Organ transplant solutions and method for transplanting organs

Abstract

A preservation solution for organs waiting to be transplanted is disclosed; the method of using the solution in a transplantation procedure is also disclosed. The preservation solutions comprise a balanced isotonic aqueous solution comprising sodium, potassium, calcium, magnesium and bicarbonate ions in a physiologically acceptable amount, together with an effective amount of a mutein of the C5 a anaphylatoxin which is a C5 a receptor antagonist wherein the amino acid residue naturally occurring at sequence position 69 is mutated.

1 . A preservation solution for storage and reperfusion of organs for implantation comprising: 
 (a) a balanced isotonic solution comprising sodium, potassium, calcium, magnesium and bicarbonate ions in a physiologically acceptable amount;    (b) a safe and effective amount of a mutein of the C5a anaphylatoxin which is a C5a receptor antagonist, wherein the amino acid residue naturally occurring at sequence position 69 is mutated; and    (c) water.    
     
     
         2 . The preservation solution according to  claim 1  which contains from about 0.1 μM to about 10 μM per liter of the C5a mutein.  
     
     
         3 . The preservation solution according to  claim 2  wherein, in the mutein, the amino acid residue at sequence position 69 is replaced by leucine or a positively charged amino acid residue.  
     
     
         4 . The preservation solution according to  claim 2  wherein, in the mutein, the amino acid residue naturally occurring at sequence position 67 is mutated.  
     
     
         5 . The preservation solution according to  claim 2  wherein, in the mutein, at least one of the amino acid residues at sequence positions 70 to 74 of the natural amino sequence is mutated and/or wherein at least one of the amino acid residues at said sequence positions 70 to 74 is deleted.  
     
     
         6 . The preservation solution according to  claim 3  wherein, in the mutein, the positively charged amino acid residue at sequence position 69 is Arg or Lys.  
     
     
         7 . The preservation solution according to  claim 4  wherein the mutein comprises at sequence position 67 an aromatic amino acid selected from the group consisting of Phe, Trp and Tyr.  
     
     
         8 . The preservation solution according to  claim 5  wherein the mutein comprises a hydrophobic amino acid residue at sequence position 70.  
     
     
         9 . The preservation solution according to  claim 8  wherein the mutein comprises Leu or Ala at sequence position 70.  
     
     
         10 . The preservation solution according to  claim 5  wherein the mutein comprises Ser at sequence position 70.  
     
     
         11 . The preservation solution according to  claim 4  wherein the mutein comprises a hydrophobic amino acid residue at sequence position 71 or 72.  
     
     
         12 . The preservation solution according to  claim 11  wherein the mutein comprises Leu at sequence position 71.  
     
     
         13 . The preservation solution according to  claim 5  wherein the mutein comprises Leu at sequence position 72.  
     
     
         14 . The preservation solution according to  claim 2  wherein the mutein comprises a terminal sequence selected from group consisting of 67-FKRSLLR-73, 67-FKRLLLR-73, 67-FKRSC-71, 67-FKRSLLC-73, 67-FKRLLLY-73, 67-FKKALLR-73, 67-FKRS-70, and 67-FKLLLLR-73.  
     
     
         15 . The preservation solution according to  claim 2  wherein the mutein further comprises Arg at sequence position 27.  
     
     
         16 . The preservation solution according to  claim 2  wherein the mutein has an amino acid sequence selected from 
 SEQ ID NO: 14 (C5a-(1-66, Cys27Ala-)A8B);    SEQ ID NO: 15 (C5a-(1-66, Cys27Ala)-A8B-Leu 70);    SEQ ID NO: 16 (C5a-(1-66, Cys27Ala)-Ab8-Cys7l);    SEQ ID NO: 17 (C5a-(1-66, Cys27Ala)-A8B-Cys73);    SEQ ID NO: 18 (C5a-(1-66, Cys27Ala)-A8B-Leu7O-Tyr73);    SEQ ID NO: 19 (C5a-(1-66, Cys27Ala)-A8B-Lys69-Ala7O);    SEQ ID NO: 20 (C5a-(1-66, Cys27Arg)-A8B);    SEQ ID NO: 21 (C5a-(1-66, Cys27Ala)-A8B-Del.71-73);    SEQ ID NO: 22 (C5a-(1-66, Cys-3, Gly-2, -1,Cys27Ala)-A8B); and    SEQ ID NO: 39 (C5a-(1-66, Cys27Ala)A5a).    
     
     
         17 . The preservation solution according to  claim 2  wherein the mutein further comprises an N-terminal linker sequence which is capable of dimerizing the C5a mutein.  
     
     
         18 . The preservation solution according to  claim 17  wherein, in the mutein, the N-terminal linker sequence comprises the sequence Cys-Gly-Gly or Cys-(Gly-Gly-Gly-Gly-Ser) 2 .  
     
     
         19 . The preservation solution according to  claim 2  wherein the mutein is fused to a protein or a peptide tag with the proviso that the fusion proteins of the mutein A8B with Jun/Fos alone or with Jun/Fos and the minor coat protein (pIII) of the filamentous M13 phage mutein A8B, wherein the Jun/Fos moiety is fused to the N-terminus of the mutein A8B, as well as the mutein A8B having a hexahistidine tag directly fused to the N-terminus are excluded.  
     
     
         20 . The preservation solution according to  claim 2  wherein the mutein is conjugated to a moiety which enhances the in vivo half-life of the mutein.  
     
     
         21 . A method of preservation, storage and reperfusion of an organ intended for implantation, said method comprising perfusing said organ with a solution comprising: 
 (a) a balanced isotonic solution comprising sodium, potassium, calcium, magnesium and bicarbonate ions in a physiologically acceptable amount;    (b) a safe and effective amount of a mutein of the C5a anaphylatoxin which is a C5a receptor antagonist wherein the amino acid residue naturally occurring at sequence position 69 is mutated; and    (c) water.    
     
     
         22 . The method of preservation according to  claim 21  wherein the C5a mutein is present at from about 0.1 to about 10 μM/liter of the solution.  
     
     
         23 . The method of preservation according to  claim 22  wherein the C5a mutein has an amino acid sequence selected from 
 SEQ ID NO: 14 (C5a-(1-66, Cys27Ala-)A8B);    SEQ ID NO: 15 (C5a-(1-66, Cys27Ala)-A8B-Leu 70);    SEQ ID NO: 16 (C5a-(1-66, Cys27Ala)-Ab8-Cys71);    SEQ ID NO: 17 (C5a-(1-66, Cys27Ala)-A8B-Cys73);    SEQ ID NO: 18 (C5a-(1-66, Cys27Ala)-A8B-Leu7O-Tyr73);    SEQ ID NO: 19 (C5a-(1-66, Cys27Ala)-A8B-Lys69-Ala7O);    SEQ ID NO: 20 (C5a-(1-66, Cys27Arg)-A8B);    SEQ ID NO: 21 (C5a-(1-66, Cys27Ala)-A8B-Del.71-73);    SEQ ID NO: 22 (C5a-(1-66, Cys-3, Gly-2, -1,Cys27Ala)-A8B); and    SEQ ID NO: 39 (C5a-(1-66, Cys27Ala)A5a).    
     
     
         24 . A method of preservation of organs intended for implantation, said method comprising perfusing the body of the dead organ donor, prior to removal of the organs, with a solution comprising: 
 (a) a balanced isotonic solution comprising sodium, potassium, calcium, magnesium and bicarbonate ions in a physiologically acceptable amount;    (b) a safe and effective amount of a mutein of the C5a anaphylatoxin which is a C5a receptor antagonist wherein the amino acid residue naturally occurring at sequence position 69 is mutated; and    (c) water.    
     
     
         25 . The method of preservation according to  claim 24  wherein the C5a mutein has an amino acid sequence selected from 
 SEQ ID NO: 14 (C5a-(1-66, Cys27Ala-)A8B);    SEQ ID NO: 15 (C5a-(1-66, Cys27Ala)-A8B-Leu 70);    SEQ ID NO: 16 (C5a-(1-66, Cys27Ala)-Ab8-Cys71);    SEQ ID NO: 17 (C5a-(1-66, Cys27Ala)-A8B-Cys73);    SEQ ID NO: 18 (C5a-(1-66, Cys27Ala)-A8B-Leu7O-Tyr73);    SEQ ID NO: 19 (C5a-(1-66, Cys27Ala)-A8B-Lys69-Ala7O);    SEQ ID NO: 20 (C5a-(1-66, Cys27Arg)-A8B);    SEQ ID NO: 21 (C5a-(1-66, Cys27Ala)-A8B-Del.71-73);    SEQ ID NO: 22 (C5a-(1-66, Cys-3, Gly-2, -1,Cys27Ala)-A8B); and    SEQ ID NO: 39 (C5a-(1-66, Cys27Ala)A5a).    
     
     
         26 . The method of preservation according to  claim 25  wherein the C5a mutein is present at from about 0.1 to about 10 μM/liter of the solution.