The present invention relates to very short heavily modified oligonucleotides which target and inhibit microRNAs in vivo, and their use in medicaments and pharmaceutical compositions.
1. An oligomer of a contiguous sequence of 7, 8, 9, or 10 nucleotide units in length, wherein at least 70% of the nucleotide units of the oligomer are selected from the group consisting of LNA (Locked Nucleic Acid) units and 2′ substituted nucleotide analogues, and wherein at least 50% of the nucleotide units of the oligomer are LNA units, and wherein at least one of the internucleoside linkages present between the nucleotide units of the contiguous nucleotide sequence is a phosphorothioate internucleoside linkage, wherein the contiguous nucleotide sequence is 100% complementary to the seed sequence of miR-155 (SEQ ID NO: 315), and wherein the seed sequence of miR-155 consists of nucleotides 2 to 8 of SEQ ID NO: 315) counting from the sequence's 5′ end.
2. A pharmaceutical composition comprising the oligomer according to claim 1 , and a pharmaceutically acceptable diluent, carrier, salt or adjuvant.
3. The pharmaceutical composition according to claim 2 , further comprising a second independent active ingredient that is a chemotherapeutic agent.
4. A method for the treatment of a disease or medical disorder associated with the presence or over-expression of a microRNA, comprising the step of administering the pharmaceutical composition according to claim 2 to a patient who is suffering from, or is likely to suffer from said disease or medical disorder.
5. The oligomer according to claim 1 , wherein at least 75% of the internucleoside linkages present between the nucleotide units of the contiguous nucleotide sequence are phosphorothioate internucleoside linkages.
6. The oligomer according to claim 1 , wherein all the internucleoside linkages present between the nucleotide units of the contiguous nucleotide sequence are phosphorothioate internucleoside linkages.
7. The oligomer according to claim 5 , wherein the nucleotide units are selected from the group consisting of DNA, 2′-O-alkyl-RNA unit, 2′-OMe-RNA unit, 2′-amino-DNA unit, 2′-fluoro-DNA unit, LNA unit, PNA unit, HNA unit, INA unit, and a 2′-MOE RNA unit.
8. The oligomer according to claim 5 which comprises a 3′ terminal LNA unit and a 5″ terminal LNA unit.
9. The oligomer according to claim 5 , wherein the length of the oligomer is 7, 8 or 9 contiguous nucleotides, wherein each one of the contiguous nucleotides independently consists of a LNA unit or a 2′ substituted nucleotide analogue unit, and wherein none of the contiguous nucleotides is a DNA unit.
10. The oligomer according to claim 6 , wherein all of the nucleotide units of the contiguous nucleotide sequence are LNA units.
11. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer is 7 nucleotide units in length, all the nucleotide units are LNA units, and all the internucleoside linkages are phosphorothioate.
12. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer is 8 nucleotide units in length, all the nucleotide units are LNA units, and all the internucleoside linkages are phosphorothioate.
13. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer is 9 nucleotide units in length, all the nucleotide units are LNA units, and all the internucleoside linkages are phosphorothioate.
14. The oligomer according to claim 10 , wherein the contiguous nucleotide sequence of the oligomer does not comprise a nucleotide which corresponds to the first nucleotide present in the microRNA sequence of miR-155 (SEQ ID NO:315) counted from the 5′ end.
15. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer comprises a sequence selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
16. The oligomer according to claim 5 , wherein the contiguous nucleotide sequence of the oligomer comprises a sequence selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
17. The oligomer according to claim 6 , wherein the contiguous nucleotide sequence of the oligomer comprises a sequence selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
18. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer is selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
19. The oligomer according to claim 5 , wherein the contiguous nucleotide sequence of the oligomer is selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
20. The oligomer according to claim 6 , wherein the contiguous nucleotide sequence of the oligomer is selected from the group consisting of:
(a) 5′-TTAGCATTA-3′
(SEQ ID NO: 986)
(b) 5′-TAGCATTA-3′;
(SEQ ID NO: 4)
and
(c) 5′-AGCATTA-3′.
(SEQ ID NO: 987)
21. The oligomer according to claim 10 , wherein the contiguous nucleotide sequence of the oligomer is 7 nucleotide units in length, all the nucleotide units are LNA units, all the internucleoside linkages are phosphorothioate, and all cytosine LNA units are 5-methylcytosines.
22. The oligomer according to claim 21 , wherein the oligomer sequence comprises 5′-AGCATTA-3′ (SEQ ID NO: 987).
23. The oligomer according to claim 21 , wherein the oligomer sequence is 5′-AGCATTA-3′ (SEQ ID NO: 987).
24. The oligomer according to claim 10 , wherein the contiguous nucleotide sequence of the oligomer is 8 nucleotide units in length, all the nucleotide units are LNA units, all the internucleoside linkages are phosphorothioate, and all cytosine LNA units are 5-metlhylcytosines.
25. The oligomer according to claim 24 , wherein the oligomer sequence comprises 5′-TAGCATTA-3′ (SEQ ID NO: 4).
26. The oligomer according to claim 24 , wherein the oligomer sequence is 5′-TAGCATTA-3′ (SEQ ID NO: 4).
27. The oligomer according to claim 10 , wherein the contiguous nucleotide sequence of the oligomer is 9 nucleotide units in length, all the nucleotide units are LNA units, all the internucleoside linkages are phosphorothioate, and all cytosine LNA units are 5-methyl cytosines.
28. The oligomer according to claim 27 , wherein the oligomer sequence comprises 5′-TTAGCATTA-3′ (SEQ ID NO: 986).
29. The oligomer according to claim 27 , wherein the oligomer sequence is 5′-TTAGCATTA-3′ (SEQ ID NO: 986).
30. The method according to claim 4 , wherein the disease or medical, disorder is cancer.
31. The oligomer according to any of claims 1 , 2 , 3 and 5 - 29 , wherein the oligomer is conjugated with at least one non-nucleotide or non-polynucleotide moiety.
32. The oligomer according to claim 31 , wherein the non-nucleotide or non-polynucleotide moiety is selected from a protein, a fatty acid chain, a sugar residue, a glycoprotein, a polymer, or any combination thereof.
33. The oligomer according to claim 32 , wherein the protein is an antibody.
34. The oligomer according to claim 32 , wherein the polymer is polyethylene glycol.
35. The oligomer according to claim 1 , wherein at least one of the internucleoside linkages present between the nucleotide units of the contiguous nucleotide sequence is not a phosphorothioate or a phosphodiester internucleoside linkage.
36. The oligomer according to claim 1 , wherein at least one cytosine LNA units is not 5-methylcytosine.
37. The oligomer according to claim 1 , wherein at least one cytosine LNA unit is 5-methylcytosine.
38. The oligomer according to claim 1 , wherein all cytosine LNA units are 5-methylcytosines.
39. The oligomer according to claim 5 , wherein at least one cytosine LNA units is not 5-methylcytosine.
40. The oligomer according to claim 5 , wherein at least one cytosine LNA unit is 5-methylcytosine.
41. The oligomer according to claim 5 , wherein all cytosine LNA units are 5-methylcytosines.
42. The method of claim 4 wherein the disorder is selected from the group consisting of: lymphoma, pancreatic cancer, breast cancer, and lung cancer.