Ligands to enhance cellular uptake of biomolecules
The present invention relates to the design and synthesis of homogeneous A-L-P constructs, which contain a hepatic ligand to direct an oligomer or “payload” to a hepatocyte intracellularly via a receptor-mediated, ligand-directed pathway.
1 . A construct comprising a homogeneous conjugate of formula A-L-P, wherein
A represents a hepatic ligand that specifically binds to a hepatic receptor, thereby facilitating the entrance of said conjugate into cells having said receptor;
L represents a bifunctional linker that is covalently linked to A in a regiospecific manner to form A-L; A-L is covalently linked to P in a regiospecific manner to form A-L-P;
P represents a biologically stable oligomer comprising the nucleotide sequence of SEQ ID NO:27 or SEQ ID NO:28, wherein P is released from the conjugate following hydrolysis or reduction of at least one specific biochemical linkage, and contains internucleotide linkages resistant to enzymatic hydrolysis of biodegradation upon release from the conjugate.
2 . The construct of claim 1 , wherein said oligomer is an oligonucleotide, an oligonucleotide analog or an oligonucleoside.
4 .- 17 . (canceled)
18 . The construct of claim 2 , wherein said oligomer comprises deoxyribose methylphosphonate internucleotide linkages.
19 . The construct of claim 2 , wherein said oligomer comprises deoxyribose phosphorothioate internucleotide linkages.
20 . The construct of claim 2 , wherein said oligomer comprises phosphodiester linkages.
21 . The construct of claim 2 , wherein said oligomer comprises a combination of deoxyribose methylphosphonate/phosphorothioate internucleotide linkages.
22 . The construct of claim 2 , wherein said oligomer comprises a combination of deoxyribose methylphosphonate/phosphodiester internucleotide linkages.
23 . The construct of claim 2 , wherein said oligomer comprises deoxyribose phosphorothioate/phosphodiester internucleotide linkages.
24 . The construct of claim 2 , wherein said oligomer comprises 2′-O-methylribose methylphosphonate internucleotide linkages.
25 . The construct of claim 2 , wherein said oligomer comprises 2′-O-methylribose phosphorothioate internucleotide linkages.
26 . The construct of claim 2 , wherein said oligomer comprises 2′-O-methylribose phosphodiester internucleotide linkages.
27 . The construct of claim 2 , wherein said oligomer comprises a combination of 2′-O-methylribose methylphosphonate/2′-O-methylribose phosphodiester internucleotide linkages.
28 . The construct of claim 2 , wherein said oligomer comprises a combination of 2′-O-methylribose methylphosphonate/2′-O-methylribose phosphorothionate internucleotide linkages.
29 . The construct of claim 2 , wherein said oligomer comprises a combination of 2′-O-methylribose phosphorothioate/2′-O-methylribose phosphodiester internucleotide linkages.
30 .- 63 . (canceled)
64 . A pharmaceutical composition comprising a construct according to claim 1 and at least one pharmaceutically acceptable excipient or carrier.
65 .- 69 . (canceled)
70 . The pharmaceutical composition of claim e 64 wherein the A-L moiety of said construct is YEE(ahGalNAc) 3 -SMCC.
71 . (canceled)
72 . The construct of claim 1 , wherein the A-L moiety of said construct is YEE(ahGalNAc) 3 -SMCC.
73 . (canceled)