Methods and compositions for preventing ischemia reperfusion injury in organs

Abstract

The invention, in some embodiments, relates to compounds and methods for the prevention of ischemia reperfusion injury (IRI) in organs, and in particular to IRI in organs aged 35 years and older. Specific uses include prevention of IRI in native organs in vivo, in reimplantations and in transplantations of donor organs aged 35 years and older. Additional embodiments include the prophylaxis of delayed graft function (DGF) and reduction in the frequency, amount and duration of dialysis in recipients of deceased donor kidney transplantations. The methods entail contacting the organ in vivo or ex vivo with a temporary p53 inhibitor. Novel temporary dsNA p53 inhibitors are further provided.

1 . A method of prophylaxis of ischemic reperfusion injury (IRI) in an organ at risk of IRI, comprising contacting the organ with a temporary inhibitor of a p53 gene in an amount effective to provide prophylaxis of IRI in the organ; wherein the organ is 35 years or older. 
     
     
         2 . A temporary inhibitor of a p53 gene for use in prophylaxis of ischemic reperfusion injury (IRI), wherein the inhibitor is for contacting an organ that is 35 years or older at risk of IRI. 
     
     
         3 . Use of a temporary inhibitor of a p53 gene for the manufacture of a medicament for providing prophylaxis of ischemic reperfusion injury (IRI), wherein the inhibitor is for contacting an organ that is 35 years or older at risk of IRI. 
     
     
         4 . The method, inhibitor or use of any of  claims 1  to  3 , wherein the organ at risk of IRI is selected from the group consisting of an organ native to a subject, a reimplanted organ and a transplanted organ. 
     
     
         5 . The method, inhibitor or use of  claim 4 , wherein the risk of IRI in the native organ is imposed by temporary cessation of blood flow to the organ or by temporary global hypoxia of the organ. 
     
     
         6 . The method, inhibitor or use of  claim 5 , wherein the temporary cessation of blood flow is due to at least one of thrombosis, vasoconstriction, pressure on blood vessels or removal of the organ from the body of a subject with subsequent reimplantation. 
     
     
         7 . The method, inhibitor or use of  claim 4 , wherein the transplant organ originates from a deceased donor. 
     
     
         8 . The method, inhibitor or use of any of  claims 1 - 7 , wherein contacting the organ with the temporary inhibitor comprises administering the temporary inhibitor to a subject possessing the organ at risk of IRI. 
     
     
         9 . The method, inhibitor or use of  claim 4 , wherein the organ at risk is a native organ of a subject and has never been removed from the body of the subject. 
     
     
         10 . The method, inhibitor or use of  claim 9 , wherein the organ at risk has been reimplanted to a subject. 
     
     
         11 . The method, inhibitor or use of  claim 7  wherein the organ at risk has been transplanted to a subject. 
     
     
         12 . The method, inhibitor or use of  claims 10  and  11 , wherein contacting the organ with the temporary inhibitor comprises contacting the organ with the temporary inhibitor ex vivo prior to transplantation or reimplantation of the organ to a recipient. 
     
     
         13 . The method, inhibitor or use of any of  claims 1  to  12  wherein the organ at risk of IRI is 45 years old or older. 
     
     
         14 . The method, inhibitor or use of  claim 9  wherein prophylaxis of IRI results in prophylaxis of IRI-associated organ dysfunction. 
     
     
         15 . The method, inhibitor or use of  claim 10  or  11 , wherein prophylaxis of IRI results in prophylaxis of IRI-associated delayed graft function. 
     
     
         16 . The method, inhibitor or use of any of  claims 1  to  15 , wherein the organ is selected from the group consisting of a kidney, a liver, a pancreas, a heart, a lung, an intestine, skin, a blood vessel, a brain, a retina, composite tissue, a blood vessel, an ear, a limb; or a part thereof. 
     
     
         17 . The method, inhibitor or use of any of  claim 16 , wherein the organ is a kidney. 
     
     
         18 . The method, inhibitor or use of any of  claim 17 , wherein the organ is a kidney graft and wherein prophylaxis of IRI results in prophylaxis of delayed graft function. 
     
     
         19 . The method, inhibitor or use of  claim 18 , wherein the prophylaxis of DGF results in the reduction of the amount, intensity and duration of dialytic support during at least the first 7 days post-transplant in a dialysis-dependent end stage renal disease (ESRD) patient undergoing deceased donor renal transplantation. 
     
     
         20 . The method, inhibitor or use of  claim 18  or  19 , wherein prophylaxis of DGF results in at least one of a longer time interval between transplantation and the first dialysis treatment post-transplant, a shorter mean duration of initial post-transplantation course of dialysis and a higher measured glomerular filtration rate (mGFR) at the end of the first post-transplant month. 
     
     
         21 . The method, inhibitor or use of  claim 17 , wherein the prophylaxis of IRI results in the reduction of the amount, intensity and/or duration of dialytic support during the first 30 days post-transplant in a dialysis-dependent end stage renal disease (ESRD) patient undergoing deceased donor renal transplantation. 
     
     
         22 . The method, inhibitor or use of  claim 17 , wherein the prophylaxis of IRI results in the reduction of the amount, intensity and/or duration of dialytic support during the first 180 days post-transplant in a dialysis-dependent end stage renal disease (ESRD) patient undergoing deceased donor renal transplantation. 
     
     
         23 . The method, inhibitor or use of any of  claims 7  to  22 , wherein the organ is preserved entirely by cold storage following removal from the donor and prior to implantation in the recipient. 
     
     
         24 . The method, inhibitor or use of any of  claims 7  to  22 , wherein the organ is preserved by machine-perfusion for at least a portion of time following removal from the donor and prior to implantation in the recipient. 
     
     
         25 . The method, inhibitor or use of any of  claims 7  to  24 , further comprising the steps of (a) selecting a recipient having a kidney from a deceased Expanded Criteria Donor, and (b) administering to the recipient a temporary inhibitor of a p53 gene in an amount effective to provide prophylaxis of DGF in the recipient. 
     
     
         26 . The method, inhibitor or use of any of  claims 7  to  24 , wherein the kidney is from a donor that is not a deceased Expanded Criteria Donor. 
     
     
         27 . The method, inhibitor or use of  claim 26 , wherein the kidney is from a donor that is between the ages of 50 and 59 (inclusive) who does not have at least two of the following: a history of high blood pressure, terminal serum creatinine level greater than 1.5 mg/dl, or cardiovascular cause of brain death. 
     
     
         28 . The method, inhibitor or use of  claim 26 , wherein the kidney is from a donor that is not over the age of 60. 
     
     
         29 . The method, inhibitor or use of any of  claims 4  to  6 , wherein the prophylaxis of IRI provides prophylaxis of acute kidney injury (AKI). 
     
     
         30 . The method, inhibitor or use of  claim 29 , wherein the AKI results from cardiovascular surgery, cardiopulmonary surgery, renal surgery, acute ureteral obstruction, shock, global hypoxia and/or exposure to a nephrotoxin. 
     
     
         31 . A method of prophylaxis of ischemic reperfusion injury (IRI) in a donor kidney from a deceased donor, comprising contacting the kidney with a temporary inhibitor of p53 in an amount effective to provide prophylaxis of IRI in the kidney. 
     
     
         32 . A temporary inhibitor of p53 for use in prophylaxis of ischemic reperfusion injury (IRI) in a donor kidney from a deceased donor, wherein the inhibitor is for contacting the kidney. 
     
     
         33 . Use of a temporary inhibitor of a p53 gene for the manufacture of a medicament for providing prophylaxis of ischemic reperfusion injury (IRI) in donor kidney from a deceased donor, wherein the inhibitor is for contacting the kidney. 
     
     
         34 . The method, inhibitor or use of any of  claims 31  to  33 , wherein the prophylaxis of IRI results in the reduction of the amount, intensity and duration of dialytic support during the first 180 days post-transplant in a dialysis-dependent end stage renal disease (ESRD) patient undergoing deceased donor renal transplantation. 
     
     
         35 . The method, inhibitor or use of any of the preceding claims, wherein the temporary inhibitor of a p53 gene is selected from the group consisting of a small organic molecule, a protein, an antibody or fragment thereof, a peptide, a polypeptide, a peptidomimetic and a nucleic acid molecule; or a pharmaceutically acceptable salt or prodrug thereof. 
     
     
         36 . The method, inhibitor or use of  claim 32 , wherein the temporary inhibitor of a p53 gene is a nucleic acid molecule selected from the group consisting of a single stranded antisense nucleic acid (ssNA), a double-stranded NA (dsNA), a small interfering NA (siNA), a short hairpin NA (shNA), a micro RNA (miRNA) or its mimetic, an aptamer, and a ribozyme, or a pharmaceutically acceptable salt or prodrug thereof. 
     
     
         37 . The method, inhibitor or use of  claim 36 , wherein the nucleic acid molecule is an ssNA or a dsNA, comprising none or at least one of a modified nucleotide, a nucleotide analogue and/or an unconventional moiety. 
     
     
         38 . The method, inhibitor or use of any of  claims 35  to  37  wherein the nucleic acid molecule is a dsNA selected from the group consisting of an unmodified dsNA or a chemically modified dsNA; or a salt or prodrug thereof. 
     
     
         39 . The method, inhibitor or use of  claim 38 , wherein the dsNA comprises an antisense strand having a nucleic acid sequence set forth in Table 2 (SEQ ID NOS:21-33, 35 37). 
     
     
         40 . The method, inhibitor or use of any of  claims 36  to  39 , wherein the dsNA comprises an antisense strand sequence 5′ UGAAGGGUGAAAUAUUCUC (SEQ ID NO:37) 3′ and a sense strand sequence 5′ GAGAAUAUUUCACCCUUCA 3′ (SEQ ID NO:36). 
     
     
         41 . The method, inhibitor or use of  claim 40 , wherein the dsNA is a synthetic small interfering ribonucleic acid (siRNA) having the structure: 
       
         
           
                 
                 
               
                     
                   (antisense strand) 
                 
                     
                   (SEQ ID NO: 37) 
                 
                     
                   5′ UGAAGGGUGAAAUAUUCUC 3′ 
                 
                     
                     
                 
                     
                   (sense strand) 
                 
                     
                   (SEQ ID NO: 36) 
                 
                     
                   3′ ACUUCCCACUUUAUAAGAG 5′ 
                 
             
                
                
                
                
                
                
                
               
            
           
         
         wherein each of A, C, U and G is a ribonucleotide and each consecutive ribonucleotide is joined to the next ribonucleotide by a covalent bond; and 
         wherein alternating ribonucleotides in both the antisense strand and the sense strand are 2′-O-methyl sugar modified ribonucleotides and a 2′-O-methyl sugar modified ribonucleotide is present at both the 5′ terminus and the 3′ terminus of the antisense strand and an unmodified ribonucleotide is present at both the 5′ terminus and the 3′ terminus of the sense strand. 
       
     
     
         42 . The method, inhibitor or use of  claim 41 , wherein the 5′ termini and the 3′ termini are unphosphorylated. 
     
     
         43 . The method, inhibitor or use of any of  claims 38  to  42 , wherein the double-stranded NA compound is in the form of a pharmaceutically acceptable salt. 
     
     
         44 . The method, inhibitor or use of  claim 43 , wherein the pharmaceutically acceptable salt is a sodium salt. 
     
     
         45 . The method, inhibitor or use of any of  claims 39  to  44 , wherein the temporary inhibitor of a p53 gene is administered at a dose of about 1 to about 50 mg/kg. 
     
     
         46 . The method, inhibitor or use of  claim 45 , wherein the temporary inhibitor of a p53 gene is administered at a dose of about 10 mg/kg. 
     
     
         47 . The method, inhibitor or use of any of  claims 4  to  46 , wherein the temporary inhibitor of a p53 gene is to be administered as a liquid composition comprising a pharmaceutically acceptable carrier. 
     
     
         48 . The method, inhibitor or use of any of  claims 4  to  47 , wherein the temporary inhibitor is administered as an injectable composition comprising a pharmacologically acceptable aqueous excipient. 
     
     
         49 . The method, inhibitor or use of  claim 48 , wherein the temporary inhibitor is administered by intravenous (IV) injection. 
     
     
         50 . The method, inhibitor or use of  claim 49 , wherein the intravenous (IV) injection is administered in a single treatment, wherein the single treatment comprises a single dose or multiple doses. 
     
     
         51 . The method, inhibitor or use of  claim 50 , wherein the single treatment is a single dose. 
     
     
         52 . The method, inhibitor or use of  claim 50  or  51 , wherein the single treatment is a single intravenous push (IVP). 
     
     
         53 . The method, inhibitor or use of any of  claims 49  to  52 , wherein the intravenous (IV) injection is administered intraoperatively following autograft/reimplantation or allograft/transplantation reperfusion. 
     
     
         54 . The method, inhibitor or use of  claim 53 , wherein the intravenous (IV) injection is administered directly into a proximal port of a central venous line or through a peripheral line. 
     
     
         55 . The method, inhibitor or use of  claim 48 , wherein the inhibitor is administered systemically, subcutaneously, topically, by inhalation, by instillation (lungs). 
     
     
         56 . The method, inhibitor or use of any of the preceding claims, wherein the temporary inhibitor is conjugated or formulated in liposomes or nanoparticles. 
     
     
         57 . The method, inhibitor or use of any of  claims 7  to  28  and  31  to  56 , wherein the recipient is further administered a medication selected from the group consisting of an antiviral agent, an antifungal agent, an antimicrobial agent, an immunosuppressant agent, and any combination thereof. 
     
     
         58 . The method, inhibitor or use of  claim 57 , wherein the medication is an immunosuppressant agent that is a calcineurin inhibitor. 
     
     
         59 . The method, inhibitor or use of  claim 57  or  58 , wherein the immunosuppressant agent is selected from the group consisting of tacrolimus (TAC), mycophenolate mofetil (MMF), mycophenolic acid (MPA), a corticosteroid, a cyclosporine, an azathioprine, a sirolimus, and any combination thereof. 
     
     
         60 . The method, inhibitor or use of  claim 59 , wherein the immunosuppressant agent is tacrolimus (TAC). 
     
     
         61 . The method, inhibitor or use of any of  claims 57  to  60 , wherein the recipient is further administered an antibody induction therapy agent. 
     
     
         62 . The method, inhibitor or use of  claim 61 , wherein the antibody induction therapy agent is administered peri-operatively and prior to transplant reperfusion. 
     
     
         63 . The method, inhibitor or use of  claim 61  or  62 , wherein the antibody induction therapy agent comprises a polyclonal anti-thymocyte globulin (ATG) or an anti-CD25 (anti-IL-2R) monoclonal antibody. 
     
     
         64 . The method, inhibitor or use of any of the preceding claims, wherein the inhibitor is present in a kit comprising the inhibitor and instructions for use. 
     
     
         65 . The method, inhibitor or use of any of the preceding claims, wherein the inhibitor is a dsRNA molecule set forth in Table P.