Compositions comprising non-immunogenic RNA encoding immunogenic epitopes and interferon alpha
The present disclosure relates to methods and agents for antigen vaccination and inducing effective antigen-specific immune effector cell responses such as T cell responses. Specifically, the present disclosure relates to methods comprising administering to a subject (i) non-immunogenic RNA encoding a peptide or protein comprising an epitope for inducing an immune response against an antigen in the subject, i.e., non-immunogenic RNA encoding vaccine antigen; and (ii) an immunostimulant or RNA encoding an immunostimulant. Administering to the subject non-immunogenic RNA encoding vaccine antigen may provide (following expression of the RNA by appropriate target cells) vaccine antigen for stimulation, priming and/or expansion of immune effector cells and, thus, may induce an immune response against vaccine antigen (and disease-associated antigen) in the subject.
1 . A medical preparation comprising:
(i) non-immunogenic RNA encoding a peptide or protein comprising an epitope for inducing an immune response against an antigen in a subject; and
(ii) non-immunogenic RNA encoding interferon α, wherein the non-immunogenic RNA is rendered non-immunogenic by the incorporation of modified nucleosides and the removal of double-stranded RNA (dsRNA).
2 . The medical preparation of claim 1 , which is for treating a disease, disorder or condition associated with expression or elevated expression of an antigen.
3 . The medical preparation of claim 1 , wherein
(i) the antigen is a tumor-associated antigen; or
(ii) the disease, disorder or condition is cancer.
4 . The medical preparation of claim 1 wherein:
a) the non-immunogenic RNA when administered results in reduced activation of dendritic cells, activation of T cells and/or secretion of IFN-alpha compared to standard RNA; or
b) the non-immunogenic RNA encoding interferon α, is mRNA or in vitro transcribed RNA.
5 . The medical preparation of claim 1 , wherein the modified nucleosides suppress RNA-mediated activation of innate immune receptors and/or comprise a replacement of one or more uridines with a nucleoside comprising a modified nucleobase, such as a modified uracil, wherein optionally the nucleoside comprising a modified nucleobase is selected from the group consisting of 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), 5-aza-uridine, 6-aza-uridine, 2-thio-5-aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridineor 5-bromo-uridine), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl-uridine (cm5U), 1-carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uridine (chm5U), 5-carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5-methoxycarbonylmethyl-2-thio-uridine (mcm5s2U), 5-aminomethyl-2-thio-uridine (nm5s2U), 5-methylaminomethyl-uridine (mnm5U), 1-ethyl-pseudouridine, 5-methylaminomethyl-2-thio-uridine (mnm5s2U), 5-methylaminomethyl-2-seleno-uridine (mnm5se2U), 5-carbamoylmethyl-uridine (ncm5U), 5-carboxymethylaminomethyl-uridine (cmnm5U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyl-uridine (τm5U), 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine (tm5s2U), 1-taurinomethyl-4-thio-pseudouridine, 5-methyl-2-thio-uridine (m5s2U), 1-methyl-4-thio-pseudouridine (m1s4ψ), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m3w), 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6-dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxy-uridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine, 3-(3-amino-3-carboxypropyl) uridine (acp3U), 5-1-methyl-3-(3-amino-3-carboxypropyl) pseudouridine (acp3 ψ), (isopentenylaminomethyl) uridine (inm5U), 5-(isopentenylaminomethyl)-2-thio-uridine (inm5s2U), α-thio-uridine, 2′-O-methyl-uridine (Um), 5,2′-O-dimethyl-uridine (m5Um), 2′-O-methyl-pseudouridine (ψm), 2-thio-2′-O-methyl-uridine (s2Um), 5-methoxycarbonylmethyl-2′-O-methyl-uridine (mcm5Um), 5-carbamoylmethyl-2′-O-methyl-uridine (ncm5Um), 5-carboxymethylaminomethyl-2′-O-methyl-uridine (cmnm5Um), 3,2′-O-dimethyl-uridine (m3Um), 5-(isopentenylaminomethyl)-2′-O-methyl-uridine (inm5Um), 1-thio-uridine, deoxythymidine, 2′-F-ara-uridine, 2′-F-uridine, 2′-OH-ara-uridine, 5-(2-carbomethoxyvinyl) uridine, and 5-[3-(1-E-propenylamino) uridine, the nucleoside comprising a modified nucleobase preferably being pseudouridine (ψ), N1-methyl-pseudouridine (m 1 ψ) or 5-methyl-uridine (m5U), the nucleoside comprising a modified nucleobase most preferably being 1-methyl-pseudouridine.
6 . The medical preparation of claim 1 , wherein
(i) the non-immunogenic RNA encoding a peptide or protein comprising an epitope is present in a formulation for targeting the lymphatic system and/or dendritic cells, such as immature dendritic cells; and/or
(ii) wherein the RNA encoding an immunostimulant is administered in a formulation for targeting the lymphatic system, the liver and or secondary lymphoid organs, in particular spleen.
7 . The medical preparation of claim 1 , wherein:
(i) the non-immunogenic RNA encoding a peptide or protein comprising an epitope and the non-immunogenic RNA encoding interferon α are present in a common or separate formulation;
(ii) the medical preparation is a kit;
(iii) the medical preparation comprises the non-immunogenic RNA encoding a peptide or protein comprising an epitope and the non-immunogenic RNA encoding interferon α in a pharmaceutical composition;
(iv) the medical preparation comprises the non-immunogenic RNA encoding a peptide or protein comprising an epitope and non-immunogenic RNA encoding interferon α in separate containers;
(v) the medical preparation further comprises instructions for using the medical preparation; and/or
(vi) the medical preparation is a pharmaceutical composition.
8 . The medical preparation of claim 1 for pharmaceutical use.
9 . The medical preparation of claim 8 , wherein the pharmaceutical use comprises a therapeutic or prophylactic treatment of a disease or disorder.
10 . The medical preparation of claim 9 , wherein the disease or disorder is cancer.