Inhibition of Bcl-2 protein expression by liposomal antisense oligodeoxynucleotides
View Patent ↗The present invention provides novel compositions and methods for use in the treatment of Bcl-2-associated diseases like cancer, specifically, in the treatment of follicular lymphoma (FL). The compositions contain antisense oligonucleotides that hybridize to Bcl-2 nucleic acids, the gene products of which are known to interact with the tumorigenic protein Bcl-2. Used alone, or in conjunction with other antisense oligonucleotides, these compositions inhibit the proliferation of FL cancer cells.
1. A method of inhibiting proliferation of a Bcl-2-associated disease cell comprising obtaining a polynucleotide that hybridizes to Bcl-2 mRNA under intracellular conditions, mixing the first polynucleotide with a neutral phospholipid to form a composition comprising a polynucleotide/phospholipid association, and administering said composition to a human having a Bcl-2-associated disease to inhibit the proliferation of said disease cells, wherein said disease cells have a t(14;18) translocation, wherein said composition is delivered to said human in an amount of from about 5 to about 30 mg polynucleotide per m 2 .
2. The method of claim 1 , wherein the cell is a cancer cell.
3. The method of claim 2 , wherein said cancer cell is a follicular lymphoma cell.
4. The method of claim 1 , wherein said first polynucleotide is an oligonucleotide having a length of between about 8 and about 50 bases.
5. The method of claim 1 , comprising a liposome formed from the phospholipid.
6. The method of claim 5 , wherein the liposome encapsulates the first polynucleotide.
7. The method of claim 1 , wherein said composition is delivered to said human in a volume of 0.50-10.0 ml per dose.
8. The method of claim 1 , wherein said composition is administered three times per week for eight weeks.
9. A method of inhibiting proliferation of a Bcl-2-associated disease cell having a t(14;18) translocation comprising:
(a) obtaining an oligonucleotide of from about 8 to about 50 bases that hybridizes to a Bcl-2-encoding polynucleotide under intracellular condition;
(b) mixing the oligonucleotide with a neutral phospholipid to form a composition comprising a neutral oligonucleotide/phospholipid association; and
(c) administering said composition to said Bcl-2-associated disease cell to inhibit the proliferation of said disease cell,
wherein said cell is in a human, and wherein said composition is delivered to said human in an amount of from about 5 to about 30 mg polynucleotide per m 2 .
10. The method of claim 9 , wherein the cell is a cancer cell.
11. The method of claim 10 , wherein said cancer cell is a follicular lymphoma cell.
12. The method of claim 9 , comprising a liposome formed from the phospholipid.
13. The method of claim 12 , wherein the liposome encapsulates the polynucleotide.
14. The method of claim 9 , wherein said composition is delivered to said human in a volume of 0.50-10.0 ml per dose.
15. The method of claim 9 , wherein said composition is administered three times per week for eight weeks.
16. The method of claim 5 , wherein said liposome consists essentially of neutral phospholipids.
17. The method of claim 12 , wherein said liposome consists essentially of neutral phospholipids.
18. A composition comprising a first antisense polynucleotide that hybridizes to a second, Bcl-2-encoding polynucleotide under intracellular conditions and a neutral phospholipid associated with said first polynucleotide, to form a Bcl-2 polynucleotide/neutral phospholipid association, wherein said first polynucleotide comprises at least 8 nucleotides of the sequence CAGCGTGCGCCATCCTTC (SEQ ID NO:1), wherein said polynucleotide is complementary to the translation initiation site of Bcl-2, said composition further comprising a charged phospholipid.
19. The composition of claim 18 , wherein said first polynucleotide is an oligonucleotide having a length of between about 8 and about 50 bases.
20. The composition of claim 18 , wherein the first polynucleotide is complementary to the translation initiation site of Bcl-2 mRNA.
21. The composition of claim 20 , wherein the polynucleotide is an oligonucleotide comprising the sequence CAGCGTGCGCCATCCTTC (SEQ ID NO:1).
22. The composition of claim 18 , comprising a liposome formed from the phospholipid.
23. The composition of claim 22 , wherein the first polynucleotide is encapsulated in the liposome.
24. The composition of claim 18 , wherein the phospholipid is a phosphatidylcholine, a phosphatidylglycerol, or a phosphatidylethanolamine.
25. The composition of claim 24 , wherein the phospholipid is dioleoylphosphatidylcholine.
26. A composition comprising an expression construct that encodes a first antisense polynucleotide that hybridizes to a second, Bcl-2-encoding polynucleotide under intracellular conditions, wherein said construct is under the control of a promoter that is active in eukaryotic cells and associated with a neutral phospholipid, wherein said first polynucleotide comprises at least 8 nucleotides of the sequence CAGCGTGCGCCATCCTTC (SEQ ID NO:1), wherein said polynucleotide is complementary to the translation initiation site of Bcl-2, further comprising a charged phospholipid.
27. A composition comprising a neutral phospholipid associated with an expression construct that encodes an oligonucleotide of from about 8 to about 50 bases and which hybridizes to Bcl-2 mRNA under intracellular conditions, wherein the construct is under the control of a promoter that is active in eukaryotic cells, further comprising a charged phospholipid.
28. The composition of claim 18 , wherein said first polynucleotide is a P-ethoxy oligonucleotide.
29. The composition of claim 22 , wherein said liposome consists essentially of neutral and charged phospholipids.
30. The composition of claim 26 , comprising a liposome formed from said neutral phospholipid.
31. The composition of claim 30 , wherein said liposome consists essentially of neutral and charged phospholipids.
32. The composition of claim 27 , comprising a liposome formed from the phospholipid.
33. The composition of claim 32 , wherein said liposome consists essentially of neutral and charged phospholipids.
34. A composition comprising a first antisense polynucleotide that hybridizes to a second, Bcl-2-encoding polynucleotide under intracellular conditions and a primary phosphatide associated with said first polynucleotide, wherein said primary phosphatide is a neutral phospholipid, and wherein said first polynucleotide comprises at least 8 nucleotides of the sequence CAGCGTGCGCCATCCTTC (SEQ ID NO:1), and wherein said polynucleotide is complementary to the translation initiation site of Bcl-2, further comprising a charged phospholipid.
35. The composition of claim 34 , comprising a liposome formed from the primary phosphatide.
36. The composition of claim 35 , wherein said liposome consists essentially of neutral and charged phospholipids.
37. The composition of claim 34 , wherein said first polynucleotide is a P-ethoxy oligonucleotide.
38. The composition of claim 18 , wherein said at least 8 nucleotides are consecutive nucleotides.
39. The composition of claim 18 , wherein the charged phospholipid is a positively charged phospholipid.
40. A method of inhibiting proliferation of a Bcl-2-associated disease cell comprising obtaining a polynucleotide that hybridizes to Bcl-2 mRNA under intracellular conditions, mixing the first polynucleotide with a neutral phospholipid to form a composition comprising a polynucleotide/phospholipid association, and administering said composition to a human having a Bcl-2-associated disease to inhibit the proliferation of said disease cells, wherein said disease cells have a t(14;18) translocation, the composition further comprising a charged phospholipid.
41. The method of claim 40 , wherein the charged phospholipid is a positively charged phospholipid.
42. The method of claim 1 , wherein said first polynucleotide is a ethoxy oligonucleotide.
43. The method of claim 9 , wherein said first oligonucleotide is a P-ethoxy oligonucleotide.