Granted Patent B2

US 12,460,204 · App. 16/920,701 · Granted Nov 4, 2025

Inventor: Andreas Thess (Kusterdingen, DE)

Assignee: CureVac SE

C12N15/113A61K39/00A61K48/0008C12N15/67C12N15/85A61K2039/5156A61K2039/5256A61K2039/53A61K48/00C12N2310/531C12N2320/52C12N2840/105

Abstract

Claims (41)

1 . An artificial nucleic acid molecule selected from the group consisting of

(i) a DNA molecule comprising, from 5′ to 3′:

a) at least one 5-untranslated region element (5′ UTR element) which comprises a nucleic acid sequence having an identity of at least 90% to the nucleic acid sequence according to SEQ ID NO: 1455; and

b) at least one open reading frame (ORF), wherein the ORF is heterologous relative to the 5′ UTR element; and

(ii) a RNA molecule encoded by the DNA molecule.

2 . The artificial nucleic acid molecule according to claim 1 , further comprising:

c) at least one histone stem-loop.

3 . The artificial nucleic acid molecule according to claim 1 , further comprising at least one 3′ UTR element of a mammalian gene positioned 3′ relative to the ORF.

4 . The artificial nucleic acid molecule according to claim 3 , wherein the 3′ UTR element comprises a 3′ UTR element of a gene selected from the group consisting of an albumin gene, an α-globin gene, a β-globin gene, a tyrosine hydroxylase gene, a lipoxygenase gene, and a collagen alpha gene.

5 . The artificial nucleic acid molecule according to claim 4 , wherein the 3′ UTR element comprises a 3′ UTR element of a human albumin gene.

6 . The artificial nucleic acid molecule according to claim 3 , further comprising a poly(A) sequence and/or polyadenylation signal, wherein the poly(A) sequence and/or polyadenylation signal is located within the 3′ UTR element.

7 . The artificial nucleic acid molecule according to claim 6 , wherein the poly(A) sequence has a length of 20 to 300 adenine nucleotides.

8 . The artificial nucleic acid molecule according to claim 3 , wherein the at least one 3′ UTR element comprises an mRNA-stabilizing element.

9 . The artificial nucleic acid molecule according to claim 8 , further comprising a poly(C) sequence positioned 3′ relative to the ORF.

10 . The artificial nucleic acid molecule according to claim 1 , wherein the molecule is a DNA and further comprises a promoter containing-sequence operably linked to the ORF.

11 . The artificial nucleic acid molecule according to claim 1 , wherein the molecule is a RNA.

12 . The artificial nucleic acid molecule according to claim 11 , wherein the RNA is a mRNA and comprises a 5′ cap.

13 . The artificial nucleic acid molecule according to claim 1 , wherein the RNA comprises at least one nucleotide position that is substituted with an analogue of the nucleotide selected from the group consisting of 2-amino-6-chloropurineriboside-5′ triphosphate, 2-aminoadenosine-5′-triphosphate, 2-thiocytidine-5′-triphosphate, 2-thiouridine-5′-triphosphate, 4-thiouridine-5′-triphosphate, 5-aminoallylcytidine-5′ triphosphate, 5-aminoallyluridine-5′-triphosphate, 5-bromocytidine-5′-triphosphate, 5-bromouridine-5′-triphosphate, 5-iodocytidine-5′-triphosphate, 5-iodouridine-5′ triphosphate, 5-methylcytidine-5′-triphosphate, 5-methyluridine-5′-triphosphate, 6-azacytidine-5′-triphosphate, 6-azauridine-5′-triphosphate, 6-chloropurineriboside-5′-triphosphate, 7-deazaadenosine-5′-triphosphate, 7-deazaguanosine-5′-triphosphate, 8-azaadenosine-5′-triphosphate, 8-azidoadenosine-5′-triphosphate, benzimidazoleriboside-5′-triphosphate, N1-methyladenosine-5′-triphosphate, N1-methylguanosine-5′-triphosphate, triphosphate, N6-methyladenosine-5′-triphosphate, 06-methylguanosine-5′-pseudouridine-5′-triphosphate, puromycin-5′-triphosphate and xanthosine-5′-triphosphate.

14 . The artificial nucleic acid molecule according to claim 1 , wherein one or more codon of the open reading frame is modified to a codon encoding the same amino acid but having an increased G/C content compared to the wild-type codon, thereby increasing the G/C content of the open reading frame compared to the wild type version of the open reading frame.

15 . The artificial nucleic acid molecule according to claim 1 , wherein the ORF encodes a reporter polypeptide, a human polypeptide, a tumour antigen or an infectious disease antigen.

16 . The artificial nucleic acid molecule according to claim 15 , wherein the ORF encodes an infectious disease antigen.

17 . The artificial nucleic acid molecule according to claim 16 , wherein the ORF encodes a viral antigen.

18 . A pharmaceutical composition comprising a RNA molecule in accordance with claim 11 in a pharmaceutically acceptable carrier.

19 . The artificial nucleic acid molecule according to claim 1 , wherein the at least one ORF is at least partially codon-optimized.

20 . The artificial nucleic acid molecule according to claim 19 , wherein the at least one 5′ UTR element comprises a nucleic acid sequence having an identity of at least 95% to the DNA sequence or the RNA sequence encoded by the nucleic acid sequence according to SEQ ID NO: 1455.

21 . An artificial RNA molecule comprising, from 5′ to 3′:

i) a 5′-cap structure;

ii) at least one 5′-untranslated region element (5′ UTR element) which comprises a nucleic acid sequence having an identity of at least 90% to the RNA sequence encoded by the nucleic acid sequence according to SEQ ID NO: 1455; and

iii) at least one open reading frame (ORF), wherein the ORF is heterologous relative to the 5′ UTR element.

22 . The artificial RNA molecule of claim 21 , wherein the at least one ORF is at least partially codon-optimized.

23 . The artificial RNA molecule of claim 21 , further comprising at least one histone stem-loop.

24 . The artificial RNA molecule of claim 21 , further comprising at least one 3′-untranslated region element (3′ UTR element).

25 . The artificial RNA molecule of claim 24 , further comprising a poly(A) sequence.

26 . The artificial RNA molecule of claim 25 , wherein the poly(A) sequence has a length of 20 to 300 adenine nucleotides.

27 . The artificial RNA molecule of claim 26 , wherein the at least one ORF encodes a reporter polypeptide, a human polypeptide, a tumour antigen or an infectious disease antigen.

28 . The artificial RNA molecule of claim 27 , wherein the ORF encodes an infectious disease antigen.

29 . The artificial RNA molecule of claim 28 , wherein the ORF encodes a viral antigen.

30 . The artificial RNA molecule of claim 28 , wherein the artificial RNA is a mRNA.

31 . The artificial RNA molecule of claim 28 , wherein the at least one 5′ UTR element comprises a nucleic acid sequence at least 95% identical to the RNA sequence encoded by the nucleic acid sequence according to SEQ ID NO: 1455.

32 . The artificial RNA molecule of claim 31 , wherein the at least one 5′ UTR element comprises a nucleic acid sequence identical to the RNA sequence encoded by the nucleic acid sequence according to SEQ ID NO: 1455.

33 . A pharmaceutical composition comprising an artificial RNA molecule in accordance with claim 30 in a pharmaceutically acceptable carrier.

Assignments (1)

Priority Claims (2)

Continuity (3)

References Cited (400)