Granted Patent B2

US 12,653,867 · App. 17/514,906 · Granted Jun 16, 2026

Inventors: Kurt Schnecker (Vienna, AT); Eva Haidweger (Vienna, AT); Peter Turecek (Klosterneuberg, AT)

Assignee: Takeda Pharmaceutical Company Limited

A61K38/36A61K9/08A61K9/19A61K38/16A61K38/17A61K47/12A61K47/183A61K47/22A61K47/26

Abstract

Claims (25)

1 . A method of preparing a stable lyophilized pharmaceutical formulation of a recombinant von Willebrand Factor (rVWF) wherein the method comprises:

(a) adding a buffer, one or more amino acids, one or more stabilizing agents, and one or more surfactants to a rVWF to obtain a solution; and

(b) lyophilizing said solution from step (a);

wherein said rVWF comprises a polypeptide selected from the group consisting of:

i) the amino acid sequence set out in SEQ ID NO: 3; and

ii) a biologically active fragment comprising the region of SEQ ID NO:3 from Val449 to Asn730 which causes agglutination of stabilized platelets in the presence of ristocetin, which has the activity to stabilize Factor VIII, or which binds to glycoprotein 1b (GP1b), collagen or heparin;

wherein said buffer comprises a pH buffering agent selected from the group consisting of citrate and HEPES at 15 mM and said pH is in a range of about 2.0 to about 12.0;

wherein said one or more amino acids is selected from the group consisting of glycine, lysine, and histidine at a concentration of about 1 to about 500 mM;

wherein said one or more stabilizing agents is at a concentration of about 0.1 to about 1000 mM and is selected from the group consisting of mannitol, lactose, sorbitol, xylitol, sucrose, trehalose, mannose, maltose, glucose, raffinose, cellobiose, gentiobiose, isomaltose, arabinose, glucosamine, fructose and combinations of these stabilizing agents; and

wherein said one or more surfactants is at a concentration of about 0.01 g/L to about 0.5 g/L.

2 . The method of claim 1 wherein the buffering agent is citrate.

3 . The method of claim 1 wherein pH is in the range of about 6.0 to about 8.0.

4 . The method of claim 3 wherein pH is in the range of about 6.5 to about 7.5.

5 . The method of claim 1 wherein pH is about 7.3.

6 . The method of claim 1 wherein the buffering agent is a citrate buffer and the pH is about 7.3.

7 . The method of claim 1 wherein the one or more amino acids is at a concentration range of about 0.5 mM to about 300 mM.

8 . The method of claim 7 wherein the one or more amino acids is glycine at a concentration of about 15 mM.

9 . The method of claim 1 wherein the rVWF comprises the amino acid sequence set out in SEQ ID NO: 3, or the biologically active fragment comprising the region of SEQ ID NO:3 from Val449 to Asn730 which causes agglutination of stabilized platelets in the presence of ristocetin, or binds to Factor VIII; wherein the buffering agent is a citrate buffer and the pH is about 7.3; and wherein the one or more amino acids is glycine at a concentration of about 15 mM.

10 . The method of claim 1 wherein the one or more stabilizing agents are trehalose at a concentration of about 10 g/L and mannitol at a concentration of about 20 g/L.

11 . The method of claim 1 wherein the surfactant is selected from the group consisting of digitonin, 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol, p-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol, polysorbate 20, polysorbate 80 and combinations of these surfactants.

12 . The method of claim 11 wherein the one or more surfactants is polysorbate 80 at about 0.01 g/L.

13 . The method of claim 1 wherein the rVWF comprises the amino acid sequence set out in SEQ ID NO: 3, or the biologically active fragment comprising the region of SEQ ID NO:3 from Val449 to Asn730; wherein the buffering agent is a citrate buffer at a concentration of about 15 mM and the pH is about 7.3; wherein the one or more amino acids is glycine at a concentration of about 15 mM; wherein the one or more stabilizing agents are trehalose at a concentration of about 10 g/L and mannitol at a concentration of about 20 g/L; and wherein the one or more surfactants is polyoxyethylene (80) sorbitan monooleate at about 0.1 g/L.

14 . The method of claim 1 wherein an amount of VWF which causes agglutination of stabilized platelets in the presence of ristocetin (VWF:RCo) in the lyophilized pharmaceutical formulation after reconstitution is from 83 U/ml to 142 U/ml.

15 . The method of claim 1 wherein the lyophilized pharmaceutical formulation after reconstitution maintains VWF:RCo activity for up to 24 months of storage at 5° C.±3° C.

16 . The method of claim 1 wherein VWF:RCo activity of the lyophilized pharmaceutical formulation after reconstitution is characterized with no more than a decrease of 0.0344 U/ml VWF:RCo per month.

Assignments (4)

Continuity (11)

References Cited (195)