5′ CpG nucleic acids and methods of use
View Patent ↗The invention relates to a class of CpG immunostimulatory oligonucleotides containing a 5′TCG motif or a CG at or near the 5′ end that are useful for stimulating an immune response.
1. An isolated oligonucleotide comprising:
5′TCGX 1 X 2 N 1 3′
wherein X 1 is any nucleotide, X 2 is A, T, or C when X 1 is C or A, X 2 is A or G when X 1 is T, X 2 is any nucleotide when X 1 is G, N 1 is 2-95 nucleotides, wherein the CG dinucleotide is an unmethylated CG dinucleotide, wherein 5′ designates the 5′ end of the oligonucleotide and 3′ designates the 3′ end of the oligonucleotide, and wherein N 1 does not include an unmethylated CG motif, wherein the oligonucleotide is 13-100 nucleotides in length.
2. The oligonucleotide of claim 1 , wherein the oligonucleotide includes at least 1 modified internucleotide linkage.
3. The oligonucleotide of claim 1 , wherein the oligonucleotide includes at least 50% modified internucleotide linkage.
4. The oligonucleotide of claim 1 , wherein all internucleotide linkages of the oligonucleotide are modified.
5. The oligonucleotide of claim 2 , wherein the stabilized internucleotide linkage is a phosphorothioate linkage.
6. The oligonucleotide of claim 1 , wherein N 1 is N 2 N 3 and wherein N 2 is 8-40 nucleotides and N 3 is 2-5 pyrimidines.
7. The oligonucleotide of claim 6 , wherein N 3 is TTTTT.
8. The oligonucleotide of claim 6 , wherein N 3 is TT.
9. The oligonucleotide of claim 1 , wherein N 1 is at least 50% pyrimidine.
10. The oligonucleotide of claim 1 , wherein N 1 is at least 80% pyrimidine.
11. The oligonucleotide of claim 1 , wherein N 1 is free of Poly-A and Poly-G sequences.
12. The oligonucleotide of claim 1 , wherein N 1 is TN 2 and wherein N 2 is 8-94 nucleotides.
13. The oligonucleotide of claim 1 , wherein the oligonucleotide has a 3′-3′ linkage with one or two accessible 5′ ends.
14. The oligonucleotide of claim 13 , wherein the oligonucleotide has two accessible 5′ ends, each of which are 5′TCG.
15. A method for inducing cytokine production, comprising:
administering to a subject an oligonucleotide of claim 1 in an effective amount to induce a cytokine selected from the group consisting of, IFN-α and IL-10.