US 12,442,005 B2
Artificial nucleic acid molecules for improved protein expression
Andreas Thess, Kusterdingen (DE); Thomas Schlake, Gundelfingen (DE); and Stefanie Grund, Stuttgart (DE)
Assigned to CureVac SE, Tübingen (DE)
Filed by CureVac SE, Tübingen (DE)
Filed on Jun. 6, 2023, as Appl. No. 18/330,356.
Application 18/330,356 is a division of application No. 17/713,533, filed on Apr. 5, 2022, granted, now 11,761,009.
Application 17/713,533 is a division of application No. 17/542,430, filed on Dec. 5, 2021, granted, now 11,345,920, issued on May 31, 2022.
Application 17/542,430 is a division of application No. 17/408,332, filed on Aug. 20, 2021, granted, now 11,286,492, issued on Mar. 29, 2022.
Application 17/408,332 is a division of application No. 15/534,496, granted, now 11,149,278, issued on Oct. 19, 2021, previously published as PCT/EP2015/002501, filed on Dec. 11, 2015.
Claims priority of application No. PCT/EP2014/003334 (WO), filed on Dec. 12, 2014.
Prior Publication US 2023/0313207 A1, Oct. 5, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/67 (2006.01); A61K 39/00 (2006.01); A61K 39/12 (2006.01); A61K 39/145 (2006.01); A61K 39/205 (2006.01); A61K 48/00 (2006.01); C12N 15/68 (2006.01); C12N 15/85 (2006.01)
CPC C12N 15/67 (2013.01) [A61K 39/12 (2013.01); A61K 39/145 (2013.01); A61K 39/205 (2013.01); C12N 15/68 (2013.01); C12N 15/85 (2013.01); A61K 2039/53 (2013.01); A61K 2039/54 (2013.01); A61K 2039/575 (2013.01); A61K 48/00 (2013.01); C12N 2760/16134 (2013.01); C12N 2760/16171 (2013.01); C12N 2760/20134 (2013.01); C12N 2760/20171 (2013.01); C12N 2830/50 (2013.01)] 41 Claims
 
1. A method for stimulating an immune response in an organism, the method comprising administering to the organism, by intramuscular injection, a RNA molecule comprising:
a) a 5′-cap structure;
b) at least one open reading frame (ORF) encoding an antigen; and
c) a heterologous 3′-untranslated region (3′-UTR) comprising at least a first and a second poly(A) sequence, wherein:
(i) the first poly(A) sequence comprises at least 20 adenine nucleotides; and
(ii) the second poly(A) sequence comprises at least 60 adenine nucleotides,
wherein the first and the second poly(A) sequences are separated by a linker nucleic acid sequence comprising at least 3 consecutive nucleotides that are not adenine nucleotides,
wherein when the RNA molecule is administered intramuscularly to the organism, the RNA molecule yields increased expression of the antigen encoded by the at least one open reading frame in comparison to the expression from a reference nucleic acid molecule comprising an identical nucleic acid sequence as the RNA molecule but lacking the linker nucleic acid sequence.