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 . Use of a modified mRNA coding for at least one biologically active 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, for the preparation of a medicament for gene therapy. 
     
     
         2 . Use of a 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 . Use of a modified mRNA according to  claim 1 , characterised in that the region of the modified mRNA coding for the peptide or polypeptide is altered in such a way as to produce a maximum G/C content in conjunction with the codons that code for the relatively frequent tRNAs. 
     
     
         4 . Use of a 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. 
     
     
         5 . Use of a modified mRNA according to  claim 1 , characterised in that the modified mRNA comprises at least one analogue of naturally occurring nucleotides. 
     
     
         6 . Use of a modified mRNA according to  claim 5 , 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. 
     
     
         7 . Use of a modified mRNA according to  claim 1 , characterised in that the mRNA codes for a polypeptide which is not formed or is only insufficiently or defectively formed in the patient to be treated. 
     
     
         8 . Use of a modified mRNA according to  claim 1 , characterised in that the modified mRNA codes for dystrophin, enzymes that are lacking or defective in metabolic diseases, or enzymes that are involved in the synthesis of neurotransmitters. 
     
     
         9 . Use of a modified mRNA according to  claim 1 , characterised in that the mRNA codes for a peptide or protein that binds to cell surface receptors. 
     
     
         10 . Use of a modified mRNA according to  claim 1 , characterised in that the mRNA codes for growth factors or growth hormones. 
     
     
         11 . Use of a modified mRNA according to  claim 7 , characterised in that the mRNA codes for the tissue plasminogen activator, insulin, interferones, GM-CSF, erythropoietin, tyrosine hydroxylase, DOPA decarboxylase, or α-1 antitrypsin. 
     
     
         12 . Use of a modified mRNA according to  claim 1 , characterised in that the mRNA is associated with a cationic peptide or protein or is bound thereto, in particular protamine, poly-L lysine, or histone. 
     
     
         13 . Use of a modified mRNA according to  claim 1 , characterised in that the mRNA in addition codes for at least one cytokine. 
     
     
         14 . Use of a modified mRNA according to  claim 1  in combination with a pharmaceutically acceptable carrier and/or vehicle. 
     
     
         15 . Use of a modified mRNA according to  claim 1  for tissue regeneration or for the treatment of degenerative, in particular neurodegenerative, diseases. 
     
     
         16 . Use of a modified mRNA according to  claim 1  for the treatment of Alzheimer's disease, Parkinson's disease or arthrosis. 
     
     
         17 . Use of a modified mRNA according to  claim 15 , characterised in that the mRNA codes for growth factors of the TGF-β family. 
     
     
         18 . Use of a modified mRNA according to  claim 15 , characterised in that the mRNA codes for EGF, FGF, PDGF, BMP, GDNF, BDNF, GDF and neuthrophic factors, such as NGF or neutrophines. 
     
     
         19 . A modified mRNA coding for at least one biologically active peptide or polypeptide which is not formed or is only insufficiently or defectively formed in the patient to be treated, 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. 
     
     
         20 . A modified mRNA according to  claim 19 , wherein the modified mRNA codes for enzymes that are lacking or are defective in metabolic diseases, or enzymes that are involved in the synthesis of neurotransmitters. 
     
     
         21 . A modified mRNA according to  claim 19 , wherein the modified mRNA codes for cell surface receptors. 
     
     
         22 . Modified mRNA according to  claim 19 , wherein the modified mRNA codes for proteins that bind to cell surface receptors or act in an extracellular manner, selected from the group consisting of tissue plasminogen activator, growth hormones, insulin, interferones, granulocyte macrophage colony stimulating factor, growth factors, and erythropoietin. 
     
     
         23 . Pharmaceutical composition containing a modified mRNA according to  claim 19  in combination with a pharmaceutically acceptable carrier and/or vehicle.