INJECTION SOLUTION FOR RNA
The invention relates to the use of RNA and an aqueous injection buffer containing a sodium salt, a calcium salt and optionally a potassium salt and optionally lactate, in the preparation of a RNA injection solution for increasing RNA transfer and/or RNA translation into/in a host organism. The invention relates further to a RNA injection solution and to a method for increasing the RNA transfer and/or RNA translation of RNA in vivo and in vitro.
1 - 21 . (canceled)
22 . A method for increasing the expression of a mRNA in an organism comprising:
a) obtaining an aqueous mRNA injection solution comprising a calcium salt and a purified mRNA encoding a selected antigen, wherein said mRNA is capped and comprises a poly-A tail; and
b) injecting the aqueous mRNA injection solution into an organism intradermally, thereby expressing the mRNA in the organism.
23 . The method of claim 22 , wherein the aqueous mRNA injection solution comprises salts with the anions selected from the group consisting of chlorides, iodides, bromides, hydroxides, carbonates, hydrogen carbonates or sulfates
24 . The method of claim 22 , wherein the aqueous mRNA injection solution contains at least 50 mM sodium chloride (NaCl), at least 0.01 mM calcium chloride (CaCl 2 ) and optionally at least 3 mM potassium chloride (KCl).
25 . The method of claim 24 , wherein the aqueous mRNA injection solution contains from 50 mM to 800 mM sodium chloride (NaCl), from 0.01 mM to 100 mM calcium chloride (CaCl 2 ), and optionally from 3 mM to 500 mM potassium chloride (KCl).
26 . The method of claim 22 , wherein the aqueous mRNA injection solution further comprises lactate.
27 . The method of claim 26 , wherein the aqueous mRNA injection solution contains at least 50 mM sodium chloride (NaCl), at least 0.01 mM calcium chloride (CaCl 2 ), at least 15 mM lactate, and optionally at least 3 mM potassium chloride (KCl).
28 . The method of claim 26 , wherein the aqueous mRNA injection solution contains from 15 mM to 500 mM lactate.
29 . The method of claim 22 , wherein the mRNA is complexed with a polycationic polypeptide.
30 . The method of claim 29 , wherein the polycationic polypeptide is poly-L-lysine, poly-L-arginine, or protamine.
31 . The method of claim 30 , wherein the poly cationic polypeptide is protamine.
32 . The method of claim 22 , wherein the mRNA is a modified mRNA comprising at least one modification selected from the group consisting of: a) having an increased G/C content in the coding region of the mRNA relative to the wild-type (WT) mRNA, b) having at least one analog of a naturally occurring ribonucleotide, c) free of one or more destabilizing sequences in the WT sequence, d) having an increased A/U content in the ribosome binding site of the mRNA relative to the WT sequence, e) having one or more codon substitutions, f) having a heterologous poly(A) tail, g) having an IRES sequence, h)having a 3′ or 5′ stabilization sequence, and i) having a sequence encoding a signal peptide.
33 . The method of claim 22 , wherein the mRNA does not comprise a chemical modification.
34 . The method of claim 22 , wherein the mRNA encodes a tumor antigen.
35 . The method of claim 34 , wherein the tumor antigen is selected from the group consisting of MAGE, NY-ESO-1, PSA, and mucin.
36 . The method of claim 22 , wherein the aqueous mRNA injection solution is a Ringer's Lactate solution.
37 . The method of claim 22 , wherein the aqueous mRNA injection solution does not contain mono-, di- or polysaccharides.
38 . The method of claim 22 , wherein the aqueous mRNA injection solution comprises from 0.5 mM to 100 mM calcium salt.
39 . The method of claim 38 , wherein the aqueous mRNA injection solution comprises from 1.5 mM to 40 mM calcium salt.
40 . The method of claim 22 , wherein the organism suffers from a disease or disorder selected from cancer, allergies, autoimmune diseases, viral infection, and bacterial infection.
41 . A method for eliciting an immune response in an organism comprising injecting an aqueous mRNA injection solution into an organism intradermally, thereby expressing the mRNA within the organism, wherein the aqueous mRNA injection solution comprises a calcium salt and a purified mRNA encoding a selected antigen.