Claims for Patent: 6,566,140

Summary for Patent: 6,566,140

Title:	Inhibition of coagulation in blood and blood products
Abstract:	The present invention provides methods for inhibiting blood clotting. In general, the methods include adding corn trypsin inhibitor (CTI) to blood or a blood product in an amount sufficient to inhibit the clotting. The CTI can be used alone or in combination with other anti-coagulants. In one aspect, the invention features plasma clotting assays featuring substantially prolonged clotting times. Clotting assays using whole or minimally altered blood are also provided. Further provided are methods for storing blood or blood products at low temperature with the CTI.
Inventor(s):	Mann; Kenneth G. (Grand Isle, VT), Rand; Mathew D. (Danvers, MA), Cawthern; Kevin M. (Haverstraw, NY)
Assignee:	University of Vermont and State Agriculture College (Burlington, VT)
Application Number:	09/844,209
Patent Claims:	1. A method for inhibiting clotting of a blood product, the method comprising adding corn trypsin inhibitor (CTI) to the blood product in an amount sufficient to inhibit the clotting, wherein the blood product is platelet deficient plasma. 2. The method of claim 1, wherein clot time of the blood product is prolonged between about 10 seconds to about 3600 seconds when compared to a control without CTI. 3. The method of claim 1, wherein clot time of the blood product has an International Normalized Ratio (INR) of between about 1 to 6.5. 4. The method of claim 1, wherein the method is responsive to a concentration of Factor VIII or IX in the range of between about 0 units/ml to about 2 units/ml. 5. The method of claim 1, wherein the method includes adding the plasma to a reaction tube comprising CTI. 6. The method of claim 5, wherein the reaction tube further includes a buffered citrate salt. 7. The method of claim 1, wherein the method further comprises adding relipidated tissue factor (TF) to the plasma in an amount sufficient to provide between about 0.001 nM to 5 nM TF. 8.The method of claim 1, wherein the plasma is treated with at least one anti-coagulant prior to adding the corn trypsin inhibitor (CTI). 9. The method of claim 8, wherein the anti-coagulant is citrate and the citrate-treated plasma is recalcified before adding the CTI with a coagulation-effective amount of a calcium salt. 10. The method of claim 9, wherein the amount of corn trypsin inhibitor (CTI) is sufficient to prolong clotting time by between about 10 to about 3600 seconds when compared to a control without CTI. 11. The method of claim 10, wherein the amount of corn trypsin inhibitor (CTI) is between about 20 .mu.g to about 500 .mu.g per milliliter of the recalcified plasma. 12. The method of claim 1 further comprising adding an antibody to the blood product, wherein the antibody is capable of specifically binding a blood coagulation factor, and the antibody is added in an amount sufficient to inhibit clotting. 13. A method for assaying clotting in plasma, the method comprising: a) treating blood with a solution comprising an amount of a calcium-chelating agent sufficient to inhibit clotting, b) subjecting the blood to conditions conducive to producing plasma, c) freezing the plasma, d) contacting the frozen plasma with an amount of corn trypsin inhibitor (CTI) sufficient to inhibit clotting, e) thawing the frozen plasma, f) recalcifying the thawed plasma; and g) assaying the clotting in the plasma. 14. A method for assaying clotting in plasma, the method comprising the steps of: a) treating blood with a solution comprising an anti-coagulation effective amount of corn trypsin inhibitor (CTI) and a calcium-chelating agent, b) subjecting the blood to conditions conducive to producing plasma, c) recalcifying the plasma; and d) assaying the clotting in the recalcified plasma. 15. A method for assaying clotting in plasma, the method comprising: a) treating blood with a solution comprising an amount of a calcium-chelating agent sufficient to inhibit clotting, b) subjecting the blood to conditions conducive to making plasma, c) contacting the plasma with an amount of corn trypsin inhibitor (CTI) sufficient to inhibit clotting, d) recalcifying the plasma; and e) assaying the recalcified plasma, wherein step a) of the method further comprises adding corn trypsin inhibitor (CTI)to the blood. 16. A method for assaying clotting in plasma, the method comprising: a) treating blood with a solution comprising an amount of a calcium-chelating agent sufficient to inhibit clotting, b) subjecting the blood to conditions conducive to producing plasma, c) freezing the plasma, d) contacting the frozen plasma with an amount of corn trypsin inhibitor (CTI) sufficient to inhibit cloning, e) thawing the frozen plasma, f) recalcifying the thawed plasma; and g) assaying the clotting in the plasma, wherein the amount of corn trypsin inhibitor (CTI) added at step d) of the method is sufficient to prolong clotting time in excess of about 1000 seconds. 17. The method of claim 16, wherein the amount of corn trypsin inhibitor (CTI) is from between about 0.02 to about 1 mg CTI per milliliter of blood. 18. The method of claim 16, wherein step g) of the method further comprises adding to the plasma between about 1 mM and about 10 mM of a mixture of phosphotidylserine (25 mol. %) and phoshotidylcholine (75% mol. %) (PSPC) in an amount sufficient to provide from between about 0.01 .mu.M to 500 .mu.M of lipid in the plasma. 19. The method of claim 16, wherein step g) of the method further comprises adding to the plasma tissue factor (TF) in an amount sufficient to provide between about 0.001 to 5 nM TF in the plasma. 20. A method for assaying clotting in plasma, the method comprising the steps of: a) treating blood with a solution comprising an anti-coagulation effective amount of corn trypsin inhibitor (CTI) and a calcium-chelating agent, b) subjecting the blood to conditions conducive to producing plasma, c) recalcifying the plasma; and d) assaying the clotting in the recalcified plasma, wherein the amount of corn trypsin inhibitor (CTI) added at step a) of the method is sufficient to prolong cloning time in excess of about 10 seconds to about 3600 seconds. 21. The method of claim 20, wherein the amount of corn trypsin inhibitor (CTI) is from between about 0.02 to 1 mg CTI per milliliter of blood. 22. The method of claim 20, wherein the method further comprises adding tissue factor to the blood after the CTI in step a) and step d) of the method further comprises adding to the plasma between about 500 to about 2000 mole excess of phoshphotidylserine (25 mol. %) and phosphatidyicholine (75 mol. %) when compared to the amount of tissue factor added in step a). 23. The method of claim 20, wherein step d) of the method further comprises adding tissue factor (TF) to the plasma in an amount sufficient to provide between about 0.001 to 5 nM TF in the plasma. 24. A method for monitoring blood coagulation, the method comprising contacting a blood sample with an amount of corn trypsin inhibitor (CTI) sufficient to reduce or eliminate blood coagulation; and monitoring the coagulation of the blood sample, wherein the blood sample is plasma contacted with lipidated tissue factor, an anti-thrombotic agent, or an anti-platelet agent, and further wherein the monitoring comprises measuring coagulation. 25. The method of claim 24, wherein the anti-thrombotic or anti-platelet agent is one of hirudin, recombinant tick anti-coagulant, enoxaparin, or abciximab. 26. A method for monitoring blood coagulation, the method comprising contacting a blood sample with an amount of corn trypsin inhibitor (CTI) sufficient to reduce or eliminate blood coagulation; and monitoring the coagulation of the blood sample, wherein the blood sample is plasma, wherein the method further comprises adding to the plasma an antibody capable of specifically binding a blood coagulation factor, and wherein the antibody is added to the plasma in an amount sufficient to inhibit the coagulation.

Details for Patent 6,566,140

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Janssen Biotech, Inc.	REOPRO	abciximab	Injection	103575	12/22/1994		2018-06-08