US2016053264A1PendingUtilityA1
Synthetic mimics of mir-34
Est. expiryFeb 3, 2031(~4.6 yrs left)· nominal 20-yr term from priority
C12N 2310/141C12N 2310/321A61P 35/00C12N 15/113C12N 2310/317A61K 48/00C12N 2310/11A61K 45/06A61K 31/713A61P 43/00
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Claims
Abstract
Embodiments concern methods and compositions involving miR-34 mimics, including miR-34a and miR-34c mimics. In some embodiments, there are double-stranded RNA molecules with modified nucleotides having an active strand with a miR-34a sequence and a complementary passenger strand. In additional embodiments, there are double-stranded RNA molecules with modified nucleotides having an active strand with a miR-34c sequence and a complementary passenger strand.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . A double-stranded, blunt-ended RNA molecule 22 or 23 basepairs in length comprising:
a) an active strand comprising SEQ ID NO:1 from 5′ to 3′ and b) a fully complementary passenger strand comprising
i) modified nucleotides in the first and last two nucleotides of the passenger strand; and
ii) a terminal modification of the nucleotide at the 5′ end.
2 . The RNA molecule of claim 1 , wherein the molecule is 23 basepairs in length and the active strand comprises SEQ ID NO:2.
3 . The RNA molecule of claim 1 , wherein the passenger strand comprises a modified nucleotide at positions 1, 2, 3, 4, 5, 6, 9, 10, 11, 12, 13, 14, 17, 18, 19, 20, 21, 22, and/or 23 relative to SEQ ID NO:4, wherein fewer than half of the total number of nucleotides in the passenger strand are modified nucleotides.
4 . The RNA molecule of any of claims 1 - 3 , wherein the passenger strand comprises modified nucleotides located at positions i) 1 and 2 and/or ii) 22 and 23 relative to SEQ ID NO:4.
5 . The RNA molecule of any of claims 2 - 3 , wherein the passenger strand comprises modified nucleotides located at positions i) 1 and 2 and ii) 22 and 23 relative to SEQ ID NO:4.
6 . The RNA molecule of any of claims 1 - 5 , wherein the passenger strand comprises a modified nucleotide located at positions i) 3 and 4 and/or ii) 19 and 20 relative to SEQ ID NO:4.
7 . The RNA molecule of claim 6 , wherein the passenger strand comprises a modified nucleotide located at positions i) 3 and 4 or ii) 19 and 20 relative to SEQ ID NO:4, but not at both positions i) and ii).
8 . The RNA molecule of any of claims 4 - 7 , wherein the passenger strand comprises modified nucleotides at positions 5 and 6 relative to SEQ ID NO:4.
9 . The RNA molecule of any of claims 4 - 7 , wherein the passenger strand comprises a modified nucleotide at positions 9 and 10 relative to SEQ ID NO:4.
10 . The RNA molecule of any of claims 4 - 7 , wherein the passenger strand comprises a modified nucleotide at positions 11 and 12 relative to SEQ ID NO:4.
11 . The RNA molecule of any of claims 4 - 7 , wherein the passenger strand comprises a modified nucleotide at positions 13 and 14 relative to SEQ ID NO:4.
12 . The RNA molecule of any of claims 4 - 7 , wherein the passenger strand comprises a modified nucleotide at positions 17 and 18 relative to SEQ ID NO:4.
13 . The RNA molecule of claim 4 , wherein the number of modified nucleotides in the passenger strand is four to eight.
14 . The RNA molecule of claim 13 , wherein the number of modified nucleotides in the passenger strand is five to seven.
15 . The RNA molecule of claim 14 , wherein the number of modified nucleotides in the passenger strand is six.
16 . The RNA molecule of any of claims 1 - 15 , wherein the passenger strand does not have a modified nucleotide located at positions 7, 8, 9, 15, and 16 relative to SEQ ID NO:4.
17 . The RNA molecule of any of claims 1 - 16 , wherein the passenger strand comprises a modified nucleotide at positions 1, 2, 21, 22, and 23 relative to SEQ ID NO:4.
18 . The RNA molecule of any of claims 1 - 16 , wherein the passenger strand comprises a modified nucleotide at positions 1, 2, 3, 4, 22, and 23 relative to SEQ ID NO:4.
19 . The RNA molecule of any of claims 1 - 16 , wherein the passenger strand comprises a modified nucleotide at positions 1, 2, 19, 20, 22, and 23 relative to SEQ ID NO:4.
20 . The RNA molecule of any of claims 1 - 19 , wherein the active strand comprises at least two modified nucleotides, wherein the modified nucleotides are not in the first two positions at the 5′ end.
21 . The RNA molecule of any of claims 1 - 19 , wherein the active strand does not have a modified nucleotide in the first two positions at the 5′ end.
22 . The RNA molecule of claim 1 , wherein the active strand does not comprise a modified nucleotide in the first or last two positions from the ends.
23 . The RNA molecule of any of claims 20 - 22 , wherein the active strand has modified nucleotides at positions 3, 4, 11, 12, 13, 14, 15, 16, 17, and/or 18 relative to SEQ ID NO:2.
24 . The RNA molecule of claim 23 , wherein the active strand comprises modified nucleotides at positions 15 and 16 relative to SEQ ID NO:2.
25 . The RNA molecule of claim 23 , wherein the active strand comprises modified nucleotides at positions 3 and 4 relative to SEQ ID NO:2.
26 . The RNA molecule of claim 23 , wherein the active strand comprises modified nucleotides at positions 11 and 12 relative to SEQ ID NO:2.
27 . The RNA molecule of claim 23 , wherein the active strand comprises modified nucleotides at positions 11, 12, 13, and 14 relative to SEQ ID NO:2.
28 . The RNA molecule of claim 23 , wherein the active strand comprises modified nucleotides at positions 17 and 18 relative to SEQ ID NO:2.
29 . The RNA molecule of any of claims 23 - 28 , wherein the active strand comprises modified nucleotides at positions 11, 12, 15, and 16 relative to SEQ ID NO:2.
30 . The RNA molecule of claim 29 , wherein the active strand further comprises modified nucleotides at positions 13, 14, 17, and 18 relative to SEQ ID NO:2.
31 . The RNA molecule of any of claims 23 - 28 , wherein the active strand comprises modified nucleotides at positions 3, 4, 15, and 16 relative to SEQ ID NO:2.
32 . The RNA molecule of any of claims 1 - 19 , wherein the active strand does not have any modified nucleotides.
33 . The RNA molecule of claim 29 , wherein the passenger strand does not comprise any modified nucleotides except a 5′ terminal modification.
34 . The RNA molecule of claim 29 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 20, 21, 22, and 23 relative to SEQ ID NO:4.
35 . The RNA molecule of claim 29 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 3, 4, 22, and 23 relative to SEQ ID NO:4.
36 . The RNA molecule of any of claim 29 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 19, 20, 22, and 23 relative to SEQ ID NO:4.
37 . The RNA molecule of claim 30 , wherein the passenger strand does not comprise any modified nucleotides except a 5′ terminal modification.
38 . The RNA molecule of claim 30 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 20, 21, 22, and 23 relative to SEQ ID NO:4.
39 . The RNA molecule of claim 30 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 3, 4, 22, and 23 relative to SEQ ID NO:4.
40 . The RNA molecule of any of claim 30 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 19, 20, 22, and 23 relative to SEQ ID NO:4. [A-ID 206/P-ID 215]
41 . The RNA molecule of claim 31 , wherein the passenger strand does not comprise any modified nucleotides except a 5′ terminal modification.
42 . The RNA molecule of claim 31 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 21, 22, and 23 relative to SEQ ID NO:4.
43 . The RNA molecule of claim 31 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 3, 4, 22, and 23 relative to SEQ ID NO:4.
44 . The RNA molecule of any of claim 31 , wherein the passenger strand comprises SEQ ID NO:4 and modified nucleotides at positions 1, 2, 19, 20, 22, and 23 relative to SEQ ID NO:4.
45 . The RNA molecule of any of claims 1 - 44 , wherein the terminal modification of the passenger strand comprises a lower alkylamine group.
46 . The RNA molecule of any of claims 1 - 45 , wherein the modified nucleotides have sugar modifications.
47 . The RNA molecule of claim 46 , wherein the sugar modification is 2′-OMe.
48 . A double-stranded RNA molecule of 20-23 basepairs in length, wherein the RNA molecule is blunt-ended at both ends, comprising an active strand having the sequence of SEQ ID NO:2 and a separate and fully complementary passenger strand with a modified nucleotide at the 5′ end, wherein the active strand comprises at least one modified internal nucleotide and wherein the double-stranded RNA molecule is more stable compared to a double-stranded, blunt-ended RNA molecule lacking any modification of an internal nucleotide.
49 . A pharmaceutical composition comprising the RNA molecule of any of claims 1 - 48 .
50 . A method for providing miR-34a activity to a cell comprising administering to the cell the RNA molecule of any of claims 1 - 48 .
51 . A method for decreasing cell proliferation comprising administering to the cell an effective amount of the RNA molecule of any of claims 1 - 48 .
52 . A method for inducing apoptosis in a cell comprising administering to the cell an effective amount of the RNA molecule of any of claims 1 - 48 .
53 . A method for treating cancer in a patient comprising administering to the patient a pharmaceutical composition comprising the RNA molecule of any of claims 1 - 48 .
54 . The method of claim 54 , further comprising administering to the patient an additional cancer therapy.
55 . The method of claim 53 , wherein the patient has been diagnosed with cancer.
56 . A double-stranded, blunt-ended RNA molecule 23 or 24 basepairs in length comprising:
a) an active strand comprising
i) SEQ ID NO:5 from 5′ to 3′ and
ii) at least one modified internal nucleotide; and,
b) a fully complementary passenger strand comprising a terminal modification of the nucleotide at the 5′ end.
57 . The RNA molecule of claim 56 , wherein the molecule is 23 basepairs in length.
58 . The RNA molecule of claim 56 , wherein the passenger strand comprises a modified nucleotide at positions 1 (G), 2 (C), 3 (A), 21 (C), 22 (C), and/or 23 (U) relative to SEQ ID NO:6.
59 . The RNA molecule of any of claims 56 - 58 , wherein the passenger strand comprises modified nucleotides located at a subset of positions, wherein the subset is i) 1 (A), 2 (A), and 3 (C) and/or ii) 22 (C), and 23 (A) relative to SEQ ID NO:6.
60 . The RNA molecule of any of claims 56 - 59 , wherein the passenger strand comprises a modified nucleotide located at positions i) 1 (A), 2 (A), and 3 (C) relative to SEQ ID NO:6.
61 . The RNA molecule of any of claims 56 - 60 , wherein the passenger strand comprises a modified nucleotide located at positions ii) 22 (C), and 23 (A) relative to SEQ ID NO:6.
62 . The RNA molecule of claim 59 , wherein the passenger strand comprises a modified nucleotide located at positions 1 (A), 2 (A), 3 (C), 22 (C), and 23 (A) relative to SEQ ID NO:6.
63 . The RNA molecule of claim 59 , wherein the passenger strand comprises a modified nucleotide located at positions i) or ii) relative to SEQ ID NO:6, but not at both positions i) and ii).
64 . The RNA molecule of any of claims 56 - 63 , wherein the passenger strand further comprises a modified nucleotide located at positions 4 (A), 7 (A), 8 (G), 9 (C), 10 (U), 11 (A), 12 (A), 13 (C), 14 (U), 19 (U), and/or 21 (C) relative to SEQ ID NO:6.
65 . The RNA molecule of any of claim 56 , wherein the passenger strand does not have a modified nucleotide located at positions 5 (U), 6 (C), 15 (A), and/or 16 (C) relative to SEQ ID NO:6.
66 . The RNA molecule of any of claim 56 , wherein the number of modified nucleotides in the passenger strand is two to eight.
67 . The RNA molecule of claim 66 , wherein the number of modified nucleotides in the passenger strand is three to five.
68 . The RNA molecule of any of claim 56 , wherein the passenger strand comprises modified nucleotides at positions 1 (A), 2 (A), 3 (C), 17 (A), 18 (C), 21 (C), 22 (C), and 23 (A) relative to SEQ ID NO:6.
69 . The RNA molecule of any of claim 56 , wherein the passenger strand comprises modified nucleotides at positions 1 (A), 2 (A), 3 (C), 11 (A), 12 (A), 13 (C), 14 (U), 21 (C), 22 (C), and 23 (A) relative to SEQ ID NO:6.
70 . The RNA molecule of any of claim 56 , wherein the passenger strand comprises modified nucleotides at positions 1 (A), 2 (A), 3 (C), 11 (A), 12 (A), 13 (C), 14 (U), 17 (A), 18 (C), 21 (C), 22 (C), and 23 (A) relative to SEQ ID NO:6.
71 . The RNA molecule of any of claims 56 - 70 , wherein the active strand comprises at least two modified nucleotides, wherein the modified nucleotides are not in the first two positions at the 5′ end.
72 . The RNA molecule of any of claims 56 - 70 , wherein the active strand does not have a modified nucleotide in the first two positions at the 5′ end.
73 . The RNA molecule of claim 56 , wherein the active strand does not comprise a modified nucleotide in the first or last two positions from the ends.
74 . The RNA molecule of any of claims 71 - 73 , wherein the active strand comprises modified nucleotides at positions 3 (G), 4 (C), 11 (G), 12 (U), 13 (A), 14 (G), 15 (C), 16 (C), 17 (U), and/or 18 (G) relative to SEQ ID NO:5.
75 . The RNA molecule of claim 74 , wherein the active strand comprises a modified nucleotide at positions 11 (G) and 12 (U) relative to SEQ ID NO:5.
76 . The RNA molecule of claim 74 , wherein the active strand comprises a modified nucleotide at positions 17 (U) and 18 (G) relative to SEQ ID NO:5.
77 . The RNA molecule of claim 74 , wherein the active strand comprises a modified nucleotide at positions 11 (G), 12 (U), 17 (U), and 18 (G) relative to SEQ ID NO:5.
78 . The RNA molecule of claim 77 , wherein the active strand further comprises a modified nucleotide at positions 3 (G), 4 (C), 13 (A), 14 (G), 15 (C), and/or 16 (C) relative to SEQ ID NO:5.
79 . The RNA molecule of any of claims 74 - 78 , wherein the passenger strand does not comprise any modified nucleotides.
80 . The RNA molecule of claim 68 , wherein the active strand comprises a modified nucleotide at positions 11 (G), 12 (U), 17 (U), and 18 (G) relative to SEQ ID NO:5.
81 . The RNA molecule of claim 69 , wherein the active strand comprises a modified nucleotide at positions 11 (G), 12 (U), 17 (U), and 18 (G) relative to SEQ ID NO:5.
82 . The RNA molecule of any of claim 70 , wherein the active strand comprises a modified nucleotide at positions 11 (G), 12 (U), 17 (U), and 18 (G) relative to SEQ ID NO:5.
83 . The RNA molecule of any of claims 56 - 82 , wherein the terminal modification of the passenger strand comprises a lower alkylamine group.
84 . The RNA molecule of any of claims 56 - 83 , wherein the modified nucleotides have modified sugars.
85 . The RNA molecule of claim 84 , wherein the sugar modification is 2′-OMe.
86 . A double-stranded, blunt-ended RNA molecule 23 or 24 basepairs in length comprising:
a) an active strand comprising
i) SEQ ID NO:5 from 5′ to 3′ and
ii) at least one modified internal nucleotide; and,
b) a fully complementary passenger strand comprising a terminal modification of the nucleotide at the 5′ end and at least one sugar modification at positions 1 (G), 2 (C), 3 (A), 21 (C), 22 (C) and/or 23 (U) relative to SEQ ID NO:6.
87 . A double-stranded, blunt-ended RNA molecule 23 or 24 basepairs in length comprising:
a) an active strand comprising at least bases 1-23 of SEQ ID NO:5 from 5′ to 3′; and, b) a fully complementary passenger strand comprising:
i) a terminal modification of the nucleotide at the 5′ end;
ii) a sugar modification in the first three and the last three nucleotides; and,
iii) a sugar modification of the nucleotides at one or more of positions 11 (A), 12 (A), 13 (C), 14 (U), 17 (A), and/or 18 (C) relative to SEQ ID NO:6.
88 . A pharmaceutical composition comprising the RNA molecule of any of claims 56 - 87 .
89 . A method for providing miR-34c activity to a cell comprising administering to the cell the RNA molecule of any of claims 56 - 86 .
90 . A method for decreasing cell proliferation comprising administering to the cell an effective amount of the RNA molecule of any of claims 56 - 86 .
91 . A method for inducing apoptosis in a cell comprising administering to the cell an effective amount of the RNA molecule of any of claims 56 - 86 .
92 . A method for treating cancer in a patient comprising administering to the patient a pharmaceutical composition comprising the RNA molecule of any of claims 56 - 86 .
93 . The method of claim 92 , further comprising administering to the patient an additional cancer therapy.
94 . The method of claim 92 , wherein the patient has been diagnosed with cancer.
95 . A double-stranded, blunt-ended RNA molecule 22 or 23 basepairs in length comprising:
a) an active strand comprising
i) SEQ ID NO:7 from 5′ to 3′ and
ii) at least one modified internal nucleotide; and,
b) a fully complementary passenger strand comprising a terminal modification of the nucleotide at the 5′ end.
96 . The RNA molecule of claim 95 , wherein the molecule is 23 basepairs in length.
97 . The RNA molecule of claims 95 - 96 , wherein the active strand comprises a modified nucleotide at position 3, 4, 11, 12, 13, 14, 15, 16, 17, and/or 18 in the active strand.
98 . The RNA molecule of any of claims 95 - 97 , wherein the active strand comprises at least two modified internal nucleotides.
99 . The RNA molecule of claim 98 , wherein the active strand comprises no more than 11 modified internal nucleotides.
100 . The RNA molecule of any of claims 95 - 99 , wherein the passenger strand comprises at least one modified internal nucleotide.
101 . The RNA molecule of claim 100 , wherein the passenger strand comprises a modified nucleotide at position 1, 2, 3, 4, 9, 10, 11, 12, 13, 14, 19, 21, and/or 22 in the passenger strand.
102 . The RNA molecule of any of claims 95 - 101 , wherein the passenger strand does not have a modified nucleotide at position 15 and/or 16.
103 . The RNA molecule of any of claims 95 - 102 , wherein the passenger strand comprises SEQ ID NO:3.
104 . The RNA molecule of claim 103 , wherein the passenger strand comprises a modified nucleotide at position 1, 2, 3, 5, 6, 9, 10, 11, 12, 13, 14, 17, 18, 19, 20, 21, and/or 22 in the passenger strand.
105 . The RNA molecule of any of claims 95 - 102 , wherein the active strand comprises SEQ ID NO:5.
106 . The RNA molecule of claim 105 , wherein the passenger strand comprises a modified nucleotide at position 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 19, 21, 22, and/or 23 in the passenger strand.
107 . The RNA molecule of any of claims 95 - 106 , wherein the active strand comprises the sequence of SEQ ID NO:9.
108 . A double-stranded, blunt-ended RNA molecule 22 or 23 basepairs in length comprising:
a) an active strand comprising
i) SEQ ID NO:9 from 5′ to 3′ and
ii) a modified nucleotide at position 3, 4, 11, 12, 13, 14, 15, 16, 17, and/or 18;
b) a fully complementary passenger strand comprising a terminal modification of the nucleotide at the 5′ end.Cited by (0)
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