Preparation and composition of inter-alpha inhibitor proteins from human plasma for therapeutic use

Abstract

The invention relates to Inter-alpha inhibitor proteins (IαIp). The invention further relates to processes for purification of IαIp compositions and their use for treatment of human diseases such as sepsis and septic shock, rheumatoid arthritis, cancer and infectious diseases.

What is claimed is: 
     
       1. A process for producing a blood plasma-derived IαIp composition comprising a mixture of inter-alpha inhibitor protein (IαI) and pre-alpha protein (PαI), wherein the IαI and the PαI are present in said mixture in a physiological proportion, the process comprising:
 isolating from blood plasma a plasma fraction containing IαI and PαI, wherein the IαI and PαI are present in a physiological proportion; and 
 purifying the plasma fraction to obtain an IαIp composition with a purity of IαIp ranging from about 85% to about 100% pure, wherein the purifying comprises hydroxylapatite chromatography. 
 
     
     
       2. The process of  claim 1 , wherein the isolating comprises solid phase extraction or chromatographing blood plasma. 
     
     
       3. The process of  claim 1 , wherein the plasma fraction comprises a side fraction obtained from the purification of clotting factor IX or from the purification of a prothrombin complex concentrate. 
     
     
       4. The process of  claim 1 , wherein the plasma fraction is isolated as a cryosupernatant resulting from cryoprecipitation of blood plasma. 
     
     
       5. The process of  claim 1 , wherein the plasma fraction is cryo-poor plasma. 
     
     
       6. The process of  claim 1 , wherein the plasma fraction is human, primate, bovine, porcine, feline, or canine. 
     
     
       7. The process of  claim 1 , further comprising obtaining blood, obtaining blood plasma, obtaining a side fraction obtained from the purification of clotting factor IX, obtaining a side fraction from the purification of a prothrombin complex concentrate, obtaining a cryosupernatant resulting from cryoprecipitation of blood plasma or obtaining cryo-poor plasma. 
     
     
       8. The process of  claim 1 , wherein the purifying further comprises affinity chromatography. 
     
     
       9. The process of  claim 1 , wherein the IαI and PαI present in the plasma fraction have an apparent molecular weight of between about 60,000 to about 280,000 kDa. 
     
     
       10. The process of  claim 1 , further comprising:
 purifying the plasma fraction; virus inactivating the plasma fraction and/or the purified IαIp; the addition of stabilizers; pasteurization of the purified IαIp;   or anion-exchange chromatography of the purified IαIp.   
     
     
       11. The process of  claim 10 , wherein the further purifying the plasma fraction is by passing through heparin affinity column and collecting the flow through (unbound) fraction; the virus inactivating is by a solvent/detergent treatment or thermal inactivation; and the anion-exchange chromatography of the purified IαIp is diethylaminoethyl (DEAE) Sepharose. 
     
     
       12. The process of  claim 11 , wherein the thermal inactivation comprises pasteurization at a temperature of between about 55 to about 65° C. or dry heat at 70 to 120° C. omega-amino acids, sugar, or combinations thereof. 
     
     
       13. A composition of IαIp comprising a mixture of inter-alpha inhibitor protein (IαI) and pre-alpha protein (PαI), wherein the IαI and the PαI are present in said mixture in a physiological proportion and: have a high trypsin inhibitory specific activity between about 1000 to about 2000 IU/mg; have a half life of greater than one hour; comprise a light chain of inter-alpha inhibitor protein associated with at least one of three heavy chains H1, H2 and H3; or comprise a light chain of inter-alpha inhibitor protein associated with at least one of three heavy chains H1, H2, H3 and H4. 
     
     
       14. The composition of  claim 13 , wherein trypsin inhibitory specific activity is between about 1400 to about 2000 IU/mg. 
     
     
       15. The composition of  claim 13 , wherein the IαIp composition has a half life of at least 5 hours. 
     
     
       16. The composition of  claim 13 , wherein the IαIp composition has a half life of at least 10 hours. 
     
     
       17. A composition of IαIp comprising a mixture of inter-alpha inhibitor protein (IαI) and pre-alpha protein (PαI), wherein the IαI and the PαI are present in said mixture in a physiological proportion, said composition having been prepared by the process according to  claim 1 . 
     
     
       18. The composition of  claim 17 , further comprising an additional therapeutic agent. 
     
     
       19. The composition of  claim 18 , wherein the additional therapeutic agent is an anti-inflammatory agent, an anti-coagulant or an immunomodulator. 
     
     
       20. A pharmaceutical composition comprising a therapeutically effective amount of the composition of  claim 17 , and a pharmaceutically acceptable carrier. 
     
     
       21. A method of treating an inflammation related disorder comprising administering to a subject in need thereof a therapeutically effective amount of the composition of  claim 17 , wherein the inflammation related disorder is selected from an acute inflammatory disease, sepsis, septic shock, rheumatoid arthritis, meningitis, Crohn's Disease, chronic obstructed pulmonary disease and rhinitis. 
     
     
       22. The method of  claim 21 , wherein the IαIp is administered as a tablet, capsule, or injectables. 
     
     
       23. A method of treating an acute inflammatory disease, sepsis, septic shock, rheumatoid arthritis, meningitis, Crohn's Disease, chronic obstructed pulmonary disease and rhinitis in a subject, comprising:
 (a) determining the pre-treatment level of one or more of the following levels in a subject:   (i) the level of IαI;   (ii) the level of PαI;   (iii) the level of IαIp;   (iv) the level of H3;   (v) the level of H4;   (vi) the level of H1;   (vii) the level of H2; and   (viii) the level of LC; and   (b) administering a therapeutically effective amount of the composition of  claim 17  to the subject.   
     
     
       24. A method of monitoring the progress of a subject being treated with an IαIp therapy, comprising:
 (a) determining the pre-treatment level of one or more of the following levels, in a subject: 
 (i) the level of IαI; 
 (ii) the level of PαI; 
 (iii) the level of IαIp; 
 (iv) the level of H3; 
 (v) the level of H4; 
 (vi) the level of H1; 
 (vii) the level of H2; and 
 (viii) the level of LC; 
 (b) administering a therapeutically effective amount of the composition of  claim 17  to the subject; and 
 (c) determining the level of one or more of the levels in the subject after an initial period of treatment with the composition, 
 wherein an increase of the level in the subject following treatment with the composition indicates that the subject is likely to have a favorable clinical response to treatment with IαIp. 
 
     
     
       25. A kit comprising a composition according to  claim 17  and instructions for therapeutic use. 
     
     
       26. A method of treating sepsis or septic shock in a human in need thereof comprising administering to the human more than one dose of a pharmaceutical composition comprising 1 to 50 milligrams (mg) inter-alpha inhibitor proteins (IαIps) per kilogram (kg) body weight of the human, wherein the IαIps comprise 5% to 95% by weight of the composition and comprise a mixture of 60% to 80% inter-alpha inhibitor (IαI) and 40% to 20% pre-alpha inhibitor (PαI), the composition is suitable for administration to the human, and each dose of the composition is administered every 4 to 120 hours from about 1 to about 6 times per day.  
     
     
       27. The method of claim 26, wherein said IαI and PαI comprise a light chain (L) associated with at least one heavy chain selected from H1, H2, H3, and H4.  
     
     
       28. The method of claim 27, wherein the composition has a high trypsin inhibitory specific activity of 1000 to 2000 IU/mg.  
     
     
       29. The method of claim 28, wherein the trypsin inhibitory specific activity of said composition is 1400 to 2000 IU/mg.  
     
     
       30. The method of claim 26, wherein the composition has a half-life of greater than one hour.  
     
     
       31. The method of claim 30, wherein the composition has a half-life of at least 5 hours.  
     
     
       32. The method of claim 31, wherein the composition has a half-life of at least 10 hours.  
     
     
       33. The method of claim 26, wherein the IαI and PαI present in said composition have an apparent molecular weight of 60,000 to 280,000 Da.  
     
     
       34. The method of 26 further comprising, prior to said administering, determining the pre-treatment level of one or more of the following IαIps in the human: IαI, PαI, H3, H4, H1, H2, and LC.  
     
     
       35. The method of claim 34 further comprising determining the level of one or more of IαI, PαI, H3, H4, H1, H2, and LC in the human after an initial period of treatment with the composition.  
     
     
       36. The method of claim 26, wherein the composition further comprises a stabilizer.  
     
     
       37. The method of claim 26, wherein the composition is administered as a tablet, capsule, or injectable.  
     
     
       38. The method of claim 26, wherein the composition is administered by intravenous infusion.  
     
     
       39. The method of claim 26, wherein the composition is administered at least twice.  
     
     
       40. The method of claim 26, wherein the composition is administered until symptoms of the sepsis or septic shock improve.