Modified oligonucleotides for telomerase inhibition
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.
1. A telomerase inhibitory compound comprising the structure:
O-(x-L)n,
wherein:
O is an oligonucleotide comprising a sequence consisting of
GTTAGGGTTAG;
GTTAGGGTTAGAC;
GTTAGGGTTAGACAA;
GGGTTAGAC;
CAGTTAGGG; or
CAGTTAGGGTTAG;
wherein the internucleoside linkages of the oligonucleotide O are N3′→P5′ thiophosphoramidate linkages;
x is an optional linker;
L is a lipid moiety; and
n=1 or 2 wherein each (x-L) component is independently covalently conjugated to the 5′ terminus or the 3′ terminus of the oligonucleotide O and wherein if n>1, each (x-L) component is independently selected;
or a pharmaceutically acceptable salt thereof.
2. A compound of claim 1 wherein x comprises an amide bond, a glycerol or an aminoglycerol linker optionally linked to the oligonucleotide O through a 5′- or 3′-linked phosphate group.
3. A compound of claim 2 wherein L is a lipid selected from the group consisting of substituted and unsubstituted fatty acids and sterols.
4. A compound of claim 3 wherein L is a fatty acid substituted with fluorine.
5. A compound of claim 2 wherein L is a substituted or unsubstituted hydrocarbon.
6. A compound of claim 5 wherein L is a hydrocarbon substituted with fluorine.
7. A compound of claim 1 wherein n=1 and the x-L component is covalently conjugated to the 5′ terminus of the oligonucleotide O.
8. A compound of claim 1 wherein n=1 and the (x-L) component is covalently conjugated to the 3′ terminus of the oligonucleotide O.
9. A compound of claim 1 wherein n=2, one independently selected (x-L) component is covalently conjugated to the 5′ terminus and one independently selected (x-L) component is covalently conjugated to the 3′ terminus.
10. A compound of claim 9 wherein each (x-L) is palmitoylamido-aminoglycerol-thiophosphate.
11. A telomerase inhibitory compound comprising the structure:
O-(x-L)n,
wherein:
O is TAGGGTTAGACAA
wherein the internucleoside linkages of the oligonucleotide O are N3′→P5′ thiophosphoramidate linkages;
wherein n=1 or 2, wherein if n>1, each (x-L) component is independently selected, and
wherein (x-L) is selected from the group consisting of 3′-myristoylamide, 3′-palmitoylamide, 3′-stearoylamide, 3′-palmitoylamido-propyl-thiophosphate, 3′-oleinylamide, 3′-linoleylamide, 5′-cholesterylamido-aminoglycerol-thiophosphate, 5′-C11-teflon-thiophosphate, 5′-C13-teflon-thiophosphate, 5′-batyl-thiophosphate, 3′-palmitoylamido-aminoglycerol-thiophosphate, 5′-palmitoylamido-bis-aminoglycerol-thiophosphate, 3′-cholesterylamido-aminoglycerol-thiophosphate, 5′-stearoylamido-aminoglycerol-thiophosphate; or a pharmaceutically acceptable salt thereof.
12. A compound of claim 11 wherein n=1.
13. A compound of claim 12 wherein L is directly linked to the 3′ terminus of the oligonucleotide L through an amide bond.
14. A compound of claim 12 , comprising the following structure:
or a pharmaceutically acceptable salt thereof.
15. A compound of claim 1 wherein each (x-L) component is independently selected from the group consisting of 3′-myristoylamide, 3′-palmitoylamide, 3′-stearoylamide, 5′-cholesterylamido-aminoglycerol-thiophosphate, 5′-palmitoylamido-aminoglycerol-thiosphosphate, 3′-palmitoylamido-aminoglycerol-thiophosphate, 3′-palmitoylamido-propyl-thiophosphate, 3′-oleinylamide, 3′-linoleylamide, 5′-C11-teflon-thiophosphate, 5′-C13-teflon-thiophosphate, 5′-palmitoylamido-bis-aminoglycerol-thiophosphate, 3′-cholesterylamido-aminoglycerol-thiophosphate, 5′-stearoylamido-aminoglycerol-thiophosphate and 5′-batyl-thiophosphate.