siRNA conjugates carrying sequentially assembled trivalent N-acetylgalactosamine linked through nucleosides elicit robust gene silencing in vivo in hepatocytes.
Asialoglycoprotein receptor (ASGPR) mediated delivery of triantennary N-acetylgalactosamine (GalNAc) conjugated short interfering RNAs (siRNAs) to hepatocytes is a promising paradigm for RNAi therapeutics. Robust and durable gene silencing upon subcutaneous administration at therapeutically acceptable dose levels resulted in the advancement of GalNAc-conjugated oligonucleotide-based drugs into preclinical and clinical developments. To systematically evaluate the effect of display and positioning of the GalNAc moiety within the siRNA duplex on ASGPR binding and RNAi activity, nucleotides carrying monovalent GalNAc were designed. Evaluation of clustered and dispersed incorporation of GalNAc units to the sense (S) strand indicated that sugar proximity is critical for ASGPR recognition, and location of the clustered ligand impacts the intrinsic potency of the siRNA. An array of nucleosidic GalNAc monomers resembling a trivalent ligand at or near the 3' end of the S strand retained in vitro and in vivo siRNA activity, similar to the parent conjugate design. This work demonstrates the utility of simple, nucleotide-based, cost-effective siRNA-GalNAc conjugation strategies.
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Shigeo
S
Matsuda
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Kristofer
K
Keiser
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Jayaprakash K
JK
Nair
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Klaus
K
Charisse
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Rajar M
RM
Manoharan
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Philip
P
Kretschmer
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Chang G
CG
Peng
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Alexander
A
V Kel'in
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Pachamuthu
P
Kandasamy
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Jennifer L S
JL
Willoughby
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Abigail
A
Liebow
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
William
W
Querbes
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Kristina
K
Yucius
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Tuyen
T
Nguyen
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Stuart
S
Milstein
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Martin A
MA
Maier
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Kallanthottathil G
KG
Rajeev
Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States.
Muthiah
M
Manoharan
D000116:Acetylgalactosamine / Q000378:metabolism*; D000818:Animals; D020868:Gene Silencing; D022781:Hepatocytes / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009705:Nucleosides / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism
D009705:Nucleosides; D034741:RNA, Small Interfering; D000116:Acetylgalactosamine