PHARMACEUTICAL COMPOSITION CONTAINING A STABILISED mRNA OPTIMISED FOR TRANSLATION IN ITS CODING REGIONS
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.
1 . A method for stimulating an anti-influenza immune response in a subject comprising administering an effective amount of a pharmaceutical composition comprising a mRNA that encodes an influenza virus antigen to the subject.
2 . The method of claim 1 , wherein the influenza virus antigen is an influenza virus surface antigen.
3 . The method of claim 1 , wherein the pharmaceutical composition is administered by injection.
4 . The method of claim 1 , wherein the pharmaceutical composition is administered intravenously, intradermally, subcutaneously, intramuscularly, topically or orally
5 . The method of claim 4 , wherein the pharmaceutical composition is administered intradermally.
6 . The method of claim 1 , wherein the mRNA that encodes the influenza virus antigen comprises an increased G/C content relative to a wild type RNA encoding the influenza virus antigen.
7 . The method of claim 1 , wherein the mRNA encoding the influenza virus antigen comprises a sequence wherein at least one codon of a wild-type sequence recognized by a rare cellular tRNA is replaced with a codon recognized by an abundant cellular tRNA, and wherein said rare cellular tRNA and said abundant cellular tRNA recognize the same amino acid.
8 . The method of claim 1 , wherein the mRNA encoding the influenza virus antigen comprises at least one nucleotide position replaced with a nucleotide analogue selected from the group consisting of phosphorus amidates, phosphorus thioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine.
9 . The method of claim 1 , wherein the mRNA further encodes a secretory leader.
10 . The method of claim 1 , wherein the mRNA is dissolved in the aqueous carrier.
11 . The method of claim 10 , wherein the aqueous carrier is water for injection (WFI), a buffered solution or a salt solution.
12 . The method of claim 11 , wherein the salt solution comprises sodium chloride or potassium chloride solution.
13 . The method of claim 1 , 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.
14 . The method of claim 13 , wherein pharmaceutical composition comprises a polycationic protein.
15 . The method of claim 14 , wherein the polycationic protein comprises protamine.
16 . The method of claim 15 , wherein the mRNA is in complex with protamine.
17 . The method of claim 1 , wherein the pharmaceutical composition further comprises an adjuvant.
18 . The method of claim 1 , further comprising administering the composition to the subject two or more times.
19 . The method of claim 1 , further comprising administering a cytokine to the subject.
20 . The method of claim 19 , wherein the cytokine comprises GM-CSF.
21 . The method of claim 1 , wherein the composition is administered intradermally and wherein the mRNA is provided in complex with protamine and encodes an influenza virus surface antigen.