Means for inhibiting the expression of ANG2
The present invention is related to an siRNA comprising an antisense strand and a sense strand, wherein all or a portion of said antisense strand comprises an antisense duplex region, wherein all or a portion of said sense strand comprises a sense duplex region, wherein said antisense duplex region is at least partially complementary to said sense duplex region, wherein said siRNA comprises a duplex region consisting of said antisense duplex region and said sense duplex region, and wherein: a) said antisense strand comprises a nucleotide sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102 or 104; or b) said antisense strand comprises an antisense duplex region, all or a portion of which, is complementary to a portion of SEQ ID NO: 1 or 70.
1. A siRNA comprising an antisense strand between 19 and 25 nucleotides in length and a sense strand between 19 and 25 nucleotides in length, wherein one or more alternating nucleotides on the sense and/or antisense strands are modified, wherein said antisense strand and said sense strand have a duplex region length consisting of 16, 17, 18, or 19 nucleotides, wherein the duplex region is perfectly complementary or contains 1, 2, 3, 4, or 5 nucleotides that do not base pair and said siRNA comprises the antisense and sense combination of SEQ ID NOs: 14 and 15.
2. The siRNA of claim 1 , wherein said duplex region is 16 nucleotides in length and is perfectly complementary.
3. The siRNA of claim 1 , wherein said duplex region is 17 nucleotides in length and is perfectly complementary.
4. The siRNA of claim 1 , wherein said duplex region is 18 nucleotides in length and is perfectly complementary.
5. The siRNA of claim 1 , wherein said duplex region is 19 nucleotides in length and is perfectly complementary.
6. The siRNA of claim 1 , wherein said siRNA:
a) is blunt ended at both ends;
b) has an overhang at one end and a blunt end at the other; or
c) has an overhang at both ends.
7. The siRNA of claim 1 , wherein said alternating nucleotides on both the sense and antisense strands are modified with an O-methyl group at the 2′ position.
8. The siRNA of claim 7 , wherein each of the odd numbered nucleotides is modified in the antisense strand and each of the even numbered nucleotides is modified in the sense strand.
9. A lipoplex comprising a siRNA of claim 1 and a liposome.
10. The lipoplex of claim 9 , 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-polyethylen-glycol.
11. A vector comprising a promoter operably linked to a nucleic acid encoding or comprising an antisense strand between 19 and 25 nucleotides in length or a sense strand between 19 and 25 nucleotides in length, said antisense and sense strands comprising SEQ ID NOs: 14 and 15.
12. A cell comprising a vector according to claim 11 , provided that if the cell is a human cell, said human cell is an isolated cell.
13. A composition comprising a siRNA of claim 1 and a physiologically acceptable excipient.
14. A method for inhibiting angiopoietin (ANG2) expression in a subject comprising the administration of a composition comprising an siRNA molecule of claim 1 to a subject in an amount effective to inhibit the expression of ANG2.
15. A method for improving pulmonary vascular barrier function in a subject comprising the administration of a therapeutically effective amount of a composition comprising an siRNA molecule of claim 1 to a subject.
16. The siRNA of claim 2 , wherein said siRNA:
a) is blunt ended at both ends;
b) has an overhang at one end and a blunt end at the other; or
c) has an overhang at both ends.
17. The siRNA of claim 3 , wherein said siRNA:
a) is blunt ended at both ends;
b) has an overhang at one end and a blunt end at the other; or
c) has an overhang at both ends.
18. The siRNA of claim 4 , wherein said siRNA:
a) is blunt ended at both ends;
b) has an overhang at one end and a blunt end at the other; or
c) has an overhang at both ends.
19. The siRNA of claim 5 , wherein said siRNA:
a) is blunt ended at both ends;
b) has an overhang at one end and a blunt end at the other; or
c) has an overhang at both ends.
20. The siRNA of claim 1 , wherein said duplex region is 16 nucleotides in length and has 1, 2, 3, 4 or 5 nucleotides that do not base pair.
21. The siRNA of claim 1 , wherein said duplex region is 17 nucleotides in length and has 1, 2, 3, 4 or 5 nucleotides that do not base pair.
22. The siRNA of claim 1 , wherein said duplex region is 18 nucleotides in length and has 1, 2, 3, 4 or 5 nucleotides that do not base pair.
23. The siRNA of claim 1 , wherein said duplex region is 19 nucleotides in length and has 1, 2, 3, 4 or 5 nucleotides that do not base pair.
24. The vector of claim 11 , wherein said promoter is pol III, U6, H1 or 7SK.