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 selected antigen in an organism comprising:
a) obtaining an aqueous mRNA injection solution comprising a calcium salt and a purified mRNA encoding the selected antigen, wherein said mRNA comprises (i) a 5′ cap; (ii) a poly-A tail of at least 25 nucleotides; and (iii) a 5′ and/or 3′ untranslated region (UTR); and
b) injecting the aqueous mRNA injection solution into the organism intradermally or intranodally,
thereby expressing the selected antigen 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, and 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 polycationic 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) being 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 mRNA, 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 or 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 , comprising injecting the aqueous mRNA injection solution into the organism intranodally.
41 . 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.
42 . A method for eliciting an immune response in an organism comprising injecting an aqueous mRNA injection solution into an organism intradermally or intranodally, wherein the aqueous mRNA injection solution comprises a calcium salt and a purified mRNA comprising: (i) a 5′ cap; (ii) a poly-A tail of at least 25 nucleotides; and (iii) a sequence encoding a selected antigen, thereby eliciting an immune response against the selected antigen within the organism.
43 . An injection syringe comprising an aqueous mRNA injection solution comprising a calcium salt and a purified mRNA comprising: (i) a 5′ cap; (ii) a poly-A tail of at least 25 nucleotides; and (iii) a sequence encoding a selected antigen, said aqueous mRNA injection solution comprised in the injection syringe.