METHOD FOR INHIBITING THROMBOSIS IN A PATIENT WHOSE BLOOD IS SUBJECTED TO EXTRACORPOREAL CIRCULATION

Abstract: This invention is a method for inhibiting thrombosis in a patient whose blood is subjected to extra-corporeal blood circulation which comprises contacting the extra-corporeal circulating blood with a Factor IXa compound in an amount effective to inhibit thrombosis in the patient. The patient may be subjected to extra-corporeal blood circulation during transplant surgery or cardiopulmonary bypass surgery or any surgery in which obligate clamping of a blood vessel is required. Further, a pharmaceutical composition includes an effective amount of a Factor IXa compound, and a pharmaceutically acceptable carrier.

Description: METHOD FOR INHIBITING THROMBOSIS IN A PATIENT WHOSE BLOOD IS SUBJECTED TO EXTRACORPOREAL CIRCULATION This application claims priority of U.S. Serial No. 09/053,872, filed April 1, 1998 which is a contmuation-m- part of PCT International application No. PCT/US97/08282 , filed May 15, 1997 which is a continuation- m-part of United States Application Serial No. 08/648,561, filed May 16, 1996 the contents of each of which are incorporated by referenence m their entireties into the present application. Background of the Invention Throughout th s application, various publications are referenced by author and date. Full citations for these publications may be found listed alphabetically at the end of the specification immediately preceding the claims. The disclosures of these publications m their entireties are hereby incorporated by reference into this application m order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein. In cardiopulmonary bypass surgery, one of the critical requirements is the maintenance of blood fluidity and the absence of thrombosis. The cardiopulmonary bypass circuit presents a unique combination of factors favoring the development of a prothrombotic environment. The contact of blood with numerous devices which are associated with this procedure, such as membrane oxygenators and filters, has been implicated m the activation of the intrinsic (contact) pathway of coagulation. Since the bypass circuitry generates a highly-thrombogenic environment, high levels of anticoagulation therapies are required (Edmunds, 1995; Edmunds, 1993; Gravlee et al . , 1990; alenga et al . , 1991; DeAnda et al . , 1994; Brister et al . , 1994; and Chomiak et al . , 1993) . Traditional intervention to prevent thrombosis m this setting has been the use of heparin. However, the use of heparin causes unacceptable side effects m certain patients. Such side effects may include the development of bleeding, heparin resistance or arterial/venous thrombosis. Despite investigations which have attempted to provide alternatives to the use of heparin, such as thrombm inhibitors (such as hirudm) and dermatan sulfate, no agent has yet been identified to replace or modify its use m clinical cardiac surgery ( alenga et al . , 1991; DeAnda et al . , 1994; Brister et al . , 1994; and Chomiak et al . , 1993). In addition, Benedict et al . (1994) Texas Heart Institute Journal Vol 21, No. 1, pp 85-90 disclose that infusion of Factor IXai at concentrations sufficient to inhibit intravenous coagulation did not produce bleeding significantly different from that m control animals. Therefore, the invention disclosed herein was unexpected m view of this report . Suπimary of the Invention This invention provides a method for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting the extracorporeal circulating blood with a Factor IXa compound m an amount effective to inhibit thrombosis m the patient. The Factor IXa compound may include an active site-blocked Factor IXa compound or Glu-Gly-Arg chloromethyl ketone- inactivated human factor IXa compound. This invention also provides that the effective amount may be from about 0.01 μg/ml plasma to about 250 μg/ml plasma or from about 0.05 μg/ml plasma to about 25 μg/ml plasma. The patient may be subjected to extracorporeal blood circulation during transplant surgery or cardiopulmonary bypass surgery. This invention further provides for a pharmaceutical composition which includes an effective amount of a Factor IXa compound and a pharmaceutically acceptable carrier. Brief Description of the Figures Figure 1. Blood Loss: Factor IXai vs Heparin Blood loss m the thoracic cavity was assessed m dogs treated with Factor IXai and dogs treated with heparin. After 1 hour of bypass followed by up to three hours of postoperative observation, blood loss was significantly less m the group receiving Factor IXai compared with the group receiving heparin (**, p<0.05) . Figures 2A, 2B, 2C, 2D, 2E and 2F. Scanning Electron Microscopic Analysis of Arterial Filter in Factor IXai vs Heparin Scanning electron microscopic analysis of the arterial filters is depicted. The Figures 2A, 2C and 2E show serial magnification of the arterial filters m Factor IXai treated dogs (50 x (top) , 200x (middle) , 2000x (bottom) ) . The Figures 2B, 2D and 2F show similar views of the heparin treated dogs. As demonstrated m these pictures, cardiopulmonary bypass performed with Factor IXai was associated with similar amounts of clinically mapparent fibrin deposition compared with the use of traditional heparin. Figures 3A, 3B, 3C, 3D, 3E, 3F 3G, and 3H. Histologic Examination of End Organs Hematoxylm/eosm staining of heart, lung, kidney, and liver dogs treated with Factor IXai are shown m Figures 3A, 3C, 3E, 3G. Figures 3B, 3D, 3F and 3H show samples evaluated dogs treated w th heparin. These studies reveal the absence of fibrin deposition and micro emboli m both groups . Figures 4A, 4B, 4C, 4D and 4E. Analysis of Blood Samples in Dogs Treated with Factor IXai and Heparin Blood samples were drawn at intervals throughout the surgical procedure and evaluated. Similar dilutional decreases m white blood cells, hematocπt, and platelet count were observed in each group. Levels of prothrombin time and partial thromboplastin time remained baseline Factor IXai treated dogs while there was significant elevation in dogs anticoagulated with heparin. (**p<0.05) {closed diamonds represent Factor IXai; open squares represent heparin} . Figures 5A and 5B. Factor IX based Clotting Assay. A clotting assay was developed to rapidly and reproducibly assess the level of anticoagulation during cardiopulmonary bypass. As shown, this is an assay based on Factor IX deficient plasma and an optimized dose of cephalin to determine the functional anticoagulant effect of Factor IXai. Figures 6A and 6B. Blood Loss and Hemostasis. Active Site-blocked Factor IXa was used in peripheral vascular surgery in rabbits and dogs. The blood loss at the aortotomy suture site was substantially less and the time to achieve hemostasis was decreased in animals treated with Factor IXa. Figure 7. Clotting Time vs. Concentration of Factor IXai By performing a series of dilutions of administered Factor IXai and determining the clotting time, it was determined that the limit of detection of Factor IXai in the assay is 0.08 μg/ml. Figure 8. Modified Cephalin Clotting Time (MCCT) vs Time Determination of MCCT after single dose of Factor IXai in canine cardiopulmonary bypass. After a single clinically- effective dose of Factor IXai (460 μg/kg) in canine cardiopulmonary bypass, the MCCT rises to 80 seconds and is maintained at that level through at least 1.5 hrs of cardiopulmonary bypass. Detailed Description of the Invention This invention provides for a method for inhibiting thrombosis a patient whose blood is subjected to extracorporeal blood circulation which includes contacting the extracorporeal circulating blood with a Factor IXa compound m an amount effective to inhibit thrombosis the patient . The Factor IXa compound may include an active site-blocked Factor IXa compound or a Glu-Gly-Arg chloromethyl ketone- inactivated human factor IXa compound. The effective amount may include from about 0.01 μg/ml plasma to about 250 μg/ml plasma or from about 0.05 μg/ml plasma to about 25 μg/ml plasma or preferably from 0.1 μg/ml plasma to about 5 μg/ml plasma. This invention further provides that the patient may be subjected to extracorporeal blood circulation during transplant surgery or cardiopulmonary bypass surgery or any surgery which obligate clamping of a blood vessel is required. The patient may be subjected to extracorporeal blood circulation during any kind of cardiac surgery, including bypass grafting, valve replacement, congential repair heart surgery and heart transplantation. The patient may be a human being. The patient may also be subjected to extracorporeal life support. The patient may be a cardiogenic shock patient. The patient may be undergoing hemodialysis, continuous arterio-venus hemoflltration (CAVH) , continuous veno-venous hemoflltration (CWH) , extracorporeal circulatory membrane oxygenation (ECMO) , brain surgery, vascular surgery, abdominal surgery, orthopaedic surgery, hip replacement surgery, transplant surgery, or any surgery requiring cardio-pulmonary bypass. The subject may be any patient requiring a mechanical circulatory assistance or ventricle assist device (i.e. LVAD) . The subject may be a patient requiring new devices as described m Wickelgren, 1996 such as implantable defibrillators . The subject may also be a patient suffering with symptoms of systemic lupus erythematosus or TTP (thrombotic thrombocytopenic purpura) . The subject may also be a patient requiring plasmapharesis . One embodiment of the present invention is a pharmaceutical composition which may include an effective amount of a Factor IXa compound and a pharmaceutically acceptable carrier. The carrier may include a diluent. Further, the carrier may include an appropriate adjuvant, a herpes virus, a liposome, a microencapsule, a polymer encapsulated cell or a retroviral vector. The carrier may include an aerosol, intravenous, oral or topical carrier. The Factor IXa compound may be attached to a solid support. The Factor IXa compound may be linked to or bonded to tubing. The tubing may be part of an extracorporeal life support system. As used herein "thrombosis" encompasses formation, development or presence of a blood clot or a blood coagulation which is located inside of a patient or inside of an extracorporeal life support system which circulates blood of the patient. Thrombosis also encompasses the presence of a thrombus which includes a blood clot occluding a blood vessel or formed m a heart cavity. Thrombosis also encompasses the activation of a plasmatic coagulation system m a patient which includes the production of cross-linked fibrin degradation product, protein C, free protein S, coagulation factor II, immunoglobulin G or albumin m the patient. "Thrombosis" also encompasses the formation of a white thrombus which may be composed of platelets and fibrin and is relatively poor erythrocytes , a disseminated fibrin deposit thrombus or a red thrombus which may be composed of red cells and fibr . Thrombosis may also include a thromboembolism which is the blocking of a blood vessel by a thrombus which may have been dislodged from a vein. Thrombosis may occur m areas of retarded blood flow m the patient, at a site of injury or at an abnormal vessel wall conjunction with an initiating platelet plug. Initiation of clot formation response to tissue injury is carried out by the extrinsic pathway of clotting. Formation of a pure red thrombus m an area of restricted blood flow or response to an abnormal vessel wall without tissue injury is carried out by the intrinsic pathway. Intrinsic and extrinsic pathways may converge m a final common pathway characterized by the activation of prothromb to thrombm and the thrombm-catalyzed conversion of fibr ogen to the fibrm clot . In the intrinsic pathway, Factor XIa cleaves Factor IX between Argl45 and Alal46 and between Arg 180-Vall81, releasing a 35 ammo acid peptide and producing Factor IXa having a 145 ammo acid light chain (ammo acids 1-145) and a 235 ammo acid heavy chain (ammo acids 181-415) joined by a disulfide bond between cysteme residues at positions 132 and 289. Factor IXa is a serine protease which, when complexed with Factor Villa on membrane surfaces, converts Factor X to its active form Factor Xa . The enzyme active site of Factor IXa is located on the heavy chain. Three ammo acids the heavy chain are principally responsible for the catalytic activity, Hιs221, Asp269 and Ser365 (H221, D269 and S365, the catalytic triad) . If the ammo acids of the heavy chain are numbered from 1 to 235, the catalytic triad is Hιs41, Asp89 and Serl85, and the disulfide bond joining the heavy chain to the light chain is at Cysl09 on the heavy chain. As used herein "a Factor IXa compound" means a compound which inhibits or reduces the conversion of Factor X to Factor Xa by naturally occurring Factor IX. As used herein, a Factor IXa compound may be chosen from one of several subsets. One subset is a chemically modified form of naturally occurring Factor IXa which chemical modification results m the mactivation of Factor IXa (e.g., inactivated Factor IXa, active-site blocked Factor IXa or Factor IXai) . Another subset of a Factor IXa compound is any recombinant mutated form of Factor IXa (e.g., a mutem form of Factor IXa, a recombmant Factor IXa with a deletion or Factor IXami) . In addition, there are other subsets of a Factor IXa compound which include but are not limited to, for example: (1) nucleic acids, (2) anti-Factor IXa antibodies or fragments thereof, (3) saccharides, (4) ribozymes, (5) small organic molecules, or (6) peptidomimetics . Thus, a Factor IXa compound may encompass the following: a Glu-Gly-Arg chloromethyl ketone- inactivated human factor IXa, an inactive Christmas factor, a Glu-Aly-Arg chloromethyl ketone- inactivated factor IXa, a glutamyl- glycyl-argmyl -Factor IXa, a dansyl Glu-Gly-Arg chloromethyl ketone-inactlvated bovine factor IXa (IXai), a Factor IXai, a competitive inhibitor of Factor IXa, a peptide mimetic of Factor IXa, a carboxylated Christmas factor, a competitive inhibitor of the formation of a Factor IXa/VIIIa/X complex, a des-γ-carboxyl Factor IX, Factor IX lacking a calcium- dependent membrane binding function, inactive Factor IX including only ammo acids 1-47, apoFactor IX including ammo acids 1-47, Factor IX Bm Kiryu, a Val-313 -to-Asp substitution m the catalytic domain of Factor IX, a Gly- 311-to-Glu substitution m the catalytic domain of Factor IX, a Gly-311 to Arg-318 deletion mutant of Factor IX, an anti -Factor IXa antibody, an anti-Factor IXa monoclonal or polyclonal antibody. The Factor IXa compound may also include inactive species of Factor IX described m the references provided herein, especially Freedman et al . , 1995; Furie and Furie, 1995; Miyata et al . , 1994 and acey et al. , 1994. Thus, a Factor IXa compound may be Factor IXa m which the active site is blocked and may be prepared as described m Experimental Details below. The Factor IXa compound may be a Factor IXa which includes post-translational modifications including glycosylation, β-hydroxylation of aspartic acid, γ- carboxylation of glutamic acid and propeptide cleavage. The Factor IXa compound may be concentrated via heparin affinity chromatography or hydrophobic interaction chromatography. The Factor IXa compound may be a genetically engineered, a recombinant Factor IXa m which ammo acids at the active site, especially the serine ammo acid at the active site, have been altered to render the recombinant Factor IXa functionally inactive, but still capable of competing with intact, native Factor IXa for cell surface binding. In one embodiment of the present invention the Factor IXa compound is a form of Factor IXa inactivated by the standard methods known to one of skill m the art, such as mutation of the gene which encodes Factor IXa. In a preferred embodiment of the present invention the Factor IXa compound is an active site-blocked Factor IXa or a Glu-Gly-Arg chloromethyl ketone-mactivated human Factor IXa. In a preferred embodiment, the effective amount is from about 0.1 μg/ml plasma to about 250 μg/ml plasma or from about 0.5 μg/ml plasma to about 25 μg/ml plasma or preferably from 0.7 μg/ml plasma to about 5 μg/ml plasma. In an embodiment of the present invention the Factor IXa compound is an inactive mutem form of Factor IXa which is useful as selective antithrombotic agent. As used herein, "mutem form" of Factor IXa means a protein which differs from natural factor IXa by the presence of one or more ammo acid additions, deletions, or substitutions which reduce or eliminate the ability of the protein to participate m the conversion of Factor X to Factor Xa . In another embodiment of the present invention the Factor IXa compound is a proteolytically inactive, recombinant mutem form of Factor IX, which has substantially the same ammo acid sequence as normal or native human Factor IX but m which a different ammo acid has been substituted for one or more of Hιs221, Asp269 and Ser365. The present invention also provides a proteolytically inactive, recombinant mutem form of Factor IXa, which has substantially the same ammo acid sequence as normal or native human factor IXa but m which a different ammo acid has been substituted for one or more of Hιs41, Asp89 or Serl85 the heavy chain of Factor IXa. The term "proteolytically inactive" means that the mutems are incapable of converting Factor X to Factor Xa . Recombinant mutems of Factor IX useful m this invention are referred to collectively as Factor Ixmi (i.e., Factor IX mutationally inactivated) . Recombinant mutems of Factor IXa useful m this invention are referred to collectively as Factor IXami . Examples of Factor IXa compounds which are recombinant mutems are as follows: Factor IXmi (Ser365-Xxx) Factor IXmi (Asp269-Yyy) Factor IXmi (Hιs221-Zzz) Factor IXmi (Ser365-Xxx, Asp269->Yyy) Factor IXmi (Ser 365-Xxx, Hιs221-Zzz) Factor IXmi (Asp269→Yyy, His→Zzz) Factor IXmi (Ser365-Xxx, Asp269-Yyy, His-Zzz) Factor IXami (Ser365-^Xxx) Factor IXami (Asp269-Yyy) Factor IXami (Hιs221-Zzz) Factor IXami (Ser365-Xxx, Asp269-Yyy) Factor IXami (Ser365-Xxx, Hιs221→Zzz) Factor IXami (Asp269-Yyy, His→Zzz) Factor IXami (Ser365-Xxx, Asp269-Yyy, His-Zzz) wherein Xxx is any one of the standard ammo acids other than serine, Yyy is any one of the standard ammo acids other than aspartic acid, and Zzz is any of the standard ammo acids other than histidme. Preferred recombinant mutems are Factor IXmi (Ser365^Ala) and Factor IXami (Ser365-Ala) . Factor IXmi and Factor IXami are functionally similar to Factor IXai m terms of their ability to establish effective anti -coagulation mtravascularly and m ex vivo equipment connected to the blood stream while permitting retention of effective hemostasis. The advantages of Factor IXmi and Factor IXami over Factor IXai are the following: -Factor IXmi and Factor IXami can be produced directly a genetically engineered organism, thus avoiding several processing and purification steps with their attendant losses, thereby improving product yield. -The cost of production of Factor IXmi and Factor IXami an appropriate genetically engineered organism is lower than the cost of production of Factor IXai from human plasma. -Factor IXmi and Factor IXami, produced m a genetically engineered organism, will not be subject to the risk of contamination with various infectious agents such as viruses or pπons (for example, the agents for HIV disease and for bovine and/or human spongiform encephalopathies) . -Factor IXmi and Factor IXami, being less different from wild-type human Factor IX and Factor IXa than chemically modified Factor IXai, will have a lower probability of eliciting an immune response m patients who are dosed with the modified protein for extended periods of time, thereby reducing the risk of delayed type hypersensitivity reactions and improving patient safety for indications such as anticoagulation m hemodialysis that will require repeated, long-term use. The recombinant mutems of this invention can be produced by known genetic engineering techniques, using as the starting material recombinant cDNA encoding Factor IX m an appropriate cloning vector. For example, a starting material which may be used the production of a Factor IXa compound is the product of Example 5 of U.S. Patent No. 4,770,999, i.e., a recombinant plaque of E. coll infected with a bacteriophage M12mpll Pst vector containing cDNA corresponding to the entire sequence of recombinant Factor IX ligated to Pst adapters. The recombinant plaques are used to prepare single-stranded DNA by either the small - scale or large-scale method described in Sambrook et al . , Molecular Cloning, A Laboratory Manual, Second Edition, Cold Spring Harbor Press, 1989, pages 4.29-4.30 and 4.32. The single-stranded M13mpll containing Factor IX cDNA is then used to carry out oligonucleotide-mediated mutagenesis using the double primer method of Zoller and Smith as described in Sambrook et al . , 1989, pages 15.51-15.73. Mutagenic primers which can be used include the following: 1) Oligonucleotides for producing Factor IXmi (Ser365-Xxx) 3'-W ACA GTT CCT CTA XXX CCC CCT GGG GTA V-5 ' where is T, 3 ' -GT or 3 ' -AGT V is C, 3'-CA, or 3 ' -CAA XXX is the complement to a DNA codon for any one of the standard amino acids other than serine . 2) Oligonucleotides for producing FACTOR IXmi (Asp269-Yyy) 3 ' - TTC ATG TTA GTA YYY TAA CGC GAA GAC V-5' where IS A, 3'=TA, OR 3 ' -TTA V is C, 3 ' -CT, or 3 ' -CTT YYY is the complement to a DNA codon for any one of the standard amino acids other than aspartic acid and cysteine . 3) Oligonucleotides for producing Factor IXmi (His221-Zzz) 3 ' -TTA CAT TGA CGA CGG ZZZ ACA CAA CTT TGA CCA-5 ' where W is A, 3'-AA, or 3 ' -TAA V is C, 3 ' -CC, or 3 ' -CCA ZZZ is the complement to a DNA codon for any one of the standard ammo acids other than histidme and cysteme. Oligonucleotide primers for producing the preferred Factor IXmi of this invention, Factor IXmi (Ser365-Ala) , are those of No. 1 above wherein XXX is the complement of a codon for alamne, i.e., 3 ' -CGA, 3'-CGC, 3 ' -CGT or 3'-CGC. A specific primer for producing Factor IXmi (Ser365-Ala) is: 3 ' -GT ACA GTT CCT CTA CGA CCC CCT GGG GTA C-5' A skilled artisan would recognize and know how to carry out the remaining steps of oligonucleotide-mediated mutagenesis as follows: -Hybridization of mutagenic oligonucleotides to the target DNA. -Extension of the hybridized ol igonucleotides to the target DNA . -Transfection of susceptible bacteria. -Screening of plaques for the desired mutation. -Preparation of single- stranded DNA from a mutant plaque. -Sequencing the single- stranded DNA. -Recovery of double-stranded Factor IXmi cDNA. -Inserting the double-stranded Factor IXmi cDNA into the expression vector used by Kaufman (for example) . -Expression of Factor IXmi. -Treating the Factor IXmi with Factor XIa to produce Factor IXami . Another embodiment of the present invention is an assay to monitor antithrombic activity of a Factor IXa compound infused into circulation of a patient which includes: (a) obtaining Factor IXa-deflcient plasma; (b) mixing the plasma from step (a) with diatomaceous earth (e.g. CELITE ® ) and with plasma from the patient; (c) mcubatmg the mixture with a source of phospholipid and calcium chloride under conditions suitable for clot formation; and (d) measuring time necessary for clot formation m the incubate, thereby monitormg the antithrombic activity of the Factor IXa compound infused into the circulation of the patient . Another embodiment of the present invention is a method for evaluating the ability of an agent to inhibit an active site of a Factor IXa compound which includes: (a) contacting the Factor IXa compound with the agent to form a protein complex; (b) mcubatmg the protein complex under conditions suitable for clot formation; (c) measuring time necessary for clot formation m the incubate, and (d) comparing the time measured step (c) with the time measured the absence of the agent, thus evaluating the ability of the agent to inhibit the active site of the Factor IXa compound. Another embodiment of the present invention is an agent capable of inhibiting the active site of Factor IXa obtained from the methods described herein. The agent may be a peptide, a peptidomimetic, a nucleic acid or a small molecule. The agent may be an antibody or portion thereof. The antibody may be a monoclonal antibody or a polyclonal antibody. The portion of the antibody may include a Fab. The present invention provides a method for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which includes contacting the extracorporeal circulating blood with an agent capable of inhibiting a step m the intrinsic pathway of coagulation m an amount effective to inhibit thrombosis m the patient. The agent may be an active site-blocked Factor XII compound or an active site-blocked Factor XI compound. The present invention provides a proteolytically inactive recombinant mutem of Factor IX, which has substantially the same ammo acid sequence as normal Factor IX but which has an ammo acid substitution for one or more of Hιs221, Asp269 or Ser365. In one embodiment, the mutem has a Ser365 to Ala substitution. The present invention also provides a proteolytically inactive recombinant mutem of Factor IXa which has substantially the same ammo acid sequence as normal human Factor IXa but which has an ammo acid substitution for one or more of Hιs41, Asp89 or Serl85 m the heavy chain of Factor IXa. In one embodiment, the mutem has a Serl85 to Ala substitution. In another embodiment, the mutem is encoded by isolated cDNA. In another embodiment, a replicable vector comprises the isolated cDNA. In another embodiment, a microorganism is transfected with the replicable vector. The transfection may be a stable transfection or a transient transfection. In another embodiment, an expression vector comprises DNA which encodes the mutem. In another embodiment, a microorganism is transfected with such vector. The present invention provides a method of inhibiting clot formation m extracorporeal human blood which comprises adding to the blood an amount of an inactive recombinant mutem m an amount effective to inhibit clot formation m the subject but which does not significantly interfere with hemostasis when the blood is administered to a patient. In one embodiment, the patient has experienced an ischemic event . The present invention also provides a method of inhibiting thrombosis a human patient which comprises administering to the patient, or adding to blood which is to be administered to the patient, a Factor IXa compound m an amount which is effective to inhibit thrombosis but which does not significantly interfere with hemostasis m the patient. In one embodiment, the Factor IXa compound is a mutem. The present invention provides a method for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting the extracorporeal circulating blood with a Factor IXa compound m an amount effective to inhibit thrombosis m the patient. In one embodiment, the Factor IXa compound is an active site-blocked Factor Ixa. In another embodiment, the Factor IXa compound is Glu-Gly-Arg chloromethyl ketone- mactivated human factor IXa. In another embodiment, the effective amount comprises from about 0.1 μg/ml plasma to about 250 μg/ml plasma. In another embodiment, the effective amount comprises from about 0.5 μg/ml plasma to about 25 μg/ml plasma. In another embodiment, the effective amount comprises from about 0.7 μg/ml plasma to about 5 μg/ml plasma. In another embodiment, the patient is subjected to extracorporeal blood circulation during transplant surgery, abdominal surgery, vascular surger or cardiopulmonary bypass surgery. In another embodiment, the patient is a human being. The present invention provides a pharmaceutical composition which comprises an effective amount of a Factor IXa compound and a pharmaceutically acceptable carrier. In one embodiment, the carrier comprises a diluent. In another embodiment, the carrier comprises an appropriate adjuvant, a herpes virus, a liposome, a microencapsule, a polymer encapsulated cell or a retroviral vector. Ir another embodiment, the carrier is an aerosol, intravenous, oral or topical carrier. The present invention provides an assay to monitor antithrombic activity of a Factor IXa compound infused into circulation of a patient which comprises: (a) obtaining Factor IXa-deflcient plasma; (b) mixing the plasma from step (a) with diatomaceious earth and with plasma from the patient; (c) mcubatmg the mixture with a source of lipid and calcium chloride under conditions suitable for clot formation; and (d) measuring time necessary for clot formation m the incubate, thereby monitoring the antithrombic activity of the Factor IXa compound infused mto the circulation of the patient. The present invention also provides a method for evaluating the ability of an agent to inhibit an active site of a Factor IXa compound which comprises: (a) contacting the Factor IXa compound with the agent to form a protein complex; (b) cubatmg the protein complex under conditions suitable for clot formation; (c) measuring time necessary for clot formation m the incubate, and (d) comparing the time measured m step (c) with the time measured m the absence of the agent, thus evaluating the ability of the agent to inhibit the active site of the Factor IXa compound. In one embodiment, the agent comprises a peptide, a peptidomimetic, a nucleic acid or a small molecule. In another embodiment, the agent is an antibody or portion thereof. In another embodiment, the antibody is a monoclonal antibody or a polyclonal antibody. In another embodiment, the portion of the antibody comprises a Fab. The present invention provides for an agent obtained from such method which agent is capable of inhibiting the active site of Factor IX. The present invention provides a method for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting the extracorporeal circulating blood with an agent capable of inhibiting a step of the intrinsic pathway of coagulation m an amount effective to inhibit thrombosis m the patient. In another embodiment, the agent is an active site-blocked Factor XII compound. In another embodiment, the agent is an active site-blocked Factor XI compound. The present mvention provides an assay to determine the anticoagulant activity of a Factor IXa compound on a subject's blood which comprises: (a) mixing Factor IX deficient plasma, diatomaceous earth and plasma containing the Factor IXa compound derived from the subject's blood; (b) mcubatmg the resulting mixture from step (a) with an effective dose of a source of phospholipid and calcium chloride under conditions such that clot formation results; and (c) measuring the time necessary for clot formation m the incubate of step (b) so as to thereby determine the anticoagulant activity of the Factor IXa compound. In one embodiment, the assay further comprises comparing the time necessary for clot formation measured m step (c) with the time necessary for clot formation measured m the absence of the Factor IXa compound. In another embodiment, the subject is a human patient. In another embodiment, the Factor IXa compound is a functionally inactive form of Factor Ixa . In another embodiment, the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. The present invention provides a method for monitoring the anticoagulant activity of a Factor IXa compound which is being infused into the circulation of a subject's blood during surgery which comprises measuring the anticoagulant activity of the Factor IXa compound at different times during the surgery using the assay of claim 23 and comparing the activities so measured. In another embodiment, the subject is a human patient. In another embodiment, the surgery is cardiopulmonary by pass surgery. In another embodiment, wherein the Factor IXa compound is a functionally inactive form of Factor IXa. In another embodiment, the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. The present invention provides a method for inhibiting thrombosis a human patient which comprises administering to the patient, or adding to blood which is to be administered to the patemt, a Factor IXa compound m an amount which is effective to inhibit thrombosis but which does not significantly interfere with henostasis m the patient . The present invention provides a method of inhibiting clot formation m extracorporeal human blood which comprises adding to the blood an amount of a Factor IXa compound m an amount effective to inhibit clot formation but which does not significantly interfere with hemostasis when the blood is administered to a patient. In another embodiment, the Factor IXa compound is a mutem. In another embodiment, the mutem is an inactive recombinant Factor IXa. In another embodiment, the mutem is a Factor IX having a Ser365 to Ala substitution or a Factor IXa having a Serl85 to Ala substitution. One embodiment of the present invention is use of a Factor IXa compound for the manufacture of a pharmaceutical composition for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises admixing an amount of a Factor IXa compound effective to inhibit thrombosis m the patient and a pharmaceutically acceptable carrier. Another embodiment of the present invention is use of an agent capable of inhibiting a step of the intrinsic pathway of coagulation for the manufacture of a pharmaceutical composition for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises admixing an amount of the agent effective to inhibit thrombosis the patient. Another embodiment of the present invention is use of a Factor IXa compound for the manufacture of a pharmaceutical composition for inhibiting clot formation m extracorporeal human blood which comprises admixing to the blood an amount of a Factor IXa compound effective to inhibit clot formation but which does not significantly interfere with hemostasis when the blood is administered to the patient. One embodiment of the present invention is a peptidomimetic compound having the biological activity of a Factor IXa compound or a Glu-Gly-Arg chloromethyl ketone- inactivated human Factor IXa compound wherein the compound has a bond, a peptide backbone or an ammo acid component replaced with a suitable mimic. Examples of unnatural ammo acids which may be suitable ammo acid mimics include β-alanme, L- - ammo butyric acid, L-γ-ammo butyric acid, L- -ammo lsobutyric acid, L-e-ammo caproic acid, 7-ammo heptanoic acid, L-aspartic acid, L-glutamic acid, cyste e (acetam domethyl) , N-e-Boc-N-α-CBZ-L-lysme, N-e-Boc-N-α- Fmoc-L-lysme, L-methiomne sulfone, L-norleucme , L- norvalme, N- -Boc-N-δCBZ-L-ormthme, N-δ-Boc-N- -CBZ-L- ormthme, Boc-p-nitro-L-phenylalanme, Boc-hydroxyprolme, Boc-L-thioprolme . (Blondelle, et al . 1994; Pmilla, et al . 1995) . Also provided by the invention are pharmaceutical compositions comprising therapeutically effective amounts of polypeptide products of the invention together with suitable diluents, preservatives, solubilizers , emulsifiers, adjuvants and/or carriers. An "effective amount" as used herein refers to that amount which provides a therapeutic effect for a given condition and administration regimen. Such compositions are liquids or lyophilized or otherwise dried formulations and include diluents of various buffer content (e.g., Tπs-HCl., acetate, phosphate), pH and ionic strength, additives such as albumin or gelatin to prevent absorption to surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), solubilizmg agents (e.g., glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic acid, sodium etabisulfite) , preservatives (e.g., Thi erosal , benzyl alcohol, parabens) , bulking substances or tonicity modifiers (e.g., lactose, mannitol), covalent attachment of polymers such as polyethylene glycol to the protein, complexation with metal ions, or mcorporation of the material into or onto particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, etc, or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts . Such compositions will influence the physical state, solubility, stability, rate of m vivo release, and rate of m vivo clearance. The choice of compositions will depend on the physical and chemical properties of the protein having the activity of a Factor IXa compound. For example, a product which includes a controlled or sustained release composition may include formulation m lipophilic depots (e.g., fatty acids, waxes, oils) . Also comprehended by the invention are particulate compositions coated with polymers (e.g., poloxamers or poloxammes) and the compound coupled to antibodies directed against tissue-specific receptors, ligands or antigens or coupled to ligands of tissue-specific receptors. Other embodiments of the compositions of the invention incorporate particulate forms protective coatings, protease inhibitors or permeation enhancers for various routes of administration, including parenteral, pulmonary, nasal, oral, injection or infusion by intravenous, mtrapeπtoneal , mtracerebral , intramuscular, intraocular, mtraarteπal or mtralesional . The present invention incorporates U.S. Patent Nos . 5,446,128, 5,422,426 and 5,440,013 m their entireties as references which disclose the synthesis of peptidomimetic compounds and methods related thereto. The compounds of the present invention may be synthesized using these methods. The present invention provides for peptidomimetic compounds which have substantially the same three-dimensional structure as those compounds described herein. In addition to the compounds disclosed herein having naturally-occurring ammo acids with peptide or unnatural linkages, the present invention also provides for other structurally similar compounds such as polypeptide analogs with unnatural amino acids in the compound. Such compounds may be readily synthesized on a peptide synthesizer available from vendors such as Applied Biosystems, Dupont and Millipore. As used herein, the term "pharmaceutically acceptable carrier" encompasses any of the standard pharmaceutically accepted carriers, such as phosphate buffered saline solution, water, emulsions such as an oil/water emulsion or a triglyceπde emulsion, various types of wetting agents, tablets, coated tablets and capsules. An example of an acceptable triglyceride emulsion useful in intravenous and traperitoneal administration of the compounds is the triglyceride emulsion commercially known as Intralipid ® . Typically such carriers contain excipients such as starch, milk, sugar, certain types of clay, gelatin, stearic acid, talc, vegetable fats or oils, gums, glycols, or other known excipients. Such carriers may also include flavor and color additives or other ingredients. When administered, compounds are often cleared rapidly from the circulation and may therefore elicit relatively shortlived pharmacological activity. Consequently, frequent injections of relatively large doses of bioactive compounds may by required to sustain therapeutic efficacy. Compounds modified by the covalent attachment of water-soluble polymers such as polyethylene glycol, copoly ers of polyethylene glycol and polypropylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone or polyproline are known to exhibit substantially longer half-lives m blood following intravenous injection than do the corresponding unmodified compounds (Abuchowski et al . , 1981; Newmark et al . , 1982; and Katre et al . , 1987). Such modifications may also increase the compound's solubility in aqueous solution, eliminate aggregation, enhance the physical and chemical stability of the compound, and greatly reduce the immunogenicity and reactivity of the compound. As a result, the desired m vi vo biological activity may be achieved by the administration of such polymer-compound adducts less frequently or m lower doses than with the unmodified compound. Attachment of polyethylene glycol (PEG) to compounds is particularly useful because PEG has very low toxicity m mammals (Carpenter et al . , 1971). For example, a PEG adduct of adenosine deammase was approved m the United States for use m humans for the treatment of severe combined immunodeficiency syndrome. A second advantage afforded by the conjugation of PEG is that of effectively reducing the immunogenicity and antigemcity of heterologous compounds. For example, a PEG adduct of a human protein might be useful for the treatment of disease m other mammalian species without the risk of triggering a severe immune response. The compound of the present invention capable of alleviating symptoms of a cognitive disorder of memory or learning may be delivered m a microencapsulation device so as to reduce or prevent an host immune response against the compound or against cells which may produce the compound. The compound of the present invention may also be delivered microencapsulated m a membrane, such as a liposome. Polymers such as PEG may be conveniently attached to one or more reactive ammo acid residues m a protein such as the alpha-ammo group of the ammo terminal ammo acid, the epsilon ammo groups of lysme side chains, the sulfhydryl groups of cysteme side chains, the carboxyl groups of aspartyl and glutamyl side chains, the alpha-carboxyl group of the carboxy-terminal ammo acid, tyrosme side chains, or to activated derivatives of glycosyl chains attached to certain asparagme, serine or threonme residues. Numerous activated forms of PEG suitable for direct reaction with proteins have been described. Useful PEG reagents for reaction with protein ammo groups include active esters of carboxylic acid or carbonate derivatives, particularly those m which the leaving groups are N-hydroxysuccmimide, p- nitrophenol, lmidazole or l-hydroxy-2 -nιtrobenzene-4- sulfonate. PEG derivatives containing maleimido or haloacetyl groups are useful reagents for the modification of protein free sulfhydryl groups. Likewise, PEG reagents containing ammo hydrazme or hydrazide groups are useful for reaction with aldehydes generated by peπodate oxidation of carbohydrate groups m proteins. The present invention provides an assay to determine the anticoagulant activity of a Factor IXa compound on a subject's blood which comprises: (a) mixing Factor IX deficient plasma, diatomaceous earth and plasma containing the Factor IXa compound derived from the subject's blood; (b) mcubatmg the resulting mixture from step (a) with an effective dose of a source of phospholipid and calcium chloride under conditions such that clot formation results; and (c) measuring the time necessary for clot formation m the incubate of step (b) so as to thereby determine the anticoagulant activity of the Factor IXa compound. In one embodiment, the assay further comprises comparing the time necessary for clot formation measured m step (c) with the time necessary for clot formation measured m the absence of the Factor IXa compound. Another embodiment of the assay is wherein the subject is a human patient . Another embodiment of the assay is wherein the Factor IXa compound is a functionally inactive form of Factor Ixa . Another embodiment of the assay is wherein the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. One embodiment of the present invention is a method for monitoring the anticoagulant activity of a which is being Factor IXa compound infused into the circulation of a subject's blood during surgery which comprises measuring the anticoagulant activity of the Factor IXa compound at different times during the surgery using the assay and comparing the activities so measured. The subject may be a human patient. In one embodiment, the surgery is cardiopulmonary by pass surgery. In another embodiment, the Factor IXa compound is a functionally inactive form of Factor Ixa. In another embodiment, the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. This mvention is illustrated m the Experimental Detail section which follows. These sections are set forth to aid an understanding of the invention but are not intended to, and should not be construed to, limit any way the invention as set forth m the claims which follow thereafter. EXPERIMENTAL DETAILS Selective inhibition of the intrinsic pathway of coagulation is one possible way to avoid the use of heparin m extracorporeal circulation. Leaving intact the tissue factor-mediated extrinsic pathway of coagulation (initiated by tissue factor-VIIa) may obviate both adverse bleeding and potential prothrombotic side effects of heparin. These side effects are found m certain sensitized patients such as patients with heparin- induced thrombocytopenia . In these cases, heparin may be associated with a prothrombic situation. The coagulation Factor IX/lXa is a single chain vitamin K- dependent coagulation protein. Limited proteolysis of Factor IX results a cleavage product which is a two-chain serine protease, Factor IXa, which requires association with the cell surface and the cofactor Villa m order to express and exhibit procoagulant activity (Gurewich et al . , 1979; Gitel et al . , 1977; and Benedict et al . , 1991). Factor IXa has an important role m coagulation. When clotting is triggered m the mtravascular space, (i.e., thrombosis), it is initiated m the presence of low amounts of tissue factor. In this case, Factor IX becomes preferentially activated to Factor IXa, which then feeds into the rest of the procoagulant cascade, leading to fibrm formation. In contrast, activation of Factor X by tissue factor is less favored by at least an order of magnitude under these conditions. Extravascular coagulation, especially protective hemostasis, occurs m the presence of large amounts of tissue factor on mesenchymal cells and the role of Factor IXa is much less important when direct activation of Factor X occurs. Factor IXa is essentially bypassed. Thus, it may be hypothesized that inhibition of Factor IXa participation m coagulation could provide a selective means of anticoagulation by inhibiting mtravascular thrombosis without impairing extravascular hemostasis . Studies have shown rapid clearance of Factor IX from the intravascular space, the association of infused and endogenous Factor IX with the vessel wall (Thompson, 1986 and Stern et al . , 1987) and in vitro studies have demonstrated Factor IX binding to endothelium and platelets. (Heimark and Schwartz, 1983; Stern et al . , 1983; and Ahmad et al . , 1989) Studies have been performed to characterize the molecular basis of this coagulation protein-cell surface interaction. At the level of the ligand, the amino-terminal gamma-carboxyglutamic acid-containing domain of Factor IX has been shown to be essential for cell surface binding (Toomey et al . , 1992; Astermark et al . , 1991; Deπan et al . , 1989; and Ryan et al . , 1989). At the level of the cell surface site, previous studies have demonstrated that Factor IX binding involves a protease-sensitive polypeptide. This polypeptide m endothelial cell studies appears to have an Mr~150 kDa (Rimon et al . , 1987) and on platelets appears to have an Mr~250 kDa (London and Walsh, 1992) . Factor IXa may be capable of binding to cellular interaction sites in the vessel wall. It is possible that such sites may be a target for therapeutic intervention in certain thrombotic disorders such as cardiopulmonary bypass. Studies have demonstrated that Factor IX/lXa may contribute to thrombosis. It was found that Factor IXa resulted in formation of thrombi (Gurewich et al . , 1979) and that Factor IXa has potent thrombogenic properties in the Wessler stasis model of thrombosis (Gitel et al . , 1977) . Factor IXa participates in procoagulant pathways as a component of the Factor IXa-VIIIa-X activation complex (intrinsic pathway of coagulation) . Multiple studies in vitro demonstrated that use of active site-blocked IXa (dansyl-glutamyl-glycyl- arginyl Factor IXa, or IXai) prevented the assembly of IXa into this critical complex (Chomiak et al . , 1993; Thompson, 1986; Lollar and Fass, 1984; Stern et al . , 1985; and Ahmad et al . , 1989) . Studies have demonstrated the functionally- active site of Factor IXa (Astermark et al . , 1992 and Ahmad et al . , 1992). A role for active site-blocked Factor IXa (Factor IXai) has been demonstrated m preventing coronary artery thrombosis m a canine model m which thrombosis is initiated by the introduction of electric current. Extravascular hemostasis was secured, as no untoward bleeding was detectable m an mcisional wound model (Benedict et al . , 1991). Therefore, Factor IXai may be an ideal antithrombotic agent for use m cardiopulmonary bypass surgery. Specifically, the extrinsic pathway of coagulation would be unaffected and thus the patient would not be predisposed to excess bleeding. Furthermore, other limiting side effects of heparin would be precluded. In the studies described below, active site-blocked Factor IXa is shown to be a safe and effective antithrombotic agent m a canine model of coronary artery bypass and surgery. An aortotomy was performed m order to stimulate typical cardiac procedures. Pathways regulating control extravascular hemostasis appeared intact because significantly less bleeding was noted m dogs treated with Factor IXai compared with dogs treated with heparin. Furthermore, model studies of cardiopulmonary bypass utilizing active site-blocked Factor IXa further support the efficacy and safety of the present mvention. MATERIALS AND METHODS Factor IX/lXa was purified from human plasma according to previously-published methods (Benedict et al . , 1991 and Stern et al . , 1987) and inactivated using glu-gly-arg- chloromethylketone as described (Benedict et al . , 1991 and Lollar and Fass, 1984) . Purity of the reagents was then ascertained using SDS-PAGE and standard clotting assays (Benedict et al . , 1991 and Lollar and Fass, 1984). Animal studies Cardiopulmonary bypass m dogs (each dog weighing approximately 15 kgs) and baboons (each baboon weighing approximately 11 kgs) (Daly et al . , 1988 and Bernabei et al . , 1995) was instituted and maintained for one hour with cooling to 32°C. An aortotomy was performed m order to simulate cardiac surgery procedures. Animals were then weaned from bypass and blood loss was monitored for up to 3 hours . Animals received either heparin (at a standard dose of 300 IU/kg and protamme (2 mg/kg) reversal) or active site- blocked Factor IXa (at different doses as indicated below) . Multiple parameters were assessed m all animals (receiving either heparin or Factor IXai) as follows: 1. In order to test for evidence of fibrm deposition m the bypass circuitry, the cardiopulmonary bypass tubing and filters were removed at the end of bypass and subjected to analysis by scanning electron microscopy m order to detect possible evidence of fibrm deposition. 2. After sacrifice of the dogs or the baboons, necropsy was performed and the heart, lungs, liver and kidneys were removed. These organs were fixed m formalin and examined by Hemotoxylm & Eosm staining and lrnmunofluorescence for evidence of clot formation or fibrm deposition as well as for the presence of microemboli . 3. Routine hematologic analysis was performed prior to initiation of cardiopulmonary bypass and at every 30 minute interval during the cardiopulmonary bypass m order to determine hemoglobin levels, hematocπt levels, levels of platelets and fibrmogen, white blood cell count, prothrombm time, activated partial thromboplastm time and activated clotting time. 4. Continuous hemodynamic measurements were performed prior to, during and for up to 3 hours after the institution of cardiopulmonary bypass. 5. Blood loss was quantitated m the thoracic cavity by collecting all blood m the area during the cardiopulmonary bypass itself and for up to three hours after completion of cardiopulmonary bypass. 6. The extent of activation of coagulation m this model was determined m order to determine the contribution of thrombm generation and fibrmolysis, which occurs the setting of treatment with heparin, and the contribution of Factor IXai to coagulation. For the canine studies, measurement of thrombm-antI -thrombm complex (or TAT, commercially available from Behrmg Diagnostics, Boston MA) was utilized as a measure of thrombm generation. This assay was cross-reactive with dog plasma. TAT was measured m animals/group of heparin or Factor IX-ai treated dogs prior to institution of cardiopulmonary bypass/lXai treatment and every 30 minutes during cardiopulmonary bypass and every 60 minutes after cardiopulmonary bypass terminated until the animal was sacrificed at 3 hours. In baboon studies, TAT was measured as described herein and prothrombm fragment 1+2 (F 1+2 ; Behrmg Diagnostics) was measured. This assay cross-reacts with standards obtained from human plasma. 7. Markers of fibrmolysis were assessed to identify the extent to which excess fibrmolysis generated during treatment with heparin likely contributed to increased bleeding. These parameters were directly compared with those obtained dogs treated with Factor IXai . Levels of d-dimers were assessed at the same time points measured for TAT. RESULTS Example 1; Canine model of cardiopulmonary bypass Four dogs were treated with standard doses of heparin (300 IU/kg followed by protamine 2 mg/kg) and five dogs were treated with Factor IXai. In dogs (3 total) treated with 460 μg/kg (single intravenous infusion just prior to the initiation of cardiopulmonary bypass) , there was no evidence of excess pressure in the cardiopulmonary bypass circuit or gross clot formation m the tubing. These results were similar to those observed in the dogs treated with heparin. Similarly, systemic hemodynamic profiles were similar in both groups throughout the procedure suggestive of the absence of clinically-relevant thromboses within the bypass circuitry. Blood loss in the thoracic cavity was significantly less in the group receiving Factor IXai compared with the dogs receiving heparin (Figure 1) . After four hours of observation (one hour during cardiopulmonary bypass itself) and three hours following the termination of cardiopulmonary bypass, dogs receiving Factor IXai accumulated 600 ml of blood within the thoracic cavity. This was m marked contrast to dogs receiving standard doses of heparin in which 1000 ml of blood was quantitated m the thoracic cavity (p<0.01) . Consistent with these data and the hypothesis that selective inhibition of the intrinsic pathway of coagulation would leave unaffected the tissue factor-mediated extrinsic pathway of coagulation, dogs that received Factor IXai were observed to have hemostatic clot along the cut surface of the sternum and in surgical tissue planes. However, the dogs treated with heparin did not have these hemostatic markers . In order to detect pathological quantities of deposited fibr within the bypass circuits, the tubing and filters were immediately removed after cardiopulmonary bypass and analyzed by scanning electron microscopy. As shown m Figure 2, cardiopulmonary bypass performed with Factor IXai resulted m similar amounts of fibrm deposition (left panel) compared with cardiopulmonary bypass performed with the use of traditional heparin (opposite) . The amount of fibrm deposition m both cases was clinically-mapparent . In addition to microscopic examination of the bypass material, organs removed at necropsy were examined by microscopy with Hematoxylm/eosm staining. These studies revealed the absence of fibrm deposition and microemboli m the liver, lungs, kidneys and myocardium as shown m Figure 3. Analysis of blood samples revealed similar dilutional decreases m hematocrit, platelet count and fibrmogen levels m both groups of animals treated with either Factor IXai or heparin (Figure 4) . As demonstrated m Figure 4, Prothrombm time (PT) remained at about the level of the control the Factor IXai -treated group (7.9 + 0.1 sec) and activated Partial thromboplastm time (PTT) was mildly elevated (30.4 ± 11 sees). As expected, heparm-treated dogs had significant elevation of PT and PTT (11.9±0.6 sees and >90 sees, respectively). ACT (Activated clotting time) used m cardiopulmonary bypass to quickly assess the level of anticoagulation with heparin (ideal > 480 sees m human subjects) was >400 sees m the heparm-treated group, but unchanged m the Factor IXai- treated group. In order to determine the optimal dose of Factor IXai that was necessary to safely prevent thrombosis while securing extravascular hemostasis, different doses of Factor IXai were utilized m the canine model. These data revealed that a dose of 600 μg/kg (one dog) produced no evidence of clotting m the bypass circuitry. However, there was evidence of increased bleeding m this group (900 ml of blood m dogs treated with Factor IXai) compared with dogs that received a lower dose (460 μg/kg) of Factor IXai. A third group of dogs (1100 ml of blood m dogs) treated with heparin also had evidence of increased bleeding. These data suggested that the 600 μg/ml dose of Factor IXai was excessive since the lower dose (at least 460 μg) was successful . In order to determine the minimal dose of Factor IXai needed to prevent clotting m the bypass circuitry, one dog was treated with Factor IXai at a dose of 300 μg/kg. This experiment demonstrated that there was no evidence of improper clotting of the bypass circuitry. In addition, blood loss m the thoracic cavity was 600 ml of blood compared with 1100 ml of blood the thoracic cavity of dogs treated with standard doses of heparin. As indicated above, there was an inability to assess the effectiveness of anticoagulation via measurement of activated clotting time dogs treated with Factor IXa. Thus, a clotting assay was developed m order to determine the functional effectiveness of Factor IXai rapidly and reproducibly, thereby providing a means to assess the level of anticoagulation frequently during cardiopulmonary bypass. It is necessary to be able to frequently determine the ability of blood of a human patient to coagulate throughout the performance of cardiopulmonary bypass surgery or any majory surgery. The sensitivity of the assay was dependent on the use of Factor IX-deficient plasma and an optimized dose of cephalin (source of phospholipid) as shown Figure 5. Comparison of plasma from dogs treated with Factor IXai compared with control dog plasma revealed a four-fold increase m clotting time. This assay therefore provided a means of rapidly determining the extent of anticoagulation m animals treated with Factor IXai. Further studies may be performed to determine the desired extent of increased clotting time m this assay m order to achieve maximal antithrombotic effects, while maintaining intact the pathways of extravascular hemostasis. Example 2 : Baboon model of cardiopulmonary bypass Studies m baboon models of cardiopulmonary bypass are effective and predictive for future testing of active site- blocked Factor IXa m human subjects. Adequate pre- clmical data m primates is expected to more closely predict expected results m humans. Experiments have been performed m baboons. In one experiment, a baboon received Factor IXai (460 μg/kg) and a second baboon received heparm/protamme (300 IU/hepar followed by protamme [2 mg/kg] ) . The data from these trials revealed that there was no evidence of pathologic clotting m the bypass circuitry observed m either group of animals. Similar to the results from the case of dogs treated with Factor IXai, there was evidence of significantly decreased blood loss m the thoracic cavity of baboons treated with Factor IXa (320 ml of blood) compared with baboons treated with standard doses of heparin (600 ml of blood) . Preliminary hematologic analysis m the baboons shows that there are similar dilutional decreases m hematocrit, white blood cell counts and platelets as seen m the dog trials. Other studies were completed to investigate the effects of Factor IXai m baboons and to characterize the hemostatic findings. DISCUSSION The use of heparin has been associated with multiple side effects m certain patients undergoing cardiopulmonary bypass. The side effects include excessive bleeding (at least m part due to excess fibrmolysis) , heparin resistance (potentially requiring use of anti-thrombm III to achieve desire heparin effects) , heparin- induced thrombocytopenia (with potential for either excess bleeding or clotting m the arterial and/or venous system) and need for reversal with protamme . Data from studies of the canine and baboon models of cardiopulmonary bypass suggest that m patients m whom heparin is relatively contramdicated, the use of active site-blocked Factor IXa or (Factor IXai) as an antithrombotic agent may be useful to prevent pathologic clotting m the circuitry. This clotting may be lessened by the inhibition of the intrinsic pathway of coagulation. However, securing the functional responsiveness of the extrinsic (extravascular) pathway of coagulation is critical m minimizing excess blood loss due to cut vessels of the sternum, etc. Furthermore, obviating the use of heparin will be useful to minimize the excessive fibrmolysis generated by its use at doses required to maintain an activated clotting time > 480 sees during cardiopulmonary bypass. The use of Factor IXai has been shown to be a safe means of inhibition of thrombosis during cardiopulmonary bypass and a range of other cardiac and surgical procedures . In canine and baboon cardiopulmonary bypass models, the selective inhibition of the intrinsic/contact system of coagulation has been demonstrated. Maintenance of the extrmsic/tissue factor mediated pathway allows for the successful maintenance of patency of the cardiopulmonary bypass circuit while allowing extravascular hemostasis. The possible applications of this therapeutic intervention extend well beyond the cardiopulmonary bypass setting. For example, extracorporeal life support with extracorporeal membrane oxygenation (ECMO) (Magovern, 1995) is being used more frequently to support adult patients who develop cardiogenic shock. Studies have demonstrated that this mtervention provides excellent oxygenation and hemodynamic support m this critically ill population. However, the morbidity and mortality associated with this procedure remains high. Central to the life-threatening complications associated with extracorporeal membrane oxygenation is the unavoidable need for systemic heparmization with its attendant hemorrhagic complications (Muehrcke et al . , 1995). Heparm-bonded circuits have provided an alternative to full heparmization with some success, but this model still carries the significant risk of bleeding and thrombosis (Perchmisky et al . , 1995 and Atkinson et al . , 1992). Selective mtravascular anticoagulation with Factor IXai would logically be an ideal alternative to traditional heparmization . Baboons may be maintained on an extracorporeal membrane oxygenation circuit for up to one week with anticoagulation initiated by Factor IXai, m lieu of heparin. Similarly, active site-blocked Factor IXa may also be useful m any setting requiring the contact of blood with extracorporeal circuitry, such as plasmapheresis , renal hemodialysis, continuous anteπo-venus hemoflltration (CAVH) , veno-venus hemoflltration (CWH) , extracorporeal circulatory membrane oxygenation (ECMO) , brain surgery, vascular surgery, abdominal surgery, transplant surgery, any procedure which systemic anticoagulation is routinely required, and any procedure which a patient requires a mechanical circulatory assistance, ventricle assist device, artificial heart, left/right ventricle assist device or a similar biomedical device. Another potential indication for the use of active site- blocked Factor IXa as an anticoagulant involves the surgical intervention of mtracramal aneurysms . Surgery for aneurysm correction must be performed without the use of systematic anticoagulation because the risk of hemorrhagic complications associated with rhe use of heparin during this procedure are unacceptable. A "watershed" portion of the bram matter immediately adjacent to the aneurysm is therefore sacrificed with the surgical clipping of the aneurysm as tributary vessels thrombose and thus mfarct adjacent bram tissue. This procedure may be performed with local infusion of Factor IXai to allow mtravascular anticoagulation, thus maintaining blood flow through tributaries of the clipped vessel to the bram matter around the aneurysm. Meanwhile, extravascular/tissue factor mediated hemostasis would ensure that a hemorrhagic infarction did not occur. A murme model of stroke may be studied to determine if local infusion of Factor IXai will limit the "watershed" region around a clipped cerebral vessel while preventing hemorrhage. Example 3 : Applications of Active site-blocked Factor IXa (IXai) as an antithrombotic agent in cardiopulmonary bypass One of the potential uses of Factor IXai might be vascular surgery. At the present time, when vascular surgery is performed, a polytetrafluoroethylene (PTFE) graft is used to reconstitute an injured native blood vessel. At the time of surgery, heparin therapy is indicated m order to prevent thrombosis at the site of the graft. One of the consequences of this intervention, however, is increased bleeding at the needle hole sites (where graft connects with the native vessel) . This may increase duration of anesthesia since these bleeding sites must be secured and hemostasis confirmed prior to conclusion of the operation. These interventions, by increasing duration of anesthesia, may actually increase morbidity, especially m predisposed patients . Studies have been performed with a standing protocol m which a graft (PTFE) is created and then tests of the effects of certain interventions are performed on the graft. One intervention, use of Factor IXai, resulted m no evidence of thrombosis and greatly minimized bleeding about the sites of the needle holes. To follow are results of data : Studies to test the role of Factor IXai as a novel antithrombotic agent this setting have been performed m a rabbit model of vascular repair. An mfrarenal abdominal aortotomy was performed and reconstructed with a Polytetrafluoroethylene (PTFE) graft (2X6 mm). Methods : A PTFE graft was placed as above and preweighed gauze pad placed near the needle holes. IXai was administered as an intravenous bolus dose at the time of the graft placement. Time to bleeding was observed and weight of gauze after bleeding ceased determined to quantitate the amount of blood loss : Results of studies; Factor IXai: 360 microgram/kg: Time to stop bleeding: 3 minutes Weight of gauze after tare: 5.54 grams Factor IXai: 260 microgram/kg: Time to stop bleeding; 3 minutes Weight of gauze after tare : 2.09 grams Heparin: 25 U/kg: Time to stop bleeding: 4 minutes Weight of gauze after tare : 8 .61 grams Even at the lowest effective dose of Factor IXai, there was no evidence of thrombus m the PTFE graft. These results suggest that use of Factor IXai may be beneficial m vascular surgery by preventing thrombosis peπ-PTFE graft and, yet, at the same time, securing vascular hemostasis. Regarding interpretation of the results of the PTFE graft placement m rabbits with either Factor IXai or standard heparin therapy: The effectiveness of Factor IXai preventing mtravascular thrombosis at the site of PTFE graft placement is dose dependent. At the lowest dose studied to date, 260 microgram/kg of IXai, there is NO evidence of thrombosis m the setting of minimal bleeding complication - which is clearly superior to the bleeding observed using the standard dose of heparin. However, as the dose of Factor IXai approaches 360 microgram/kg, there is similar prevention of thrombosis, but increased bleeding. These data suggest that at higher doses of Factor IXai, the anticoagulant effect is more potent than the selective antithrombotic effect at lower doses (of Factor IXai) . Use of Active Site-blocked Factor Ixa(IXai in peripheral vascular surgery) . A. Rabbit aortotomy To date, 37 rabbits have been completed using the New Zealand rabbit model. 21 rabbits have been treated with Factor IXai 300 microgram/kg and 16 rabbits received standard doses of heparin, m this case (50 U/kg) . The results of these studies show that the blood loss at the aortotomy suture site was substantially less m the rabbits treated with Factor IXai vs. heparin : Factor IXai 2.7+2.5 gms vs heparm 6.9+4.4 gms p<0.05 Similarly, the time to achieve hemostasis was decreased m animals treated with Factor IXai: Factor IXai 120±38 seconds heparin 176+46 seconds p<0.05 (See Figures 6A and 6B) . 12 rabbits were allowed to survive up to 2 months postoperatively . Of these, 6 had been treated with Factor IXai for aortotomy/PTFE graft placement and 6 were treated with heparin. At sacrifice, 100% of the grafts were patent, with no evidence of mtimal hyperplasia by hematoxylm and eosm staining, and no evidence of systemic thrombm generation, as measured by Thrombm-antlthrombm III complex (TAT) . B. Canine carotid 2x8 mm PTFE patch repair of Right carotid aortotomy To date, 14 dogs have undergone this procedure, 7 dogs received Factor IXai (300 microgram/kg) and 7 dogs received heparin (50 IU/kg) . The data demonstrate that there is substantially decreased blood loss m dogs treated with Factor IXai vs. those treated with heparin : Factor IXai 20.8±12.9 gms heparin 39.1±5.5 gms p<0.05 In addition, time to achieve hemostasis was lower the animals treated with Factor IXai: Factor IXai 162+34 seconds vs heparin 228+17 seconds p<0.05 See Figures 6A and 6B . One of the important concerns such a model is whether or not the agent increased the incidence of graft occlusion or mtimal hyperplasia. Similar to the results obtained m rabbits, ultrasound at one month revealed all grafts to be patent; there were no difference observed m dogs treated with Factor IXai or heparm. Also, serial measurement of TAT revealed that there was no evidence of thrombm generation from one week postoperatively, through two months postoperatively . II. Modified Cephalin Clotting Time (MCCT) MCCT has also been characterized. A. Limit of detection. By performing a series of dilutions of added Factor IXai and determining the clotting time as otherwise noted m the original application, it has been determined that the limit of detection of Factor IXai m the assay is 0.4μg/ml. See Figure 7. B. Determination of MCCT after single dose of Factor IXai in canine cardiopulmonary bypass . After a single clinically-effective dose of Factor IXai (460μg/kg) m canine CPB, the MCCT rises to 80 sees, and is maintained at the level through at least 1.5 hours of cardiopulmonary bypass (CPB) . Post -CPB, MCCT remained elevated through at least 2 hours, and normalized by 3 hours after initiation of CPB. These data suggested that m uncomplicated CPB (at least 1.5 hr) , one dose of Factor IXai is likely sufficient. See Figure 8. Summary: Active-site Blocked Factor IXa (IX ai) : A Novel Selective Anticoagulant for Use in Cardiopulmonary Bypass The use of heparin m cardiopulmonary bypass (CPB) prevents mtravascular/extracorporeal circulation thrombosis but also interferes with protective extravascular hemostasis due to its multiple sites of blockade the coagulation cascade and its adverse effect on platelet function. In addition, the presence of heparin-associated antibodies can result in paradoxical thrombosis with severe consequences. Furthermore, protamine reversal may be accompanied by allergy, hemodynamic instability, or pathologic thrombosis. An alternative to heparin would have great clinical implications. The prothrombotic stimuli associated with CPB are complex with early activation likely to occur through activation of the contact (intrinsic) pathway. Later in CPB(>2 hrs) , activated monocytes on the bypass circuitry express low levels of tissue factor (TF), resulting m activation of Factor IX by Factor VIIa-TF pathway. The role of Factor IXa (IXa) was studied since it is activated upon stimulation of the contact/intrinsic system; as well as m the presence of low amounts of TF, such as that observed early in CPB or later in CPB with low numbers of activated monocytes on the bypass circuitry. It was hypothesized that blockade of IXa would inhibit intravascular/extracorporeal thrombosis while preserving extravascular hemostasis (where high amounts of Factor VIIa-TF will directly activate Factor X and promote clot formation) . A baboon model of CPB was established using active site-blocked IXa, (dansyl-glutamyl , glycyl-argmyl chlormethylketone IXai) , a competitive inhibitor of the assembly of IXa in the Factor X activation complex, as an anticoagulant. Standard single-stage CPB was performed in 10 baboons; 6 received Ixai(300 to 600 μg/kg) /no reversal and 4 received heparin(300 IU/kg) /protamine (2 mg/kg) . CPB was maintained for one hour with cooling to 32°c. An aortotomy was performed and repaired. Baboons were weaned from CPB and blood loss monitored for 3 hours postoperatively. At doses from 400 μg/kg to 460 μg/kg systemic hemodynamic profiles were similar in both groups throughout the procedure, with no evidence of excess pressures m the CPB circuit or gross clot formation in the tubing. Scanning electron microscopy of arterial filters revealed no differences in fibrin deposition in the animals treated with IXai vs. those treated with heparin. At necropsy, there was no evidence of pathologic clot formation or bleeding withm the abdominal or thoracic cavities nor fibr deposition/microemboli m the heart, lung, liver or kidney m animals treated with IXai or heparin. In contrast to heparin, animals treated with IXai (at doses < 600 μg/kg) had hemostatic clot along the cut surface of the sternum and m surgical tissue planes during the entire procedure. Blood samples revealed similar dilutional decreases m hematocπt, platelet count, and fibrmogen levels. Prothrombm time and partial thromboplastm time were only slightly elevated m baboons receiving IXai (13.2 ± 1.2 and 27.1 ± 4.5, respectively). Similarly, activated clotting time (ACT) was unchanged m the IXai group. In order to rapidly and reproducibly measure the effective antithrombotic level of IXai m plasma, a cephalin- IXa-based clotting assay was developed (modified cephalin clotting time; or MCCT) . At 1:32 dilution of cephalin, normal test plasma had clotting time of 21 sees. After a single infusion of IXai (400 to 460 μg/kg) , MCCT rose to 80 sees and remained at that level for up to 2 hours postoperatively; thereby potentially protecting bypass graft patency into the immediate postoperative period. Taken together, these data suggest that IXai may be a safe and effective alternative anticoagulant for selected patients undergoing CPB, preventing mtravscular-extracorporeal circulation thrombosis while preserving extravascular hemostasis . References Ahmad, S., et al . (1989) J. Clin. Invest. 84:824-828. Ahmad, S., et al . (1989) J. Biol. Chem. 264:3244-3251. Ahmad, S.D., et al . (1992) J. Biol. Chem. 267 (12) :8571-6. Astermark, J., et al . (1991) J. Biol. Chem. 266:2430-2437. Astermark, J. , et al . (1992) J. Of Biol. Chem. 267(5): 3249- 56. Atkinson, J.B., et al. (1992) J. Of Pediatric Surgery 27 (9) :1197-1198. Benedict, C, et al . (1991) J. Clin. Invest. 88:1760-1765. Bernabei, A., et al . (1995) J. Thorac. & Cardiovasc. Surg . 109(4) :765-71. Berntorp, E., et al . (1993) Thrombosis and Haemostasis 70(5) :768-773. Blondelle et al . (1994). Brister, S.J., et al . (1994) Thrombosis & Haemostasis 71:468-473. Chomiak, P.N., et al . (1993) Circ. 88:407-412. Daly, R.C., et al . (1988) J. Thorac. & Cardiovasc. Surg. 96 (1) :19-29. DeAnda, A., et al . (1994) Ann. Thorac. Surg. 58:344-350. Derian, C, et al . (1989) Biol. Chem. 264:6615-6618. Edmunds, L. H. (1993) J. Card. Surg. 8:404-410. Edmunds, L. H. (1995) Adv. In Cardiac Surg. 6:131-167. Freedman, S.J., et al . (1995) J. Biol. Chem. 270 (14) : 7980- 7987. Furie, B.C. and Furie, B. (1995) Thrombosis and Haemostasis 74(1) :274-277. Gitel, S., et al . (1977) Proc. Natl. Acad. Sci. 74:3028- 3032. Gravlee, G.P., et al . (1990) J. Thorac, Cardiovasc. Surg. 99:518-527. Gurewich, V., et al . (1979) Thromb. Res. 14:931-940. Heimark, R. , and Schwartz, S. (1983) Biochem. Biophys. Res. Commun. 111:723-731. Iberti, T.J., et al . (1994) Neurosurgery 34 (3) : 389-395. Keating, M.T. and Sanguinetti, M.C. (1996) Science 272:681- 685. Kuwabara, K. , et al . (1995) J. Biol. Chem. 270 (14) : 8179- 8187. Lollar, P., and Fass, D. (1984) Arch. Biohcem. Biophys. 233:673-682. London, F. and P. Walsh. (1992) Circulation 86:(suppl 1), 1855. Magovern, G.L. (1995) Advances in Cardiac Surgery 6:169- 193. Miyata, T., et al . (1994) British J. of Haematology 88:156- 165. Muehrcke, D.D., et al . (1995) J. Thorac. Cardiovasc. Surg. 110:843-51. Perchinisky, M.J., et al.(1995) Am. J. Surg. 169:488-491. Rimon, S., et al . (1987) J. Biol. Chem. 262:6023-6031. Ryan, J., et al . (1989) J. Biol. Chem. 264:20283-20287. Santagostino, E., et al . (1994) Thrombosis and Haemostasis 71(6) :737-740. Stern, D., et al . (1983) Proc. Natl. Acad. Sci. 81:913-917. Stern, D., et al . (1987) Br. J. Hematol . 66:227-232. Stern, D., et al . (1985) J. Biol. Chem. 260:6717-6722. Thompson, A. (1986) Blood 67:565-572. Toomey, J., et al . (1991) J. Biol. Chem. 266:19198-19202. 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Claims: What is claimed is: 1. A method for inhibiting thrombosis m a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting or admixing with the extracorporeal circulating blood with a Factor IXa compound m an amount effective to inhibit thrombosis m the patient . 2. The method of claim 1, wherein the Factor IXa compound is an active site-blocked Factor IXa. 3. The method of claim 1, wherein the Factor IXa compound is Glu-Gly-Arg chloromethyl ketone-mactivated human factor IXa. 4. The method of claim 1, wherein the effective amount comprises from about 0.01 μg/ml plasma to about 250 μg/ml plasma. 5. The method of claim 1, wherein the effective amount comprises from about 0.05 μg/ml plasma to about 25 μg/ml plasma. 6. The method of claim 1, wherein the effective amount comprises from about 0.1 μg/ml plasma to about 5 μg/ml plasma . 7. The method of claim 1, wherein the patient is subjected to extracorporeal blood circulation during transplant surgery, abdominal surgery, vascular surger or cardiopulmonary bypass surgery. 8. The method of claim 1, wherein the patient is a human being. 9. A pharmaceutical composition which comprises an effective amount of a Factor IXa compound and a pharmaceutically acceptable carrier. 10. The pharmaceutical composition of claim 9, wherein the carrier comprises a diluent . 11. The pharmaceutical composition of claim 9, wherein the carrier comprises an appropriate adjuvant, a herpes virus, a liposome, a microencapsule, a polymer encapsulated cell or a retroviral vector. 12 The pharmaceutical composition of claim 9, wherein the carrier is an aerosol, intravenous, oral or topical carrier . 13. An assay to monitor antithrombic activity of a Factor IXa compound infused into circulation of a patient which comprises : (a) obtaining Factor IXa-deflcient plasma; (b) mixing the plasma from step (a) with diatomaceious earth and with plasma from the patient; (c) incubating the mixture with a source of lipid and calcium chloride under conditions suitable for clot formation; and (d) measuring time necessary for clot formation m the incubate, thereby monitoring the antithrombic activity of the Factor IXa compound infused into the circulation of the patient. 14. A method for evaluating the ability of an agent to inhibit an active site of a Factor IXa compound which comprises : (a) contacting the Factor IXa compound with the agent to form a protein complex; (b) mcubatmg the protein complex under conditions suitable for clot formation; (c) measuring time necessary for clot formation m the incubate, and (d) comparing the time measured m step (c) with the time measured m the absence of the agent, thus evaluating the ability of the agent to inhibit the active site of the Factor IXa compound. 15. The method of claim 14, wherein the agent comprises a peptide, a peptidomimetic, a nucleic acid or a small molecule . 16. The method of claim 14, wherein the agent is an antibody or portion thereof. 17. The method of claim 16, wherein the antibody is a monoclonal antibody or a polyclonal antibody. 18. The method of claim 16, wherein the portion of the antibody comprises a Fab. 19. An agent obtained from the method of claim 14, which agent is capable of inhibiting the active site of Factor IX. 20. A method for inhibiting thrombosis a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting or admixing with the extracorporeal circulating blood with an agent capable of inhibiting a step of the intrinsic pathway of coagulation m an amount effective to inhibit thrombosis m the patient. 21. The method of claim 20, wherein the agent is an active site-blocked Factor XII compound. 22. The method of claim 20, wherein the agent is an active site-blocked Factor XI compound. 23. An assay to determine the anticoagulant activity of a Factor IXa compound on a subject's blood which comprises : (a) mixing Factor IX deficient plasma, diatomaceous earth and plasma containing the Factor IXa compound derived from the subject's blood; (b) mcubatmg the resulting mixture from step (a) with an effective dose of a source of phospholipid and calcium chloride under conditions such that clot formation results; and (c) measuring the time necessary for clot formation m the incubate of step (b) so as to thereby determine the anticoagulant activity of the Factor IXa compound. 24. An assay of claim 23, further comprising comparing the time necessary for clot formation measured m step (c) with the time necessary for clot formation measured m the absence of the Factor IXa compound. 25. An assay of claim 23, wherein the subject is a human patient . 26. An assay of claim 23, wherein the Factor IXa compound is a functionally inactive form of Factor IXa. 27. An assay of claim 26, wherein the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. 28 A method for monitoring the anticoagulant activity of a Factor IXa compound which is being infused into the circulation of a subject's blood during surgery which comprises measuring the anticoagulant activity of the Factor IXa compound at different times during the surgery using the assay of claim 23 and comparing the activities so measured. 29. The method of claim 28, wherein the subject is a human patient . 30. The method of claim 28, wherein the surgery is cardiopulmonary by pass surgery. 31. The method of claim 28, wherein the Factor IXa compound is a functionally inactive form of Factor IXa. 32. The method of claim 31, wherein the functionally inactive form of Factor IXa is a Factor IXa m which the active serine ammo acid site has been altered. 33. A method for inhibiting thrombosis m a human patient which comprises administering to the patient, or adding to blood which is to be administered to the patient, a Factor IXa compound m an amount which is effective to inhibit thrombosis but which does not significantly interfere with henostasis the patient. 34. A method of inhibiting clot formation m extracorporeal human blood which comprises adding to the blood an amount of a Factor IXa compound m an amount effective to inhibit clot formation but which does not significantly interfere with hemostasis when the blood is administered to a patient. 35. The method of claim 33 or 34, wherein the Factor IXa compound is a mutein. 36. The method of claim 35, wherein the mutein is an inactive recombinant Factor IXa. 37. The method of claim 35, wherein the mutein is a Factor IX having a Ser365 to Ala substitution or a Factor IXa having a Serl85 to Ala substitution.

Inventors: ROSE ERIC, SCHMIDT ANN MARIE, SPANIER TALIA, STERN DAVID, ROSE, ERIC, SCHMIDT, ANN, MARIE, SPANIER, TALIA, STERN, DAVID
Applicants: UNIV COLUMBIA, THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
Application year: 1999
Main classification: A61B19
IPC codes: A61K38, A61M1, C12Q1, G01N33