Claims for Patent: 4,395,396

Summary for Patent: 4,395,396

Title:	Blood-coagulation-promoting preparation based on human proteins and a method of producing the same
Abstract:	In a method of producing a blood-coagulation-promoting preparation based on human proteins and having a content of coagulation factors II, VII, IX and X and factor-VIII-inhibitor-bypassing-activity (FEIBA), human plasma is treated with sulphated high-polymer carbohydrates and/or with basic ion exchangers, the protein mixture with generated FEIB-activity is adsorbed on the ion exchanger, and the preparation is gained by elution and concentration.
Inventor(s):	Eibl; Johann (Vienna, AT), Schwarz; Otto (Vienna, AT), Elsinger; Fritz (Vienna, AT), Philapitsch; Anton (Ebenfurt, AT)
Assignee:	Immuno Aktiengesellschaft fur Chemisch-medizinische Produkte (Vienna, AT)
Application Number:	06/283,143
Patent Claims:	1. A blood-coagulation-promoting preparation based on human proteins, having a content of coagulation factors II, VII, IX and X and factor-VIII-inhibitor-by-passing-activity (FEIBA), which preparation is characterized in that it is free of thrombogenic activity up to at least 2 units of FEIBA per kg rabbit in the thrombosis inducing activity test according to Wessler, it is free of kallikrein activity and free of prekallikrein activator activity, measured in an aqueous solution of said preparation with a FEIBA concentration of up to at least 10 units per ml, it is affinity-chromatographically separable on dextran sulphate agarose by means of an NaCl gradient so as to obtain an eluate containing a protein with factor IX activity and an eluate containing a protein with FEIB-activity, said protein with factor IX activity eluting at a lower NaCl concentration than said protein with FEIB-activity, said eluate containing said protein with factor IX activity and said eluate containing said protein with FEIB-activity contain .alpha.- and .beta.-globulins when electrophoretically separated thus obtaining a separation curve having a main peak in the .alpha.-globulin region corresponding to a content of 60 to 80% of the total protein, a shoulder of 10 to 20% of the total protein following thereupon, and a slightly pronounced peak in the .beta.-globulin region corresponding to a content of 10 to 20% of the total protein following upon the shoulder-like course of said separation curve. 2. A preparation as set forth in claim 1, wherein said protein with factor IX activity elutes at an NaCl concentration of 0.1 to 0.5 molar and said protein with FEIB-activity elutes at an NaCl concentration of 0.3 to 0.5 molar, the maximum factor IX activity eluting at 0.3 molar and the maximum FEIB-activity eluting at 0.4 molar. 3. A preparation as set forth in claim 1, wherein said FEIB-activity, after a one-hour incubation in factor VIII inhibitor plasma, is preserved by at least 50%. 4. A preparation as set forth in claim 1, wherein said factor IX activity, after a one-hour incubation in factor IX deficient plasma, is preserved by at least 50%. 5. A preparation as set forth in claim 1, further comprising a content of inter-alpha-trypsin inhibitor (ITI) of 0.05 to 5 mg per FEIBA unit. 6. A method of producing a blood-coagulation-promoting preparation as set forth in claim 1, which method comprises treating human plasma with sulphated high-polymer carbohydrates and/or with basic ion exchangers so as to obtain a protein mixture with generated FEIB-activity, adsorbing said protein mixture, and recovering said preparation by elution and concentration. 7. A method as set forth in claim 6, wherein said human plasma at first is briefly treated with sulphated high-polymer carbohydrates, said protein mixture with generated FEIB-activity is then adsorbed on an ion exchanger on dextran basis and is then immediately eluted and concentrated. 8. A method as set forth in claim 6, wherein said human plasma is treated with an ion exchanger on dextran basis, and after at least two hours of exposure said protein mixture with generated FEIB-activity, adsorbed on said ion exchanger, is eluted and concentrated. 9. A method for promoting the coagulation of blood containing an inhibitor to factor VIII which comprises administering to a patient having blood which contains an inhibitor to factor VIII, an amount, effective to provide factor-VIII-inhibitor-by-passing-activity, of a blood coagulation-promoting preparation, which is produced according to the method described in claim 6. 10. A blood-coagulation-promoting preparation based on human proteins, having a content of coagulation factors II, VII, IX and X and a factor-VIII-inhibitor-by-passing-activity (FEIBA), which preparation is characterized in that it is free of thrombogenic activity up to at least 2 units of FEIBA per kg rabbit in the thrombosis inducing activity test according to Wessler; it is free of kallikrein activity and free of prekallikrein activator activity, measured in an aqueous solution of said preparation with a FEIBA concentration of up to at least 10 units per ml; and it is affinity-chromatographically separable on dextran sulphate agarose by means of an NaCl gradient so as to obtain an eluate containing a protein with factor IX activity and an eluate containing a protein with FEIB-activity, said protein with factor IX activity eluting at a lower NaCl concentration than said protein with FEIB-activity. 11. A preparation as set forth in claim 10, wherein said FEIB-activity, after a one-hour incubation in factor VIII inhibitor plasma, is preserved by at least 50%. 12. A preparation as set forth in claim 10,further comprising a content of inter-alpha-trypsin inhibitor (ITI) OF 0.05 to 5 mg per FEIBA unit. 13. A method for promoting the coagulation of blood containing an inhibitor to Factor VIII, which comprises administering to a patient having blood which contains an inhibitor to factor VIII, an amount, effective to provide factor-VIII-inhibitor-bypassing activity, of a blood coagulation-promoting preparation based on human proteins, having a content of coagulation factors II, VII, IX and X and factor-VIII-inhibitor-bypassing-activity (FEIBA), which preparation is characterized in that it is free of thrombogenic activity up to at least 2 units of FEIBA per kg rabbit in the thrombosis inducing activity test according to WEssler; it is free of kallikrein activity and free of prekallikrein activator activity, measured in an aqueous solution of said preparation with a FEIBA concentration of up to at least 10 units per ml; and it is affinity-chromatographically separable on dextran sulphate agarose by means of an NaCl gradient so as to obtain an eluate containing a protein with factor IX activity and an elute containing a protein with FEIB-activity, said protein with factor IX activity eluting at a lower NaCl concentration than said protein with FEIB-activity, said eluate containing said protein with factor IX activity and said eluate containing said protein with FEIB-activity contain .alpha.- and .beta.-globulins when electrophoretically separated thus obtaining a separation curve having a main peak in the .alpha.-globulin region corresponding to a content of 60 to 80% of the total protein, a shoulder of 10 to 20% of the total protein following thereupon, and a slightly pronounced peak in the .beta.-globulin region corresponding to a content of 10 to 20% of the total protein following upon the shoulderlike course of said separation curve. 14. A method for promoting the coagulation of blood containing an inhibitor to factor VIII, which comprises administering to a patient having blood which contains an inhibitor to factor VIII, an amount, effective to provide factor-VIII-inhibitor-bypassing-activity, of a blood coagulation-promoting preparation based on human proteins, having a content of coagulation factors II, VII, IX and X and factor-VIII-inhibitor-by-passing-activity (FEIBA), which preparation is characterized in that it is free of thrombogenic activity up to at least 2 units of FEIBA per kg rabbit in the thrombosis inducing activity test according to Wessler; it is free of kallikrein activity and free of prekallikrein activator activity, measured in an aqueous solution of said preparation with a FEIBA concentration of up to at least 10 units per ml; and it is affinity-chromatographically separable on dextran sulphate agarose by means of a NaCl gradient so as to obtain an eluate containing a protein with factor IX activity and an elute containing a protein with FEIB-activity, said protein with factor IX activity eluting at a lower NaCl concentration than said protein with FEIB-activity.

Details for Patent 4,395,396

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Baxalta Us Inc.	AUTOPLEX, FEIBA NF, FEIBA VH	anti-inhibitor coagulant complex	For Injection	101447	12/21/1979		2000-07-26
Baxalta Us Inc.	AUTOPLEX, FEIBA NF, FEIBA VH	anti-inhibitor coagulant complex	For Injection	101447	07/31/2000		2000-07-26
Baxalta Us Inc.	AUTOPLEX, FEIBA NF, FEIBA VH	anti-inhibitor coagulant complex	For Injection	101447	08/11/2005		2000-07-26