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, or 9 nucleotide units in length, for use in reducing the effective amount of a microRNA target in a cell or an organism, 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-21 (SEQ ID NO: 410), and wherein the seed sequence of miR-21 consists of nucleotides 2 to 8 of SEQ ID NO: 410 counting from the sequence's 5′ end.
2. The oligomer according to claim 1 , for use in the treatment of a medical disorder or disease wherein the medical disease or disorder is glioblastoma, breast cancer, hepatocellular carcinoma, colorectal cancer, sensitization of gliomas to cytotoxic drugs, or cardiac hypertrophy.
3. A pharmaceutical composition comprising the oligomer according to claim 1 , and a pharmaceutically acceptable diluent, carrier, salt or adjuvant.
4. The pharmaceutical composition according to claim 3 , further comprising a second independent active ingredient that is a chemotherapeutic agent.
5. 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 3 to a patient who is suffering from, or is likely to suffer from said disease or medical disorder.
6. 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.
7. 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.
8. The oligomer according to claim 6 , 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.
9. The oligomer according to claim 6 which comprises a 3′ terminal LNA unit and a 5′ terminal LNA unit.
10. The oligomer according to claim 6 , 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.
11. The oligomer according to claim 7 , wherein all of the nucleotide units of the contiguous nucleotide sequence are LNA units.
12. 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.
13. 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.
14. 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.
15. The oligomer according to claim 11 , wherein said contiguous nucleotide sequence of the oligomer does not comprise a nucleotide which corresponds to the first nucleotide present in the microRNA sequence of miR-21 (SEQ ID NO: 410) counted from the 5′ end.
16. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer comprises the sequence:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
17. The oligomer according to claim 6 , wherein the contiguous nucleotide sequence of the oligomer comprises the sequence:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
18. The oligomer according to claim 7 , wherein the contiguous nucleotide sequence of the oligomer comprises the sequence:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
19. The oligomer according to claim 1 , wherein the contiguous nucleotide sequence of the oligomer is:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
20. The oligomer according to claim 6 , wherein the contiguous nucleotide sequence of the oligomer is:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
21. The oligomer according to claim 7 , wherein the contiguous nucleotide sequence of the oligomer is:
(a) 5′-TGATAAGCT-3′ (SEQ ID NO: 9);
(b) 5′-GATAAGCT-3′ (SEQ ID NO: 2); or,
(c) 5′-ATAAGCT-3′ (SEQ ID NO: 8).
22. The oligomer according to claim 11 , 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.
23. The oligomer according to claim 22 , wherein the oligomer sequence comprises 5′-ATAAGCT-3′ (SEQ ID NO: 8).
24. The oligomer according to claim 22 , wherein the oligomer sequence is 5′-ATAAGCT-3′ (SEQ ID NO: 8).
25. The oligomer according to claim 11 , 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-methylcytosines.
26. The oligomer according to claim 25 , wherein the oligomer sequence comprises 5′-GATAAGCT-3′ (SEQ ID NO: 2).
27. The oligomer according to claim 25 , wherein the oligomer sequence is 5′-GATAAGCT-3′ (SEQ ID NO: 2).
28. The oligomer according to claim 11 , 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-methylcytosines.
29. The oligomer according to claim 28 , wherein the oligomer sequence comprises 5′-TGATAAGCT-3′ (SEQ ID NO: 9).
30. The oligomer according to claim 28 , wherein the oligomer sequence is 5′-TGATAAGCT-3′ (SEQ ID NO: 9).
31. The method according to claim 5 , wherein the disease or medical disorder is cancer.
32. The oligomer according to claim 1 , wherein the oligomer is conjugated with at least one non-nucleotide or non-polynucleotide moiety.
33. The oligomer according to claim 32 , 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.
34. The oligomer according to claim 33 , wherein the protein is an antibody.
35. The oligomer according to claim 33 , wherein the polymer is polyethylene glycol.
36. 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.
37. The oligomer according to claim 1 , wherein at least one cytosine LNA units is not 5-methylcytosine.
38. The oligomer according to claim 1 , wherein at least one cytosine LNA unit is 5-methylcytosine.
39. The oligomer according to claim 1 , wherein all cytosine LNA units are 5-methylcytosines.
40. The oligomer according to claim 6 , wherein at least one cytosine LNA units is not 5-methylcytosine.
41. The oligomer according to claim 6 , wherein at least one cytosine LNA unit is 5-methylcytosine.
42. The oligomer according to claim 6 , wherein all cytosine LNA units are 5-methylcytosines.