Var. 2147


About 15
Var. 2148


About 20
Var. 2149


About 25
Var. 2150


About 30
Var. 2151


About 35
Var. 2152


About 40
Var. 2153


About 45
Var. 2154


About 50
Var. 2155


About 55
Var. 2156


About 60
Var. 2157


About 65
Var. 2158


About 70
Var. 2159


About 75
Var. 2160


About 80
Var. 2161


About 85
Var. 2162


About 90
Var. 2163


About 95
Var. 2164


About 100
Var. 2165


About 105
Var. 2166


About 110
Var. 2167


About 115
Var. 2168


About 120
Var. 2169


About 125
Var. 2170


About 130
Var. 2171


About 135
Var. 2172


About 140
Var. 2173


About 145
Var. 2174


About 150
Var. 2175


About 155
Var. 2176


About 160
Var. 2177


About 165
Var. 2178


About 170
Var. 2179


About 175
Var. 2180


About 180
Var. 2181


About 185
Var. 2182


About 190
Var. 2183


About 195
Var. 2184


About 200
Var. 2185


 0.5-200
Var. 2186


 0.5-175
Var. 2187


 0.5-150
Var. 2188


 0.5-125
Var. 2189


 0.5-100
Var. 2190


0.5-75 
Var. 2191


0.5-50 
Var. 2192


0.5-40 
Var. 2193


0.5-30 
Var. 2194


0.5-25 
Var. 2195


0.5-20 
Var. 2196


0.5-15 
Var. 2197


0.5-10 
Var. 2198


0.5-7.5
Var. 2199


0.5-5  
Var. 2200


0.5-2.5
Var. 2201


0.5-1  
Var. 2202


 1-200
Var. 2203


 1-175
Var. 2204


 1-150
Var. 2205


 1-125
Var. 2206


 1-100
Var. 2207


 1-75
Var. 2208


 1-50
Var. 2209


 1-40
Var. 2210


 1-30
Var. 2211


 1-25
Var. 2212


 1-20
Var. 2213


 1-15
Var. 2214


 1-10
Var. 2215


  1-7.5
Var. 2216


1-5
Var. 2217


  1-2.5
Var. 2218


 2.5-200
Var. 2219


 2.5-175
Var. 2220


 2.5-150
Var. 2221


 2.5-125
Var. 2222


 2.5-100
Var. 2223


2.5-75 
Var. 2224


2.5-50 
Var. 2225


2.5-40 
Var. 2226


2.5-30 
Var. 2227


2.5-25 
Var. 2228


2.5-20 
Var. 2229


2.5-15 
Var. 2230


2.5-10 
Var. 2231


2.5-7.5
Var. 2232


2.5-5  
Var. 2233


 5-200
Var. 2234


 5-175
Var. 2235


 5-150
Var. 2236


 5-125
Var. 2237


 5-100
Var. 2238


 5-75
Var. 2239


 5-50
Var. 2240


 5-40
Var. 2241


 5-30
Var. 2242


 5-25
Var. 2243


 5-20
Var. 2244


 5-15
Var. 2245


 5-10
Var. 2246


  5-7.5
Var. 2247


 7.5-200
Var. 2248


 7.5-175
Var. 2249


 7.5-150
Var. 2250


 7.5-125
Var. 2251


 7.5-100
Var. 2252


7.5-75 
Var. 2253


7.5-50 
Var. 2254


7.5-40 
Var. 2255


7.5-30 
Var. 2256


7.5-25 
Var. 2257


7.5-20 
Var. 2258


7.5-15 
Var. 2259


7.5-10 
Var. 2260


 10-200
Var. 2261


 10-175
Var. 2262


 10-150
Var. 2263


 10-125
Var. 2264


 10-100
Var. 2265


10-75
Var. 2266


10-50
Var. 2267


10-40
Var. 2268


10-30
Var. 2269


10-25
Var. 2270


10-20
Var. 2271


10-15
Var. 2272


 15-200
Var. 2273


 15-175
Var. 2274


 15-150
Var. 2275


 15-125
Var. 2276


 15-100
Var. 2277


15-75
Var. 2278


15-50
Var. 2279


15-40
Var. 2280


15-30
Var. 2281


15-25
Var. 2282


15-20
Var. 2283


 20-200
Var. 2284


 20-175
Var. 2285


 20-150
Var. 2286


 20-125
Var. 2287


 20-100
Var. 2288


20-75
Var. 2289


20-50
Var. 2290


20-40
Var. 2291


20-30
Var. 2292


20-25
Var. 2293


 25-200
Var. 2294


 25-175
Var. 2295


 25-150
Var. 2296


 25-125
Var. 2297


 25-100
Var. 2298


25-75
Var. 2299


25-50
Var. 2300


25-40
Var. 2301


25-30
Var. 2302


 30-200
Var. 2303


 30-175
Var. 2304


 30-150
Var. 2305


 30-125
Var. 2306


 30-100
Var. 2307


30-75
Var. 2308


30-50
Var. 2309


30-40
Var. 2310


 40-200
Var. 2311


 40-175
Var. 2312


 40-150
Var. 2313


 40-125
Var. 2314


 40-100
Var. 2315


40-75
Var. 2316


40-50
Var. 2317


 50-200
Var. 2318


 50-175
Var. 2319


 50-150
Var. 2320


 50-125
Var. 2321


 50-100
Var. 2322


50-75
Var. 2323


 75-200
Var. 2324


 75-175
Var. 2325


 75-150
Var. 2326


 75-125
Var. 2327


 75-100
Var. 2328


100-200
Var. 2329


100-175
Var. 2330


100-150
Var. 2331


100-125
Var. 2332


125-200
Var. 2333


125-175
Var. 2334


125-150
Var. 2335


150-200
Var. 2336


150-200
Var. 2337


175-200
Var. 2338





Var. = Variation






In still further embodiments, rVWF is administered at a dose such that it increases half-life of plasma FVIII by about 1.0-4.5, 1.5-4.0, 2.0-3.5, 2.5-3.0 fold. In still further embodiments, the dose and/or frequency of rVWF administration is such that the increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In yet further embodiments, the dose and/or frequency of rVWF administration is such that the increase in FVIII half-life is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF. In one embodiment, rVWF is administered at a dose such that it increases the half-life of plasma FVIII by a value selected from variations 1046 to 1089 found in Table 7.
As discussed above, the rVWF of the invention shows an increased effect on the stability of FVIII as compared to the effect of pdVWF. In certain aspects, rVWF is administered at a dose and/or with a frequency such that it increases average FVIII half-life by about 1-15, 2-14, 3-13, 4-12, 5-11, 6-10, 5-9, 6-8 hours. In still further embodiments, rVWF is administered at a dose and/or frequency such that it increases FVIII half-life by about 10% to about 75% as compared to pdVWF. In yet further embodiments, rVWF increases FVIII half-life by about 10-80%, 15-65%, 20-60%, 25-55%, 30-50%, 35-45% as compared to pdVWF. In yet further embodiments, the average or percentage increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In yet further embodiments, the increase in FVIII half-life is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF. As will be appreciated, the increase in FVIII stability, half-life and/or activity can be assessed using methods known in the art, including without limitation coagulation assays. In other embodiments, administration of rVWF increases the half-life of FVIII by an amount selected from variations 1300 to 1643 found in Table 9, as compared to administration of plasma-derived VWF.
As discussed herein, the stabilization of FVIII activity by rVWF as compared to pdVWF can be measured by metrics in addition to FVIII half-life, including mean residence time (MRT) and area under curve (AUC). In exemplary embodiments, rVWF is administered at a dose and/or frequency such that it increases MRT by about 1-15 hours as compared to pdVWF. In further embodiments, rVWF increases MRT by about 1-25, 2-20, 3-15, 4-10, 5-9, 6-8 hours as compared to pdVWF. In still further embodiments, rVWF increases FVIII MRT by about 10-80%, 15-65%, 20-60%, 25-55%, 30-50%, 35-45% as compared to pdVWF. In yet further embodiments, the average or percentage increase in FVIII half-life is maintained at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 hours after administration of rVWF. In yet further embodiments, the increase in MRT is maintained at least 5-125, 10-115, 15-105, 20-95, 25-85, 30-75, 35-65, 40-55 hours after administration of rVWF. In other embodiments, administration of rVWF increases the MRT of FVIII by an amount selected from variations 1300 to 1643 found in Table 9, as compared to administration of plasma-derived VWF.
In further aspects, the doses of rVWF administered to patients are comparable to doses used in administration of pdVWF/pdFVIII.
Compositions of rVWF with or without FVIII can be contained in pharmaceutical formulations, as described herein. Such formulations can be administered orally, topically, transdermally, parenterally, by inhalation spray, vaginally, rectally, or by intracranial injection. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intracisternal injection, or infusion techniques. Administration by intravenous, intradermal, intramuscular, intramammary, intraperitoneal, intrathecal, retrobulbar, intrapulmonary injection and or surgical implantation at a particular site is contemplated as well. Generally, compositions are essentially free of pyrogens, as well as other impurities that could be harmful to the recipient.
In one aspect, formulations of the invention are administered by an initial bolus followed by a continuous infusion to maintain therapeutic circulating levels of drug product. As another example, the inventive compound is administered as a one-time dose. Those of ordinary skill in the art will readily optimize effective dosages and administration regimens as determined by good medical practice and the clinical condition of the individual patient. The route of administration can be, but is not limited to, by intravenous, intraperitoneal, subcutaneous, or intramuscular administration. The frequency of dosing depends on the pharmacokinetic parameters of the agents and the route of administration. The optimal pharmaceutical formulation is determined by one skilled in the art depending upon the route of administration and desired dosage. See for example, Remington's Pharmaceutical Sciences, 18th Ed., 1990, Mack Publishing Co., Easton, Pa. 18042 pages 1435-1712, the disclosure of which is hereby incorporated by reference in its entirety for all purposes and in particular for all teachings related to formulations, routes of administration and dosages for pharmaceutical products. Such formulations influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the administered agents. Depending on the route of administration, a suitable dose is calculated according to body weight, body surface area or organ size. Appropriate dosages may be ascertained through use of established assays for determining blood level dosages in conjunction with appropriate dose-response data. The final dosage regimen is determined by the attending physician, considering various factors which modify the action of drugs, e.g. the drug's specific activity, the severity of the damage and the responsiveness of the patient, the age, condition, body weight, sex and diet of the patient, the severity of any infection, time of administration and other clinical factors. By way of example, a typical dose of a recombinant VWF of the present invention is approximately 50 U/kg, equal to 500 μg/kg. As studies are conducted, further information will emerge regarding the appropriate dosage levels and duration of treatment for various diseases and conditions.
In some embodiments, rVWF is administered to a subject alone. In some embodiments, rVWF is administered to a subject in combination with one or more coagulation factors. In further embodiments, the coagulation factor administered with rVWF is FVIII. In still further embodiments rVWF is administered prior to, subsequent to, or simultaneously with a coagulation factor such as FVIII. In certain embodiments, rVWF and FVIII are administered together in a single composition. As will be appreciated, the FVIII that is co-administered with rVWF can be either recombinant FVIII or plasma derived.
In further embodiments, rVWF (with or without FVIII) is administered to a subject no more than once daily. In further embodiments, rVWF (with or without FVIII) is administered to a subject: no more than once every other day, no more than once every third day, no more than once every fourth day, no more than once every fifth day, no more than once a week, no more than once every two weeks, no more than once a month. In still further embodiments, rVWF (with or without FVIII) is administered to a subject no more than twice a day.
In further embodiments, rVWF and FVIII are administered together to a subject in an initial dose, and then subsequent re-dosing is conducted with rVWF alone. In other embodiments, re-dosing is conducted with both rVWF and FVIII.
In still further embodiments, rVWF (with or without rFVIII) is administered at a dose such that plasma FVIII activity is stabilized for about 10 to about 90 hours. In further embodiments, plasma FVIII activity is stabilized for at least 12, 24, 36, 48 or 72 hours. As will be appreciated, the stabilized plasma FVIII activity may be that of endogenous FVIII, co-administered FVIII (plasma-derived or recombinant) or a combination of both endogenous and co-administered FVIII.
In some embodiments, rVWF and FVIII are administered together at a dose such that extension of in vivo half-life of plasma FVIII activity is stabilized for at least 12, 24, 36, 48 or 72 hours. In further embodiments, the plasma FVIII activity is stabilized for about 10 to about 90 hours. In still further embodiments, the increase in half-life of plasma FVIII is maintained for at least 24, 36, 48, 72, 90, 120, or 168 hours in a patient. The co-administered FVIII can be rFVIII or pdFVIII. In some embodiments, plasma FVIII activity is stabilized for a time selected from variations 1090 to 1299 found in Table 8, after co-administration of rVWF and FVIII.
In preferred aspects, the present invention provides methods for treating coagulation disease, including hemophilia and von Willebrand Disease (VWD).
As used herein, the terms “hemophilia” or “haemophilia” refer to a group of disease states broadly characterized by reduced blood clotting or coagulation. Hemophilia may refer to Type A, Type B, or Type C hemophilia, or to the composite of all three diseases types. Type A hemophilia (hemophilia A) is caused by a reduction or loss of factor VIII (FVIII) activity and is the most prominent of the hemophilia subtypes. Type B hemophilia (hemophilia B) results from the loss or reduction of factor IX (FIX) clotting function. Type C hemophilia (hemophilia C) is a consequence of the loss or reduction in factor XI (FXI) clotting activity. Hemophilia A and B are X-linked diseases, while hemophilia C is autosomal. Common treatments for hemophilia include both prophylactic and on-demand administration of clotting factors, such as FVIII, FIX, including Bebulin®-VH, and FXI, as well as FEIBA-VH, desmopressin, and plasma infusions.
As used herein “von Willebrand Disease” refers to the group of diseases caused by a deficiency of von Willebrand factor. Von Willebrand factor helps blood platelets clump together and stick to the blood vessel wall, which is necessary for normal blood clotting. There are several types of Von Willebrand disease. The following table summarizes the characteristics of different types of VWD:











Type
Characteristics








Quantitative forms of VWD


Type 1
Partial quantitative deficiencies of VWF



VWF plasma levels that are 5% to 30% of normal



approximately 60% to 80% of patients


Type 3
Virtually complete deficiency of VWF



Approximately 1% to 5% of patients



Qualitative forms of VWD


Type 2
Approximately 10% to 30% of patients


Type 2A
Decreased VWF-dependent platelet adhesion



Selective deficiency of HMW-VMF multimers



Platelet binding functions of VWF are impaired


Type 2B
Gain of function mutation with increased VWF binding to



platelets



Loss of HMW-VWF multimers from plasma, but not from



platelets



Loss of both VWF and platelets through a clearance



mechanism


Type 2M
Decreased VWF-dependent platelet adhesion without selective



deficiency of HMW-VWF multimers


Type 2N
Markedly decreased binding affinity for factor VIII





Adopted from Sadler, J Thromb Haemost 2006; 4, 2103






Methods for treating coagulation disease include administering rVWF or a combination of rVWF rFVIII to subjects in need thereof in accordance with any of the methods of administration described herein and known in the art. Such subjects may be suffering from any coagulation disease, including without limitation von Willebrand Disease or hemophilia. As will be appreciated, any type of von Willebrand Disease, including any of the types listed in the above table, can be treated in accordance with any of the methods and compositions described herein.
In some embodiments, rVWF (with or without rFVIII) is administered to a subject such that the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24, 36, 48 or more hours post-administration is at least 90% of the level of FVIII:C activity present in the plasma 1 hour post-administration. In further embodiments, the level of the FVIII:C in the plasma of the subject 24, 36, 48 or more hours post-administration is at least between 50% and 100% of the level of FVIII:C activity present in the plasma 1 hour post-administration. In still further embodiments, the level of the FVIII:C in the plasma of the subject 24, 36, 48 or more hours post-administration is at least 50%, 55%, 60%, 65%. 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%. 99%, 100% of the level of FVIII:C activity present in the plasma 1 hour post-administration.
Administration of rVWF
In one aspect, the present disclosure provides method for treating Von Willebrand Disease (VWD) or Hemophilia A in a subject in need thereof, which includes administering a composition of recombinant Von Willebrand Factor (rVWF) such that Factor VIII (FVIII) stability is increased, as compared to FVIII half-life in a subject administered a composition of plasma derived Von Willebrand Factor (pdVWF). In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF such that FVIII stability is extended by at least 10%, 20%, 30%, 2 hr, 4 hr, 6 hr, or by an amount selected from variations 1300 to 1643 found in Table 9, as compared to FVIII stability in a subject administered a composition of pdVWF. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF such that FVIII stability is extended by at least 10% as compared to FVIII stability in a subject administered a composition of pdVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF such that FVIII stability is extended by at least 20% as compared to FVIII stability in a subject administered a composition of pdVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF such that FVIII stability is extended by at least 30% as compared to FVIII stability in a subject administered a composition of pdVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF. wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.







TABLE 13







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved,


as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)
















at least 1 hr
at least 2 hr
at least 3 hr
at least 4 hr
at least 5 hr
at least 6 hr
at least 7 hr
at least 8 hr



















(mU/μg)
at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



20
2339
2385
2431
2477
2523
2569
2615
2661



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



30
2340
2386
2432
2478
2524
2570
2616
2662



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



40
2341
2387
2433
2479
2525
2571
2617
2663



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



50
2342
2388
2434
2480
2526
2572
2618
2664



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



60
2343
2389
2435
2481
2527
2573
2619
2665



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



70
2344
2390
2436
2482
2528
2574
2620
2666



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



80
2345
2391
2437
2483
2529
2575
2621
2667



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



90
2346
2392
2438
2484
2530
2576
2622
2668



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



100
2347
2393
2439
2485
2531
2577
2623
2669



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



125
2348
2394
2440
2486
2532
2578
2624
2670



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



150
2349
2395
2441
2487
2533
2579
2625
2671



20-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2350
2396
2442
2488
2534
2580
2626
2672



20-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2351
2397
2443
2489
2535
2581
2627
2673



20-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2352
2398
2444
2490
2536
2582
2628
2674



20-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2353
2399
2445
2491
2537
2583
2629
2675



20-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2354
2400
2446
2492
2538
2584
2630
2676



20-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2355
2401
2447
2493
2539
2585
2631
2677



20-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2356
2402
2448
2494
2540
2586
2632
2678



20-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2357
2403
2449
2495
2541
2587
2633
2679



20-40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2358
2404
2450
2496
2542
2588
2634
2680



40-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2359
2405
2451
2497
2543
2589
2635
2681



40-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2360
2406
2452
2498
2544
2590
2636
2682



40-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2361
2407
2453
2499
2545
2591
2637
2683



40-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2362
2408
2454
2500
2546
2592
2638
2684



40-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2363
2409
2455
2501
2547
2593
2639
2685



40-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2364
2410
2456
2502
2548
2594
2640
2686



40-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2365
2411
2457
2503
2549
2595
2641
2687



40-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2366
2412
2458
2504
2550
2596
2642
2688



60-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2367
2413
2459
2505
2551
2597
2643
2689



60-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2368
2414
2460
2506
2552
2598
2644
2690



60-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2369
2415
2461
2507
2553
2599
2645
2691



60-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2370
2416
2462
2508
2554
2600
2646
2692



60-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2371
2417
2463
2509
2555
2601
2647
2693



60-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2372
2418
2464
2510
2556
2602
2648
2694



70-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2373
2419
2465
2511
2557
2603
2649
2695



70-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2374
2420
2466
2512
2558
2604
2651
2696



70-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2375
2421
2467
2513
2559
2605
2651
2697



70-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2376
2422
2468
2514
2560
2606
2652
2698



70-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2377
2423
2469
2515
2561
2607
2653
2699



80-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2378
2424
2470
2516
2562
2608
2654
2700



80-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2379
2425
2471
2517
2563
2609
2655
2701



80-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2380
2426
2472
2518
2564
2610
2656
2702



80-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2381
2427
2473
2519
2565
2611
2657
2703



90-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2382
2428
2474
2520
2566
2612
2658
2704



90-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2383
2429
2475
2521
2567
2613
2659
2705



90-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2384
2430
2476
2522
2568
2614
2660
2706





Var. = Variation













TABLE 14







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)
















1-8 hr
1-7 hr
1-6 hr
1-5 hr
1-4 hr
1-3 hr
1-2 hr
2-8 hr



















(mU/μg)
at least 20
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2707
2753
2799
2845
2891
2937
2983
3029



at least 30
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2708
2754
2800
2846
2892
2938
2984
3030



at least 40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2709
2755
2801
2847
7893
2939
2985
3031



at least 50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2710
2756
2802
2848
2894
2940
2986
3032



at least 60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2711
2757
2803
2849
2895
2941
2987
3033



at least 70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2712
2758
2804
2850
2896
2942
2988
3034



at least 80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2713
2759
2805
2851
2897
2943
2989
3035



at least 90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2714
2760
2806
2852
2898
2944
2990
3036



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



100
2715
2761
2807
2853
2899
2945
2991
3037



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



125
2716
2762
2808
2854
2900
2946
2992
3038



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



150
2717
2763
2809
2855
2901
2947
2993
3039



20-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2718
2764
2810
2856
2902
2948
2994
3040



20-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2719
2765
2811
2857
2903
2949
2995
3041



20-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2720
2766
2812
2858
2904
2950
2996
3042



20-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2721
2767
2813
2859
2905
2951
2997
3043



20-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2722
2768
2814
2860
2906
2952
2998
3044



20-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2723
2769
2815
2861
2907
2953
2999
3045



20-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2724
2770
2816
2862
2908
2954
3000
3046



20-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2725
2771
2817
2863
2909
2955
3001
3047



20-40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2726
2772
2818
2864
2910
2956
3002
3048



40-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2727
2773
2819
2865
2911
2957
3003
3049



40-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2728
2774
2820
2866
2912
2958
3004
3050



40-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2779
2775
2821
2867
2913
2959
3005
3051



40-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2730
2776
2822
2868
2914
2960
3006
3052



40-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2731
2777
2823
2869
2915
2961
3007
3053



40-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2732
2778
2824
2870
2916
2962
3008
3054



40-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2733
2779
2825
2871
2917
2963
3009
3055



40-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2734
2780
2826
2872
2918
2964
3010
3056



60-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2735
2781
2827
2873
2919
2965
3011
3057



60-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2736
2782
2828
2874
2920
2966
3012
3058



60-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2737
2783
2829
2875
2921
2967
3013
3059



60-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2738
2784
2830
7876
2922
2968
3014
3060



60-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2739
2785
2831
2877
2923
2969
3015
3061



60-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2740
2786
2832
2878
2924
2970
3016
3062



70-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2741
2787
2833
2879
2925
2971
3017
3063



70-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2742
2788
2834
2880
2926
2972
3018
3064



70-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2743
2789
2835
2881
2927
2973
3019
3065



70-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2744
2790
2836
2887
2928
2974
3020
3066



70-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2745
7791
2837
2883
2929
2975
3021
3067



80-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2746
2792
2838
2884
2930
2976
3022
3068



80-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2747
2793
2839
2885
2931
2977
3023
3069



80-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2748
2794
2840
2886
2932
2978
3024
3070



80-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2749
2795
2841
2887
2933
2979
3025
3071



90-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2750
2796
2842
2888
2934
2980
3026
3072



90-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2751
2797
2843
2889
2935
2981
3027
3073



90-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




2752
2798
2844
2890
2936
2982
3028
3074





Var. = Variation













TABLE 15







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)
















2-7 hr
2-6 hr
2-5 hr
2-4 hr
2-3 hr
3-8 hr
3-7 hr
3-6 hr



















(mU/μg)
at least 20
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3075
3121
3167
3213
3259
3305
3351
3397



at least 30
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3076
3122
3168
3214
3260
3306
3352
3398



at least 40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3077
3123
3169
3215
3261
3307
3353
3399



at least 50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3078
3124
3170
3216
3262
3308
3354
3400



at least 60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3079
3125
3171
3217
3263
3309
3355
3401



at least 70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3080
3126
1177
3218
3264
3310
3356
3402



at least 80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3081
3127
3173
3219
3265
3311
3357
3403



at least 90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3082
3128
3174
3220
3266
3312
3358
3404



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



100
3083
3129
3175
3221
3267
3313
3359
3405



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



125
3084
3130
3176
3222
3268
3314
3360
3406



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



150
3085
3131
3177
3223
3269
3315
3361
3407



20-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3086
3132
3178
3224
3270
3316
3362
3408



20-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3087
3133
3179
3225
3271
3317
3363
3409



20-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3088
3134
3180
3226
3272
3318
3364
3410



20-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3089
3135
3181
3227
3273
3319
3365
3411



20-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3090
3136
3182
3228
3274
3320
3366
3412



20-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3091
3137
3183
3229
3275
3321
3367
3413



20-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3092
3138
3184
3230
3276
3322
3368
3414



20-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3093
3139
3185
3231
3277
3323
3369
3415



20-40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3094
3140
3186
3232
3278
3324
3370
3416



40-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3095
3141
3187
3233
3279
3325
3371
3417



40-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3096
3142
3188
3234
3280
3326
3372
3418



40-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3097
3143
3189
3235
3281
3327
3373
3419



40-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3098
3144
3190
3236
3282
3328
3374
3420



40-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3099
3145
3191
3237
3283
3329
3375
3421



40-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3100
3146
3192
3238
3284
3330
3376
3422



40-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3101
3147
3193
3239
3285
3331
3377
3423



40-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3102
3148
3194
3240
3286
3332
3378
3424



60-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3103
3149
3195
3241
3287
3333
3379
3425



60-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3104
3150
3196
3242
3288
3334
3380
3426



60-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3105
3151
3197
3243
3289
3335
3381
3427



60-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3106
3152
3198
3244
3290
3336
3382
3428



60-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3107
3153
3199
3245
3291
3337
3383
3429



60-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3108
3154
3200
3246
3292
3338
3384
3430



70-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3109
3155
3201
3247
3293
3339
3385
3431



70-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3110
3156
3202
3248
3294
3340
3386
3432



70-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3111
3157
3203
3249
3295
3341
3387
3433



70-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3112
3158
3204
3250
3296
3342
3388
3434



70-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3113
3159
3205
3251
3297
3343
3389
3435



80-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3114
3160
3206
3252
3298
3344
3390
3436



80-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3115
3161
3207
3253
3299
3345
3391
3437



80-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3116
3162
3208
3254
3300
3346
3392
3438



80-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3117
3163
3209
3255
3301
3347
3393
3439



90-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3118
3164
3210
3256
3302
3348
3394
3440



90-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3119
3165
3211
3257
3303
3349
3395
3441



90-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3120
3166
3212
3258
3304
3350
3396
3442





Var. = Variation













TABLE 16







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)
















3-5 hr
3-4 hr
4-8 hr
4-7 hr
4-6 hr
4-5 hr
5-8 hr
5-7 hr



















(mU/μg)
at least 20
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3443
3489
3535
3581
3627
3673
3719
3765



at least 30
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3444
3490
3536
3582
3628
3674
3720
3766



at least 40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3445
3491
3537
3583
3629
3675
3721
3767



at least 50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3446
3492
3538
3584
3630
3676
3722
3768



at least 60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3447
3493
3539
3585
3631
3677
3723
3769



at least 70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3448
3494
3540
3586
3632
3678
3724
3770



at least 80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3449
3495
3541
3587
3633
3679
3725
3771



at least 90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3450
3496
3542
3588
3634
3680
3726
3772



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



100
3451
3497
3543
3589
3635
3681
3727
3773



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



125
3452
3498
3544
3590
3636
3682
3728
3774



at least
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.



150
3453
3499
3545
3591
3637
3683
3729
3775



20-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3454
3500
3546
3592
3638
3684
3730
3776



20-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3455
3501
3547
3593
3639
3685
3731
3777



20-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3456
3502
3548
3594
3640
3686
3732
3778



20-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3457
3503
3549
3595
3641
3687
3733
3779



20-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3458
3504
3550
3596
3642
3688
3734
3780



20-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3459
3505
3551
3597
3643
3689
3735
3781



20-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3460
3506
3552
3598
3644
3690
3736
3782



20-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3461
3507
3553
3599
3645
3691
3737
3783



20-40
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3462
3508
3554
3600
3646
3692
3738
3784



40-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3463
3509
3555
3601
3647
3693
3739
3785



40-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3464
3510
3556
3602
3648
3694
3740
3786



40-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3465
3511
3557
3603
3649
3695
3741
3787



40-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3466
3512
3558
3604
3650
3696
3742
3788



40-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3467
3513
3559
3605
3651
3697
3743
3789



40-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3468
3514
3560
3606
3652
3698
3744
3790



40-60
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3469
3515
3561
3607
3653
3699
3745
3791



40-50
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3470
3516
3562
3608
3654
3700
3746
3792



60-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3471
3517
3563
3609
3655
3701
3747
3793



60-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3472
3518
3564
3610
3656
3702
3748
3794



60-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3473
3519
3565
3611
3657
3703
3749
3795



60-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3474
3520
3566
3612
3658
3704
3750
3796



60-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3475
3521
3567
3613
3659
3705
3751
3797



60-70
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3476
3522
3568
3614
3660
3706
3752
3798



70-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3477
3523
3569
3615
3661
3707
3753
3799



70-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3478
3524
3570
3616
3662
3708
3754
3800



70-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3479
3525
3571
3617
3663
3709
3755
3801



70-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3480
3526
3572
3618
3664
3710
3756
3802



70-80
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3481
3527
3573
3619
3665
3711
3757
3803



80-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3482
3528
3574
3620
3666
3712
3758
3804



80-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3483
3529
3575
3621
3667
3713
3759
3805



80-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3484
3530
3576
3622
3668
3714
3760
3806



80-90
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3485
3531
3577
3623
3669
3715
3761
3807



90-150
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3486
3532
3578
3624
3670
3716
3762
3808



90-125
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3487
3533
3579
3625
3671
3717
3763
3809



90-100
Var.
Var.
Var.
Var.
Var.
Var.
Var.
Var.




3488
3534
3580
3626
3672
3718
3764
3810





Var. = Variation













TABLE 17







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Percent)













Increased Stability (Hours)
at least
at least
at least
at least
















5-6 hr
6-8 hr
6-7 hr
7-8 hr
10%
20%
30%
40%



















(mU/μg)
at least
Var.
Var.
Var.
Var.
Var. 3995
Var. 4041
Var. 4087
Var. 4133



20
3811
3857
3903
3949







at least
Var.
Var.
Var.
Var.
Var. 3996
Var. 4042
Var. 4088
Var. 4134



30
3812
3858
3904
3950







at least
Var.
Var.
Var.
Var.
Var. 3997
Var. 4043
Var. 4089
Var. 4135



40
3813
3859
3905
3951







at least
Var.
Var.
Var.
Var.
Var. 3998
Var. 4044
Var. 4090
Var. 4136



50
3814
3860
3906
3952







at least
Var.
Var.
Var.
Var.
Var. 3999
Var. 4045
Var. 4091
Var. 4137



60
3815
3861
3907
3953







at least
Var.
Var.
Var.
Var.
Var. 4000
Var. 4046
Var. 4092
Var. 4138



70
3816
3862
3908
3954







at least
Var.
Var.
Var.
Var.
Var. 4001
Var. 4047
Var. 4093
Var. 4139



80
3817
3863
3909
3955







at least
Var.
Var.
Var.
Var.
Var. 4002
Var. 4048
Var. 4094
Var. 4140



90
3818
3864
3910
3956







at least
Var.
Var.
Var.
Var.
Var. 4003
Var. 4049
Var. 4095
Var. 4141



100
3819
3865
3911
3957







at least
Var.
Var.
Var.
Var.
Var. 4004
Var. 4050
Var. 4096
Var. 4142



125
3820
3866
3912
3958







at least
Var.
Var.
Var.
Var.
Var. 4005
Var. 4051
Var. 4097
Var. 4143



150
3821
3867
3913
3959







20-150
Var.
Var.
Var.
Var.
Var. 4006
Var. 4052
Var. 4098
Var. 4144




3822
3868
3914
3960







20-125
Var.
Var.
Var.
Var.
Var. 4007
Var. 4053
Var. 4099
Var. 4145




3823
3869
3915
3961







20-100
Var.
Var.
Var.
Var.
Var. 4008
Var. 4054
Var. 4100
Var. 4146




3824
3870
3916
3962







20-90
Var.
Var.
Var.
Var.
Var. 4009
Var. 4055
Var. 4101
Var. 4147




3825
3871
3917
3963







20-80
Var.
Var.
Var.
Var.
Var. 4010
Var. 4056
Var. 4102
Var. 4148




3826
3872
3918
3964







20-70
Var.
Var.
Var.
Var.
Var. 4011
Var. 4057
Var. 4103
Var. 4149




3827
3873
3919
3965







20-60
Var.
Var.
Var.
Var.
Var. 4012
Var. 4058
Var. 4104
Var. 4150




3828
3874
3920
3966







20-50
Var.
Var.
Var.
Var.
Var. 4013
Var. 4059
Var. 4105
Var. 4151




3829
3875
3921
3967







20-40
Var.
Var.
Var.
Var.
Var. 4014
Var. 4060
Var. 4106
Var. 4152




3830
3876
3922
3968







40-150
Var.
Var.
Var.
Var.
Var. 4015
Var. 4061
Var. 4107
Var. 4153




3831
3877
3923
3969







40-125
Var.
Var.
Var.
Var.
Var. 4016
Var. 4062
Var. 4108
Var. 4154




3832
3878
3924
3970







40-100
Var.
Var.
Var.
Var.
Var. 4017
Var. 4063
Var. 4109
Var. 4155




3833
3879
3925
3971







40-90
Var.
Var.
Var.
Var.
Var. 4018
Var. 4064
Var. 4110
Var. 4156




3834
3880
3926
3972







40-80
Var.
Var.
Var.
Var.
Var. 4019
Var. 4065
Var. 4111
Var. 4157




3835
3881
3927
3973







40-70
Var.
Var.
Var.
Var.
Var. 4020
Var. 4066
Var. 4112
Var. 4158




3836
3882
3928
3974







40-60
Var.
Var.
Var.
Var.
Var. 4021
Var. 4067
Var. 4113
Var. 4159




3837
3883
3929
3975







40-50
Var.
Var.
Var.
Var.
Var. 4022
Var. 4068
Var. 4114
Var. 4160




3838
3884
3930
3976







60-150
Var.
Var.
Var.
Var.
Var. 4023
Var. 4069
Var. 4115
Var. 4161




3839
3885
3931
3977







60-125
Var.
Var.
Var.
Var.
Var. 4024
Var. 4070
Var. 4116
Var. 4162




3840
3886
3932
3978







60-100
Var.
Var.
Var.
Var.
Var. 4025
Var. 4071
Var. 4117
Var. 4163




3841
3887
3933
3979







60-90
Var.
Var.
Var.
Var.
Var. 4026
Var. 4072
Var. 4118
Var. 4164




3842
3888
3934
3980







60-80
Var.
Var.
Var.
Var.
Var. 4027
Var. 4073
Var. 4119
Var. 4165




3843
3889
3935
3981







60-70
Var.
Var.
Var.
Var.
Var. 4028
Var. 4074
Var. 4120
Var. 4166




3844
3890
3936
3982







70-150
Var.
Var.
Var.
Var.
Var. 4029
Var. 4075
Var. 4121
Var. 4167




3845
3891
3937
3983







70-125
Var.
Var.
Var.
Var.
Var. 4030
Var. 4076
Var. 4122
Var. 4168




3846
3892
3938
3984







70-100
Var.
Var.
Var.
Var.
Var. 4031
Var. 4077
Var. 4123
Var. 4169




3847
3893
3939
3985







70-90
Var.
Var.
Var.
Var.
Var. 4032
Var. 4078
Var. 4124
Var. 4170




3848
3894
3940
3986







70-80
Var.
Var.
Var.
Var.
Var. 4033
Var. 4079
Var. 4125
Var. 4171




3849
3895
3941
3987







80-150
Var.
Var.
Var.
Var.
Var. 4034
Var. 4080
Var. 4126
Var. 4172




3850
3896
3942
3988







80-125
Var.
Var.
Var.
Var.
Var. 4035
Var. 4081
Var. 4127
Var. 4173




3851
3897
3943
3989







80-100
Var.
Var.
Var.
Var.
Var. 4036
Var. 4082
Var. 4128
Var. 4174




3852
3898
3944
3990







80-90
Var.
Var.
Var.
Var.
Var. 4037
Var. 4083
Var. 4129
Var. 4175




3853
3899
3945
3991







90-150
Var.
Var.
Var.
Var.
Var. 4038
Var. 4084
Var. 4130
Var. 4176




3854
3900
3946
3992







90-125
Var.
Var.
Var.
Var.
Var. 4039
Var. 4085
Var. 4131
Var. 4177




3855
3901
3947
3993







90-100
Var.
Var.
Var.
Var.
Var. 4040
Var. 4086
Var. 4132
Var. 4178




3856
3902
3948
3994





Var. = Variation













TABLE 18







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Percent)
















at least 50%
10-50%
10-40%
10-30%
10-20%
15-50%
15-40%
15-30%



















(mU/μg)
at least 20
Var. 4179
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4225
4271
4317
4363
4409
4455
4501



at least 30
Var. 4180
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4226
4272
4318
4364
4410
4456
4502



at least 40
Var. 4181
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4227
4273
4319
4365
4411
4457
4503



at least 50
Var. 4182
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4228
4274
4320
4366
4412
4458
4504



at least 60
Var. 4183
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4229
4275
4321
4367
4413
4459
4505



at least 70
Var. 4184
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4230
4276
4322
4368
4414
4460
4506



at least 80
Var. 4185
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4231
4277
4323
4369
4415
4461
4507



at least 90
Var. 4186
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4232
4278
4324
4370
4416
4462
4508



at least
Var. 4187
Var.
Var.
Var.
Var.
Var.
Var.
Var.



100

4233
4279
4325
4371
4417
4463
4509



at least
Var. 4188
Var.
Var.
Var.
Var.
Var.
Var.
Var.



125

4234
4280
4326
4372
4418
4464
4510



at least
Var. 4189
Var.
Var.
Var.
Var.
Var.
Var.
Var.



150

4235
4281
4327
4373
4419
4465
4511



20-150
Var. 4190
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4236
4282
4328
4374
4420
4466
4512



20-125
Var. 4191
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4237
4283
4329
4375
4421
4467
4513



20-100
Var. 4192
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4238
4284
4330
4376
4422
4468
4514



20-90
Var. 4193
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4239
4285
4331
4377
4423
4469
4515



20-80
Var. 4194
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4240
4286
4332
4378
4424
4470
4516



20-70
Var. 4195
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4241
4287
4333
4379
4425
4471
4517



20-60
Var. 4196
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4242
4288
4334
4380
4426
4472
4518



20-50
Var. 4197
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4243
4289
4335
4381
4427
4473
4519



20-40
Var. 4198
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4244
4290
4336
4382
4428
4474
4520



40-150
Var. 4199
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4245
4291
4337
4383
4429
4475
4521



40-125
Var. 4200
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4246
4292
4338
4384
4430
4476
4522



40-100
Var. 4201
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4247
4293
4339
4385
4431
4477
4523



40-90
Var. 4202
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4248
4294
4340
4386
4432
4478
4524



40-80
Var. 4203
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4249
4295
4341
4387
4433
4479
4525



40-70
Var. 4204
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4250
4296
4342
4388
4434
4480
4526



40-60
Var. 4205
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4251
4297
4343
4389
4435
4481
4527



40-50
Var. 4206
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4252
4298
4344
4390
4436
4482
4528



60-150
Var. 4207
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4253
4299
4345
4391
4437
4483
4529



60-125
Var. 4208
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4254
4300
4346
4392
4438
4484
4530



60-100
Var. 4209
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4255
4301
4347
4393
4439
4485
4531



60-90
Var. 4210
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4256
4302
4348
4394
4440
4486
4532



60-80
Var. 4211
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4257
4303
4349
4395
4441
4487
4533



60-70
Var. 4212
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4258
4304
4350
4396
4442
4488
4534



70-150
Var. 4213
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4259
4305
4351
4397
4443
4489
4535



70-125
Var. 4214
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4260
4306
4352
4398
4444
4490
4536



70-100
Var. 4215
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4261
4307
4353
4399
4445
4491
4537



70-90
Var. 4216
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4262
4308
4354
4400
4446
4492
4538



70-80
Var. 4217
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4263
4309
4355
4401
4447
4493
4539



80-150
Var. 4218
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4264
4310
4356
4402
4448
4494
4540



80-125
Var. 4219
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4265
4311
4357
4403
4449
4495
4541



80-100
Var. 4220
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4266
4312
4358
4404
4450
4496
4542



80-90
Var. 4221
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4267
4313
4359
4405
4451
4497
4543



90-150
Var. 4222
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4268
4314
4360
4406
4452
4498
4544



90-125
Var. 4223
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4269
4315
4361
4407
4453
4499
4545



90-100
Var. 4224
Var.
Var.
Var.
Var.
Var.
Var.
Var.





4270
4316
4362
4408
4454
4500
4546





Var. = Variation













TABLE 19







Exemplary embodiments for the combination of rVWF specific activity in a


composition used herein and increase in FVIII stability achieved, as compared


to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Percent)















15-20%
20-50%
20-40%
20-30%
30-50%
30-40%
40-50%


















(mU/μg)
at least 20
Var. 4547
Var. 4593
Var. 4639
Var. 4685
Var. 4731
Var. 4777
Var. 4823



at least 30
Var. 4548
Var. 4594
Var. 4640
Var. 4686
Var. 4732
Var. 4778
Var. 4824



at least 40
Var. 4549
Var. 4595
Var. 4641
Var. 4687
Var. 4733
Var. 4779
Var. 4825



at least 50
Var. 4550
Var. 4596
Var. 4642
Var. 4688
Var. 4734
Var. 4780
Var. 4826



at least 60
Var. 4551
Var. 4597
Var. 4643
Var. 4689
Var. 4735
Var. 4781
Var. 4827



at least 70
Var. 4552
Var. 4598
Var. 4644
Var. 4690
Var. 4736
Var. 4782
Var. 4828



at least 80
Var. 4553
Var. 4599
Var. 4645
Var. 4691
Var. 4737
Var. 4783
Var. 4829



at least 90
Var. 4554
Var. 4600
Var. 4646
Var. 4692
Var. 4738
Var. 4784
Var. 4830



at least 100
Var. 4555
Var. 4601
Var. 4647
Var. 4693
Var. 4739
Var. 4785
Var. 4831



at least 125
Var. 4556
Var. 4602
Var. 4648
Var. 4694
Var. 4740
Var. 4786
Var. 4832



at least 150
Var. 4557
Var. 4603
Var. 4649
Var. 4695
Var. 4741
Var. 4787
Var. 4833



20-150
Var. 4558
Var. 4604
Var. 4650
Var. 4696
Var. 4742
Var. 4788
Var. 4834



20-125
Var. 4559
Var. 4605
Var. 4651
Var. 4697
Var. 4743
Var. 4789
Var. 4835



20-100
Var. 4560
Var. 4606
Var. 4652
Var. 4698
Var. 4744
Var. 4790
Var. 4836



20-90
Var. 4561
Var. 4607
Var. 4653
Var. 4699
Var. 4745
Var. 4791
Var. 4837



20-80
Var. 4562
Var. 4608
Var. 4654
Var. 4700
Var. 4746
Var. 4792
Var. 4838



20-70
Var. 4563
Var. 4609
Var. 4655
Var. 4701
Var. 4747
Var. 4793
Var. 4839



20-60
Var. 4564
Var. 4610
Var. 4656
Var. 4702
Var. 4748
Var. 4794
Var. 4840



20-50
Var. 4565
Var. 4611
Var. 4657
Var. 4703
Var. 4749
Var. 4795
Var. 4841



20-40
Var. 4566
Var. 4612
Var. 4658
Var. 4704
Var. 4750
Var. 4796
Var. 4842



40-150
Var. 4567
Var. 4613
Var. 4659
Var. 4705
Var. 4751
Var. 4797
Var. 4843



40-125
Var. 4568
Var. 4614
Var. 4660
Var. 4706
Var. 4752
Var. 4798
Var. 4844



40-100
Var. 4569
Var. 4615
Var. 4661
Var. 4707
Var. 4753
Var. 4799
Var. 4845



40-90
Var. 4570
Var. 4616
Var. 4662
Var. 4708
Var. 4754
Var. 4800
Var. 4846



40-80
Var. 4571
Var. 4617
Var. 4663
Var. 4709
Var. 4755
Var. 4801
Var. 4847



40-70
Var. 4572
Var. 4618
Var. 4664
Var. 4710
Var. 4756
Var. 4802
Var. 4848



40-60
Var. 4573
Var. 4619
Var. 4665
Var. 4711
Var. 4757
Var. 4803
Var. 4849



40-50
Var. 4574
Var. 4620
Var. 4666
Var. 4712
Var. 4758
Var. 4804
Var. 4850



60-150
Var. 4575
Var. 4621
Var. 4667
Var. 4713
Var. 4759
Var. 4805
Var. 4851



60-125
Var. 4576
Var. 4622
Var. 4668
Var. 4714
Var. 4760
Var. 4806
Var. 4852



60-100
Var. 4577
Var. 4623
Var. 4669
Var. 4715
Var. 4761
Var. 4807
Var. 4853



60-90
Var. 4578
Var. 4624
Var. 4670
Var. 4716
Var. 4762
Var. 4808
Var. 4854



60-80
Var. 4579
Var. 4625
Var. 4671
Var. 4717
Var. 4763
Var. 4809
Var. 4855



60-70
Var. 4580
Var. 4626
Var. 4672
Var. 4718
Var. 4764
Var. 4810
Var. 4856



70-150
Var. 4581
Var. 4627
Var. 4673
Var. 4719
Var. 4765
Var. 4811
Var. 4857



70-125
Var. 4582
Var. 4628
Var. 4674
Var. 4720
Var. 4766
Var. 4812
Var. 4858



70-100
Var. 4583
Var. 4629
Var. 4675
Var. 4721
Var. 4767
Var. 4813
Var. 4859



70-90
Var. 4584
Var. 4630
Var. 4676
Var. 4722
Var. 4768
Var. 4814
Var. 4860



70-80
Var. 4585
Var. 4631
Var. 4677
Var. 4723
Var. 4769
Var. 4815
Var. 4861



80-150
Var. 4586
Var. 4632
Var. 4678
Var. 4724
Var. 4770
Var. 4816
Var. 4862



80-125
Var. 4587
Var. 4633
Var. 4679
Var. 4725
Var. 4771
Var. 4817
Var. 4863



80-100
Var. 4588
Var. 4634
Var. 4680
Var. 4726
Var. 4772
Var. 4818
Var. 4864



80-90
Var. 4589
Var. 4635
Var. 4681
Var. 4727
Var. 4773
Var. 4819
Var. 4865



90-150
Var. 4590
Var. 4636
Var. 4682
Var. 4728
Var. 4774
Var. 4820
Var. 4866



90-125
Var. 4591
Var. 4637
Var. 4683
Var. 4729
Var. 4775
Var. 4821
Var. 4867



90-100
Var. 4592
Var. 4638
Var. 4684
Var. 4730
Var. 4776
Var. 4822
Var. 4868





Var. = Variation






In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF composition containing from 10 IU/kg to 40 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 20 IU/kg to 30 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF composition containing from 25 IU/kg to 75 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 40 IU/kg to 60 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF composition containing from 75 IU/kg to 125 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 75 IU/kg to 100 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the subject is administered a dose of rVWF selected from variations 2141 to 2338 in Table 12, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF. In one embodiment. FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific emobidment, the method is for treating Type 3 VWD.







TABLE 20







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII


stability achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)















at least 1 hr
at least 2 hr
at least 3 hr
at least 4 hr
at least 5 hr
at least 6 hr
at least 7 hr


















Dosage
 0.5-200
Var. 4869
Var. 4926
Var. 4983
Var. 5040
Var. 5097
Var. 5154
Var. 5211


(IU/kg
 0.5-150
Var. 4870
Var. 4927
Var. 4984
Var. 5041
Var. 5098
Var. 5155
Var. 5212


rVWF:
 0.5-100
Var. 4871
Var. 4928
Var. 4985
Var. 5042
Var. 5099
Var. 5156
Var. 5213


RCo 
 0.5-75
Var. 4872
Var. 4929
Var. 4986
Var. 5043
Var. 5100
Var. 5157
Var. 5214


activity)
 0.5-50
Var. 4873
Var. 4930
Var. 4987
Var. 5044
Var. 5101
Var. 5158
Var. 5215



 0.5-25
Var. 4874
Var. 4931
Var. 4988
Var. 5045
Var. 5102
Var. 5159
Var. 5216



 0.5-10
Var. 4875
Var. 4932
Var. 4989
Var. 5046
Var. 5103
Var. 5160
Var. 5217



 0.5-5
Var. 4876
Var. 4933
Var. 4990
Var. 5047
Var. 5104
Var. 5161
Var. 5218



 0.5-2.5
Var. 4877
Var. 4934
Var. 4991
Var. 5048
Var. 5105
Var. 5162
Var. 5219



 0.5-1
Var. 4878
Var. 4935
Var. 4992
Var. 5049
Var. 5106
Var. 5163
Var. 5220



 2.5-200
Var. 4879
Var. 4936
Var. 4993
Var. 5050
Var. 5107
Var. 5164
Var. 5221



 2.5-150
Var. 4880
Var. 4937
Var. 4994
Var. 5051
Var. 5108
Var. 5165
Var. 5222



 2.5-100
Var. 4881
Var. 4938
Var. 4995
Var. 5052
Var. 5109
Var. 5166
Var. 5223



 2.5-75
Var. 4882
Var. 4939
Var. 4996
Var. 5053
Var. 5110
Var. 5167
Var. 5224



 2.5-50
Var. 4883
Var. 4940
Var. 4997
Var. 5054
Var. 5111
Var. 5168
Var. 5225



 2.5-25
Var. 4884
Var. 4941
Var. 4998
Var. 5055
Var. 5112
Var. 5169
Var. 5226



 2.5-10
Var. 4885
Var. 4942
Var. 4999
Var. 5056
Var. 5113
Var. 5170
Var. 5227



 2.5-5
Var. 4886
Var. 4943
Var. 5000
Var. 5057
Var. 5114
Var. 5171
Var. 5228



 5-200
Var. 4887
Var. 4944
Var. 5001
Var. 5058
Var. 5115
Var. 5172
Var. 5229



 5-175
Var. 4888
Var. 4945
Var. 5002
Var. 5059
Var. 5116
Var. 5173
Var. 5230



 5-150
Var. 4889
Var. 4946
Var. 5003
Var. 5060
Var. 5117
Var. 5174
Var. 5231



 5-125
Var. 4890
Var. 4947
Var. 5004
Var. 5061
Var. 5118
Var. 5175
Var. 5232



 5-100
Var. 4891
Var. 4948
Var. 5005
Var. 5062
Var. 5119
Var. 5176
Var. 5233



 5-75
Var. 4892
Var. 4949
Var. 5006
Var. 5063
Var. 5120
Var. 5177
Var. 5234



 5-50
Var. 4893
Var. 4950
Var. 5007
Var. 5064
Var. 5121
Var. 5178
Var. 5235



 5-25
Var. 4894
Var. 4951
Var. 5008
Var. 5065
Var. 5122
Var. 5179
Var. 5236



 5-10
Var. 4895
Var. 4952
Var. 5009
Var. 5066
Var. 5123
Var. 5180
Var. 5237



 10-200
Var. 4896
Var. 4953
Var. 5010
Var. 5067
Var. 5124
Var. 5181
Var. 5238



 10-150
Var. 4897
Var. 4954
Var. 5011
Var. 5068
Var. 5125
Var. 5182
Var. 5239



 10-100
Var. 4898
Var. 4955
Var. 5012
Var. 5069
Var. 5126
Var. 5183
Var. 5240



 10-75
Var. 4899
Var. 4956
Var. 5013
Var. 5070
Var. 5127
Var. 5184
Var. 5241



 10-50
Var. 4900
Var. 4957
Var. 5014
Var. 5071
Var. 5128
Var. 5185
Var. 5242



 10-25
Var. 4901
Var. 4958
Var. 5015
Var. 5072
Var. 5129
Var. 5186
Var. 5243



 25-200
Var. 4902
Var. 4959
Var. 5016
Var. 5073
Var. 5130
Var. 5187
Var. 5244



 25-150
Var. 4903
Var. 4960
Var. 5017
Var. 5074
Var. 5131
Var. 5188
Var. 5245



 25-100
Var. 4904
Var. 4961
Var. 5018
Var. 5075
Var. 5132
Var. 5189
Var. 5246



 25-75
Var. 4905
Var. 4962
Var. 5019
Var. 5076
Var. 5133
Var. 5190
Var. 5247



 25-50
Var. 4906
Var. 4963
Var. 5020
Var. 5077
Var. 5134
Var. 5191
Var. 5248



 50-200
Var. 4907
Var. 4964
Var. 5021
Var. 5078
Var. 5135
Var. 5192
Var. 5249



 50-150
Var. 4908
Var. 4965
Var. 5022
Var. 5079
Var. 5136
Var. 5193
Var. 5250



 50-100
Var. 4909
Var. 4966
Var. 5023
Var. 5080
Var. 5137
Var. 5194
Var. 5251



 50-75
Var. 4910
Var. 4967
Var. 5024
Var. 5081
Var. 5138
Var. 5195
Var. 5252



 75-200
Var. 4911
Var. 4968
Var. 5025
Var. 5082
Var. 5139
Var. 5196
Var. 5253



 75-175
Var. 4912
Var. 4969
Var. 5026
Var. 5083
Var. 5140
Var. 5197
Var. 5254



 75-150
Var. 4913
Var. 4970
Var. 5027
Var. 5084
Var. 5141
Var. 5198
Var. 5255



 75-125
Var. 4914
Var. 4971
Var. 5028
Var. 5085
Var. 5142
Var. 5199
Var. 5256



 75-100
Var. 4915
Var. 4972
Var. 5029
Var. 5086
Var. 5143
Var. 5200
Var. 5257



100-200
Var. 4916
Var. 4973
Var. 5030
Var. 5087
Var. 5144
Var. 5201
Var. 5258



100-175
Var. 4917
Var. 4974
Var. 5031
Var. 5088
Var. 5145
Var. 5202
Var. 5259



100-150
Var. 4918
Var. 4975
Var. 5032
Var. 5089
Var. 5146
Var. 5203
Var. 5260



100-125
Var. 4919
Var. 4976
Var. 5033
Var. 5090
Var. 5147
Var. 5204
Var. 5261



125-200
Var. 4920
Var. 4977
Var. 5034
Var. 5091
Var. 5148
Var. 5205
Var. 5262



125-175
Var. 4921
Var. 4978
Var. 5035
Var. 5092
Var. 5149
Var. 5206
Var. 5263



125-150
Var. 4922
Var. 4979
Var. 5036
Var. 5093
Var. 5150
Var. 5207
Var. 5264



150-200
Var. 4923
Var. 4980
Var. 5037
Var. 5094
Var. 5151
Var. 5208
Var. 5265



150-200
Var. 4924
Var. 4981
Var. 5038
Var. 5095
Var. 5152
Var. 5209
Var. 5266



175-200
Var. 4925
Var. 4982
Var. 5039
Var. 5096
Var. 5153
Var. 5210
Var. 5267





Var. = Variation













TABLE 21







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII


stability achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)















at least 8 hr
1-8 hr
1-7 hr
1-6 hr
1-5 hr
1-4 hr
1-3 hr


















Dosage
 0.5-200
Var. 5268
Var. 5325
Var. 5382
Var. 5439
Var. 5496
Var. 5553
Var. 5610


(IU/kg
 0.5-150
Var. 5269
Var. 5326
Var. 5383
Var. 5440
Var. 5497
Var. 5554
Var. 5611


rVWF:
 0.5-100
Var. 5270
Var. 5327
Var. 5384
Var. 5441
Var. 5498
Var. 5555
Var. 5612


RCo 
 0.5-75
Var. 5271
Var. 5328
Var. 5385
Var. 5442
Var. 5499
Var. 5556
Var. 5613


activity)
 0.5-50
Var. 5272
Var. 5329
Var. 5386
Var. 5443
Var. 5500
Var. 5557
Var. 5614



 0.5-25
Var. 5273
Var. 5330
Var. 5387
Var. 5444
Var. 5501
Var. 5558
Var. 5615



 0.5-10
Var. 5274
Var. 5331
Var. 5388
Var. 5445
Var. 5502
Var. 5559
Var. 5616



 0.5-5
Var. 5275
Var. 5332
Var. 5389
Var. 5446
Var. 5503
Var. 5560
Var. 5617



 0.5-2.5
Var. 5276
Var. 5333
Var. 5390
Var. 5447
Var. 5504
Var. 5561
Var. 5618



 0.5-1
Var. 5277
Var. 5334
Var. 5391
Var. 5448
Var. 5505
Var. 5562
Var. 5619



 2.5-200
Var. 5278
Var. 5335
Var. 5392
Var. 5449
Var. 5506
Var. 5563
Var. 5620



 2.5-150
Var. 5279
Var. 5336
Var. 5393
Var. 5450
Var. 5507
Var. 5564
Var. 5621



 2.5-100
Var. 5280
Var. 5337
Var. 5394
Var. 5451
Var. 5508
Var. 5565
Var. 5622



 2.5-75
Var. 5281
Var. 5338
Var. 5395
Var. 5452
Var. 5509
Var. 5566
Var. 5623



 2.5-50
Var. 5282
Var. 5339
Var. 5396
Var. 5453
Var. 5510
Var. 5567
Var. 5624



 2.5-25
Var. 5283
Var. 5340
Var. 5397
Var. 5454
Var. 5511
Var. 5568
Var. 5625



 2.5-10
Var. 5284
Var. 5341
Var. 5398
Var. 5455
Var. 5512
Var. 5569
Var. 5626



 2.5-5
Var. 5285
Var. 5342
Var. 5399
Var. 5456
Var. 5513
Var. 5570
Var. 5627



 5-200
Var. 5286
Var. 5343
Var. 5400
Var. 5457
Var. 5514
Var. 5571
Var. 5628



 5-175
Var. 5287
Var. 5344
Var. 5401
Var. 5458
Var. 5515
Var. 5572
Var. 5629



 5-150
Var. 5288
Var. 5345
Var. 5402
Var. 5459
Var. 5516
Var. 5573
Var. 5630



 5-125
Var. 5289
Var. 5346
Var. 5403
Var. 5460
Var. 5517
Var. 5574
Var. 5631



 5-100
Var. 5290
Var. 5347
Var. 5404
Var. 5461
Var. 5518
Var. 5575
Var. 5632



 5-75
Var. 5291
Var. 5348
Var. 5405
Var. 5462
Var. 5519
Var. 5576
Var. 5633



 5-50
Var. 5292
Var. 5349
Var. 5406
Var. 5463
Var. 5520
Var. 5577
Var. 5634



 5-25
Var. 5293
Var. 5350
Var. 5407
Var. 5464
Var. 5521
Var. 5578
Var. 5635



 5-10
Var. 5294
Var. 5351
Var. 5408
Var. 5465
Var. 5522
Var. 5579
Var. 5636



 10-200
Var. 5295
Var. 5352
Var. 5409
Var. 5466
Var. 5523
Var. 5580
Var. 5637



 10-150
Var. 5296
Var. 5353
Var. 5410
Var. 5467
Var. 5524
Var. 5581
Var. 5638



 10-100
Var. 5297
Var. 5354
Var. 5411
Var. 5468
Var. 5525
Var. 5582
Var. 5639



 10-75
Var. 5298
Var. 5355
Var. 5412
Var. 5469
Var. 5526
Var. 5583
Var. 5640



 10-50
Var. 5299
Var. 5356
Var. 5413
Var. 5470
Var. 5527
Var. 5584
Var. 5641



 10-25
Var. 5300
Var. 5357
Var. 5414
Var. 5471
Var. 5528
Var. 5585
Var. 5642



 25-200
Var. 5301
Var. 5358
Var. 5415
Var. 5472
Var. 5529
Var. 5586
Var. 5643



 25-150
Var. 5302
Var. 5359
Var. 5416
Var. 5473
Var. 5530
Var. 5587
Var. 5644



 25-100
Var. 5303
Var. 5360
Var. 5417
Var. 5474
Var. 5531
Var. 5588
Var. 5645



 25-75
Var. 5304
Var. 5361
Var. 5418
Var. 5475
Var. 5532
Var. 5589
Var. 5646



 25-50
Var. 5305
Var. 5362
Var. 5419
Var. 5476
Var. 5533
Var. 5590
Var. 5647



 50-200
Var. 5306
Var. 5363
Var. 5420
Var. 5477
Var. 5534
Var. 5591
Var. 5648



 50-150
Var. 5307
Var. 5364
Var. 5421
Var. 5478
Var. 5535
Var. 5592
Var. 5649



 50-100
Var. 5308
Var. 5365
Var. 5422
Var. 5479
Var. 5536
Var. 5593
Var. 5650



 50-75
Var. 5309
Var. 5366
Var. 5423
Var. 5480
Var. 5537
Var. 5594
Var. 5651



 75-200
Var. 5310
Var. 5367
Var. 5424
Var. 5481
Var. 5538
Var. 5595
Var. 5652



 75-175
Var. 5311
Var. 5368
Var. 5425
Var. 5482
Var. 5539
Var. 5596
Var. 5653



 75-150
Var. 5312
Var. 5369
Var. 5426
Var. 5483
Var. 5540
Var. 5597
Var. 5654



 75-125
Var. 5313
Var. 5370
Var. 5427
Var. 5484
Var. 5541
Var. 5598
Var. 5655



 75-100
Var. 5314
Var. 5371
Var. 5428
Var. 5485
Var. 5542
Var. 5599
Var. 5656



100-200
Var. 5315
Var. 5372
Var. 5429
Var. 5486
Var. 5543
Var. 5600
Var. 5657



100-175
Var. 5316
Var. 5373
Var. 5430
Var. 5487
Var. 5544
Var. 5601
Var. 5658



100-150
Var. 5317
Var. 5374
Var. 5431
Var. 5488
Var. 5545
Var. 5602
Var. 5659



100-125
Var. 5318
Var. 5375
Var. 5432
Var. 5489
Var. 5546
Var. 5603
Var. 5660



125-200
Var. 5319
Var. 5376
Var. 5433
Var. 5490
Var. 5547
Var. 5604
Var. 5661



125-175
Var. 5320
Var. 5377
Var. 5434
Var. 5491
Var. 5548
Var. 5605
Var. 5662



125-150
Var. 5321
Var. 5378
Var. 5435
Var. 5492
Var. 5549
Var. 5606
Var. 5663



150-200
Var. 5322
Var. 5379
Var. 5436
Var. 5493
Var. 5550
Var. 5607
Var. 5664



150-200
Var. 5323
Var. 5380
Var. 5437
Var. 5494
Var. 5551
Var. 5608
Var. 5665



175-200
Var. 5324
Var. 5381
Var. 5438
Var. 5495
Var. 5552
Var. 5609
Var. 5666





Var. = Variation













TABLE 22







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII


stability achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)















1-2 hr
2-8 hr
2-7 hr
2-6 hr
2-5 hr
2-4 hr
2-3 hr


















Dosage
 0.5-200
Var. 5667
Var. 5724
Var. 5781
Var. 5838
Var. 5895
Var. 5952
Var. 6009


(IU/kg
 0.5-150
Var. 5668
Var. 5725
Var. 5782
Var. 5839
Var. 5896
Var. 5953
Var. 6010


rVWF:
 0.5-100
Var. 5669
Var. 5726
Var. 5783
Var. 5840
Var. 5897
Var. 5954
Var. 6011


RCo 
 0.5-75
Var. 5670
Var. 5727
Var. 5784
Var. 5841
Var. 5898
Var. 5955
Var. 6012


activity)
 0.5-50
Var. 5671
Var. 5728
Var. 5785
Var. 5842
Var. 5899
Var. 5956
Var. 6013



 0.5-25
Var. 5672
Var. 5729
Var. 5786
Var. 5843
Var. 5900
Var. 5957
Var. 6014



 0.5-10
Var. 5673
Var. 5730
Var. 5787
Var. 5844
Var. 5901
Var. 5958
Var. 6015



 0.5-5
Var. 5674
Var. 5731
Var. 5788
Var. 5845
Var. 5902
Var. 5959
Var. 6016



 0.5-2.5
Var. 5675
Var. 5732
Var. 5789
Var. 5846
Var. 5903
Var. 5960
Var. 6017



 0.5-1
Var. 5676
Var. 5733
Var. 5790
Var. 5847
Var. 5904
Var. 5961
Var. 6018



 2.5-200
Var. 5677
Var. 5734
Var. 5791
Var. 5848
Var. 5905
Var. 5962
Var. 6019



 2.5-150
Var. 5678
Var. 5735
Var. 5792
Var. 5849
Var. 5906
Var. 5963
Var. 6020



 2.5-100
Var. 5679
Var. 5736
Var. 5793
Var. 5850
Var. 5907
Var. 5964
Var. 6021



 2.5-75
Var. 5680
Var. 5737
Var. 5794
Var. 5851
Var. 5908
Var. 5965
Var. 6022



 2.5-50
Var. 5681
Var. 5738
Var. 5795
Var. 5852
Var. 5909
Var. 5966
Var. 6023



 2.5-25
Var. 5682
Var. 5739
Var. 5796
Var. 5853
Var. 5910
Var. 5967
Var. 6024



 2.5-10
Var. 5683
Var. 5740
Var. 5797
Var. 5854
Var. 5911
Var. 5968
Var. 6025



 2.5-5
Var. 5684
Var. 5741
Var. 5798
Var. 5855
Var. 5912
Var. 5969
Var. 6026



 5-200
Var. 5685
Var. 5742
Var. 5799
Var. 5856
Var. 5913
Var. 5970
Var. 6027



 5-175
Var. 5686
Var. 5743
Var. 5800
Var. 5857
Var. 5914
Var. 5971
Var. 6028



 5-150
Var. 5687
Var. 5744
Var. 5801
Var. 5858
Var. 5915
Var. 5972
Var. 6029



 5-125
Var. 5688
Var. 5745
Var. 5802
Var. 5859
Var. 5916
Var. 5973
Var. 6030



 5-100
Var. 5689
Var. 5746
Var. 5803
Var. 5860
Var. 5917
Var. 5974
Var. 6031



 5-75
Var. 5690
Var. 5747
Var. 5804
Var. 5861
Var. 5918
Var. 5975
Var. 6032



 5-50
Var. 5691
Var. 5748
Var. 5805
Var. 5862
Var. 5919
Var. 5976
Var. 6033



 5-25
Var. 5692
Var. 5749
Var. 5806
Var. 5863
Var. 5920
Var. 5977
Var. 6034



 5-10
Var. 5693
Var. 5750
Var. 5807
Var. 5864
Var. 5921
Var. 5978
Var. 6035



 10-200
Var. 5694
Var. 5751
Var. 5808
Var. 5865
Var. 5922
Var. 5979
Var. 6036



 10-150
Var. 5695
Var. 5752
Var. 5809
Var. 5866
Var. 5923
Var. 5980
Var. 6037



 10-100
Var. 5696
Var. 5753
Var. 5810
Var. 5867
Var. 5924
Var. 5981
Var. 6038



 10-75
Var. 5697
Var. 5754
Var. 5811
Var. 5868
Var. 5925
Var. 5982
Var. 6039



 10-50
Var. 5698
Var. 5755
Var. 5812
Var. 5869
Var. 5926
Var. 5983
Var. 6040



 10-25
Var. 5699
Var. 5756
Var. 5813
Var. 5870
Var. 5927
Var. 5984
Var. 6041



 25-200
Var. 5700
Var. 5757
Var. 5814
Var. 5871
Var. 5928
Var. 5985
Var. 6042



 25-150
Var. 5701
Var. 5758
Var. 5815
Var. 5872
Var. 5929
Var. 5986
Var. 6043



 25-100
Var. 5702
Var. 5759
Var. 5816
Var. 5873
Var. 5930
Var. 5987
Var. 6044



 25-75
Var. 5703
Var. 5760
Var. 5817
Var. 5874
Var. 5931
Var. 5988
Var. 6045



 25-50
Var. 5704
Var. 5761
Var. 5818
Var. 5875
Var. 5932
Var. 5989
Var. 6046



 50-200
Var. 5705
Var. 5762
Var. 5819
Var. 5876
Var. 5933
Var. 5990
Var. 6047



 50-150
Var. 5706
Var. 5763
Var. 5820
Var. 5877
Var. 5934
Var. 5991
Var. 6048



 50-100
Var. 5707
Var. 5764
Var. 5821
Var. 5878
Var. 5935
Var. 5992
Var. 6049



 50-75
Var. 5708
Var. 5765
Var. 5822
Var. 5879
Var. 5936
Var. 5993
Var. 6050



 75-200
Var. 5709
Var. 5766
Var. 5823
Var. 5880
Var. 5937
Var. 5994
Var. 6051



 75-175
Var. 5710
Var. 5767
Var. 5824
Var. 5881
Var. 5938
Var. 5995
Var. 6052



 75-150
Var. 5711
Var. 5768
Var. 5825
Var. 5882
Var. 5939
Var. 5996
Var. 6053



 75-125
Var. 5712
Var. 5769
Var. 5826
Var. 5883
Var. 5940
Var. 5997
Var. 6054



 75-100
Var. 5713
Var. 5770
Var. 5827
Var. 5884
Var. 5941
Var. 5998
Var. 6055



100-200
Var. 5714
Var. 5771
Var. 5828
Var. 5885
Var. 5942
Var. 5999
Var. 6056



100-175
Var. 5715
Var. 5772
Var. 5829
Var. 5886
Var. 5943
Var. 6000
Var. 6057



100-150
Var. 5716
Var. 5773
Var. 5830
Var. 5887
Var. 5944
Var. 6001
Var. 6058



100-125
Var. 5717
Var. 5774
Var. 5831
Var. 5888
Var. 5945
Var. 6002
Var. 6059



125-200
Var. 5718
Var. 5775
Var. 5832
Var. 5889
Var. 5946
Var. 6003
Var. 6060



125-175
Var. 5719
Var. 5776
Var. 5833
Var. 5890
Var. 5947
Var. 6004
Var. 6061



125-150
Var. 5720
Var. 5777
Var. 5834
Var. 5891
Var. 5948
Var. 6005
Var. 6062



150-200
Var. 5721
Var. 5778
Var. 5835
Var. 5892
Var. 5949
Var. 6006
Var. 6063



150-200
Var. 5722
Var. 5779
Var. 5836
Var. 5893
Var. 5950
Var. 6007
Var. 6064



175-200
Var. 5723
Var. 5780
Var. 5837
Var. 5894
Var. 5951
Var. 6008
Var. 6065





Var. = Variation













TABLE 23







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII stability


achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)















3-8 hr
3-7 hr
3-6 hr
3-5 hr
3-4 hr
4-8 hr
4-7 hr


















Dosage
0.5-200 
Var. 6066
Var. 6123
Var. 6180
Var. 6237
Var. 6294
Var. 6351
Var. 6408


(IU/kg rVWF:RCo
0.5-150 
Var. 6067
Var. 6124
Var. 6181
Var. 6238
Var. 6295
Var. 6352
Var. 6409


activity)
0.5-100 
Var. 6068
Var. 6125
Var. 6182
Var. 6239
Var. 6296
Var. 6353
Var. 6410



0.5-75  
Var. 6069
Var. 6126
Var. 6183
Var. 6240
Var. 6297
Var. 6354
Var. 6411



0.5-50  
Var. 6070
Var. 6127
Var. 6184
Var. 6241
Var. 6298
Var. 6355
Var. 6412



0.5-25  
Var. 6071
Var. 6128
Var. 6185
Var. 6242
Var. 6299
Var. 6356
Var. 6413



0.5-10  
Var. 6072
Var. 6129
Var. 6186
Var. 6243
Var. 6300
Var. 6357
Var. 6414



0.5-5  
Var. 6073
Var. 6130
Var. 6187
Var. 6244
Var. 6301
Var. 6358
Var. 6415



0.5-2.5 
Var. 6074
Var. 6131
Var. 6188
Var. 6245
Var. 6302
Var. 6359
Var. 6416



0.5-1  
Var. 6075
Var. 6132
Var. 6189
Var. 6246
Var. 6303
Var. 6360
Var. 6417



2.5-200 
Var. 6076
Var. 6133
Var. 6190
Var. 6247
Var. 6304
Var. 6361
Var. 6418



2.5-150 
Var. 6077
Var. 6134
Var. 6191
Var. 6248
Var. 6305
Var. 6362
Var. 6419



2.5-100 
Var. 6078
Var. 6135
Var. 6192
Var. 6249
Var. 6306
Var. 6363
Var. 6420



2.5-75  
Var. 6079
Var. 6136
Var. 6193
Var. 6250
Var. 6307
Var. 6364
Var. 6421



2.5-50  
Var. 6080
Var. 6137
Var. 6194
Var. 6251
Var. 6308
Var. 6365
Var. 6422



2.5-25  
Var. 6081
Var. 6138
Var. 6195
Var. 6252
Var. 6309
Var. 6366
Var. 6423



2.5-10  
Var. 6082
Var. 6139
Var. 6196
Var. 6253
Var. 6310
Var. 6367
Var. 6424



2.5-5  
Var. 6083
Var. 6140
Var. 6197
Var. 6254
Var. 6311
Var. 6368
Var. 6425



 5-200
Var. 6084
Var. 6141
Var. 6198
Var. 6255
Var. 6312
Var. 6369
Var. 6426



 5-175
Var. 6085
Var. 6142
Var. 6199
Var. 6256
Var. 6313
Var. 6370
Var. 6427



 5-150
Var. 6086
Var. 6143
Var. 6200
Var. 6257
Var. 6314
Var. 6371
Var. 6428



 5-125
Var. 6087
Var. 6144
Var. 6201
Var. 6258
Var. 6315
Var. 6372
Var. 6429



 5-100
Var. 6088
Var. 6145
Var. 6202
Var. 6259
Var. 6316
Var. 6373
Var. 6430



5-75
Var. 6089
Var. 6146
Var. 6203
Var. 6260
Var. 6317
Var. 6374
Var. 6431



5-50
Var. 6090
Var. 6147
Var. 6204
Var. 6261
Var. 6318
Var. 6375
Var. 6432



5-25
Var. 6091
Var. 6148
Var. 6205
Var. 6262
Var. 6319
Var. 6376
Var. 6433



5-10
Var. 6092
Var. 6149
Var. 6206
Var. 6263
Var. 6320
Var. 6377
Var. 6434



10-200
Var. 6093
Var. 6150
Var. 6207
Var. 6264
Var. 6321
Var. 6378
Var. 6435



10-150
Var. 6094
Var. 6151
Var. 6208
Var. 6265
Var. 6322
Var. 6379
Var. 6436



10-100
Var. 6095
Var. 6152
Var. 6209
Var. 6266
Var. 6323
Var. 6380
Var. 6437



10-75 
Var. 6096
Var. 6153
Var. 6210
Var. 6267
Var. 6324
Var. 6381
Var. 6438



10-50 
Var. 6097
Var. 6154
Var. 6211
Var. 6268
Var. 6325
Var. 6382
Var. 6439



10-25 
Var. 6098
Var. 6155
Var. 6212
Var. 6269
Var. 6326
Var. 6383
Var. 6440



25-200
Var. 6099
Var. 6156
Var. 6213
Var. 6270
Var. 6327
Var. 6384
Var. 6441



25-150
Var. 6100
Var. 6157
Var. 6214
Var. 6271
Var. 6328
Var. 6385
Var. 6442



25-100
Var. 6101
Var. 6158
Var. 6215
Var. 6272
Var. 6329
Var. 6386
Var. 6443



25-75 
Var. 6102
Var. 6159
Var. 6216
Var. 6273
Var. 6330
Var. 6387
Var. 6444



25-50 
Var. 6103
Var. 6160
Var. 6217
Var. 6274
Var. 6331
Var. 6388
Var. 6445



50-200
Var. 6104
Var. 6161
Var. 6218
Var. 6275
Var. 6332
Var. 6389
Var. 6446



50-150
Var. 6105
Var. 6162
Var. 6219
Var. 6276
Var. 6333
Var. 6390
Var. 6447



50-100
Var. 6106
Var. 6163
Var. 6220
Var. 6277
Var. 6334
Var. 6391
Var. 6448



50-75 
Var. 6107
Var. 6164
Var. 6221
Var. 6278
Var. 6335
Var. 6392
Var. 6449



75-200
Var. 6108
Var. 6165
Var. 6222
Var. 6279
Var. 6336
Var. 6393
Var. 6450



75-175
Var. 6109
Var. 6166
Var. 6223
Var. 6280
Var. 6337
Var. 6394
Var. 6451



75-150
Var. 6110
Var. 6167
Var. 6224
Var. 6281
Var. 6338
Var. 6395
Var. 6452



75-125
Var. 6111
Var. 6168
Var. 6225
Var. 6282
Var. 6339
Var. 6396
Var. 6453



75-100
Var. 6112
Var. 6169
Var. 6226
Var. 6283
Var. 6340
Var. 6397
Var. 6454



100-200 
Var. 6113
Var. 6170
Var. 6227
Var. 6284
Var. 6341
Var. 6398
Var. 6455



100-175 
Var. 6114
Var. 6171
Var. 6228
Var. 6285
Var. 6342
Var. 6399
Var. 6456



100-150 
Var. 6115
Var. 6172
Var. 6229
Var. 6286
Var. 6343
Var. 6400
Var. 6457



100-125 
Var. 6116
Var. 6173
Var. 6230
Var. 6287
Var. 6344
Var. 6401
Var. 6458



125-200 
Var. 6117
Var. 6174
Var. 6231
Var. 6288
Var. 6345
Var. 6402
Var. 6459



125-175 
Var. 6118
Var. 6175
Var. 6232
Var. 6289
Var. 6346
Var. 6403
Var. 6460



125-150 
Var. 6119
Var. 6176
Var. 6233
Var. 6290
Var. 6347
Var. 6404
Var. 6461



150-200 
Var. 6120
Var. 6177
Var. 6234
Var. 6291
Var. 6348
Var. 6405
Var. 6462



150-200 
Var. 6121
Var. 6178
Var. 6235
Var. 6292
Var. 6349
Var. 6406
Var. 6463



175-200 
Var. 6122
Var. 6179
Var. 6236
Var. 6293
Var. 6350
Var. 6407
Var. 6464





Var. = Variation













TABLE 24







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII stability


achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours)















4-6 hr
4-5 hr
5-8 hr
5-7 hr
5-6 hr
6-8 hr
6-7 hr


















Dosage
0.5-200 
Var. 6465
Var. 6522
Var. 6579
Var. 6636
Var. 6693
Var. 6750
Var. 6807


(IU/kg rVWF:RCo
0.5-150 
Var. 6466
Var. 6523
Var. 6580
Var. 6637
Var. 6694
Var. 6751
Var. 6808


activity)
0.5-100 
Var. 6467
Var. 6524
Var. 6581
Var. 6638
Var. 6695
Var. 6752
Var. 6809



0.5-75  
Var. 6468
Var. 6525
Var. 6582
Var. 6639
Var. 6696
Var. 6753
Var. 6810



0.5-50  
Var. 6469
Var. 6526
Var. 6583
Var. 6640
Var. 6697
Var. 6754
Var. 6811



0.5-25  
Var. 6470
Var. 6527
Var. 6584
Var. 6641
Var. 6698
Var. 6755
Var. 6812



0.5-10  
Var. 6471
Var. 6528
Var. 6585
Var. 6642
Var. 6699
Var. 6756
Var. 6813



0.5-5  
Var. 6472
Var. 6529
Var. 6586
Var. 6643
Var. 6700
Var. 6757
Var. 6814



0.5-2.5 
Var. 6473
Var. 6530
Var. 6587
Var. 6644
Var. 6701
Var. 6758
Var. 6815



0.5-1  
Var. 6474
Var. 6531
Var. 6588
Var. 6645
Var. 6702
Var. 6759
Var. 6816



2.5-200 
Var. 6475
Var. 6532
Var. 6589
Var. 6646
Var. 6703
Var. 6760
Var. 6817



2.5-150 
Var. 6476
Var. 6533
Var. 6590
Var. 6647
Var. 6704
Var. 6761
Var. 6818



2.5-100 
Var. 6477
Var. 6534
Var. 6591
Var. 6648
Var. 6705
Var. 6762
Var. 6819



2.5-75  
Var. 6478
Var. 6535
Var. 6592
Var. 6649
Var. 6706
Var. 6763
Var. 6820



2.5-50  
Var. 6479
Var. 6536
Var. 6593
Var. 6650
Var. 6707
Var. 6764
Var. 6821



2.5-25  
Var. 6480
Var. 6537
Var. 6594
Var. 6651
Var. 6708
Var. 6765
Var. 6822



2.5-10  
Var. 6481
Var. 6538
Var. 6595
Var. 6652
Var. 6709
Var. 6766
Var. 6823



2.5-5  
Var. 6482
Var. 6539
Var. 6596
Var. 6653
Var. 6710
Var. 6767
Var. 6824



 5-200
Var. 6483
Var. 6540
Var. 6597
Var. 6654
Var. 6711
Var. 6768
Var. 6825



 5-175
Var. 6484
Var. 6541
Var. 6598
Var. 6655
Var. 6712
Var. 6769
Var. 6826



 5-150
Var. 6485
Var. 6542
Var. 6599
Var. 6656
Var. 6713
Var. 6770
Var. 6827



 5-125
Var. 6486
Var. 6543
Var. 6600
Var. 6657
Var. 6714
Var. 6771
Var. 6828



 5-100
Var. 6487
Var. 6544
Var. 6601
Var. 6658
Var. 6715
Var. 6772
Var. 6829



5-75
Var. 6488
Var. 6545
Var. 6602
Var. 6659
Var. 6716
Var. 6773
Var. 6830



5-50
Var. 6489
Var. 6546
Var. 6603
Var. 6660
Var. 6717
Var. 6774
Var. 6831



5-25
Var. 6490
Var. 6547
Var. 6604
Var. 6661
Var. 6718
Var. 6775
Var. 6832



5-10
Var. 6491
Var. 6548
Var. 6605
Var. 6662
Var. 6719
Var. 6776
Var. 6833



10-200
Var. 6492
Var. 6549
Var. 6606
Var. 6663
Var. 6720
Var. 6777
Var. 6834



10-150
Var. 6493
Var. 6550
Var. 6607
Var. 6664
Var. 6721
Var. 6778
Var. 6835



10-100
Var. 6494
Var. 6551
Var. 6608
Var. 6665
Var. 6722
Var. 6779
Var. 6836



10-75 
Var. 6495
Var. 6552
Var. 6609
Var. 6666
Var. 6723
Var. 6780
Var. 6837



10-50 
Var. 6496
Var. 6553
Var. 6610
Var. 6667
Var. 6724
Var. 6781
Var. 6838



10-25 
Var. 6497
Var. 6554
Var. 6611
Var. 6668
Var. 6725
Var. 6782
Var. 6839



25-200
Var. 6498
Var. 6555
Var. 6612
Var. 6669
Var. 6726
Var. 6783
Var. 6840



25-150
Var. 6499
Var. 6556
Var. 6613
Var. 6670
Var. 6727
Var. 6784
Var. 6841



25-100
Var. 6500
Var. 6557
Var. 6614
Var. 6671
Var. 6728
Var. 6785
Var. 6842



25-75 
Var. 6501
Var. 6558
Var. 6615
Var. 6672
Var. 6729
Var. 6786
Var. 6843



25-50 
Var. 6502
Var. 6559
Var. 6616
Var. 6673
Var. 6730
Var. 6787
Var. 6844



50-200
Var. 6503
Var. 6560
Var. 6617
Var. 6674
Var. 6731
Var. 6788
Var. 6845



50-150
Var. 6504
Var. 6561
Var. 6618
Var. 6675
Var. 6732
Var. 6789
Var. 6846



50-100
Var. 6505
Var. 6562
Var. 6619
Var. 6676
Var. 6733
Var. 6790
Var. 6847



50-75 
Var. 6506
Var. 6563
Var. 6620
Var. 6677
Var. 6734
Var. 6791
Var. 6848



75-200
Var. 6507
Var. 6564
Var. 6621
Var. 6678
Var. 6735
Var. 6792
Var. 6849



75-175
Var. 6508
Var. 6565
Var. 6622
Var. 6679
Var. 6736
Var. 6793
Var. 6850



75-150
Var. 6509
Var. 6566
Var. 6623
Var. 6680
Var. 6737
Var. 6794
Var. 6851



75-125
Var. 6510
Var. 6567
Var. 6624
Var. 6681
Var. 6738
Var. 6795
Var. 6852



75-100
Var. 6511
Var. 6568
Var. 6625
Var. 6682
Var. 6739
Var. 6796
Var. 6853



100-200 
Var. 6512
Var. 6569
Var. 6626
Var. 6683
Var. 6740
Var. 6797
Var. 6854



100-175 
Var. 6513
Var. 6570
Var. 6627
Var. 6684
Var. 6741
Var. 6798
Var. 6855



100-150 
Var. 6514
Var. 6571
Var. 6628
Var. 6685
Var. 6742
Var. 6799
Var. 6856



100-125 
Var. 6515
Var. 6572
Var. 6629
Var. 6686
Var. 6743
Var. 6800
Var. 6857



125-200 
Var. 6516
Var. 6573
Var. 6630
Var. 6687
Var. 6744
Var. 6801
Var. 6858



125-175 
Var. 6517
Var. 6574
Var. 6631
Var. 6688
Var. 6745
Var. 6802
Var. 6859



125-150 
Var. 6518
Var. 6575
Var. 6632
Var. 6689
Var. 6746
Var. 6803
Var. 6860



150-200 
Var. 6519
Var. 6576
Var. 6633
Var. 6690
Var. 6747
Var. 6804
Var. 6861



150-200 
Var. 6520
Var. 6577
Var. 6634
Var. 6691
Var. 6748
Var. 6805
Var. 6862



175-200 
Var. 6521
Var. 6578
Var. 6635
Var. 6692
Var. 6749
Var. 6806
Var. 6863





Var. = Variation













TABLE 25







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII stability


achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Hours/Percent)
















at least
at least
at least
at least
at least




7-8 hr
10%
20%
30%
40%
50%
10-50%


















Dosage
0.5-200 
Var. 6864
Var. 6921
Var. 6978
Var. 7035
Var. 7092
Var. 7149
Var. 7206


(IU/kg rVWF:RCo
0.5-150 
Var. 6865
Var. 6922
Var. 6979
Var. 7036
Var. 7093
Var. 7150
Var. 7207


activity)
0.5-100 
Var. 6866
Var. 6923
Var. 6980
Var. 7037
Var. 7094
Var. 7151
Var. 7208



0.5-75  
Var. 6867
Var. 6924
Var. 6981
Var. 7038
Var. 7095
Var. 7152
Var. 7209



0.5-50  
Var. 6868
Var. 6925
Var. 6982
Var. 7039
Var. 7096
Var. 7153
Var. 7210



0.5-25  
Var. 6869
Var. 6926
Var. 6983
Var. 7040
Var. 7097
Var. 7154
Var. 7211



0.5-10  
Var. 6870
Var. 6927
Var. 6984
Var. 7041
Var. 7098
Var. 7155
Var. 7212



0.5-5  
Var. 6871
Var. 6928
Var. 6985
Var. 7042
Var. 7099
Var. 7156
Var. 7213



0.5-2.5 
Var. 6872
Var. 6929
Var. 6986
Var. 7043
Var. 7100
Var. 7157
Var. 7214



0.5-1  
Var. 6873
Var. 6930
Var. 6987
Var. 7044
Var. 7101
Var. 7158
Var. 7215



2.5-200 
Var. 6874
Var. 6931
Var. 6988
Var. 7045
Var. 7102
Var. 7159
Var. 7216



2.5-150 
Var. 6875
Var. 6932
Var. 6989
Var. 7046
Var. 7103
Var. 7160
Var. 7217



2.5-100 
Var. 6876
Var. 6933
Var. 6990
Var. 7047
Var. 7104
Var. 7161
Var. 7218



2.5-75  
Var. 6877
Var. 6934
Var. 6991
Var. 7048
Var. 7105
Var. 7162
Var. 7219



2.5-50  
Var. 6878
Var. 6935
Var. 6992
Var. 7049
Var. 7106
Var. 7163
Var. 7220



2.5-25  
Var. 6879
Var. 6936
Var. 6993
Var. 7050
Var. 7107
Var. 7164
Var. 7221



2.5-10  
Var. 6880
Var. 6937
Var. 6994
Var. 7051
Var. 7108
Var. 7165
Var. 7222



2.5-5  
Var. 6881
Var. 6938
Var. 6995
Var. 7052
Var. 7109
Var. 7166
Var. 7223



 5-200
Var. 6882
Var. 6939
Var. 6996
Var. 7053
Var. 7110
Var. 7167
Var. 7224



 5-175
Var. 6883
Var. 6940
Var. 6997
Var. 7054
Var. 7111
Var. 7168
Var. 7225



 5-150
Var. 6884
Var. 6941
Var. 6998
Var. 7055
Var. 7112
Var. 7169
Var. 7226



 5-125
Var. 6885
Var. 6942
Var. 6999
Var. 7056
Var. 7113
Var. 7170
Var. 7227



 5-100
Var. 6886
Var. 6943
Var. 7000
Var. 7057
Var. 7114
Var. 7171
Var. 7228



5-75
Var. 6887
Var. 6944
Var. 7001
Var. 7058
Var. 7115
Var. 7172
Var. 7229



5-50
Var. 6888
Var. 6945
Var. 7002
Var. 7059
Var. 7116
Var. 7173
Var. 7230



5-25
Var. 6889
Var. 6946
Var. 7003
Var. 7060
Var. 7117
Var. 7174
Var. 7231



5-10
Var. 6890
Var. 6947
Var. 7004
Var. 7061
Var. 7118
Var. 7175
Var. 7232



10-200
Var. 6891
Var. 6948
Var. 7005
Var. 7062
Var. 7119
Var. 7176
Var. 7233



10-150
Var. 6892
Var. 6949
Var. 7006
Var. 7063
Var. 7120
Var. 7177
Var. 7234



10-100
Var. 6893
Var. 6950
Var. 7007
Var. 7064
Var. 7121
Var. 7178
Var. 7235



10-75 
Var. 6894
Var. 6951
Var. 7008
Var. 7065
Var. 7122
Var. 7179
Var. 7236



10-50 
Var. 6895
Var. 6952
Var. 7009
Var. 7066
Var. 7123
Var. 7180
Var. 7237



10-25 
Var. 6896
Var. 6953
Var. 7010
Var. 7067
Var. 7124
Var. 7181
Var. 7238



25-200
Var. 6897
Var. 6954
Var. 7011
Var. 7068
Var. 7125
Var. 7182
Var. 7239



25-150
Var. 6898
Var. 6955
Var. 7012
Var. 7069
Var. 7126
Var. 7183
Var. 7240



25-100
Var. 6899
Var. 6956
Var. 7013
Var. 7070
Var. 7127
Var. 7184
Var. 7241



25-75 
Var. 6900
Var. 6957
Var. 7014
Var. 7071
Var. 7128
Var. 7185
Var. 7242



25-50 
Var. 6901
Var. 6958
Var. 7015
Var. 7072
Var. 7129
Var. 7186
Var. 7243



50-200
Var. 6902
Var. 6959
Var. 7016
Var. 7073
Var. 7130
Var. 7187
Var. 7244



50-150
Var. 6903
Var. 6960
Var. 7017
Var. 7074
Var. 7131
Var. 7188
Var. 7245



50-100
Var. 6904
Var. 6961
Var. 7018
Var. 7075
Var. 7132
Var. 7189
Var. 7246



50-75 
Var. 6905
Var. 6962
Var. 7019
Var. 7076
Var. 7133
Var. 7190
Var. 7247



75-200
Var. 6906
Var. 6963
Var. 7020
Var. 7077
Var. 7134
Var. 7191
Var. 7248



75-175
Var. 6907
Var. 6964
Var. 7021
Var. 7078
Var. 7135
Var. 7192
Var. 7249



75-150
Var. 6908
Var. 6965
Var. 7022
Var. 7079
Var. 7136
Var. 7193
Var. 7250



75-125
Var. 6909
Var. 6966
Var. 7023
Var. 7080
Var. 7137
Var. 7194
Var. 7251



75-100
Var. 6910
Var. 6967
Var. 7024
Var. 7081
Var. 7138
Var. 7195
Var. 7252



100-200 
Var. 6911
Var. 6968
Var. 7025
Var. 7082
Var. 7139
Var. 7196
Var. 7253



100-175 
Var. 6912
Var. 6969
Var. 7026
Var. 7083
Var. 7140
Var. 7197
Var. 7254



100-150 
Var. 6913
Var. 6970
Var. 7027
Var. 7084
Var. 7141
Var. 7198
Var. 7255



100-125 
Var. 6914
Var. 6971
Var. 7028
Var. 7085
Var. 7142
Var. 7199
Var. 7256



125-200 
Var. 6915
Var. 6972
Var. 7029
Var. 7086
Var. 7143
Var. 7200
Var. 7257



125-175 
Var. 6916
Var. 6973
Var. 7030
Var. 7087
Var. 7144
Var. 7201
Var. 7258



125-150 
Var. 6917
Var. 6974
Var. 7031
Var. 7088
Var. 7145
Var. 7202
Var. 7259



150-200 
Var. 6918
Var. 6975
Var. 7032
Var. 7089
Var. 7146
Var. 7203
Var. 7260



150-200 
Var. 6919
Var. 6976
Var. 7033
Var. 7090
Var. 7147
Var. 7204
Var. 7261



175-200 
Var. 6920
Var. 6977
Var. 7034
Var. 7091
Var. 7148
Var. 7205
Var. 7262





Var. = Variation













TABLE 26







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII stability


achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Percent)















10-40%
10-30%
10-20%
15-50%
15-40%
15-30%
15-20%


















Dosage
0.5-200 
Var. 7263
Var. 7320
Var. 7377
Var. 7434
Var. 7491
Var. 7548
Var. 7605


(IU/kg rVWF:RCo
0.5-150 
Var. 7264
Var. 7321
Var. 7378
Var. 7435
Var. 7492
Var. 7549
Var. 7606


activity)
0.5-100 
Var. 7265
Var. 7322
Var. 7379
Var. 7436
Var. 7493
Var. 7550
Var. 7607



0.5-75  
Var. 7266
Var. 7323
Var. 7380
Var. 7437
Var. 7494
Var. 7551
Var. 7608



0.5-50  
Var. 7267
Var. 7324
Var. 7381
Var. 7438
Var. 7495
Var. 7552
Var. 7609



0.5-25  
Var. 7268
Var. 7325
Var. 7382
Var. 7439
Var. 7496
Var. 7553
Var. 7610



0.5-10  
Var. 7269
Var. 7326
Var. 7383
Var. 7440
Var. 7497
Var. 7554
Var. 7611



0.5-5  
Var. 7270
Var. 7327
Var. 7384
Var. 7441
Var. 7498
Var. 7555
Var. 7612



0.5-2.5 
Var. 7271
Var. 7328
Var. 7385
Var. 7442
Var. 7499
Var. 7556
Var. 7613



0.5-1  
Var. 7272
Var. 7329
Var. 7386
Var. 7443
Var. 7500
Var. 7557
Var. 7614



2.5-200 
Var. 7273
Var. 7330
Var. 7387
Var. 7444
Var. 7501
Var. 7558
Var. 7615



2.5-150 
Var. 7274
Var. 7331
Var. 7388
Var. 7445
Var. 7502
Var. 7559
Var. 7616



2.5-100 
Var. 7275
Var. 7332
Var. 7389
Var. 7446
Var. 7503
Var. 7560
Var. 7617



2.5-75  
Var. 7276
Var. 7333
Var. 7390
Var. 7447
Var. 7504
Var. 7561
Var. 7618



2.5-50  
Var. 7277
Var. 7334
Var. 7391
Var. 7448
Var. 7505
Var. 7562
Var. 7619



2.5-25  
Var. 7278
Var. 7335
Var. 7392
Var. 7449
Var. 7506
Var. 7563
Var. 7620



2.5-10  
Var. 7279
Var. 7336
Var. 7393
Var. 7450
Var. 7507
Var. 7564
Var. 7621



2.5-5  
Var. 7280
Var. 7337
Var. 7394
Var. 7451
Var. 7508
Var. 7565
Var. 7622



 5-200
Var. 7281
Var. 7338
Var. 7395
Var. 7452
Var. 7509
Var. 7566
Var. 7623



 5-175
Var. 7282
Var. 7339
Var. 7396
Var. 7453
Var. 7510
Var. 7567
Var. 7624



 5-150
Var. 7283
Var. 7340
Var. 7397
Var. 7454
Var. 7511
Var. 7568
Var. 7625



 5-125
Var. 7284
Var. 7341
Var. 7398
Var. 7455
Var. 7512
Var. 7569
Var. 7626



 5-100
Var. 7285
Var. 7342
Var. 7399
Var. 7456
Var. 7513
Var. 7570
Var. 7627



5-75
Var. 7286
Var. 7343
Var. 7400
Var. 7457
Var. 7514
Var. 7571
Var. 7628



5-50
Var. 7287
Var. 7344
Var. 7401
Var. 7458
Var. 7515
Var. 7572
Var. 7629



5-25
Var. 7288
Var. 7345
Var. 7402
Var. 7459
Var. 7516
Var. 7573
Var. 7630



5-10
Var. 7289
Var. 7346
Var. 7403
Var. 7460
Var. 7517
Var. 7574
Var. 7631



10-200
Var. 7290
Var. 7347
Var. 7404
Var. 7461
Var. 7518
Var. 7575
Var. 7632



10-150
Var. 7291
Var. 7348
Var. 7405
Var. 7462
Var. 7519
Var. 7576
Var. 7633



10-100
Var. 7292
Var. 7349
Var. 7406
Var. 7463
Var. 7520
Var. 7577
Var. 7634



10-75 
Var. 7293
Var. 7350
Var. 7407
Var. 7464
Var. 7521
Var. 7578
Var. 7635



10-50 
Var. 7294
Var. 7351
Var. 7408
Var. 7465
Var. 7522
Var. 7579
Var. 7636



10-25 
Var. 7295
Var. 7352
Var. 7409
Var. 7466
Var. 7523
Var. 7580
Var. 7637



25-200
Var. 7296
Var. 7353
Var. 7410
Var. 7467
Var. 7524
Var. 7581
Var. 7638



25-150
Var. 7297
Var. 7354
Var. 7411
Var. 7468
Var. 7525
Var. 7582
Var. 7639



25-100
Var. 7298
Var. 7355
Var. 7412
Var. 7469
Var. 7526
Var. 7583
Var. 7640



25-75 
Var. 7299
Var. 7356
Var. 7413
Var. 7470
Var. 7527
Var. 7584
Var. 7641



25-50 
Var. 7300
Var. 7357
Var. 7414
Var. 7471
Var. 7528
Var. 7585
Var. 7642



50-200
Var. 7301
Var. 7358
Var. 7415
Var. 7472
Var. 7529
Var. 7586
Var. 7643



50-150
Var. 7302
Var. 7359
Var. 7416
Var. 7473
Var. 7530
Var. 7587
Var. 7644



50-100
Var. 7303
Var. 7360
Var. 7417
Var. 7474
Var. 7531
Var. 7588
Var. 7645



50-75 
Var. 7304
Var. 7361
Var. 7418
Var. 7475
Var. 7532
Var. 7589
Var. 7646



75-200
Var. 7305
Var. 7362
Var. 7419
Var. 7476
Var. 7533
Var. 7590
Var. 7647



75-175
Var. 7306
Var. 7363
Var. 7420
Var. 7477
Var. 7534
Var. 7591
Var. 7648



75-150
Var. 7307
Var. 7364
Var. 7421
Var. 7478
Var. 7535
Var. 7592
Var. 7649



75-125
Var. 7308
Var. 7365
Var. 7422
Var. 7479
Var. 7536
Var. 7593
Var. 7650



75-100
Var. 7309
Var. 7366
Var. 7423
Var. 7480
Var. 7537
Var. 7594
Var. 7651



100-200 
Var. 7310
Var. 7367
Var. 7424
Var. 7481
Var. 7538
Var. 7595
Var. 7652



100-175 
Var. 7311
Var. 7368
Var. 7425
Var. 7482
Var. 7539
Var. 7596
Var. 7653



100-150 
Var. 7312
Var. 7369
Var. 7426
Var. 7483
Var. 7540
Var. 7597
Var. 7654



100-125 
Var. 7313
Var. 7370
Var. 7427
Var. 7484
Var. 7541
Var. 7598.
Var. 7655



125-200 
Var. 7314
Var. 7371
Var. 7428
Var. 7485
Var. 7542
Var. 7599
Var. 7656



125-175 
Var. 7315
Var. 7372
Var. 7429
Var. 7486
Var. 7543
Var. 7600
Var. 7657



125-150 
Var. 7316
Var. 7373
Var. 7430
Var. 7487
Var. 7544
Var. 7601
Var. 7658



150-200 
Var. 7317
Var. 7374
Var. 7431
Var. 7488
Var. 7545
Var. 7602
Var. 7659



150-200 
Var. 7318
Var. 7375
Var. 7432
Var. 7489
Var. 7546
Var. 7603
Var. 7660



175-200 
Var. 7319
Var. 7376
Var. 7433
Var. 7490
Var. 7547
Var. 7604
Var. 7661





Var. = Variation













TABLE 27







Exemplary embodiments for the combination of rVWF dosage and increase in FVIII stability


achieved, as compared to FVIII stability in a subject administered a composition of pdVWF.









Increased Stability (Percent)














20-50%
20-40%
20-30%
30-50%
30-40%
40-50%

















Dosage
0.5-200 
Var. 7662
Var. 7719
Var. 7776
Var. 7833
Var. 7890
Var. 7947


(IU/kg rVWF:RCo
0.5-150 
Var. 7663
Var. 7720
Var. 7777
Var. 7834
Var. 7891
Var. 7948


activity)
0.5-100 
Var. 7664
Var. 7721
Var. 7778
Var. 7835
Var. 7892
Var. 7949



0.5-75  
Var. 7665
Var. 7722
Var. 7779
Var. 7836
Var. 7893
Var. 7950



0.5-50  
Var. 7666
Var. 7723
Var. 7780
Var. 7837
Var. 7894
Var. 7951



0.5-25  
Var. 7667
Var. 7724
Var. 7781
Var. 7838
Var. 7895
Var. 7952



0.5-10  
Var. 7668
Var. 7725
Var. 7782
Var. 7839
Var. 7896
Var. 7953



0.5-5  
Var. 7669
Var. 7726
Var. 7783
Var. 7840
Var. 7897
Var. 7954



0.5-2.5 
Var. 7670
Var. 7727
Var. 7784
Var. 7841
Var. 7898
Var. 7955



0.5-1  
Var. 7671
Var. 7728
Var. 7785
Var. 7842
Var. 7899
Var. 7956



2.5-200 
Var. 7672
Var. 7729
Var. 7786
Var. 7843
Var. 7900
Var. 7957



2.5-150 
Var. 7673
Var. 7730
Var. 7787
Var. 7844
Var. 7901
Var. 7958



2.5-100 
Var. 7674
Var. 7731
Var. 7788
Var. 7845
Var. 7902
Var. 7959



2.5-75  
Var. 7675
Var. 7732
Var. 7789
Var. 7846
Var. 7903
Var. 7960



2.5-50  
Var. 7676
Var. 7733
Var. 7790
Var. 7847
Var. 7904
Var. 7961



2.5-25  
Var. 7677
Var. 7734
Var. 7791
Var. 7848
Var. 7905
Var. 7962



2.5-10  
Var. 7678
Var. 7735
Var. 7792
Var. 7849
Var. 7906
Var. 7963



2.5-5  
Var. 7679
Var. 7736
Var. 7793
Var. 7850
Var. 7907
Var. 7964



 5-200
Var. 7680
Var. 7737
Var. 7794
Var. 7851
Var. 7908
Var. 7965



 5-175
Var. 7681
Var. 7738
Var. 7795
Var. 7852
Var. 7909
Var. 7966



 5-150
Var. 7682
Var. 7739
Var. 7796
Var. 7853
Var. 7910
Var. 7967



 5-125
Var. 7683
Var. 7740
Var. 7797
Var. 7854
Var. 7911
Var. 7968



 5-100
Var. 7684
Var. 7741
Var. 7798
Var. 7855
Var. 7912
Var. 7969



5-75
Var. 7685
Var. 7742
Var. 7799
Var. 7856
Var. 7913
Var. 7970



5-50
Var. 7686
Var. 7743
Var. 7800
Var. 7857
Var. 7914
Var. 7971



5-25
Var. 7687
Var. 7744
Var. 7801
Var. 7858
Var. 7915
Var. 7972



5-10
Var. 7688
Var. 7745
Var. 7802
Var. 7859
Var. 7916
Var. 7973



10-200
Var. 7689
Var. 7746
Var. 7803
Var. 7860
Var. 7917
Var. 7974



10-150
Var. 7690
Var. 7747
Var. 7804
Var. 7861
Var. 7918
Var. 7975



10-100
Var. 7691
Var. 7748
Var. 7805
Var. 7862
Var. 7919
Var. 7976



10-75 
Var. 7692
Var. 7749
Var. 7806
Var. 7863
Var. 7920
Var. 7977



10-50 
Var. 7693
Var. 7750
Var. 7807
Var. 7864
Var. 7921
Var. 7978



10-25 
Var. 7694
Var. 7751
Var. 7808
Var. 7865
Var. 7922
Var. 7979



25-200
Var. 7695
Var. 7752
Var. 7809
Var. 7866
Var. 7923
Var. 7980



25-150
Var. 7696
Var. 7753
Var. 7810
Var. 7867
Var. 7924
Var. 7981



25-100
Var. 7697
Var. 7754
Var. 7811
Var. 7868
Var. 7925
Var. 7982



25-75 
Var. 7698
Var. 7755
Var. 7812
Var. 7869
Var. 7926
Var. 7983



25-50 
Var. 7699
Var. 7756
Var. 7813
Var. 7870
Var. 7927
Var. 7984



50-200
Var. 7700
Var. 7757
Var. 7814
Var. 7871
Var. 7928
Var. 7985



50-150
Var. 7701
Var. 7758
Var. 7815
Var. 7872
Var. 7929
Var. 7986



50-100
Var. 7702
Var. 7759
Var. 7816
Var. 7873
Var. 7930
Var. 7987



50-75 
Var. 7703
Var. 7760
Var. 7817
Var. 7874
Var. 7931
Var. 7988



75-200
Var. 7704
Var. 7761
Var. 7818
Var. 7875
Var. 7932
Var. 7989



75-175
Var. 7705
Var. 7762
Var. 7819
Var. 7876
Var. 7933
Var. 7990



75-150
Var. 7706
Var. 7763
Var. 7820
Var. 7877
Var. 7934
Var. 7991



75-125
Var. 7707
Var. 7764
Var. 7821
Var. 7878
Var. 7935
Var. 7992



75-100
Var. 7708
Var. 7765
Var. 7822
Var. 7879
Var. 7936
Var. 7993



100-200 
Var. 7709
Var. 7766
Var. 7823
Var. 7880
Var. 7937
Var. 7994



100-175 
Var. 7710
Var. 7767
Var. 7824
Var. 7881
Var. 7938
Var. 7995



100-150 
Var. 7711
Var. 7768
Var. 7825
Var. 7882
Var. 7939
Var. 7996



100-125 
Var. 7712
Var. 7769
Var. 7826
Var. 7883
Var. 7940
Var. 7997



125-200 
Var. 7713
Var. 7770
Var. 7827
Var. 7884
Var. 7941
Var. 7998



125-175 
Var. 7714
Var. 7771
Var. 7828
Var. 7885
Var. 7942
Var. 7999



125-150 
Var. 7715
Var. 7772
Var. 7829
Var. 7886
Var. 7943
Var. 8000



150-200 
Var. 7716
Var. 7773
Var. 7830
Var. 7887
Var. 7944
Var. 8001



150-200 
Var. 7717
Var. 7774
Var. 7831
Var. 7888
Var. 7945
Var. 8002



175-200 
Var. 7718
Var. 7775
Var. 7832
Var. 7889
Var. 7946
Var. 8003





Var. = Variation






In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the rVWF in the composition has a specific activity of from 40 mU/μg to 60 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the rVWF in the composition has a specific activity of at least 60 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the rVWF in the composition has a specific activity of at least 80 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the rVWF in the composition has a specific activity selected from variations 1 to 133 found in Table 1, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.







TABLE 28







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)















at least 20
at least 30
at least 40
at least 50
at least 60
at least 70
at least 80


















Dosage
0.5-200 
Var. 8004
Var. 8061
Var. 8118
Var. 8175
Var. 8232
Var. 8289
Var. 8346


(IU/kg rVWF:RCo
0.5-150 
Var. 8005
Var. 8062
Var. 8119
Var. 8176
Var. 8233
Var. 8290
Var. 8347


activity)
0.5-100 
Var. 8006
Var. 8063
Var. 8120
Var. 8177
Var. 8234
Var. 8291
Var. 8348



0.5-75  
Var. 8007
Var. 8064
Var. 8121
Var. 8178
Var. 8235
Var. 8292
Var. 8349



0.5-50  
Var. 8008
Var. 8065
Var. 8122
Var. 8179
Var. 8236
Var. 8293
Var. 8350



0.5-25  
Var. 8009
Var. 8066
Var. 8123
Var. 8180
Var. 8237
Var. 8294
Var. 8351



0.5-10  
Var. 8010
Var. 8067
Var. 8124
Var. 8181
Var. 8238
Var. 8295
Var. 8352



0.5-5  
Var. 8011
Var. 8068
Var. 8125
Var. 8182
Var. 8239
Var. 8296
Var. 8353



0.5-2.5 
Var. 8012
Var. 8069
Var. 8126
Var. 8183
Var. 8240
Var. 8297
Var. 8354



0.5-1  
Var. 8013
Var. 8070
Var. 8127
Var. 8184
Var. 8241
Var. 8298
Var. 8355



2.5-200 
Var. 8014
Var. 8071
Var. 8128
Var. 8185
Var. 8242
Var. 8299
Var. 8356



2.5-150 
Var. 8015
Var. 8072
Var. 8129
Var. 8186
Var. 8243
Var. 8300
Var. 8357



2.5-100 
Var. 8016
Var. 8073
Var. 8130
Var. 8187
Var. 8244
Var. 8301
Var. 8358



2.5-75  
Var. 8017
Var. 8074
Var. 8131
Var. 8188
Var. 8245
Var. 8302
Var. 8359



2.5-50  
Var. 8018
Var. 8075
Var. 8132
Var. 8189
Var. 8246
Var. 8303
Var. 8360



2.5-25  
Var. 8019
Var. 8076
Var. 8133
Var. 8190
Var. 8247
Var. 8304
Var. 8361



2.5-10  
Var. 8020
Var. 8077
Var. 8134
Var. 8191
Var. 8248
Var. 8305
Var. 8362



2.5-5  
Var. 8021
Var. 8078
Var. 8135
Var. 8192
Var. 8249
Var. 8306
Var. 8363



 5-200
Var. 8022
Var. 8079
Var. 8136
Var. 8193
Var. 8250
Var. 8307
Var. 8364



 5-175
Var. 8023
Var. 8080
Var. 8137
Var. 8194
Var. 8251
Var. 8308
Var. 8365



 5-150
Var. 8024
Var. 8081
Var. 8138
Var. 8195
Var. 8252
Var. 8309
Var. 8366



 5-125
Var. 8025
Var. 8082
Var. 8139
Var. 8196
Var. 8253
Var. 8310
Var. 8367



 5-100
Var. 8026
Var. 8083
Var. 8140
Var. 8197
Var. 8254
Var. 8311
Var. 8368



5-75
Var. 8027
Var. 8084
Var. 8141
Var. 8198
Var. 8255
Var. 8312
Var. 8369



5-50
Var. 8028
Var. 8085
Var. 8142
Var. 8199
Var. 8256
Var. 8313
Var. 8370



5-25
Var. 8029
Var. 8086
Var. 8143
Var. 8200
Var. 8257
Var. 8314
Var. 8371



5-10
Var. 8030
Var. 8087
Var. 8144
Var. 8201
Var. 8258
Var. 8315
Var. 8372



10-200
Var. 8031
Var. 8088
Var. 8145
Var. 8202
Var. 8259
Var. 8316
Var. 8373



10-150
Var. 8032
Var. 8089
Var. 8146
Var. 8203
Var. 8260
Var. 8317
Var. 8374



10-100
Var. 8033
Var. 8090
Var. 8147
Var. 8204
Var. 8261
Var. 8318
Var. 8375



10-75 
Var. 8034
Var. 8091
Var. 8148
Var. 8205
Var. 8262
Var. 8319
Var. 8376



10-50 
Var. 8035
Var. 8092
Var. 8149
Var. 8206
Var. 8263
Var. 8320
Var. 8377



10-25 
Var. 8036
Var. 8093
Var. 8150
Var. 8207
Var. 8264
Var. 8321
Var. 8378



25-200
Var. 8037
Var. 8094
Var. 8151
Var. 8208
Var. 8265
Var. 8322
Var. 8379



25-150
Var. 8038
Var. 8095
Var. 8152
Var. 8209
Var. 8266
Var. 8323
Var. 8380



25-100
Var. 8039
Var. 8096
Var. 8153
Var. 8210
Var. 8267
Var. 8324
Var. 8381



25-75 
Var. 8040
Var. 8097
Var. 8154
Var. 8211
Var. 8268
Var. 8325
Var. 8382



25-50 
Var. 8041
Var. 8098
Var. 8155
Var. 8212
Var. 8269
Var. 8326
Var. 8383



50-200
Var. 8042
Var. 8099
Var. 8156
Var. 8213
Var. 8270
Var. 8327
Var. 8384



50-150
Var. 8043
Var. 8100
Var. 8157
Var. 8214
Var. 8271
Var. 8328
Var. 8385



50-100
Var. 8044
Var. 8101
Var. 8158
Var. 8215
Var. 8272
Var. 8329
Var. 8386



50-75 
Var. 8045
Var. 8102
Var. 8159
Var. 8216
Var. 8273
Var. 8330
Var. 8387



75-200
Var. 8046
Var. 8103
Var. 8160
Var. 8217
Var. 8274
Var. 8331
Var. 8388



75-175
Var. 8047
Var. 8104
Var. 8161
Var. 8218
Var. 8275
Var. 8332
Var. 8389



75-150
Var. 8048
Var. 8105
Var. 8162
Var. 8219
Var. 8276
Var. 8333
Var. 8390



75-125
Var. 8049
Var. 8106
Var. 8163
Var. 8220
Var. 8277
Var. 8334
Var. 8391



75-100
Var. 8050
Var. 8107
Var. 8164
Var. 8221
Var. 8278
Var. 8335
Var. 8392



100-200 
Var. 8051
Var. 8108
Var. 8165
Var. 8222
Var. 8279
Var. 8336
Var. 8393



100-175 
Var. 8052
Var. 8109
Var. 8166
Var. 8223
Var. 8280
Var. 8337
Var. 8394



100-150 
Var. 8053
Var. 8110
Var. 8167
Var. 8224
Var. 8281
Var. 8338
Var. 8395



100-125 
Var. 8054
Var. 8111
Var. 8168
Var. 8225
Var. 8282
Var. 8339
Var. 8396



125-200 
Var. 8055
Var. 8112
Var. 8169
Var. 8226
Var. 8283
Var. 8340
Var. 8397



125-175 
Var. 8056
Var. 8113
Var. 8170
Var. 8227
Var. 8284
Var. 8341
Var. 8398



125-150 
Var. 8057
Var. 8114
Var. 8171
Var. 8228
Var. 8285
Var. 8342
Var. 8399



150-200 
Var. 8058
Var. 8115
Var. 8172
Var. 8229
Var. 8286
Var. 8343
Var. 8400



150-200 
Var. 8059
Var. 8116
Var. 8173
Var. 8230
Var. 8287
Var. 8344
Var. 8401



175-200 
Var. 8060
Var. 8117
Var. 8174
Var. 8231
Var. 8288
Var. 8345
Var. 8402





Var. = Variation













TABLE 29







Exemplary embodiments for the combination of rVWF dosage and rVWF


specific activity useful in the methods described herein.









Specific Activity (mU/μg)















at least 90
at least 100
at least 125
at least 150
20-150
20-125
20-100


















Dosage
0.5-200 
Var. 8403
Var. 8460
Var. 8517
Var. 8574
Var. 8631
Var. 8688
Var. 8745


(IU/kg rVWF:RCo
0.5-150 
Var. 8404
Var. 8461
Var. 8518
Var. 8575
Var. 8632
Var. 8689
Var. 8746


activity)
0.5-100 
Var. 8405
Var. 8462
Var. 8519
Var. 8576
Var. 8633
Var. 8690
Var. 8747



0.5-75  
Var. 8406
Var. 8463
Var. 8520
Var. 8577
Var. 8634
Var. 8691
Var. 8748



0.5-50  
Var. 8407
Var. 8464
Var. 8521
Var. 8578
Var. 8635
Var. 8692
Var. 8749



0.5-25  
Var. 8408
Var. 8465
Var. 8522
Var. 8579
Var. 8636
Var. 8693
Var. 8750



0.5-10  
Var. 8409
Var. 8466
Var. 8523
Var. 8580
Var. 8637
Var. 8694
Var. 8751



0.5-5  
Var. 8410
Var. 8467
Var. 8524
Var. 8581
Var. 8638
Var. 8695
Var. 8752



0.5-2.5 
Var. 8411
Var. 8468
Var. 8525
Var. 8582
Var. 8639
Var. 8696
Var. 8753



0.5-1  
Var. 8412
Var. 8469
Var. 8526
Var. 8583
Var. 8640
Var. 8697
Var. 8754



2.5-200 
Var. 8413
Var. 8470
Var. 8527
Var. 8584
Var. 8641
Var. 8698
Var. 8755



2.5-150 
Var. 8414
Var. 8471
Var. 8528
Var. 8585
Var. 8642
Var. 8699
Var. 8756



2.5-100 
Var. 8415
Var. 8472
Var. 8529
Var. 8586
Var. 8643
Var. 8700
Var. 8757



2.5-75  
Var. 8416
Var. 8473
Var. 8530
Var. 8587
Var. 8644
Var. 8701
Var. 8758



2.5-50  
Var. 8417
Var. 8474
Var. 8531
Var. 8588
Var. 8645
Var. 8702
Var. 8759



2.5-25  
Var. 8418
Var. 8475
Var. 8532
Var. 8589
Var. 8646
Var. 8703
Var. 8760



2.5-10  
Var. 8419
Var. 8476
Var. 8533
Var. 8590
Var. 8647
Var. 8704
Var. 8761



2.5-5  
Var. 8420
Var. 8477
Var. 8534
Var. 8591
Var. 8648
Var. 8705
Var. 8762



 5-200
Var. 8421
Var. 8478
Var. 8535
Var. 8592
Var. 8649
Var. 8706
Var. 8763



 5-175
Var. 8422
Var. 8479
Var. 8536
Var. 8593
Var. 8650
Var. 8707
Var. 8764



 5-150
Var. 8423
Var. 8480
Var. 8537
Var. 8594
Var. 8651
Var. 8708
Var. 8765



 5-125
Var. 8424
Var. 8481
Var. 8538
Var. 8595
Var. 8652
Var. 8709
Var. 8766



 5-100
Var. 8425
Var. 8482
Var. 8539
Var. 8596
Var. 8653
Var. 8710
Var. 8767



5-75
Var. 8426
Var. 8483
Var. 8540
Var. 8597
Var. 8654
Var. 8711
Var. 8768



5-50
Var. 8427
Var. 8484
Var. 8541
Var. 8598
Var. 8655
Var. 8712
Var. 8769



5-25
Var. 8428
Var. 8485
Var. 8542
Var. 8599
Var. 8656
Var. 8713
Var. 8770



5-10
Var. 8429
Var. 8486
Var. 8543
Var. 8600
Var. 8657
Var. 8714
Var. 8771



10-200
Var. 8430
Var. 8487
Var. 8544
Var. 8601
Var. 8658
Var. 8715
Var. 8772



10-150
Var. 8431
Var. 8488
Var. 8545
Var. 8602
Var. 8659
Var. 8716
Var. 8773



10-100
Var. 8432
Var. 8489
Var. 8546
Var. 8603
Var. 8660
Var. 8717
Var. 8774



10-75 
Var. 8433
Var. 8490
Var. 8547
Var. 8604
Var. 8661
Var. 8718
Var. 8775



10-50 
Var. 8434
Var. 8491
Var. 8548
Var. 8605
Var. 8662
Var. 8719
Var. 8776



10-25 
Var. 8435
Var. 8492
Var. 8549
Var. 8606
Var. 8663
Var. 8720
Var. 8777



25-200
Var. 8436
Var. 8493
Var. 8550
Var. 8607
Var. 8664
Var. 8721
Var. 8778



25-150
Var. 8437
Var. 8494
Var. 8551
Var. 8608
Var. 8665
Var. 8722
Var. 8779



25-100
Var. 8438
Var. 8495
Var. 8552
Var. 8609
Var. 8666
Var. 8723
Var. 8780



25-75 
Var. 8439
Var. 8496
Var. 8553
Var. 8610
Var. 8667
Var. 8724
Var. 8781



25-50 
Var. 8440
Var. 8497
Var. 8554
Var. 8611
Var. 8668
Var. 8725
Var. 8782



50-200
Var. 8441
Var. 8498
Var. 8555
Var. 8612
Var. 8669
Var. 8726
Var. 8783



50-150
Var. 8442
Var. 8499
Var. 8556
Var. 8613
Var. 8670
Var. 8727
Var. 8784



50-100
Var. 8443
Var. 8500
Var. 8557
Var. 8614
Var. 8671
Var. 8728
Var. 8785



50-75 
Var. 8444
Var. 8501
Var. 8558
Var. 8615
Var. 8672
Var. 8729
Var. 8786



75-200
Var. 8445
Var. 8502
Var. 8559
Var. 8616
Var. 8673
Var. 8730
Var. 8787



75-175
Var. 8446
Var. 8503
Var. 8560
Var. 8617
Var. 8674
Var. 8731
Var. 8788



75-150
Var. 8447
Var. 8504
Var. 8561
Var. 8618
Var. 8675
Var. 8732
Var. 8789



75-125
Var. 8448
Var. 8505
Var. 8562
Var. 8619
Var. 8676
Var. 8733
Var. 8790



75-100
Var. 8449
Var. 8506
Var. 8563
Var. 8620
Var. 8677
Var. 8734
Var. 8791



100-200 
Var. 8450
Var. 8507
Var. 8564
Var. 8621
Var. 8678
Var. 8735
Var. 8792



100-175 
Var. 8451
Var. 8508
Var. 8565
Var. 8622
Var. 8679
Var. 8736
Var. 8793



100-150 
Var. 8452
Var. 8509
Var. 8566
Var. 8623
Var. 8680
Var. 8737
Var. 8794



100-125 
Var. 8453
Var. 8510
Var. 8567
Var. 8624
Var. 8681
Var. 8738
Var. 8795



125-200 
Var. 8454
Var. 8511
Var. 8568
Var. 8625
Var. 8682
Var. 8739
Var. 8796



125-175 
Var. 8455
Var. 8512
Var. 8569
Var. 8626
Var. 8683
Var. 8740
Var. 8797



125-150 
Var. 8456
Var. 8513
Var. 8570
Var. 8627
Var. 8684
Var. 8741
Var. 8798



150-200 
Var. 8457
Var. 8514
Var. 8571
Var. 8628
Var. 8685
Var. 8742
Var. 8799



150-200 
Var. 8458
Var. 8515
Var. 8572
Var. 8629
Var. 8686
Var. 8743
Var. 8800



175-200 
Var. 8459
Var. 8516
Var. 8573
Var. 8630
Var. 8687
Var. 8744
Var. 8801





Var. = Variation













TABLE 30







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)















20-90
20-80
20-70
20-60
20-50
20-40
40-150


















Dosage
0.5-200 
Var. 8802
Var. 8859
Var. 8916
Var. 8973
Var. 9030
Var. 9087
Var. 9144


(IU/kg rVWF:RCo
0.5-150 
Var. 8803
Var. 8860
Var. 8917
Var. 8974
Var. 9031
Var. 9088
Var. 9145


activity)
0.5-100 
Var. 8804
Var. 8861
Var. 8918
Var. 8975
Var. 9032
Var. 9089
Var. 9146



0.5-75  
Var. 8805
Var. 8862
Var. 8919
Var. 8976
Var. 9033
Var. 9090
Var. 9147



0.5-50  
Var. 8806
Var. 8863
Var. 8920
Var. 8977
Var. 9034
Var. 9091
Var. 9148



0.5-25  
Var. 8807
Var. 8864
Var. 8921
Var. 8978
Var. 9035
Var. 9092
Var. 9149



0.5-10  
Var. 8808
Var. 8865
Var. 8922
Var. 8979
Var. 9036
Var. 9093
Var. 9150



0.5-5  
Var. 8809
Var. 8866
Var. 8923
Var. 8980
Var. 9037
Var. 9094
Var. 9151



0.5-2.5 
Var. 8810
Var. 8867
Var. 8924
Var. 8981
Var. 9038
Var. 9095
Var. 9152



0.5-1  
Var. 8811
Var. 8868
Var. 8925
Var. 8982
Var. 9039
Var. 9096
Var. 9153



2.5-200 
Var. 8812
Var. 8869
Var. 8926
Var. 8983
Var. 9040
Var. 9097
Var. 9154



2.5-150 
Var. 8813
Var. 8870
Var. 8927
Var. 8984
Var. 9041
Var. 9098
Var. 9155



2.5-100 
Var. 8814
Var. 8871
Var. 8928
Var. 8985
Var. 9042
Var. 9099
Var. 9156



2.5-75  
Var. 8815
Var. 8872
Var. 8929
Var. 8986
Var. 9043
Var. 9100
Var. 9157



2.5-50  
Var. 8816
Var. 8873
Var. 8930
Var. 8987
Var. 9044
Var. 9101
Var. 9158



2.5-25  
Var. 8817
Var. 8874
Var. 8931
Var. 8988
Var. 9045
Var. 9102
Var. 9159



2.5-10  
Var. 8818
Var. 8875
Var. 8932
Var. 8989
Var. 9046
Var. 9103
Var. 9160



2.5-5  
Var. 8819
Var. 8876
Var. 8933
Var. 8990
Var. 9047
Var. 9104
Var. 9161



 5-200
Var. 8820
Var. 8877
Var. 8934
Var. 8991
Var. 9048
Var. 9105
Var. 9162



 5-175
Var. 8821
Var. 8878
Var. 8935
Var. 8992
Var. 9049
Var. 9106
Var. 9163



 5-150
Var. 8822
Var. 8879
Var. 8936
Var. 8993
Var. 9050
Var. 9107
Var. 9164



 5-125
Var. 8823
Var. 8880
Var. 8937
Var. 8994
Var. 9051
Var. 9108
Var. 9165



 5-100
Var. 8824
Var. 8881
Var. 8938
Var. 8995
Var. 9052
Var. 9109
Var. 9166



5-75
Var. 8825
Var. 8882
Var. 8939
Var. 8996
Var. 9053
Var. 9110
Var. 9167



5-50
Var. 8826
Var. 8883
Var. 8940
Var. 8997
Var. 9054
Var. 9111
Var. 9168



5-25
Var. 8827
Var. 8884
Var. 8941
Var. 8998
Var. 9055
Var. 9112
Var. 9169



5-10
Var. 8828
Var. 8885
Var. 8942
Var. 8999
Var. 9056
Var. 9113
Var. 9170



10-200
Var. 8829
Var. 8886
Var. 8943
Var. 9000
Var. 9057
Var. 9114
Var. 9171



10-150
Var. 8830
Var. 8887
Var. 8944
Var. 9001
Var. 9058
Var. 9115
Var. 9172



10-100
Var. 8831
Var. 8888
Var. 8945
Var. 9002
Var. 9059
Var. 9116
Var. 9173



10-75 
Var. 8832
Var. 8889
Var. 8946
Var. 9003
Var. 9060
Var. 9117
Var. 9174



10-50 
Var. 8833
Var. 8890
Var. 8947
Var. 9004
Var. 9061
Var. 9118
Var. 9175



10-25 
Var. 8834
Var. 8891
Var. 8948
Var. 9005
Var. 9062
Var. 9119
Var. 9176



25-200
Var. 8835
Var. 8892
Var. 8949
Var. 9006
Var. 9063
Var. 9120
Var. 9177



25-150
Var. 8836
Var. 8893
Var. 8950
Var. 9007
Var. 9064
Var. 9121
Var. 9178



25-100
Var. 8837
Var. 8894
Var. 8951
Var. 9008
Var. 9065
Var. 9122
Var. 9179



25-75 
Var. 8838
Var. 8895
Var. 8952
Var. 9009
Var. 9066
Var. 9123
Var. 9180



25-50 
Var. 8839
Var. 8896
Var. 8953
Var. 9010
Var. 9067
Var. 9124
Var. 9181



50-200
Var. 8840
Var. 8897
Var. 8954
Var. 9011
Var. 9068
Var. 9125
Var. 9182



50-150
Var. 8841
Var. 8898
Var. 8955
Var. 9012
Var. 9069
Var. 9126
Var. 9183



50-100
Var. 8842
Var. 8899
Var. 8956
Var. 9013
Var. 9070
Var. 9127
Var. 9184



50-75 
Var. 8843
Var. 8900
Var. 8957
Var. 9014
Var. 9071
Var. 9128
Var. 9185



75-200
Var. 8844
Var. 8901
Var. 8958
Var. 9015
Var. 9072
Var. 9129
Var. 9186



75-175
Var. 8845
Var. 8902
Var. 8959
Var. 9016
Var. 9073
Var. 9130
Var. 9187



75-150
Var. 8846
Var. 8903
Var. 8960
Var. 9017
Var. 9074
Var. 9131
Var. 9188



75-125
Var. 8847
Var. 8904
Var. 8961
Var. 9018
Var. 9075
Var. 9132
Var. 9189



75-100
Var. 8848
Var. 8905
Var. 8962
Var. 9019
Var. 9076
Var. 9133
Var. 9190



100-200 
Var. 8849
Var. 8906
Var. 8963
Var. 9020
Var. 9077
Var. 9134
Var. 9191



100-175 
Var. 8850
Var. 8907
Var. 8964
Var. 9021
Var. 9078
Var. 9135
Var. 9192



100-150 
Var. 8851
Var. 8908
Var. 8965
Var. 9022
Var. 9079
Var. 9136
Var. 9193



100-125 
Var. 8852
Var. 8909
Var. 8966
Var. 9023
Var. 9080
Var. 9137
Var. 9194



125-200 
Var. 8853
Var. 8910
Var. 8967
Var. 9024
Var. 9081
Var. 9138
Var. 9195



125-175 
Var. 8854
Var. 8911
Var. 8968
Var. 9025
Var. 9082
Var. 9139
Var. 9196



125-150 
Var. 8855
Var. 8912
Var. 8969
Var. 9026
Var. 9083
Var. 9140
Var. 9197



150-200 
Var. 8856
Var. 8913
Var. 8970
Var. 9027
Var. 9084
Var. 9141
Var. 9198



150-200 
Var. 8857
Var. 8914
Var. 8971
Var. 9028
Var. 9085
Var. 9142
Var. 9199



175-200 
Var. 8858
Var. 8915
Var. 8972
Var. 9029
Var. 9086
Var. 9143
Var. 9200





Var. = Variation













TABLE 31







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)















40-125
40-100
40-90
40-80
40-70
40-60
40-50


















Dosage
0.5-200 
Var. 9201
Var. 9258
Var. 9315
Var. 9372
Var. 9429
Var. 9486
Var. 9543


(IU/kg rVWF:RCo
0.5-150 
Var. 9202
Var. 9259
Var. 9316
Var. 9373
Var. 9430
Var. 9487
Var. 9544


activity)
0.5-100 
Var. 9203
Var. 9260
Var. 9317
Var. 9374
Var. 9431
Var. 9488
Var. 9545



0.5-75  
Var. 9204
Var. 9261
Var. 9318
Var. 9375
Var. 9432
Var. 9489
Var. 9546



0.5-50  
Var. 9205
Var. 9262
Var. 9319
Var. 9376
Var. 9433
Var. 9490
Var. 9547



0.5-25  
Var. 9206
Var. 9263
Var. 9320
Var. 9377
Var. 9434
Var. 9491
Var. 9548



0.5-10  
Var. 9207
Var. 9264
Var. 9321
Var. 9378
Var. 9435
Var. 9492
Var. 9549



0.5-5  
Var. 9208
Var. 9265
Var. 9322
Var. 9379
Var. 9436
Var. 9493
Var. 9550



0.5-2.5 
Var. 9209
Var. 9266
Var. 9323
Var. 9380
Var. 9437
Var. 9494
Var. 9551



0.5-1  
Var. 9210
Var. 9267
Var. 9324
Var. 9381
Var. 9438
Var. 9495
Var. 9552



2.5-200 
Var. 9211
Var. 9268
Var. 9325
Var. 9382
Var. 9439
Var. 9496
Var. 9553



2.5-150 
Var. 9212
Var. 9269
Var. 9326
Var. 9383
Var. 9440
Var. 9497
Var. 9554



2.5-100 
Var. 9213
Var. 9270
Var. 9327
Var. 9384
Var. 9441
Var. 9498
Var. 9555



2.5-75  
Var. 9214
Var. 9271
Var. 9328
Var. 9385
Var. 9442
Var. 9499
Var. 9556



2.5-50  
Var. 9215
Var. 9272
Var. 9329
Var. 9386
Var. 9443
Var. 9500
Var. 9557



2.5-25  
Var. 9216
Var. 9273
Var. 9330
Var. 9387
Var. 9444
Var. 9501
Var. 9558



2.5-10  
Var. 9217
Var. 9274
Var. 9331
Var. 9388
Var. 9445
Var. 9502
Var. 9559



2.5-5  
Var. 9218
Var. 9275
Var. 9332
Var. 9389
Var. 9446
Var. 9503
Var. 9560



 5-200
Var. 9219
Var. 9276
Var. 9333
Var. 9390
Var. 9447
Var. 9504
Var. 9561



 5-175
Var. 9220
Var. 9277
Var. 9334
Var. 9391
Var. 9448
Var. 9505
Var. 9562



 5-150
Var. 9221
Var. 9278
Var. 9335
Var. 9392
Var. 9449
Var. 9506
Var. 9563



 5-125
Var. 9222
Var. 9279
Var. 9336
Var. 9393
Var. 9450
Var. 9507
Var. 9564



 5-100
Var. 9223
Var. 9280
Var. 9337
Var. 9394
Var. 9451
Var. 9508
Var. 9565



5-75
Var. 9224
Var. 9281
Var. 9338
Var. 9395
Var. 9452
Var. 9509
Var. 9566



5-50
Var. 9225
Var. 9282
Var. 9339
Var. 9396
Var. 9453
Var. 9510
Var. 9567



5-25
Var. 9226
Var. 9283
Var. 9340
Var. 9397
Var. 9454
Var. 9511
Var. 9568



5-10
Var. 9227
Var. 9284
Var. 9341
Var. 9398
Var. 9455
Var. 9512
Var. 9569



10-200
Var. 9228
Var. 9285
Var. 9342
Var. 9399
Var. 9456
Var. 9513
Var. 9570



10-150
Var. 9229
Var. 9286
Var. 9343
Var. 9400
Var. 9457
Var. 9514
Var. 9571



10-100
Var. 9230
Var. 9287
Var. 9344
Var. 9401
Var. 9458
Var. 9515
Var. 9572



10-75 
Var. 9231
Var. 9288
Var. 9345
Var. 9402
Var. 9459
Var. 9516
Var. 9573



10-50 
Var. 9232
Var. 9289
Var. 9346
Var. 9403
Var. 9460
Var. 9517
Var. 9574



10-25 
Var. 9233
Var. 9290
Var. 9347
Var. 9404
Var. 9461
Var. 9518
Var. 9575



25-200
Var. 9234
Var. 9291
Var. 9348
Var. 9405
Var. 9462
Var. 9519
Var. 9576



25-150
Var. 9235
Var. 9292
Var. 9349
Var. 9406
Var. 9463
Var. 9520
Var. 9577



25-100
Var. 9236
Var. 9293
Var. 9350
Var. 9407
Var. 9464
Var. 9521
Var. 9578



25-75 
Var. 9237
Var. 9294
Var. 9351
Var. 9408
Var. 9465
Var. 9522
Var. 9579



25-50 
Var. 9238
Var. 9295
Var. 9352
Var. 9409
Var. 9466
Var. 9523
Var. 9580



50-200
Var. 9239
Var. 9296
Var. 9353
Var. 9410
Var. 9467
Var. 9524
Var. 9581



50-150
Var. 9240
Var. 9297
Var. 9354
Var. 9411
Var. 9468
Var. 9525
Var. 9582



50-100
Var. 9241
Var. 9298
Var. 9355
Var. 9412
Var. 9469
Var. 9526
Var. 9583



50-75 
Var. 9242
Var. 9299
Var. 9356
Var. 9413
Var. 9470
Var. 9527
Var. 9584



75-200
Var. 9243
Var. 9300
Var. 9357
Var. 9414
Var. 9471
Var. 9528
Var. 9585



75-175
Var. 9244
Var. 9301
Var. 9358
Var. 9415
Var. 9472
Var. 9529
Var. 9586



75-150
Var. 9245
Var. 9302
Var. 9359
Var. 9416
Var. 9473
Var. 9530
Var. 9587



75-125
Var. 9246
Var. 9303
Var. 9360
Var. 9417
Var. 9474
Var. 9531
Var. 9588



75-100
Var. 9247
Var. 9304
Var. 9361
Var. 9418
Var. 9475
Var. 9532
Var. 9589



100-200 
Var. 9248
Var. 9305
Var. 9362
Var. 9419
Var. 9476
Var. 9533
Var. 9590



100-175 
Var. 9249
Var. 9306
Var. 9363
Var. 9420
Var. 9477
Var. 9534
Var. 9591



100-150 
Var. 9250
Var. 9307
Var. 9364
Var. 9421
Var. 9478
Var. 9535
Var. 9592



100-125 
Var. 9251
Var. 9308
Var. 9365
Var. 9422
Var. 9479
Var. 9536
Var. 9593



125-200 
Var. 9252
Var. 9309
Var. 9366
Var. 9423
Var. 9480
Var. 9537
Var. 9594



125-175 
Var. 9253
Var. 9310
Var. 9367
Var. 9424
Var. 9481
Var. 9538
Var. 9595



125-150 
Var. 9254
Var. 9311
Var. 9368
Var. 9425
Var. 9482
Var. 9539
Var. 9596



150-200 
Var. 9255
Var. 9312
Var. 9369
Var. 9426
Var. 9483
Var. 9540
Var. 9597



150-200 
Var. 9256
Var. 9313
Var. 9370
Var. 9427
Var. 9484
Var. 9541
Var. 9598



175-200 
Var. 9257
Var. 9314
Var. 9371
Var. 9428
Var. 9485
Var. 9542
Var. 9599





Var. = Variation













TABLE 32







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)














60-150
60-125
60-100
60-90
60-80
60-70

















Dosage
0.5-200 
Var. 9600
Var. 9657
Var. 9714
Var. 9771
Var. 9828
Var. 9885


(IU/kg rVWF:RCo
0.5-150 
Var. 9601
Var. 9658
Var. 9715
Var. 9772
Var. 9829
Var. 9886


activity)
0.5-100 
Var. 9602
Var. 9659
Var. 9716
Var. 9773
Var. 9830
Var. 9887



0.5-75  
Var. 9603
Var. 9660
Var. 9717
Var. 9774
Var. 9831
Var. 9888



0.5-50  
Var. 9604
Var. 9661
Var. 9718
Var. 9775
Var. 9832
Var. 9889



0.5-25  
Var. 9605
Var. 9662
Var. 9719
Var. 9776
Var. 9833
Var. 9890



0.5-10  
Var. 9606
Var. 9663
Var. 9720
Var. 9777
Var. 9834
Var. 9891



0.5-5  
Var. 9607
Var. 9664
Var. 9721
Var. 9778
Var. 9835
Var. 9892



0.5-2.5 
Var. 9608
Var. 9665
Var. 9722
Var. 9779
Var. 9836
Var. 9893



0.5-1  
Var. 9609
Var. 9666
Var. 9723
Var. 9780
Var. 9837
Var. 9894



2.5-200 
Var. 9610
Var. 9667
Var. 9724
Var. 9781
Var. 9838
Var. 9895



2.5-150 
Var. 9611
Var. 9668
Var. 9725
Var. 9782
Var. 9839
Var. 9896



2.5-100 
Var. 9612
Var. 9669
Var. 9726
Var. 9783
Var. 9840
Var. 9897



2.5-75  
Var. 9613
Var. 9670
Var. 9727
Var. 9784
Var. 9841
Var. 9898



2.5-50  
Var. 9614
Var. 9671
Var. 9728
Var. 9785
Var. 9842
Var. 9899



2.5-25  
Var. 9615
Var. 9672
Var. 9729
Var. 9786
Var. 9843
Var. 9900



2.5-10  
Var. 9616
Var. 9673
Var. 9730
Var. 9787
Var. 9844
Var. 9901



2.5-5  
Var. 9617
Var. 9674
Var. 9731
Var. 9788
Var. 9845
Var. 9902



 5-200
Var. 9618
Var. 9675
Var. 9732
Var. 9789
Var. 9846
Var. 9903



 5-175
Var. 9619
Var. 9676
Var. 9733
Var. 9790
Var. 9847
Var. 9904



 5-150
Var. 9620
Var. 9677
Var. 9734
Var. 9791
Var. 9848
Var. 9905



 5-125
Var. 9621
Var. 9678
Var. 9735
Var. 9792
Var. 9849
Var. 9906



 5-100
Var. 9622
Var. 9679
Var. 9736
Var. 9793
Var. 9850
Var. 9907



5-75
Var. 9623
Var. 9680
Var. 9737
Var. 9794
Var. 9851
Var. 9908



5-50
Var. 9624
Var. 9681
Var. 9738
Var. 9795
Var. 9852
Var. 9909



5-25
Var. 9625
Var. 9682
Var. 9739
Var. 9796
Var. 9853
Var. 9910



5-10
Var. 9626
Var. 9683
Var. 9740
Var. 9797
Var. 9854
Var. 9911



10-200
Var. 9627
Var. 9684
Var. 9741
Var. 9798
Var. 9855
Var. 9912



10-150
Var. 9628
Var. 9685
Var. 9742
Var. 9799
Var. 9856
Var. 9913



10-100
Var. 9629
Var. 9686
Var. 9743
Var. 9800
Var. 9857
Var. 9914



10-75 
Var. 9630
Var. 9687
Var. 9744
Var. 9801
Var. 9858
Var. 9915



10-50 
Var. 9631
Var. 9688
Var. 9745
Var. 9802
Var. 9859
Var. 9916



10-25 
Var. 9632
Var. 9689
Var. 9746
Var. 9803
Var. 9860
Var. 9917



25-200
Var. 9633
Var. 9690
Var. 9747
Var. 9804
Var. 9861
Var. 9918



25-150
Var. 9634
Var. 9691
Var. 9748
Var. 9805
Var. 9862
Var. 9919



25-100
Var. 9635
Var. 9692
Var. 9749
Var. 9806
Var. 9863
Var. 9920



25-75 
Var. 9636
Var. 9693
Var. 9750
Var. 9807
Var. 9864
Var. 9921



25-50 
Var. 9637
Var. 9694
Var. 9751
Var. 9808
Var. 9865
Var. 9922



50-200
Var. 9638
Var. 9695
Var. 9752
Var. 9809
Var. 9866
Var. 9923



50-150
Var. 9639
Var. 9696
Var. 9753
Var. 9810
Var. 9867
Var. 9924



50-100
Var. 9640
Var. 9697
Var. 9754
Var. 9811
Var. 9868
Var. 9925



50-75 
Var. 9641
Var. 9698
Var. 9755
Var. 9812
Var. 9869
Var. 9926



75-200
Var. 9642
Var. 9699
Var. 9756
Var. 9813
Var. 9870
Var. 9927



75-175
Var. 9643
Var. 9700
Var. 9757
Var. 9814
Var. 9871
Var. 9928



75-150
Var. 9644
Var. 9701
Var. 9758
Var. 9815
Var. 9872
Var. 9929



75-125
Var. 9645
Var. 9702
Var. 9759
Var. 9816
Var. 9873
Var. 9930



75-100
Var. 9646
Var. 9703
Var. 9760
Var. 9817
Var. 9874
Var. 9931



100-200 
Var. 9647
Var. 9704
Var. 9761
Var. 9818
Var. 9875
Var. 9932



100-175 
Var. 9648
Var. 9705
Var. 9762
Var. 9819
Var. 9876
Var. 9933



100-150 
Var. 9649
Var. 9706
Var. 9763
Var. 9820
Var. 9877
Var. 9934



100-125 
Var. 9650
Var. 9707
Var. 9764
Var. 9821
Var. 9878
Var. 9935



125-200 
Var. 9651
Var. 9708
Var. 9765
Var. 9822
Var. 9879
Var. 9936



125-175 
Var. 9652
Var. 9709
Var. 9766
Var. 9823
Var. 9880
Var. 9937



125-150 
Var. 9653
Var. 9710
Var. 9767
Var. 9824
Var. 9881
Var. 9938



150-200 
Var. 9654
Var. 9711
Var. 9768
Var. 9825
Var. 9882
Var. 9939



150-200 
Var. 9655
Var. 9712
Var. 9769
Var. 9826
Var. 9883
Var. 9940



175-200 
Var. 9656
Var. 9713
Var. 9770
Var. 9827
Var. 9884
Var. 9941





Var. = Variation













TABLE 33







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)














70-150
70-125
70-100
70-90
70-80
80-150

















Dosage
0.5-200 
Var. 9942
Var. 9999
Var. 10056
Var. 10113
Var. 10170
Var. 10227


(IU/kg
0.5-150 
Var. 9943
Var. 10000
Var. 10057
Var. 10114
Var. 10171
Var. 10228


rVWF:
0.5-100 
Var. 9944
Var. 10001
Var. 10058
Var. 10115
Var. 10172
Var. 10229


RCo
0.5-75  
Var. 9945
Var. 10002
Var. 10059
Var. 10116
Var. 10173
Var. 10230


activity
0.5-50  
Var. 9946
Var. 10003
Var. 10060
Var. 10117
Var. 10174
Var. 10231



0.5-25  
Var. 9947
Var. 10004
Var. 10061
Var. 10118
Var. 10175
Var. 10232



0.5-10  
Var. 9948
Var. 10005
Var. 10062
Var. 10119
Var. 10176
Var. 10233



0.5-5  
Var. 9949
Var. 10006
Var. 10063
Var. 10120
Var. 10177
Var. 10234



0.5-2.5 
Var. 9950
Var. 10007
Var. 10064
Var. 10121
Var. 10178
Var. 10235



0.5-1  
Var. 9951
Var. 10008
Var. 10065
Var. 10122
Var. 10179
Var. 10236



2.5-200 
Var. 9952
Var. 10009
Var. 10066
Var. 10123
Var. 10180
Var. 10237



2.5-150 
Var. 9953
Var. 10010
Var. 10067
Var. 10124
Var. 10181
Var. 10238



2.5-100 
Var. 9954
Var. 10011
Var. 10068
Var. 10125
Var. 10182
Var. 10239



2.5-75  
Var. 9955
Var. 10012
Var. 10069
Var. 10126
Var. 10183
Var. 10240



2.5-50  
Var. 9956
Var. 10013
Var. 10070
Var. 10127
Var. 10184
Var. 10241



2.5-25  
Var. 9957
Var. 10014
Var. 10071
Var. 10128
Var. 10185
Var. 10242



2.5-10  
Var. 9958
Var. 10015
Var. 10072
Var. 10129
Var. 10186
Var. 10243



2.5-5  
Var. 9959
Var. 10016
Var. 10073
Var. 10130
Var. 10187
Var. 10244



 5-200
Var. 9960
Var. 10017
Var. 10074
Var. 10131
Var. 10188
Var. 10245



 5-175
Var. 9961
Var. 10018
Var. 10075
Var. 10132
Var. 10189
Var. 10246



 5-150
Var. 9962
Var. 10019
Var. 10076
Var. 10133
Var. 10190
Var. 10247



 5-125
Var. 9963
Var. 10020
Var. 10077
Var. 10134
Var. 10191
Var. 10248



 5-100
Var. 9964
Var. 10021
Var. 10078
Var. 10135
Var. 10192
Var. 10249



5-75
Var. 9965
Var. 10022
Var. 10079
Var. 10136
Var. 10193
Var. 10250



5-50
Var. 9966
Var. 10023
Var. 10080
Var. 10137
Var. 10194
Var. 10251



5-25
Var. 9967
Var. 10024
Var. 10081
Var. 10138
Var. 10195
Var. 10252



5-10
Var. 9968
Var. 10025
Var. 10082
Var. 10139
Var. 10196
Var. 10253



10-200
Var. 9969
Var. 10026
Var. 10083
Var. 10140
Var. 10197
Var. 10254



10-150
Var. 9970
Var. 10027
Var. 10084
Var. 10141
Var. 10198
Var. 10255



10-100
Var. 9971
Var. 10028
Var. 10085
Var. 10142
Var. 10199
Var. 10256



10-75 
Var. 9972
Var. 10029
Var. 10086
Var. 10143
Var. 10200
Var. 10257



10-50 
Var. 9973
Var. 10030
Var. 10087
Var. 10144
Var. 10201
Var. 10258



10-25 
Var. 9974
Var. 10031
Var. 10088
Var. 10145
Var. 10202
Var. 10259



25-200
Var. 9975
Var. 10032
Var. 10089
Var. 10146
Var. 10203
Var. 10260



25-150
Var. 9976
Var. 10033
Var. 10090
Var. 10147
Var. 10204
Var. 10261



25-100
Var. 9977
Var. 10034
Var. 10091
Var. 10148
Var. 10205
Var. 10262



25-75 
Var. 9978
Var. 10035
Var. 10092
Var. 10149
Var. 10206
Var. 10263



25-50 
Var. 9979
Var. 10036
Var. 10093
Var. 10150
Var. 10207
Var. 10264



50-200
Var. 9980
Var. 10037
Var. 10094
Var. 10151
Var. 10208
Var. 10265



50-150
Var. 9981
Var. 10038
Var. 10095
Var. 10152
Var. 10209
Var. 10266



50-100
Var. 9982
Var. 10039
Var. 10096
Var. 10153
Var. 10210
Var. 10267



50-75 
Var. 9983
Var. 10040
Var. 10097
Var. 10154
Var. 10211
Var. 10268



75-200
Var. 9984
Var. 10041
Var. 10098
Var. 10155
Var. 10212
Var. 10269



75-175
Var. 9985
Var. 10042
Var. 10099
Var. 10156
Var. 10213
Var. 10270



75-150
Var. 9986
Var. 10043
Var. 10100
Var. 10157
Var. 10214
Var. 10271



75-125
Var. 9987
Var. 10044
Var. 10101
Var. 10158
Var. 10215
Var. 10272



75-100
Var. 9988
Var. 10045
Var. 10102
Var. 10159
Var. 10216
Var. 10273



100-200 
Var. 9989
Var. 10046
Var. 10103
Var. 10160
Var. 10217
Var. 10274



100-175 
Var. 9990
Var. 10047
Var. 10104
Var. 10161
Var. 10218
Var. 10275



100-150 
Var. 9991
Var. 10048
Var. 10105
Var. 10162
Var. 10219
Var. 10276



100-125 
Var. 9992
Var. 10049
Var. 10106
Var. 10163
Var. 10220
Var. 10277



125-200 
Var. 9993
Var. 10050
Var. 10107
Var. 10164
Var. 10221
Var. 10278



125-175 
Var. 9994
Var. 10051
Var. 10108
Var. 10165
Var. 10222
Var. 10279



125-150 
Var. 9995
Var. 10052
Var. 10109
Var. 10166
Var. 10223
Var. 10280



150-200 
Var. 9996
Var. 10053
Var. 10110
Var. 10167
Var. 10224
Var. 10281



150-200 
Var. 9997
Var. 10054
Var. 10111
Var. 10168
Var. 10225
Var. 10282



175-200 
Var. 9998
Var. 10055
Var. 10112
Var. 10169
Var. 10226
Var. 10283





Var. = Variation













TABLE 34







Exemplary embodiments for the combination of rVWF dosage and


rVWF specific activity useful in the methods described herein.









Specific Activity (mU/μg)














80-125
80-100
80-90
90-150
90-125
90-100

















Dosage
0.5-200 
Var. 10284
Var. 10341
Var. 10398
Var. 10455
Var. 10512
Var. 10569


(IU/kg rVWF:RCo
0.5-150 
Var. 10285
Var. 10342
Var. 10399
Var. 10456
Var. 10513
Var. 10570


activity)
0.5-100 
Var. 10286
Var. 10343
Var. 10400
Var. 10457
Var. 10514
Var. 10571



0.5-75  
Var. 10287
Var. 10344
Var. 10401
Var. 10458
Var. 10515
Var. 10572



0.5-50  
Var. 10288
Var. 10345
Var. 10402
Var. 10459
Var. 10516
Var. 10573



0.5-25  
Var. 10289
Var. 10346
Var. 10403
Var. 10460
Var. 10517
Var. 10574



0.5-10  
Var. 10290
Var. 10347
Var. 10404
Var. 10461
Var. 10518
Var. 10575



0.5-5  
Var. 10291
Var. 10348
Var. 10405
Var. 10462
Var. 10519
Var. 10576



0.5-2.5 
Var. 10292
Var. 10349
Var. 10406
Var. 10463
Var. 10520
Var. 10577



0.5-1  
Var. 10293
Var. 10350
Var. 10407
Var. 10464
Var. 10521
Var. 10578



2.5-200 
Var. 10294
Var. 10351
Var. 10408
Var. 10465
Var. 10522
Var. 10579



2.5-150 
Var. 10295
Var. 10352
Var. 10409
Var. 10466
Var. 10523
Var. 10580



2.5-100 
Var. 10296
Var. 10353
Var. 10410
Var. 10467
Var. 10524
Var. 10581



2.5-75  
Var. 10297
Var. 10354
Var. 10411
Var. 10468
Var. 10525
Var. 10582



2.5-50  
Var. 10298
Var. 10355
Var. 10412
Var. 10469
Var. 10526
Var. 10583



2.5-25  
Var. 10299
Var. 10356
Var. 10413
Var. 10470
Var. 10527
Var. 10584



2.5-10  
Var. 10300
Var. 10357
Var. 10414
Var. 10471
Var. 10528
Var. 10585



23-5
Var. 10301
Var. 10358
Var. 10415
Var. 10472
Var. 10529
Var. 10586



 5-200
Var. 10302
Var. 10359
Var. 10416
Var. 10473
Var. 10530
Var. 10587



 5-175
Var. 10303
Var. 10360
Var. 10417
Var. 10474
Var. 10531
Var. 10588



 5-150
Var. 10304
Var. 10361
Var. 10418
Var. 10475
Var. 10532
Var. 10589



 5-125
Var. 10305
Var. 10362
Var. 10419
Var. 10476
Var. 10533
Var. 10590



 5-100
Var. 10306
Var. 10363
Var. 10420
Var. 10477
Var. 10534
Var. 10591



5-75
Var. 10307
Var. 10364
Var. 10421
Var. 10478
Var. 10535
Var. 10592



5-50
Var. 10308
Var. 10365
Var. 10422
Var. 10479
Var. 10536
Var. 10593



5-25
Var. 10309
Var. 10366
Var. 10423
Var. 10480
Var. 10537
Var. 10594



5-10
Var. 10310
Var. 10367
Var. 10424
Var. 10481
Var. 10538
Var. 10595



10-200
Var. 10311
Var. 10368
Var. 10425
Var. 10482
Var. 10539
Var. 10596



10-150
Var. 10312
Var. 10369
Var. 10426
Var. 10483
Var. 10540
Var. 10597



10-100
Var. 10313
Var. 10370
Var. 10427
Var. 10484
Var. 10541
Var. 10598



10-75 
Var. 10314
Var. 10371
Var. 10428
Var. 10485
Var. 10542
Var. 10599



10-50 
Var. 10315
Var. 10372
Var. 10429
Var. 10486
Var. 10543
Var. 10600



10-25 
Var. 10316
Var. 10373
Var. 10430
Var. 10487
Var. 10544
Var. 10601



25-200
Var. 10317
Var. 10374
Var. 10431
Var. 10488
Var. 10545
Var. 10602



25-150
Var. 10318
Var. 10375
Var. 10432
Var. 10489
Var. 10546
Var. 10603



25-100
Var. 10319
Var. 10376
Var. 10433
Var. 10490
Var. 10547
Var. 10604



25-75 
Var. 10320
Var. 10377
Var. 10434
Var. 10491
Var. 10548
Var. 10605



25-50 
Var. 10321
Var. 10378
Var. 10435
Var. 10492
Var. 10549
Var. 10606



50-200
Var. 10322
Var. 10379
Var. 10436
Var. 10493
Var. 10550
Var. 10607



50-150
Var. 10323
Var. 10380
Var. 10437
Var. 10494
Var. 10551
Var. 10608



50-100
Var. 10324
Var. 10381
Var. 10438
Var. 10495
Var. 10552
Var. 10609



50-75 
Var. 10325
Var. 10382
Var. 10439
Var. 10496
Var. 10553
Var. 10610



75-200
Var. 10326
Var. 10383
Var. 10440
Var. 10497
Var. 10554
Var. 10611



75-175
Var. 10327
Var. 10384
Var. 10441
Var. 10498
Var. 10555
Var. 10612



75-150
Var. 10328
Var. 10385
Var. 10442
Var. 10499
Var. 10556
Var. 10613



75-125
Var. 10329
Var. 10386
Var. 10443
Var. 10500
Var. 10557
Var. 10614



75-100
Var. 10330
Var. 10387
Var. 10444
Var. 10501
Var. 10558
Var. 10615



100-200 
Var. 10331
Var. 10388
Var. 10445
Var. 10502
Var. 10559
Var. 10616



100-175 
Var. 10332
Var. 10389
Var. 10446
Var. 10503
Var. 10560
Var. 10617



100-150 
Var. 10333
Var. 10390
Var. 10447
Var. 10504
Var. 10561
Var. 10618



100-125 
Var. 10334
Var. 10391
Var. 10448
Var. 10505
Var. 10562
Var. 10619



125-200 
Var. 10335
Var. 10392
Var. 10449
Var. 10506
Var. 10563
Var. 10620



125-175 
Var. 10336
Var. 10393
Var. 10450
Var. 10507
Var. 10564
Var. 10621



125-150 
Var. 10337
Var. 10394
Var. 10451
Var. 10508
Var. 10565
Var. 10622



150-200 
Var. 10338
Var. 10395
Var. 10452
Var. 10509
Var. 10566
Var. 10623



150-200 
Var. 10339
Var. 10396
Var. 10453
Var. 10510
Var. 10567
Var. 10624



175-200 
Var. 10340
Var. 10397
Var. 10454
Var. 10511
Var. 10568
Var. 10625





Var. = Variation






In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein the composition of rVWF administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein FVIII in the subject is stabilized for at least 18 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein FVIII in the subject is stabilized for at least 24 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF, wherein FVIII in the subject is stabilized for at least 30 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF administered to the subject has a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
Administration of rVWF/rFVIII
In one aspect, the present disclosure provides method for treating Von Willebrand Disease (VWD) or Hemophilia A in a subject in need thereof, which includes administering a composition of recombinant Von Willebrand Factor (rVWF) and recombinant FVIII (rFVIII) such that Factor VIII (FVIII) stability is increased, as compared to FVIII half-life in a subject administered a composition of plasma derived Von Willebrand Factor (pdVWF). In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers with a higher VWF specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII such that FVIII stability is extended by at least 10%, 20%, 30%, 2 hr, 4 hr, 6 hr, or by an amount selected from variations 1300 to 1643 found in Table 9, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In yet another embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII such that FVIII stability is extended by at least 10% as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII such that FVIII stability is extended by at least 20% as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII such that FVIII stability is extended by at least 30% as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF/rFVIII composition containing from 10 IU/kg to 40 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 20 IU/kg to 30 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF/rFVIII composition containing from 25 IU/kg to 75 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 40 IU/kg to 60 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a dosage of a rVWF/rFVIII composition containing from 75 IU/kg to 125 IU/kg rVWF:RCo activity, wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the composition contains from 75 IU/kg to 100 IU/kg rVWF:RCo activity. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the subject is administered a dose of rVWF selected from variations 2141 to 2338 in Table 12, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF. In yet another embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers with a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment. FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the rVWF in the composition has a specific activity of from 40 mU/μg to 60 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the rVWF in the composition has a specific activity of at least 60 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the rVWF in the composition has a specific activity of at least 80 mU/μg, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the rVWF in the composition has a specific activity selected from variations 1 to 133 found in Table 1, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In yet another embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers with a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 30% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 50% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers in which at least 70% of rVWF molecules in the composition are present in a multimer of at least 10 subunits, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein FVIII in the subject is stabilized for at least 18 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein FVIII in the subject is stabilized for at least 24 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein FVIII in the subject is stabilized for at least 30 hours post-administration, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 4:1-3:2, and wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:1-3:2. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment. FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:1-1:2, and wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:2-2:3. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:6, and wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is selected from variations 1988 to 2140 found in Table 11, and wherein the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers having a minimal percentage of rVWF molecules present in a particular higher-order rVWF multimer or larger multimer according to any one of variations 134 to 457 found in Table 3 to Table 5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is selected from variations 1988 to 2140 found in Table 11. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 4:1-3:2, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:1-3:2. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:1-1:2, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:2-2:3. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:6, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:5. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is selected from variations 1988 to 2140 found in Table 11, and wherein the combination of rVWF specific activity in the composition and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 2339 to 4868 in Table 13 to Table 19. In one embodiment, the composition of rVWF/rFVIII administered to the subject is a composition of high molecular weight rVWF multimers. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 4:1-3:2, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:1-3:2. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:1-1:2, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:2-2:3. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:6, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is selected from variations 1988 to 2140 found in Table 11, and wherein the dose of rVWF and increase in FVIII stability, as compared to FVIII stability in a subject administered a composition of pdVWF/FVIII, is selected from variations 4869 to 8003 in Table 20 to Table 27. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 4:1-3:2, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:1-3:2. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment. FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:1-1:2, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 3:2-2:3. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:6, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In a specific embodiment, the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is from 2:3-1:5. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
In one embodiment, the method comprises administering a composition of rVWF/rFVIII, wherein the ratio of rFVIII procoagulant activity (IU rFVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) in the composition is selected from variations 1988 to 2140 found in Table 11, and wherein the dose of rVWF and specific activity of rVWF in the composition is selected from variations 8004 to 10625 in Table 28 to Table 34. In one embodiment, the composition of rVWF/rFVIII administered to the subject has a higher VWF specific activity than a composition of pdVWF/FVIII. In one embodiment, FVIII stability is characterized by the half life of FVIII. In another embodiment, FVIII stability is characterized by mean residence time (MRT) of FVIII. In a further embodiment, the method is for treating any type of VWD. In a specific embodiment, the method is for treating Type 3 VWD.
The practice of the present invention may employ, unless otherwise indicated, conventional techniques and descriptions of organic chemistry, polymer technology, molecular biology (including recombinant techniques), cell biology, biochemistry, and immunology, which are within the skill of the art. Such conventional techniques include polymer array synthesis, hybridization, ligation, and detection of hybridization using a label. Specific illustrations of suitable techniques can be had by reference to the example herein below. However, other equivalent conventional procedures can, of course, also be used. Such conventional techniques and descriptions can be found in standard laboratory manuals such as Genome Analysis: A Laboratory Manual Series (Vols. I-IV), Using Antibodies: A Laboratory Manual, Cells: A Laboratory Manual, PCR Primer: A Laboratory Manual, and Molecular Cloning: A Laboratory Manual (all from Cold Spring Harbor Laboratory Press), Stryer, L. (1995) Biochemistry (4th Ed.) Freeman, Highly stabilized York, Gait, “Oligonucleotide Synthesis: A Practical Approach” 1984, IRL Press, London, Nelson and Cox (2000), Lehninger, Principles of Biochemistry 3rd Ed., W. H. Freeman Pub., Highly stabilized York, N.Y. and Berg et al. (2002) Biochemistry, 5th Ed., W. H. Freeman Pub., Highly stabilized York, N.Y., all of which are herein incorporated in their entirety by reference for all purposes.
Note that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a polymerase” refers to one agent or mixtures of such agents, and reference to “the method” includes reference to equivalent steps and methods known to those skilled in the art, and so forth.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications mentioned herein are incorporated herein by reference for the purpose of describing and disclosing devices, compositions, formulations and methodologies which are described in the publication and which might be used in connection with the presently described invention.
Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention.
In the above description, numerous specific details are set forth to provide a more thorough understanding of the present invention. However, it will be apparent to one of skill in the art that the present invention may be practiced without one or more of these specific details. In other instances, well-known features and procedures well known to those skilled in the art have not been described in order to avoid obscuring the invention.
Although the present invention is described primarily with reference to specific embodiments, it is also envisioned that other embodiments will become apparent to those skilled in the art upon reading the present disclosure, and it is intended that such embodiments be contained within the present inventive methods.
EXAMPLES
Example 1
Study of rVWF:rFVIII Co-Administration.
The immediate tolerability and safety after single doses of rVWF:rFVIII at 2 IU/kg, 7.5 IU/kg, 20 IU/kg and 50 IU/kg VWF:RCo was assessed as a primary endpoint of the study. Secondary endpoints included PK for VWF:RCo. VWF:CB, VWF:Ag, FVIII and multimeric composition of the VWF. An additional secondary endpoint was a PK comparison with pdVWF/pdFVIII [Cohort 4 (50 IU/kg VWF:RCo)]. See FIG. 1 for a schematic illustration of the study design.
Recombinant human Von Willebrand Factor (rVWF) was expressed in CHO cells. Propeptide removal was mediated in vitro through exposure of the pro-VWF to recombinant Furin. Fully glycosylated/ABO blood groups glycans were absent. The recombinant VWF has higher specific activity than plasma-derived VWF (pdVWF) and offers the therapeutic flexibility of dosing with or without rFVIII. The rVWF used in this study was not exposed to ADAMTS13, resulting in the presence of ultra-large VWF multimers and intact VWF subunits. ADAMTS13 results in subunit cleavage at TYR1605-MET1606.
VWF:RCo PK/VWF multimer kinetics showed highly similar PK for VWF:RCo (VWF activity) between rVWF and Humate P (FIG. 2A). Humate P is human derived medium purity Factor VIII concentrates complexed to VWF. A surrogate marker was used for efficacy and dosing recommendations. The data in FIG. 2A show that rVWF shows similar activity to that of plasma derived VW. Progressive loss of high molecular weight rVWF was seen upon exposure to ADAMTS13 (FIG. 2B), showing that rVWF is present in high molecular weight multimers prior to the ADAMTS13 exposure.
Higher FVIII levels were observed in rVWF patients as compared to Humate (FIG. 3). These data show that rVWF stabilizes endogenous FVIII in vivo. There was a difference in effects seen with different VWF:FVIII ratios (1.3:1 vs.  ̃2.1 VWF/FVIII). These different ratios suggest that less rVWF can be used to stabilize FVIII than is needed when using plasma derived VWF. The study design provided a flexibility of re-dosing with rVWF alone (no rFVIII) after the initial dose.
Ultra-large molecular weight multimers are present with rVWF—these ultra-large molecular weight multimers rapidly disappear following infusion. ADAMTS13 mediated cleavage fragments were seen in all subjects. The pharmacokinetic profile of VWF:RCo was similar to that of pdVWF. There was sustained stabilization of endogenous FVIII with rVWF which was comparable to that observed with pdVWF. The adverse drug reaction profile with rVWF was similar to that of pdVWF/pdFVIII, and all related adverse events (AEs) were mild.
The overall safety and pharmacokinetic profile suggests that rVWF can be used for the treatment and preventing of bleeding episodes at doses comparable to pdVWF/pdFVIII, while enhancing the levels of FVIII as compared to the effect from the use of pdVWF. Without being limited by theory, it is possible that the high proportion of ultra-large multimers (decamers or higher) in the population of rVWF leads to the enhanced stabilization of FVIII seen with rVWF as compared to that by plasma derived VWF.
Example 2
In-Human Study Evaluating Pharmacokinetics Demonstrating Safety and Tolerability in Severe Von Willebrand Disease (VWD).
This study compared the effects of pdVWF and rVWF in patients with type 3 VWD or severe Type 1 VWD.
The pdVWF used in this study was synthesized in endothelial cells and megakaryocytes. Post-translational modification of propeptide removal occurred intracellularly during passage of the protein to the Golgi and post-Golgi compartments. Glycosylation/ABO blood group glycans were present. The pdVWF consisted of VWF subunits that had been exposed to plasma ADAMTS13. There were no ultra-large VWF multimers in the pdVWF population and subunits were cleaved at TYR1605-MET1606. pdVWF concentrates contained other proteins, including ADAMTS13 and hemagglutins.
The rVWF used in this study was expressed in CHO cells. Propeptide removal was mediated in vitro through exposure of the pro-VWF to recombinant Furin. Fully glycosylated/ABO blood group glycans were absent. The rVWF was not exposed to ADAMTS13. The subunits of the rVWF were intact and ultra-large VWF multimers were present. Subunit cleavage did occur upon ADAMTS13 exposure. The rVWF showed higher specific activity than pdVWF.
The inclusion criteria for this study were:
Type 3 VWD (VWF:Ag≦3 IU/dL)
Severe Type 1 VWD (VWF:RCo≦10 IU/dL and FVIII:C<20 IU/dL
18 to 60 years of age
Previous coagulation factor replacement therapy (≧25 ED)
Non-bleeding state
The exclusion criteria for this study were:
Other coagulation disorders
History of VWF and/or FVIII inhibitors
Cardiovascular disease
Medical history of thromboembolic event
Medical history of other immunological disorders
The immediate tolerability and safety after single doses of rVWF:rFVIII at 2 IU/kg, 7.5 IU/kg, 20 IU/kg and 50 IU/kg VWF:RCo was assessed as a primary endpoint of this study. Secondary endpoints included PK for VWF:RCo, VWF:CB, VWF:Ag, FVIII and multimeric composition of the VWF. An additional secondary endpoint was a PK comparison with pdVWF/pdFVIII [Cohort 4 (50 IU/kg VWF:RCo)]. Patient demographics for this study are shown in FIG. 4. Pharmacokinetic analysis of Cohort 4A (Type 3 VWD) VWF:RCo/VWF:Ag is shown in FIG. 5. These data show that the total activities of rVWF were comparable to those of pdVWF. Progressive loss of high molecular weight rVWF was seen upon exposure to ADAMTS13 (FIG. 6). Further pharmacokinetic analysis of Cohort 4A (Type 3 VWD) FVIII:C is shown in FIG. 7. The data in FIG. 7 suggests that rVWF was more effective at stabilizing the in vivo activity of FVIII than pdVWF, resulting in increased FVIII procoagulant activity after treatment with rVWF/rVWF as compared to treatment with pdVWF/pdFVIII.
Ultra-large molecular weight multimers were present in rVWF and rapidly disappeared following infusion. ADAMTS13 mediated cleavage fragments were seen in all subjects. The pharmacokinetic profile of VWF:RCo was similar to that of pdVWF. There was sustained stabilization of endogenous FVIII that was comparable to that observed with pdVWF. The adverse drug reaction profile was similar to that of pdVWF/pdFVIII, and all related adverse events were mild.
The overall safety and pharmacokinetic profile suggests that rVWF can be used for the treatment and preventing of bleeding episodes at doses comparable to pdVWF/pdFVIII.
Example 3
Clinical Study of rVWF:rFVIII Complex.
This study was a pharmacokinetics, immediate safety and tolerability study in congenital von Willebrand disease Type 3. FIG. 9 shows the rVWF PK parameters for three patients in the study.
FIG. 10 shows PK data for Patient 1. There was a good correlation between VWF:RCo and VWF:CBA. Activities were measurable up to 12 hours (both were below the limit of quantification at 24 hours). VWF:Ag was still measurable at 96 hours. Endogenous FVIII activity increased to a maximum of 0.13 IU/mL.
FIG. 11 shows PK data in Patient 2. As with Patient 1, there was a good correlation between VWF:RCo and VWF:CBA. Both activities were measurable (albeit at the limit of quantification) up to 12 hours. VWF:Ag was still measurable at 72 hours (the 96 hour sample was not tested). Endogenous FVIII activity increased to a maximum of 0.18 IU/mL for this patient.
FIG. 12 shows PK data in Patient 3. VWF:RCo activity was measurable up to 24 hours (albeit at the limit of quantification), while VWF:CBA was below LoQ at 24 hours. VWF:Ag was still measurable up to 48 hours. Endogenous FVIII activity increased to a maximum of 0.13 IU/mL for this patient.
FIG. 13 shows data comparing the VWF parameters and FVIII activity for the three patients. There was a good correlation see in all three patients for all parameters tested (VWF:RCo, VWF:CBA, VWF:Ag, and FVII:Chr activity).
rVWF had  ̃100% recovery in all patients. The three patients showed slightly different rVWF PK. For VWF antigen, there was a maximum 0.06 IU/ml 30 minutes after application of 2 IU VWF:RCo/kg followed by a steady decline. For FVIII activity, the baseline FVIII activity increased to approximately 0.1 IU/ml 15 minutes after application of rVWF+rFVIII. Thereafter, activity further increased to 0.16 IU/ml, reached a plateau at 3-6 hours that stayed up to 28 hours followed by a steady decline. The delayed increase in FVIII indicates that the secondary rise was induced by rVWF.
Example 4
rVWF Study-FVII PK Assessment and TA Assessment.
FIG. 14 shows data for FVIII activity across all cohorts. FIG. 15 shows data from Cohort 1 (2 IU/kg VWF:RCo/1.5 IU/kg FVIII). There was good correlation for all three patients in this cohort for all parameters tested.
FIG. 15 shows data from Cohort 2 (7.5 IU/kg VWF:RCo/5.8 IU/kg FVIII). FIG. 16 shows the pharmacokinetics data for this cohort for FVIII:C.
FIG. 17 shows data for Cohort 3 (20 IU/kg VWF:RCo/15.4 IU/kg FVIII). These data show a comparison of plasma parameters for VWF and FVIII. FIG. 18 shows the pharmacokinetics data for this cohort for FVIII:C.
FIG. 19 shows data for Cohort 4 (50 IU/kg VWF:RCo). FVIII dosing was 38.5 IU/kg rFVIII or 25 IU/kg pdFVIII. These data show that the parameters for rVWF are similar to those for pdVWF (Haemate), but that rVWF is more effective at stabilizing FVIII activity, resulting in an increase seen in FVIII activity as compared to that seen with pdVWF. FIG. 20 shows a summary of pharmacokinetics data for this cohort for FVIII:C. As can be seen in these data, the median and mean T1/2 was increased for rVWF/FVIII as compared to pdVWF/FVIII, suggesting that rVWF is more effective at stabilizing in vivo FVIII than is pdVWF resulting in increased half-life.
Example 5
Clinical Study of rVWF Effect on rFVIII Half-Life in the Treatment of Hemophilia A.
This study evaluates the immediate tolerability and safety of rVWF after single doses of 50 IU/kg rFVIII (Advate) alone or in combination with rVWF at 10 or 50 IU/kg von Willebrand Factor:Ristocetin cofactor activity (VWF:RCo). This study also evaluates the pharmacokinetics after single doses of 50 IU/kg rFVIII (Advate) alone or in combination with rVWF at 10 or 50 IU/kg (VWF:RCo).
The population for the study is selected using the following criteria:
Inclusion Criteria:
Severe Hemophilia A (FVIII:C<1 IU/dL)
18 to 60 years of age
Previous coagulation factor replacement therapy (>150 ED)
Non-bleeding state
Exclusion Criteria:
Other coagulation disorders
History of VWF and/or FVIII inhibitors
Cardiovascular disease
Medical history of thromboembolic event
Medical history of other immunological disorders (exceptions)
Subject participation is for 16 weeks with an overall study duration of 7 months.
The design of the study is a multicenter, uncontrolled, non randomized, open-label clinical study to assess safety, immediate tolerability and pharmacokinetics of rFVIII:rVWF in subjects with hereditary severe hemophilia A (FVIII:C<1%).
Each subject (N=14) participating in the triple-period PK analysis receives in a non randomized fashion 50 IU/kg rFVIII (Advate) with no, 10 IU/kg VWF:RCo, or 50 IU/kg VWF:RCo rVWF, and is expected to participate for approximately 16 weeks.
The intra-individual pharmacokinetics are spaced at 5-14 days apart to ensure a “wash-out” period of at least 5 days and a timely completion of the study. Safety analyses are performed separately once 6 subjects have been dosed with 10 IU/KG VWF:RCo rVWF and once 6 subjects have been dosed with 50 IU/KG VWF:RCo rVWF.
Subjects should not have received cryoprecipitate, fresh frozen plasma or other drugs interfering with VWF or FVIII PK for at least 5 days before either of the infusions.
Dosage of rVWF is escalated when immediate tolerability and safety are demonstrated after a minimum of 6 subjects treated with 10 IU VWF:RCo rVWF. The effects of the investigational product on vital signs, hematology, and clinical chemistry parameters will determine short-term safety.
Samples for the determination of levels of FVIII activity (FVIII:C), VWF antigen (VWF:Ag), VWF activity (VWF:RCo), VWF:CB, (and VWF multimer distribution) are taken pre-infusion (within 30 minutes prior to the start of the infusion) and after the end of the infusion at 15 minutes (±5 minutes), 30 minutes (±5 minutes), 1 hour (±5 minutes), 3 hours (±10 minutes), 6 hours (±10 minutes), 9 hours (±15 minutes), 24 hours (±2 hours) and 48 hours (+2 hours), 72 hours (±2 hours), 96 hours (±2 hours), and 120 hours (±2 hours) (or at least every 24 hours thereafter until FVIII drops below <1%). FVIII activity is determined using both the chromogenic and one-stage a PTT-based assay methods performed at the local and central laboratory.
The effects of each infusion on vital signs and other symptoms indicative of an adverse event (AE), hematology, and clinical chemistry parameters are used as indicators of short-term safety. Vital signs, clinical chemistry, and hematology parameters are assessed pre-infusion and until 120 hours post-infusion. The occurrence of AEs are continuously monitored for up to 2 hours post-infusion (rVWF treated subjects) and at various time points during the follow-up period.
The presence of inhibitors to FVIII and VWF are assessed before investigational product infusion and at the study completion visit.
If at any time a serious AE related to the investigational product occurs, an independent data monitoring committee (DMC) will advise whether to continue the study.
Pharmacokinetic primary endpoints for the study include: AUC0-∞/Dose (area under the plasma concentration/time curve from time 0 to infinity); AUC0-96h/Dose (area under the plasma concentration/time curve from time 0 to 96 hours); mean residence time (MRT); clearance (CL); T1⁄2 (elimination phase half-life); Volume of distribution at steady state (Vss) of VWF:RCo, VWF:Ag, VWF:CB, and FVIII.
Analysis for pharmacokinetic primary endpoints include AUC0-96h/Dose, AUC0-∞/Dose, MRT, CL, T1/2 and Vss summarized per treatment group (50 IU/kg rFVIII (Advate) alone, 50 IU/kg rFVIII (Advate) premixed with rVWF at 10 IU/kg VWF:RCo, 50 IU/kg rFVIII (Advate) premixed with rVWF at 50 IU/kg VWF:RCo) by median and two-sided 95% CIs for the median, mean, standard deviation, coefficient of variation and geometric mean. Descriptive statistics (medians and ranges) are used to summarize VWF:RCo, VWF:Ag, VWF:CB and FVIII levels over time.
Safety primary endpoint includes: occurrence of treatment related AEs. Analysis of the safety primary endpoint includes the number and percentage of subjects who experience a treatment related AE. The number and rate of treatment related AEs are tabulated.
Pharmacokinetic secondary endpoints include: In vivo recovery (IVR) and incremental recovery (IR) of FVIII, VWF:RCo, VWF:Ag and VWF:CB. Analysis of pharmacokinetic secondary endpoints includes determining IVR and IR of FVIII, VWF:RCo, VWF:Ag and VWF:CB by subject and summarizing by medians and ranges.
Safety secondary endpoints include: development of inhibitory and total binding anti-VWF antibodies, development of inhibitory antibodies to FVIII, development of antibodies to Chinese hamster ovary (CHO) proteins, mouse immunoglobulin G (IgG) and rFurin, occurrence of thrombotic events.
Safety secondary endpoints are analyzed by tabulating the number of subjects who experience an SAE and the number of SAEs. In addition, the number of subjects who experience a treatment related AE and the number and rate of treatment related AEs are subcategorized for thrombotic events, inhibitory and total binding anti-VWF antibodies, inhibitory antibodies to FVIII, antibodies to Chinese hamster ovary (CHO) proteins, antibodies to mouse immunoglobulin G (IgG) and antibodies to rFurin.
A listing of all AEs are presented by subject identifier, age, sex, preferred term and reported term of the AE, duration, severity, seriousness, action taken, outcome, causality assessment, onset date, stop date and medication or non-drug therapy to treat the AE. An overview table for AEs are provided, presenting the number of AEs, the number of subjects with AEs and the corresponding percent of subjects in total and by seriousness and relationship to treatment. An additional summary table presents the total number of (mild, moderate, severe) AEs by system organ class and preferred term with relationship to treatment.
No formal sample size calculation has been conducted for this study.
It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

## Claims

1. A method for treating Von Willebrand Disease or Hemophilia A in a subject in need thereof, the method comprising: intravenously administering to the subject a composition comprising recombinant Von Willebrand Factor (rVWF) wherein endogenous Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein the rVWF is not modified with a water soluble polymer, wherein the composition is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, and wherein the rVWF has a higher specific activity than plasma derived Von Willebrand Factor, and the higher order rVWF multimers are stable for at least 3 hours post-administration.

2. The method of claim 1, wherein the method comprises co-administering to the subject said composition comprising recombinant Von Willebrand Factor (rVWF) and a composition comprising recombinant Factor VIII (rFVIII).

3. The method of claim 2, wherein the rVWF and rFVIII are administered together in a single composition.

4. The method according to claim 1, wherein the subject is administered between 1.0 IU/kg VWF:RCo and 150 IU/kg VWF:RCo per dose.

5. The method of claim 4, wherein the subject is administered between 2 IU/kg VWF:RCo and 50 IU/kg VWF:RCo per dose.

6. The method of claim 4, wherein the subject is administered between 5 IU/kg VWF:RCo and 40 IU/kg VWF:RCo per dose.

7. The method of claim 4, wherein the subject is administered between 10 IU/kg VWF:RCo and 20 IU/kg VWF:RCo per dose.

8. The method according to claim 1, wherein the rVWF is matured in vitro by treatment with Furin.

9. The method according to claim 1, wherein the rVWF is produced through expression in a Chinese Hamster Ovary (CHO cell culture).

10. The method according to claim 2, wherein the rFVIII and rVWF are produced through expression in the same cell culture.

11. The method according to claim 1, wherein the subject is administered rVWF no more than once every other day.

12. The method of claim 11, wherein the subject is administered rVWF no more than twice a week.

13. The method according claim 1, wherein the Factor VIII half-life is extended by at least 5 hours.

14. The method of claim 13, wherein the Factor VIII half-life is extended by at least 12 hours.

15. The method of claim 13, wherein the Factor VIII half-life is extended by at least 24 hours.

16. The method of claim 13, wherein the Factor VIII half-life is extended by at least 36 hours.

17. The method of claim 13, wherein the Factor VIII half-life is extended by at least 48 or 72 hours.

18. The method of claim 2, wherein the ratio of procoagulant activity (IU FVIII:C) of the administered FVIII to Ristocetin cofactor activity (IU rVWF:RCo) of the administered rVWF is between 2:1 and 1:4.

19. The method of claim 18, wherein the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) is between 3:2 and 1:3.

20. The method of claim 18, wherein the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) is between 1:1 and 1:2.

21. The method of claim 18, wherein the ratio of FVIII procoagulant activity (IU FVIII:C) to rVWF Ristocetin cofactor activity (IU rVWF:RCo) is about 3:4.

22. The method of claim 1, wherein the rVWF has a specific activity of about 20-150 mU/μg.

23. The method of claim 1, wherein the high molecular weight VWF multimer composition comprises at least 30% VWF decamers or higher order multimers.

24. The method of claim 1, wherein the high molecular weight VWF multimer composition comprises at least 40% VWF decamers or higher order multimers.

25. The method of claim 1, wherein the high molecular weight VWF multimer composition comprises at least 50% VWF decamers or higher order multimers.

26. The method of claim 1, wherein the high molecular weight VWF multimer composition comprises at least 60% VWF decamers or higher order multimers.

27. The method of claim 1, wherein the high molecular weight VWF multimer composition comprises at least 70% VWF decamers or higher order multimers.

28. A method for treating Hemophilia A or Von Willebrand Disease in a subject in need thereof, the method comprising: administering to the subject a composition comprising recombinant Von Willebrand Factor (rVWF) wherein Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein: (a) the rVWF has a higher specific activity than plasma derived Von Willebrand Factor, wherein the specific activity of rVWF is about 20-150 mU/g; (b) the FVIII half-life is at least 1.5 times higher as compared to FVIII half-life in a subject administered plasma derived Von Willebrand Factor; and (c) the rVWF is not modified with a water soluble polymer; and (d) the rVWF is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, wherein the higher order rVWF multimers are stable for at least 3 hours post-administration.

29. A method for treating Hemophilia A or Von Willebrand Disease in a subject in need thereof, the method comprising: administering to the subject a composition comprising recombinant Von Willebrand Factor (rVWF) wherein Factor VIII half-life is extended as compared to a subject administered plasma derived Von Willebrand Factor, wherein: (a) the composition is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, (b) the rVWF has a higher specific activity than plasma derived Von Willebrand Factor, wherein the specific activity of rVWF is at least about 20-150 mU/g; (c) the FVIII half-life is at least 1.5 times higher as compared to FVIII half-life in a subject administered plasma derived Von Willebrand Factor; and (d) the rVWF is not modified with a water soluble polymer; and (e) the rVWF is a high molecular weight VWF multimer composition comprising at least 20% VWF decamers or higher order multimers, wherein the higher order rVWF multimers are stable for at least 3 hours post-administration.

30. The method of claim 1, 28 or 29, wherein the method is a method for treating Von Willebrand Disease Type 3.

31. The method of claim 1, 28 or 29, wherein the level of Factor VIII procoagulant activity (FVIII:C) in the plasma of the subject 24 hours post-administration is at least 90% of the level of FVIII:C activity present in the plasma 1 hour post-administration.

32. The method of claim 1, 28 or 29 wherein said water soluble polymer is PEG.