Rna-coded antibody

Abstract

The present application describes an antibody-coding, non-modified or modified RNA and the use thereof for expression of this antibody, for the preparation of a pharmaceutical composition, in particular a passive vaccine, for treatment of tumours and cancer diseases, cardiovascular diseases, infectious diseases, autoimmune diseases, virus diseases and monogenetic diseases, e.g. also in gene therapy. The present invention furthermore describes an in vitro transcription method, in vitro methods for expression of this antibody using the RNA according to the invention and an in vivo method.

What is claimed is: 
     
         1 . A method of expressing an antibody in a subject comprising administering an effective amount of a pharmaceutical composition comprising purified mRNA encoding
 (i) a first polypeptide comprising the variable domain heavy chain (VH) of the antibody; and   (ii) a second polypeptide comprising the variable domain light chain (VL) of the antibody,   wherein the mRNA of the pharmaceutical composition comprises at least one 1-methyl-pseudouridine nucleotide substitution.   
     
     
         2 . The method of  claim 1 , wherein the subject has cancer or an infection. 
     
     
         3 . The method of  claim 1 , wherein the pharmaceutical composition is administered by injection. 
     
     
         4 . The method of  claim 1 , wherein the mRNA comprises a 5′ cap structure. 
     
     
         5 . The method of  claim 1 , wherein the antibody comprises a human antibody or a humanized antibody. 
     
     
         6 . The method of  claim 1 , wherein the mRNA comprises a sequence encoding an antibody operably linked to a secretory signal sequence. 
     
     
         7 . The method of  claim 1 , wherein the composition comprises a mRNA that encodes an antibody light chain and a mRNA that encodes an antibody heavy chain. 
     
     
         8 . The method of  claim 1 , wherein the composition comprises a mRNA that encodes an antibody light chain and an antibody heavy chain, wherein the antibody light chain and an antibody heavy chain coding sequences are linked by an internal ribosomal entry site (IRES). 
     
     
         9 . The method of  claim 1 , wherein the mRNA comprises a poly-A tail of 10 to 200 adenosine nucleotides. 
     
     
         10 . The method of  claim 1 , wherein the mRNA comprises a poly-C tail of 10 to 200 cytosine nucleotides. 
     
     
         11 . The method of  claim 1 , wherein the mRNA comprises a poly-A tail of 10 to 200 adenosine nucleotides and a 5′ cap structure. 
     
     
         12 . The method of  claim 1 , wherein the mRNA is further modified by introduction of a non-native nucleotide compared with a native mRNA sequence and/or by covalent coupling of the mRNA with a further chemical moiety. 
     
     
         13 . The method of  claim 12 , wherein the mRNA comprises a G/C content in the anti-body coding region which is greater than the G/C content of the coding region of the native mRNA sequence encoding the antibody. 
     
     
         14 . The method of  claim 12 , wherein the mRNA comprises an antibody coding sequence that is modified, compared with the native mRNA encoding the antibody, such that at least one codon of the native mRNA which codes for a tRNA which is relatively rare in the cell is exchanged for a codon which codes for a tRNA which is relatively frequent in the cell. 
     
     
         15 . The method of  claim 12 , wherein the mRNA further comprises a chemical modification relative to a naturally occurring mRNA. 
     
     
         16 . The method of  claim 12 , wherein the mRNA further comprises at least a nucleotide that is substituted with a nucleotide analog selected from the group consisting of: 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N6-methyl-adenine, N6-isopentenyl-adenine, 2-thio-cytosine, 3-methylcytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine, 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, dihydro-uracil, 2-thio-uracil, 4-thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5-methoxycarbonylmethyl-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), pseudouracil, queosine, β-D-mannosyl-queosine, wybutoxosine, phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. 
     
     
         17 . The method of  claim 12 , wherein the mRNA modification further comprises at least one base-modified nucleotide chosen from the group consisting of 2-amino-6-chloropurine riboside 5′-triphosphate, 2-aminoadenosine 5′-triphosphate, 2-thiocytidine 5′-triphosphate, 2-thiouridine 5′-triphosphate, 4-thiouridine 5′-triphosphate, 5-aminoallylcytidine 5′-triphosphate, 5-aminoallyluridine 5′-triphosphate, 5-bromocytidine 5′-triphosphate, 5-bromouridine 5′-triphosphate, 5-iodocytidine 5′-triphosphate, 5-iodouridine 5′-triphosphate, 5-methylcytidine 5′-triphosphate, 5-methyluridine 5′triphosphate, 6-azacytidine 5′-triphosphate, 6-azauridine 5′-triphosphate, 6-chloropurine riboside 5′-triphosphate, 7-deazaadenosine 5′-triphosphate, 7-deazaguanosine 5′-triphosphate, 8-azaadenosine 5′-triphosphate, 8-azidoadenosine 5′-triphosphate, benzimidazole riboside 5′-triphosphate, N1-methyladenosine 5′-triphosphate, N1-methylguanosine 5′-triphosphate, N6-methyladenosine 5′-triphosphate, O6-methylguanosine 5 ‘-triphosphate, pseudouridine 5’-triphosphate, puromycin 5′-triphosphate and xanthosine 5′-triphosphate. 
     
     
         18 . The method of  claim 1 , wherein the mRNA has been purified by reverse phase chromatography. 
     
     
         19 . The method of  claim 20 , wherein the chromatography is over a porous stationary phase comprising non-alkylated polystyrene-divinylbenzene. 
     
     
         20 . A pharmaceutical composition comprising purified mRNA encoding
 (i) a first polypeptide comprising the variable domain heavy chain (VH) of the antibody; and   (ii) a second polypeptide comprising the variable domain light chain (VL) of the antibody,   wherein the mRNA of the pharmaceutical composition comprises at least one 1-methyl-pseudouridine nucleotide substitution.