Modified Oligonucleotides for Telomerase Inhibition
1 Assignment
0 Petitions
Accused Products
Abstract
Compounds comprising an oligonucleotide moiety covalently linked to a lipid moiety are disclosed. The oligonucleotide moiety comprises a sequence that is complementary to the RNA component of human telomerase. The compounds inhibit telomerase activity in cells with a high potency and have superior cellular uptake characteristics.
14 Citations
56 Claims
-
1-25. -25. (canceled)
-
26. A method of synthesizing a lipid conjugated oligonucleotide comprising at least 5 contiguous nucleoside subunits complementary to the RNA component of human telomerase each subunit joined by a N3′
- →
P5′
thiophosphoramidate linkage, the method comprising;(a) deprotecting and reacting a lipid containing solid support with a first 3′
-protected aminonucleoside-5′
-O-cyanoethyl-N,N-diisopropylaminophosphoramidite monomer to produce a first nucleoside attached to the lipid containing solid support through a phosphite linkage;(b) sulfurizing the phosphite linkage to form a thiophosphate linkage; (c) deprotecting the protected 3′
-amino group of the first aminonucleoside to form a free 3′
-amino group;(d) reacting the free 3′
-amino group with a second 3′
-protected aminonucleoside-5′
-O-cyanoethyl-N,N-diisopropylaminophosphoramidite monomer to form an internucleoside N3′
→
P5′
phosphoramidite linkage; and(e) sulfurizing the internucleoside N3′
→
P5′
phosphoramidite linkage to form a N3′
-P5′
thiophosphoramidate linkage; and(f) repeating steps (c) through (e) to produce the lipid conjugated oligonucleotide. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
- →
-
41. A method of synthesizing a lipid conjugated oligonucleotide comprising at least 5 contiguous nucleoside subunits complementary to the RNA component of human telomerase each subunit joined by a N3′
- →
P5′
thiophosphoramidate linkage, the method comprising;(a) deprotecting a protected 3′
-amino group of a first nucleoside attached to a lipid containing solid phase support, the first nucleoside having a protected 3′
amino group and a 5′
phosphate conjugated to the lipid containing solid phase support, wherein said deprotecting forms a free 3′
-amino group;(b) reacting the free 3′
-amino group with a second 3′
-protected aminonucleoside-5′
-O-cyanoethyl-N,N-diisopropylaminophosphoramidite monomer to form an internucleoside N3′
→
P5′
phosphoramidite linkage; and(c) sulfurizing the internucleoside N3′
→
P5′
phosphoramidite linkage to form a N3′
-P5′
thiophosphoramidate linkage; and(d) repeating steps (a) through (c) to produce the lipid conjugated oligonucleotide.
- →
-
42. A method of synthesizing a lipid conjugated oligonucleotide, the method comprising conjugating a terminus of an oligonucleotide comprising at least 5 contiguous nucleoside subunits complementary to the RNA component of human telomerase with a lipid conjugate comprising the structure (x-L) wherein x comprises a linker and L comprises a lipid moiety, wherein the internucleoside linkages of the oligonucleotide are N3′
- →
P5′
thiophosphoramidate linkages. - View Dependent Claims (43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53)
- →
-
54. A lipid containing solid support comprising the structure:
-
55. A method of preparing a modified lipid containing solid support, the method comprising:
-
reacting a 3-amino-1,2-propanediol that comprises a 3-amino group, a 1-hydroxy group and a 2-hydroxy group with a fatty acid;
dimethoxytritylating the 1-hydroxy group; and
succinylating the 2-hydroxy group;
to produce a 3-fatty acid amido-1-dimethoxytrityl-2-succinyloxy-propane; andreacting the 3-fatty acid amido-1-dimethoxytrityl-2-succinyloxy-propane with a support to produce the modified lipid containing solid support. - View Dependent Claims (56)
-
Specification