title: Comprehensive Structure-Activity Relationship of Triantennary N-Acetylgalactosamine Conjugated Antisense Oligonucleotides for Targeted Delivery to Hepatocytes.
abstract: The comprehensive structure-activity relationships of triantennary GalNAc conjugated ASOs for enhancing potency via ASGR mediated delivery to hepatocytes is reported. Seventeen GalNAc clusters were assembled from six distinct scaffolds and attached to ASOs. The resulting ASO conjugates were evaluated in ASGR binding assays, in primary hepatocytes, and in mice. Five structurally distinct GalNAc clusters were chosen for more extensive evaluation using ASOs targeting SRB-1, A1AT, FXI, TTR, and ApoC III mRNAs. GalNAc-ASO conjugates exhibited excellent potencies (ED50 0.5-2 mg/kg) for reducing the targeted mRNAs and proteins. This work culminated in the identification of a simplified tris-based GalNAc cluster (THA-GN3), which can be efficiently assembled using readily available starting materials and conjugated to ASOs using a solution phase conjugation strategy. GalNAc-ASO conjugates thus represent a viable approach for enhancing potency of ASO drugs in the clinic without adding significant complexity or cost to existing protocols for manufacturing oligonucleotide drugs.
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Thazha P
authors.initials: TP
authors.lastname: Prakash
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Jinghua
authors.initials: J
authors.lastname: Yu
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Migawa
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Garth A
authors.initials: GA
authors.lastname: Kinberger
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: W Brad
authors.initials: WB
authors.lastname: Wan
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Østergaard
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Recaldo L
authors.initials: RL
authors.lastname: Carty
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Vasquez
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Audrey
authors.initials: A
authors.lastname: Low
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Alfred
authors.initials: A
authors.lastname: Chappell
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Karsten
authors.initials: K
authors.lastname: Schmidt
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Mariam
authors.initials: M
authors.lastname: Aghajan
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Jeff
authors.initials: J
authors.lastname: Crosby
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Heather M
authors.initials: HM
authors.lastname: Murray
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Sheri L
authors.initials: SL
authors.lastname: Booten
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Jill
authors.initials: J
authors.lastname: Hsiao
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Armand
authors.initials: A
authors.lastname: Soriano
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Todd
authors.initials: T
authors.lastname: Machemer
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Patrick
authors.initials: P
authors.lastname: Cauntay
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Sebastien A
authors.initials: SA
authors.lastname: Burel
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Susan F
authors.initials: SF
authors.lastname: Murray
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Hans
authors.initials: H
authors.lastname: Gaus
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Graham
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Eric E
authors.initials: EE
authors.lastname: Swayze
authors.affiliation: Ionis Pharmaceuticals Inc. , 2855 Gazelle Court, Carlsbad, California 92010, United States.
authors.forename: Punit P
authors.initials: PP
authors.lastname: Seth
mesh_terms: D000116:Acetylgalactosamine / Q000138:chemical synthesis* / Q000494:pharmacology*; D000818:Animals; D053305:Apolipoprotein C-III / Q000187:drug effects; D016503:Drug Delivery Systems; D005172:Factor XI / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis* / Q000494:pharmacology*; D051122:Scavenger Receptors, Class B / Q000096:biosynthesis / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D053305:Apolipoprotein C-III; D016376:Oligonucleotides, Antisense; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; D005172:Factor XI; D000116:Acetylgalactosamine