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 method of inhibiting the activity of a telomerase enzyme, the method comprising contacting the telomerase enzyme with a 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. The method of claim 1 , wherein the telomerase enzyme is in a cell.
3. The method of claim 2 , wherein the cell is a cancer cell.
4. The method 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.
5. The method of claim 4 , wherein L is a lipid selected from the group consisting of substituted and unsubstituted fatty acids and sterols.
6. The method of claim 5 , wherein L is a fatty acid substituted with fluorine.
7. The method of claim 4 , wherein L is a substituted or unsubstituted hydrocarbon.
8. The method of claim 7 , wherein L is a hydrocarbon substituted with fluorine.
9. The method of claim 1 , wherein n=1 and the x-L component is covalently conjugated to the 5′ terminus of the oligonucleotide O.
10. The method of claim 1 , wherein n=1 and the (x-L) component is covalently conjugated to the 3′ terminus of the oligonucleotide O.
11. The method 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.
12. The method of claim 11 , wherein each (x-L) is palmitoylamido-aminoglycerol-thiophosphate.
13. The method 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-thiophosphate, 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.
14. A method of inhibiting the activity of a telomerase enzyme, the method comprising contacting the telomerase enzyme with a 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.
15. The method of claim 14 , wherein the telomerase enzyme is in a cell.
16. The method of claim 15 , wherein the cell is a cancer cell.
17. The method of claim 14 , wherein n=1.
18. The method of claim 17 , wherein L is directly linked to the 3′ terminus of the oligonucleotide L through an amide bond.
19. The method of claim 17 , wherein the compound comprises the following structure:
or a pharmaceutically acceptable salt thereof.