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 treating a subject comprising administering an effective amount of a pharmaceutical composition comprising mRNA encoding CD20-binding antibody. 
     
     
         2 . The method of  claim 1 , wherein the subject has a cancer. 
     
     
         3 . The method of  claim 2 , wherein the subject has a leukemia, lymphoma or myeloma. 
     
     
         4 . The method of  claim 1 , wherein the pharmaceutical composition is administered by injection. 
     
     
         5 . The method of  claim 1 , wherein the antibody comprises a Fab, Fab′, F(ab′)2, Fc, pFc′, Fd, FIT and scFv fragment of an antibody. 
     
     
         6 . The method of  claim 1 , wherein the antibody comprises a human antibody or a humanized antibody. 
     
     
         7 . The method of  claim 1 , wherein the mRNA comprises a sequence encoding an antibody operably linked to a heterologous secretory signal sequence. 
     
     
         8 . 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. 
     
     
         9 . 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). 
     
     
         10 . The method of  claim 1 , wherein the mRNA comprises a 5′ cap structure. 
     
     
         11 . The method of  claim 1 , wherein the mRNA additionally comprises a poly-A tail of 10 to 200 adenosine nucleotides and/or a poly-C tail of 10 to 200 cytosine nucleotides. 
     
     
         12 . The method of  claim 1 , wherein the CD20-binding antibody comprises ibritumomab, tositumomab, ofatumamab or rituximab. 
     
     
         13 . The method of  claim 12 , wherein the antibody comprises rituximab. 
     
     
         14 . The method of  claim 13 , wherein the mRNA encodes: a sequence at least 80% identical to SEQ ID NOs: 1 or 2; and/or a sequence at least 80% identical to SEQ ID NOs: 4 or 5. 
     
     
         15 . The method of  claim 1 , wherein the mRNA is 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. 
     
     
         16 . The method of  claim 15 , wherein the mRNA comprises:
 (i) a G/C content in the antibody coding region which is greater than the G/C content of the coding region of the native mRNA sequence encoding the antibody; or   (ii) 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.   
     
     
         17 . The method of  claim 15 , wherein the mRNA comprises a chemical modification relative to a naturally occurring mRNA. 
     
     
         18 . The method of  claim 15 , wherein the mRNA 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, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), pseudouracil, 1-methyl-pseudouracil, queosine, β-D-mannosyl-queosine, wybutoxosine, phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. 
     
     
         19 . The method of  claim 15 , wherein the mRNA modification 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, 06-methylguanosine 5′-triphosphate, pseudouridine 5′-triphosphate, puromycin 5′-triphosphate and xanthosine 5′-triphosphate. 
     
     
         20 . The method of  claim 19 , wherein the base-modified nucleotide is chosen from the group consisting of: 5-methylcytidine 5′-triphosphate, 1-methyl-pseudouracil and pseudouridine 5′-triphosphate. 
     
     
         21 . A pharmaceutical composition comprising an isolated mRNA comprising a coding region encoding at least one antibody variable domain of a CD20-binding antibody, wherein said coding region is linked to a heterologous secretory signal sequence.