Claims for Patent: 4,720,385

Summary for Patent: 4,720,385

Title:	Protein compositions substantially free from infectious agents
Abstract:	Compositions containing therapeutically or immunologically active proteins are rendered substantially free from infectious agents such as viable viruses and bacteria without substantial loss of therapeutic or immunologic activity by mixing the protein composition with a complex formed from transition metal ions, such as copper ions, and an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a \"cis-ortho\" relationship, such as phenanthroline, and a reducing agent such as a thiol or ascorbic acid or ascorbate salt or mixtures of ascorbic acid or ascorbate with a thiol in amounts and at a temperature and for a time sufficient to inactivate substantially all of the viruses and bacteria contained therein. Compositions containing therapeutically active proteins substantially free from viral and bacterial infectivity, which have heretofore been unattainable, can be prepared by the method of the invention.
Inventor(s):	Lembach; Kenneth J. (Danville, CA)
Assignee:	Miles Laboratories, Inc. (Elkhart, IN)
Application Number:	06/736,197
Patent Claims:	1. A method for rendering a composition, which contains a therapeutically or immunologically active protein, selected from the group consisting of blood plasma proteins, virus vaccines, bacterial vaccines, and non-infectious antigens dissolved in an aqueous medium, substantially free from infectious agents selected from viable viruses and bacteria without substantial loss of therapeutic or immunologic activity, which comprises (a) mixing an aqueous solution of the protein composition with an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of thereapeutic or immunologic activity, and (b) holding the mixture for a time and at a temperature sufficient to render said infectious agents substantially non-infective in the protein composition without substantial loss of therapeutic or immunologic activity therein. 2. The method of claim 1 wherein the mixture is held in step (b) at a temperature of about 2.degree.-60.degree. C. for a period of at least about 1 minute. 3. The method of claim 1 wherein the mixture is held in step (b) at a temperature of about 20.degree.-37.degree. C. for a period of at least about 0.25 hours. 4. The method of claim 1 wherein the mixture is held for a time and at a temperature to inactivate at least about 99% of the viruses and bacteria in the protein composition. 5. The method of claim 1 wherein the pH in step (b) is physiologically compatible. 6. The method of claim 1 wherein the pH is about 4-10. 7. The method of claim 1 wherein the amount of complex in step (a) is about 0.0001-0.1 mM in an aqueous solution containing about 0.005-10% (weight/volume) of the protein composition. 8. The method of claim 1 wherein the amount of reducing agent is at least about 1 mM in the absence of added hydrogen peroxide in an aqueous solution containing about 0.005-10% (weight/volume) of the protein composition. 9. The method of claim 1 wherein the complex is a copper/1,10-phenanthroline complex. 10. The method of claim 1 wherein the reducing agent is selected from (a) ascorbic acid, (b) ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt, and (c) a mixture of one of ascorbic acid and ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt with a thiol. 11. The method of claim 1 wherein the reducing agent is a mixture of one of ascorbic acid and ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt with a thiol. 12. The method of claim 1 wherein the reducing agent is a mixture of one of cysteine and 3-mercaptopropionic acid with ascorbic acid (sodium salt). 13. The method of claim 1 including the further step of: (c) removing the added complex formed from the angularly-fused, polynuclear heterocyclic arene and the transition metal ion, low molecular weight reaction components, and low molecular weight products of reaction components. 14. The method of claim 13 wherein the protein composition comprises a component derived from blood plasma. 15. The method of claim 13 wherein the protein composition comprises a viral vaccine. 16. The method of claim 13 wherein the protein composition comprises non-infectious antigen. 17. The method of claim 13 wherein the protein composition comprises a protein selected from the group consisting of blood plasma, partially fractionated blood plasma, plasminogen, albumin, antihemophilic factor (Factor VIII), Factor IX concentrate containing Factors II and VII and IX and X, Factor II, Factor VII, Factor IX, Factor X, Plasma Protein Fraction (human), fibronectin (cold insoluble globulin), Factor XIII, IgG, IgA, IgD, IgE, IgM, high molecular weight kininogen (90,000-106,000), an immune globulin, intravenous (modified, either chemically or enzymatically or by fractional separation) immune serum globulin, monoclonal antibodies, anti-inhibitor coagulant complex (FEIBA), antithrombin III, alpha-1-proteinase inhibitor, plasma growth hormone, somatomedin, prealbumin, plasminogen-streptokinase complex, ceruloplasmin, transferrin, haptoglobin, and prekallikrein and mixtures thereof. 18. A product produced by the method of claim 14. 19. A product produced by the method of claim 15. 20. A product produced by the method of claim 16. 21. A product produced by the method of claim 17. 22. A method for rendering non-infective and innocuous, without substantial loss of activity, a composition which contains a protein produced by means of biotechnology by protein-producing organisms in a suitable medium which comprises (a) contacting said medium containing at least one of (i) intact protein-producing organisms, and (ii) adventitious agents such as viruses, and (iii) nucleic acid genetic material from the protein-producing organisms or adventitious agents with an aqueous solution of an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from a thiol, ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of activity of the protein, and (b) holding the mixture from Step (a) for a time and at a temperature sufficient to render said protein-producing organisms, adventitious agents, and nucleic acid genetic material from said protein-producing organisms and adventitious agents substantially non-infective and innocuous without substantial loss of protein activity. 23. The method of claim 22 including the further step of: (c) removing the added complex formed from the angularly-fused, polynuclear heterocyclic arene and the transition metal ion, low molecular weight reaction components, and low molecular weight products of reaction components. 24. A product produced by the method of claim 23.

Details for Patent 4,720,385

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	PLASMANATE	plasma protein fraction (human)	Injection	101140	10/02/1958		2005-01-19
Baxalta Us Inc.	AUTOPLEX, FEIBA NF, FEIBA VH	anti-inhibitor coagulant complex	For Injection	101447	12/21/1979		2005-01-19
Baxalta Us Inc.	AUTOPLEX, FEIBA NF, FEIBA VH	anti-inhibitor coagulant complex	For Injection	101447	07/31/2000		2005-01-19