Assessment of the Drug Interaction Potential of Unconjugated and GalNAc3-Conjugated 2'-MOE-ASOs.

Colby S Shemesh,Yanfeng Wang,Noah Post,Eunju Hurh,Tanya Watanabe,Mark S Warren,Jane Huang,Daniel A Norris,Song Lin,Brandon Nichols,Michael Liu,Scott P Henry,Rosie Z Yu,Mirza Jahic

doi:10.1016/j.omtn.2017.08.012

Abstract

Antisense oligonucleotides are metabolized by nucleases and drug interactions with small drug molecules at either the cytochrome P450 (CYP) enzyme or transporter levels have not been observed to date. Herein, a comprehensive in vitro assessment of the drug-drug interaction (DDI) potential was carried out with four 2′-O-(2-methoxyethyl)-modified antisense oligonucleotides (2′-MOE-ASOs), including a single triantennary N-acetyl galactosamine (GalNAc3)-conjugated ASO. Several investigations to describe the DDI potential of a 2′-MOE-ASO conjugated to a high-affinity ligand for hepatocyte-specific asialoglycoprotein receptors are explored. The inhibition on CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4 and induction on CYP1A2, CYP2B6, and CYP3A4 were investigated in cryopreserved hepatocytes using up to 100 μM of each ASO. No significant inhibition (half maximal inhibitory concentration [IC50] > 100 μM) or induction was observed based on either enzymatic phenotype or mRNA levels. In addition, transporter interaction studies were conducted with nine major transporters per recommendations from regulatory guidances and included three hepatic uptake transporters, organic cation transporter 1 (OCT1), organic anion transporting polypeptide 1B1 (OATP1B1), and OATP1B3; three renal uptake transporters, organic anion transporter 1 (OAT1), OAT3, and OCT2; and three efflux transporters, P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), and bile salt export pump (BSEP). None of the four ASOs (10 μM) were substrates of any of the nine transporters, with uptake <2-fold compared to controls, and efflux ratios were below 2.0 for BCRP and P-gp. Additionally, neither of the four ASOs showed meaningful inhibition on any of the nine transporters tested, with the mean percent inhibition ranging from −38.3% to 24.2% with 100 μM ASO. Based on these findings, the unconjugated and GalNAc3-conjugated 2′-MOE-ASOs would have no or minimal DDI with small drug molecules via any major CYP enzyme or drug transporters at clinically relevant exposures.

Highlights

Antisense oligonucleotides (ASOs) are a growing class of versatile biomolecules, which have garnered much attention in the past decade as a mature and attractive platform for therapeutic drug development
It is well established that the major cytochrome P450 (CYP) enzymes and drug transporters make a significant contribution to the PK and pharmacodynamic (PD) properties of small molecules, yet the interaction and relation of unconjugated ASOs toward these enzymes and drug transporters are not as well characterized, with limited drug-drug interaction (DDI) data and no existing investigations of GalNAc3-ASO conjugates
Because ASOs are drastically different from small molecules in their physico-chemical properties, such as molecular weight, number of hydrogen bonds, and disposition, the ability to interact with CYP enzymes and drug transporters is significantly limited

Summary

Introduction

Antisense oligonucleotides (ASOs) are a growing class of versatile biomolecules, which have garnered much attention in the past decade as a mature and attractive platform for therapeutic drug development. As one of the most compelling successes in oligonucleotide drug development, triantennary N-acetyl galactosamine (GalNAc3)-conjugated ASOs allow for efficient delivery, with high affinity to asialoglycoprotein receptors (ASGPRs) for liver targeting.[12] Remarkably over 20- to 30-fold improved potency of GalNac3-ASO conjugates compared to unconjugated ASOs have been observed in vivo.[13]

Methods

Results

Conclusion