Means for inhibiting the expression of Orc-1
The present invention is related to a nucleic acid molecule comprising a double-stranded structure, whereby the double-stranded structure comprises a first strand and a second strand, whereby the first strand comprises a first stretch of contiguous nucleotides and said first stretch is at least partially complementary to a target nucleic acid, and whereby the second strand comprises a second stretch of contiguous nucleotides and said second stretch is at least partially complementary to the first stretch, whereby the first stretch comprises a nucleic acid sequence which is at least complementary to a nucleotide core sequence of the nucleic acid sequence according to SEQ ID NO: 1, whereby the nucleotide core sequence comprises the nucleotide sequence from nucleotide positions 1755 to 1763 of SEQ ID NO: 1; from nucleotide positions 1904 to 1912 of SEQ.ID.No.1; from nucleotide positions 1905 to 1913 of SEQ ID NO: 1; from nucleotide positions 2548 to 2556 of SEQ ID NO: 1; whereby the first stretch is additionally at least partially complementary to a region preceding the 5′ end of the nucleotide core sequence and/or to a region following the 3′ end of the nucleotide core sequence.
1. A nucleic acid molecule comprising a double-stranded structure that is optionally covalently linked and each strand of the doubled-stranded structure has a length of 18-29 nucleotides,
wherein the first stretch and/or the second stretch have a plurality of modified nucleotides having a modification at the 2′ position
wherein the first strand comprises a first stretch of contiguous nucleotides and said first stretch is at least partially complementary to a target nucleic acid, and
wherein the second strand comprises a second stretch of contiguous nucleotides and said second stretch is at least partially complementary to the first stretch,
wherein the first stretch consists of a nucleotide sequence according to SEQ ID NOS: 6, 34, 20 or 36 and the second stretch consists of a nucleotide sequence according to SEQ ID NOS: 7, 35, 21 or 37.
2. The nucleic acid according to claim 1 , wherein the first stretch and/or the second stretch are: a) 18 to 29 consecutive nucleotides, b) 19 to 25 consecutive nucleotides or c) 19 to 23 consecutive nucleotides.
3. The nucleic acid molecule according to claim 1 , wherein the first stretch and/or the second stretch have a plurality of modified nucleotides having a modification at the 2′ position, wherein within the stretch each group of modified nucleotides is flanked on one or both sides by a flanking group of nucleotides, wherein the flanking nucleotide(s) forming the flanking group of nucleotides is/are either an unmodified nucleotide or a nucleotide having a modification different from the modification of the modified nucleotides, wherein the first stretch and/or the second stretch comprises at least two groups of modified nucleotides and at least two flanking groups of nucleotides.
4. The nucleic acid according to claim 1 , wherein the first stretch and/or the second stretch contain a pattern of groups of modified nucleotides and/or a pattern of flanking groups of nucleotides, wherein the pattern is a positional pattern.
5. The nucleic acid according to claim 1 , wherein the first stretch and/or the second stretch comprise at the 3′ end a dinucleotide.
6. The nucleic acid according to claim 1 , wherein the first and/or the second stretch have an overhang of 1 to 5 nucleotides at the 3′ end.
7. The nucleic acid according to claim 1 , wherein the length of the double-stranded structure is from about 16 to 27 nucleotide pairs.
8. A lipoplex comprising a liposome and a nucleic acid molecule comprising a double-stranded structure that is optionally covalently linked and each strand of the doubled-stranded structure has a length of 18-29 nucleotides,
wherein the first strand comprises a first stretch of contiguous nucleotides and said first stretch is at least partially complementary to a target nucleic acid, and
wherein the second strand comprises a second stretch of contiguous nucleotides and said second stretch is at least partially complementary to the first stretch,
wherein the first stretch consists of a nucleotide sequence according to SEQ ID NOS: 6, 34, 20 or 36 and the second stretch consists of a nucleotide sequence according to SEQ ID NOS: 7, 35, 21 or 37.
9. The lipoplex according to claim 8 , wherein the liposome consists of
a) about 50 mol % β-arginyl-2,3-diaminopropionic acid-N-palmityl-N-oleyl-amide trihydrochloride;
b) about 48 to 49 mol % 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPhyPE); and
c) about 1 to 2 mol % 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene-glycole.
10. A vector comprising or encoding a nucleic acid according to claim 1 .
11. An isolated cell comprising a nucleic acid according to claim 1 .
12. A composition, comprising a nucleic acid according to claim 1 .
13. The composition according to claim 12 , wherein the composition is a pharmaceutical composition, said pharmaceutical composition optionally further comprising a pharmaceutically acceptable vehicle.
14. The nucleic acid molecule according to claim 3 , wherein said nucleotides are modified with amino, fluoro, methoxy, alkoxy or alkyl groups.
15. The nucleic acid molecule according to claim 14 , wherein said nucleotides are modified with methoxy groups.
16. The lipoplex according to claim 8 , wherein the nucleic acid has a first stretch and/or the second stretch that are: a) 18 to 29 consecutive nucleotides, b) 19 to 25 consecutive nucleotides or c) 19 to 23 consecutive nucleotides.
17. The lipoplex according to claim 8 , wherein the first stretch and/or the second stretch of said nucleic acid molecule have a plurality of modified nucleotides having a modification at the 2′ position, wherein within the stretch each group of modified nucleotides is flanked on one or both sides by a flanking group of nucleotides, wherein the flanking nucleotide(s) forming the flanking group of nucleotides is/are either an unmodified nucleotide or a nucleotide having a modification different from the modification of the modified nucleotides, wherein the first stretch and/or the second stretch comprises at least two groups of modified nucleotides and at least two flanking groups of nucleotides.
18. The lipoplex according to claim 8 , wherein the first stretch and/or the second stretch of said nucleic acid contain a pattern of groups of modified nucleotides and/or a pattern of flanking groups of nucleotides, wherein the pattern is a positional pattern.
19. The lipoplex according to claim 8 , wherein the first stretch and/or the second stretch of said nucleic acid comprise at the 3′ end a dinucleotide.
20. The lipoplex according to claim 8 , wherein the first and/or the second stretch of said nucleic acid molecule have an overhang of 1 to 5 nucleotides at the 3′ end.
21. The lipoplex according to claim 8 , wherein the length of the double-stranded structure is from about 16 to 27 nucleotide pairs.