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.

What is claimed is: 
     
         1 - 27 . (canceled) 
     
     
         28 . A method for stimulating an immune response to a virus surface antigen in a subject comprising administering to the subject an effective amount of a pharmaceutical composition comprising:
 1. a mRNA comprising a protein coding sequence that encodes the virus surface antigen, said mRNA comprising a 5′ Cap structure and a poly-A region of at least 50 nucleotides; and   2. an aqueous injection solution,   
       wherein the mRNA lacks any promoter sequences, wherein the protein coding sequence of the mRNA comprises an increased G/C content of at least 15% points relative to an original coding sequence encoding the virus surface antigen, wherein the protein sequence encoded by the mRNA is unchanged compared to protein encoded by the original coding sequence, and wherein the virus surface antigen is not a HIV, flavivirus or ebolavirus antigen, 
       wherein the pharmaceutical composition is administered by intramuscular injection. 
     
     
         29 . The method of  claim 28 , wherein the mRNA further encodes a secretory leader. 
     
     
         30 . The method of  claim 29 , wherein the aqueous injection solution comprises water for injection (WFI), a buffered solution or a salt solution. 
     
     
         31 . The method of  claim 30 , wherein the salt solution comprises sodium chloride or potassium chloride solution. 
     
     
         32 . The method of  claim 30 , wherein the pharmaceutical composition comprises a component selected from the group consisting of human serum albumin, a polycationic protein, polysorbate 80, a sugar and an amino acid. 
     
     
         33 . The method of  claim 30 , wherein the pharmaceutical composition comprises a sugar. 
     
     
         34 . The method of  claim 29 , wherein the mRNA encoding the virus surface antigen comprises at least one nucleotide position replaced with a nucleotide analogue. 
     
     
         35 . The method of  claim 34 , wherein the nucleotide analogue is selected from the group consisting of phosphorus amidates, phosphorus thioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. 
     
     
         36 . The method of  claim 29 , wherein the protein coding sequence of the mRNA comprises an increased G/C content of at least 20% points relative to an original coding sequence encoding the virus surface antigen. 
     
     
         37 . The method of  claim 29 , wherein the virus surface antigen is an influenza surface antigen. 
     
     
         38 . The method of  claim 29 , further comprising administering the composition to the subject two or more times. 
     
     
         39 . The method of  claim 29 , wherein the mRNA comprises a 5′ untranslated region (UTR) and a 3′UTR.