Hydrophobic charge induction chromatographic depletion of a protein from a solution

Patent metadata

Field	Value
Publication number	US-10188965-B2
Application number	US-201314649374-A
Country	US
Kind code	B2
Filing date	Dec 5, 2013
Priority date	Dec 5, 2012
Publication date	Jan 29, 2019
Grant date	Jan 29, 2019

Abstract

The present invention relates generally to a method of reducing the level of at least one protein selected from the group consisting of plasminogen, tissue plasminogen activator and other protease(s) in a solution comprising at least one protein selected from the group consisting of fibrinogen, Factor VIII and von Willebrand factor (VWF), the method comprising: (i) passing a feedstock comprising at least one protein selected from the group consisting of fibrinogen, Factor VIII and VWF through a hydrophobic charge-induction chromatographic resin under conditions selected such that at least one protein selected from the group consisting of plasminogen, tissue plasminogen activator and other protease(s) present in the feedstock is bound to the resin; and (ii) recovering a solution comprising the at least one protein selected from the group consisting of fibrinogen, Factor VIII and VWF which passes through the resin, wherein the concentration of the at least one protein selected from the group consisting of plasminogen, tissue plasminogen activator and other protease(s) in the solution is reduced by at least 50% compared to the feedstock. Also provided are solutions and pharmaceutical formulations comprising the fibrinogen and/or Factor VIII and/or VWF recovered by such methods, and uses thereof.

First claim

The invention claimed is:

1. A pharmaceutical formulation comprising a solution of fibrinogen and an effective amount of a stabilizer, wherein the fibrinogen is recovered by the method of:
(i) passing a feedstock comprising fibrinogen through a hydrophobic charge-induction chromatographic resin equilibrated at a pH from 6.5 to 8.5, wherein at least one protein selected from the group consisting of plasminogen, tissue plasminogen activator, and other protease(s) present in the feedstock binds to the resin; and
(ii) recovering a solution comprising at least 80% total protein of fibrinogen which passes through the resin,
wherein the concentration of tissue plasminogen activator is less than 50 pg/mg and the concentration of plasminogen is less than 1 μg/mg in the recovered fibrinogen solution.

2. The pharmaceutical formulation of claim 1 , wherein the recovered solution comprises at least 80% total protein of monomeric fibrinogen.

3. The pharmaceutical formulation of claim 1 , wherein the fibrinogen retains from about 90% to 100% activity after at least 4 weeks in storage of the solution at a temperature of about 0° C. to about 8° C.

4. The pharmaceutical formulation of claim 1 , wherein the fibrinogen retains from about 60% to about 70% activity after 5 weeks in storage of the solution at a temperature of about 30° C.

5. The pharmaceutical formulation of claim 1 , further comprising a pharmaceutically acceptable carrier.

6. The pharmaceutical formulation of claim 5 having a volume of at least 5 mL and comprising at least 5 mg/mL fibrinogen.

7. The pharmaceutical formulation of claim 5 having a volume of at least 5 mL and comprising at least 20 mg/mL fibrinogen.

8. A method of treating or preventing a condition associated with fibrinogen deficiency, the method comprising administering to a subject in need thereof the pharmaceutical formulation of claim 1 .

9. The method of claim 8 , wherein the condition is selected from the group consisting of afibrinogenemia, hypofibrinogenemia and dysfibrinogenemia.

10. A fibrin glue comprising a solution of fibrinogen, wherein the fibrinogen is recovered by the method of:
(i) passing a feedstock comprising fibrinogen through a hydrophobic charge-induction chromatographic resin equilibrated at a pH from 6.5 to 8.5, wherein at least one protein selected from the group consisting of plasminogen, tissue plasminogen activator, and other protease(s) present in the feedstock binds to the resin; and
(ii) recovering a solution comprising at least 80% total protein of fibrinogen which passes through the resin,
wherein the concentration of tissue plasminogen activator is less than 50 pg/mg and the concentration of plasminogen is less than 1 μg/mg in the recovered fibrinogen solution.

Assignees

• Csl Ltd
• Csl Behring Gmbh

Inventors

• Pham Hung
• Hey Jeffrey Michael
• Nguy Darren

Classifications

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K14/745

  Blood coagulation or fibrinolysis factors · CPC title

• B01D15/363 Primary

  Anion-exchange · CPC title

• C07K14/75

  Fibrinogen · CPC title

• A61J1/05

  for collecting, storing or administering blood, plasma or medical fluids {(multiple bags systems for separating or storing blood components A61M1/0209); Infusion or perfusion containers} · CPC title