PHARMACEUTICAL COMPOSITION CONTAINING A STABILISED mRNA OPTIMISED FOR TRANSLATION IN ITS CODING REGIONS

Abstract

The present invention relates to a pharmaceutical composition comprising a modified mRNA that is stabilised by sequence modifications and optimised for translation. The pharmaceutical composition according to the invention is particularly well suited for use as an inoculating agent, as well as a therapeutic agent for tissue regeneration. In addition, a process is described for determining sequence modifications that promote stabilisation and translational efficiency of modified mRNA of the invention.

1 . An isolated mRNA comprising a polypeptide coding sequence of any of SEQ ID NOs: 14-30, wherein the sequence encodes a human MUC1 polypeptide. 
     
     
         2 . The isolated mRNA of  claim 1 , wherein the mRNA is complexed with as least one polycationic agent. 
     
     
         3 . The isolated mRNA of  claim 1 , wherein the mRNA is complexed with as least one polycationic polypeptide. 
     
     
         4 . The isolated mRNA of  claim 2 , wherein the polycationic agent is chosen from the group consisting of a protamine, poly-L-lysine, and histones. 
     
     
         5 . The isolated mRNA of  claim 7 , wherein the mRNA is complexed with protamine. 
     
     
         6 . The isolated mRNA of  claim 1 , wherein the mRNA comprises at least one chemical modification of the mRNA. 
     
     
         7 . The isolated mRNA of  claim 1 , wherein the mRNA comprises at least one nucleotide of the mRNA is substituted with an analog of the naturally occurring nucleotide. 
     
     
         8 . The isolated mRNA of  claim 1 , wherein the mRNA comprises at least one nucleotide position replaced with a nucleotide analogue selected from the group consisting of phosphorus amidates, phosphorus thioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. 
     
     
         9 . The isolated mRNA of  claim 1 , wherein the mRNA comprises a 5′ cap structure. 
     
     
         10 . The isolated mRNA of  claim 1 , wherein the mRNA comprises a poly-A tail of at least 50 nucleotides. 
     
     
         11 . The isolated mRNA of  claim 1 , wherein the mRNA further comprises 5′ non-translated region and/or a 3′ non-translated region 
     
     
         12 . The method of  claim 11 , wherein the 5′ non-translated region and/or a 3′ non-translated region is/are chosen from the group consisting of untranslated sequences (UTR) of the β-globin gene and a stabilizing sequence of the general formula (C/U)CCANxCCC(U/A)PyxUC(C/U)CC. 
     
     
         13 . The method of  claim 1 , wherein the mRNA further encodes a secretion signal. 
     
     
         14 . The method of  claim 1 , wherein the mRNA is provided in a liposome complex. 
     
     
         15 . A composition comprising a isolated mRNA of  claim 1  in a pharmaceutically acceptable carrier. 
     
     
         16 . The composition of  claim 15 , wherein the mRNA is dissolved in the aqueous carrier. 
     
     
         17 . The composition of  claim 16 , wherein the aqueous carrier is water for injection (WFI), a buffered solution or a salt solution. 
     
     
         18 . The composition of  claim 17 , wherein the salt solution comprises sodium chloride or potassium chloride solution. 
     
     
         19 . The composition of  claim 15 , wherein the composition further comprises an adjuvant.