title: Comparative Characterization of Hepatic Distribution and mRNA Reduction of Antisense Oligonucleotides Conjugated with Triantennary N-Acetyl Galactosamine and Lipophilic Ligands Targeting Apolipoprotein B.
abstract: TriantennaryN-acetyl galactosamine (GalNAc, GN3) and lipophilic ligands such as cholesterol andα-tocopherol conjugations dramatically improve the distribution and efficacy of second-generation antisense oligonucleotides (ASOs) in the whole liver. To characterize ligands for delivery to liver cells based on pharmacokinetics and efficacy, we used a locked nucleic acid gapmer of ASO targeting apolipoprotein B as a model compound and evaluated the amount of ASO and apolipoprotein B mRNA in the whole liver, hepatocytes, and nonparenchymal (NP) cells as well as plasma total cholesterol after administration of ASO conjugated with these ligands to mice. Compared with unconjugated ASO, GN3 conjugation increased the amount (7-fold) and efficacy (more than 10-fold) of ASO in hepatocytes only and showed higher efficacy than the increased rate of the amount of ASO. On the other hand, lipophilic ligand conjugations led to increased delivery (3- to 5-fold) and efficacy (5-fold) of ASO to both hepatocytes and NP cells. GN3 and lipophilic ligand conjugations increased the area under the curve of ASOs and the pharmacodynamic duration but did not change the half-life in hepatocytes and NP cells compared with unconjugated ASO. In the liver, the phosphodiester bond between ASO and these ligands was promptly cleaved to liberate unconjugated ASO. These ligand conjugations reduced plasma total cholesterol compared with unconjugated ASO, although these ASOs were well tolerated with no elevation in plasma transaminases. These findings could facilitate ligand selection tailored to liver cells expressed in disease-related genes and could contribute to the discovery and development of RNA interference-based therapy.
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan ayahisa.watanabe@shionogi.co.jp.
authors.forename: Ayahisa
authors.initials: A
authors.lastname: Watanabe
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Mado
authors.initials: M
authors.lastname: Nakajima
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kasuya
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Reina
authors.initials: R
authors.lastname: Onishi
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Naohisa
authors.initials: N
authors.lastname: Kitade
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Mayumi
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Ikehara
authors.affiliation: Physicochemical and Preformulation (A.W.), Bioanalysis (R.O.), and Drug Metabolism and Pharmacokinetics (N.K., K.M.), Research Laboratory for Development, Exploratory Chemistry (M.N.) and Biotechnology-Based Medicine (T.K., T.I, A.K.), Discovery Research Laboratory for Innovative Frontier Medicines, Shionogi & Co., Ltd., Osaka, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Kugimiya
mesh_terms: D000116:Acetylgalactosamine / Q000737:chemistry*; D000818:Animals; D001055:Apolipoproteins B / Q000187:drug effects*; D002784:Cholesterol / Q000097:blood; D018014:Gene Transfer Techniques; D006207:Half-Life; D022781:Hepatocytes / Q000378:metabolism*; D008024:Ligands; D008055:Lipids / Q000737:chemistry*; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D000637:Transaminases / Q000378:metabolism
chemical_list: D001055:Apolipoproteins B; D008024:Ligands; D008055:Lipids; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D002784:Cholesterol; D000637:Transaminases; D000116:Acetylgalactosamine