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

Abstract

The present invention relates to a pharmaceutical composition containing an mRNA that is stabilised by sequence modifications in the translated region and is optimised for the translation. The pharmaceutical composition according to the invention is particularly suitable as an inoculating agent as well as a therapeutic agent for tissue regeneration. In addition a process is described for determining sequence modifications that serve for the stabilisation and translation optimisation of mRNA.

1 . Modified mRNA coding for at least one viral peptide or polypeptide, characterised in that the G/C content of the region of the modified mRNA coding for the peptide or polypeptide is increased compared to the G/C content of the coding region of the wild type mRNA coding for the peptide or polypeptide, and the encoded amino acid sequence is unchanged as compared to the wild type. 
     
     
         2 . Modified mRNA according to  claim 1 , characterised in that the G/C content of the region of the modified mRNA coding for the peptide or polypeptide is increased by at least 7% points, preferably at least 15% points, compared to the G/C content of the coding region of the wild type mRNA coding for the peptide or polypeptide. 
     
     
         3 . Modified mRNA according to  claim 1 , characterised in that the modified mRNA comprises a 5′ cap structure and/or a poly-A tail of at least 70 nucleotides and/or an IRES and/or a 5′ stabilisation sequence and/or a 3′ stabilisation sequence. 
     
     
         4 . Modified mRNA according to  claim 1 , characterised in that the modified mRNA comprises at least one analogue of naturally occurring nucleotides. 
     
     
         5 . Modified mRNA according to  claim 4 , characterised in that the analogue is selected from the group consisting of phosphorus thioates, phosphorus amidates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. 
     
     
         6 . Modified mRNA according to  claim 1 , characterised in that the viral antigen derives from the secreted form of a surface antigen. 
     
     
         7 . Modified mRNA according to  claim 1 , characterised in that the mRNA codes for a surface antigen of a pathogenic viral germ. 
     
     
         8 . Modified mRNA according to  claim 1 , characterised in that the mRNA is associated with a cationic peptide or protein or is bound thereto. 
     
     
         9 . Modified mRNA according to  claim 8 , characterised in that the cationic peptide or protein is selected from the group consisting of protamine, poly-L lysine, and histones. 
     
     
         10 . Modified mRNA according to  claim 1 , characterised in that the polypeptide is a polyepitope of viral antigens. 
     
     
         11 . Modified mRNA according to  claim 1 , characterised in that the modified mRNA is a multicistronic mRNA. 
     
     
         12 . Modified mRNA according to  claim 1 , characterised in that the mRNA in addition codes for at least one cytokine. 
     
     
         13 . Modified mRNA according to  claim 1 , characterised in that the multicistronic RNA comprises more than one IRES sequence, wherein the IRES sequences are in particular selected from picorna viruses (e.g. FMDV), plague viruses (CFFV), polio viruses (PV), encephalo myocarditis viruses (ECMV), foot-and-mouth disease viruses (FMDV), hepatitis C viruses (HCV), classic swine fever viruses (CSFV), murine leukemia virus (MLV), simian immunodefiency viruses (SIV), or cricket paralysis viruses (CrPV). 
     
     
         14 . Pharmaceutical composition characterized in that it contains a modified mRNA according to  claim 1  in combination with a pharmaceutically acceptable carrier and/or vehicle. 
     
     
         15 . Pharmaceutical composition according to  claim 14 , characterised in that the pharmaceutical composition contains at least one immune response stimulating adjuvant. 
     
     
         16 . Pharmaceutical composition according to  claim 14 , which in addition contains at least one cytokine. 
     
     
         17 . A vaccine for inoculation against viral infectious diseases comprising a modified mRNA according to  claim 1 . 
     
     
         18 . Use of a pharmaceutical composition according to  claim 17  for the preparation of a vaccine for inoculation against infections caused by AIDS, hepatitis A, B or C, Herpes, Herpes zoster, Dengue, haemorrhagic infection, Yellow fever and influenza.