A5084516005- DNA and Nucleic Acid Chemistry
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- CRISPR and Genetic Engineering
- Synthetic Organic Chemistry Methods
- Synthesis and Catalytic Reactions
- Genetic Neurodegenerative Diseases
- Asymmetric Synthesis and Catalysis
- Carbohydrate Chemistry and Synthesis
- Chemical Synthesis and Analysis
- Viral Infections and Immunology Research
- Crystallography and molecular interactions
- Biochemical and Molecular Research
- Muscle Physiology and Disorders
- Catalytic C–H Functionalization Methods
- Hepatitis C virus research
- Synthesis of Organic Compounds
- RNA regulation and disease
- Mitochondrial Function and Pathology
- Spectroscopy Techniques in Biomedical and Chemical Research
- Fluorine in Organic Chemistry
Ionis Pharmaceuticals (United States)
2015-2024
Alnylam Pharmaceuticals (United States)
2024
University of British Columbia
2013
Université de Montréal
2012
University of Bern
2012
Vanderbilt University
2010
Syracuse University
2005
University of Iowa
1999-2001
The Ohio State University
1995-1999
Abstract Triantennary N-acetyl galactosamine (GalNAc, GN3), a high-affinity ligand for the hepatocyte-specific asialoglycoprotein receptor (ASGPR), enhances potency of second-generation gapmer antisense oligonucleotides (ASOs) 6–10-fold in mouse liver. When combined with next-generation ASO designs comprised short S-cEt (S-2′-O-Et-2′,4′-bridged nucleic acid) ASOs, ∼60-fold enhancement relative to parent MOE (2′-O-methoxyethyl RNA) was observed. GN3-conjugated ASOs showed high affinity ASGPR,...
The potency of second generation antisense oligonucleotides (ASOs) in animals was increased 3- to 5 -fold (ED50 ≈ 2−5 mg/kg) without producing hepatotoxicity, by reducing ASO length (20-mer 14-mer) and employing novel nucleoside modifications that combine structural elements 2′-O-methoxyethyl residues locked nucleic acid. ability achieve this level any formulation agents is remarkable likely have a significant impact on the future design ASOs as therapeutic agents.
We have recently shown that combining the structural elements of 2'O-methoxyethyl (MOE) and locked nucleic acid (LNA) nucleosides yielded a series nucleoside modifications (cMOE, 2',4'-constrained MOE; cEt, ethyl) display improved potency over MOE an therapeutic index relative to LNA antisense oligonucleotides. In this report we present details regarding synthesis cMOE cEt phosphoramidites biophysical evaluation oligonucleotides containing these modifications. The was efficiently...
High affinity antisense oligonucleotides (ASOs) containing bicylic modifications (BNA) such as locked nucleic acid (LNA) designed to induce target RNA cleavage have been shown enhanced potency along with a higher propensity cause hepatotoxicity. In order understand the mechanism of this hepatotoxicity, transcriptional profiles were collected from livers mice treated panel highly efficacious hepatotoxic or non-hepatotoxic LNA ASOs. We observed selective transcript knockdown in ASOs, while...
Receptor-dependent productive uptake of GLP1-conjugated antisense oligonucleotides occurs selectively in pancreatic β-cells.
Phosphorothioate (PS)-modified antisense oligonucleotides (ASOs) have been extensively investigated over the past three decades as pharmacological and therapeutic agents. One second generation ASO, Kynamro™, was recently approved by FDA for treatment of homozygous familial hypercholesterolemia 35 PS ASOs are at various stages clinical development. In this report, we show that Stabilin class scavenger receptors, which were not previously thought to bind DNA, do internalize ASOs. With use...
Autosomal dominant diseases such as Huntington's disease (HD) are caused by a gain of function mutant protein and/or RNA. An ideal treatment for these is to selectively suppress expression the allele while preserving wild-type variant. RNase H active antisense oligonucleotides (ASOs) or small interfering RNAs can achieve selective suppression gene targeting single nucleotide polymorphisms (SNPs) associated with repeat expansion. ASOs have been previously shown discriminate changes in...
Antisense oligonucleotide (ASO) therapeutics show tremendous promise for the treatment of previously intractable human diseases but to exert their effects on cellular RNA processing they must first cross plasma membrane by endocytosis. The conjugation ASOs a receptor ligand can dramatically increase entry into certain cells and tissues, as demonstrated implementation N-acetylgalactosamine (GalNAc)-conjugated Asialoglycoprotein Receptor (ASGR)-mediated uptake liver hepatocytes. We compared...
Huntington disease (HD) is a dominant, genetic neurodegenerative characterized by progressive loss of voluntary motor control, psychiatric disturbance, and cognitive decline, for which there currently no disease-modifying therapy. HD caused the expansion CAG tract in huntingtin (HTT) gene. The mutant HTT protein (muHTT) acquires toxic functions, significant evidence that muHTT lowering would be therapeutically efficacious. However, wild-type (wtHTT) serves vital making allele-specific...
Bicyclic oxazaphospholidine monomers were used to prepare a series of phosphorothioate (PS)-modified gapmer antisense oligonucleotides (ASOs) with control the chirality each PS linkages within 10-base gap. The stereoselectivity was determined be 98% for coupling. objective this work study how influences biophysical and biological properties ASO including binding affinity (Tm), nuclease stability, activity in vitro vivo, RNase H activation cleavage patterns (both human E. coli) context....
The comprehensive structure-activity relationships of triantennary GalNAc conjugated ASOs for enhancing potency via ASGR mediated delivery to hepatocytes is reported. Seventeen clusters were assembled from six distinct scaffolds and attached ASOs. resulting ASO conjugates evaluated in binding assays, primary hepatocytes, mice. Five structurally chosen more extensive evaluation using targeting SRB-1, A1AT, FXI, TTR, ApoC III mRNAs. GalNAc-ASO exhibited excellent potencies (ED50 0.5-2 mg/kg)...
Interactions of chemically modified nucleic acid therapeutics with plasma proteins play an important role in facilitating distribution from the injection site to peripheral tissues by reducing renal clearance. Despite importance these interactions, analytical methods that can characterize binding constants individual a reliable and high throughput manner are not easily available. We developed fluorescence polarization (FP) based assay measured for 25 most abundant human phosphorothioate (PS)...
Abstract Antisense oligonucleotides (ASOs) are recognized therapeutic agents for the modulation of specific genes at post-transcriptional level. Similar to any medical drugs, there opportunities improve their efficacy and safety. Here we develop a short DNA/RNA heteroduplex oligonucleotide (HDO) with structure different from double-stranded RNA used interfering single-stranded DNA ASO. A DNA/locked nucleotide acid gapmer duplex an α-tocopherol-conjugated complementary (Toc-HDO) is...
Abstract Enhancing the functional uptake of antisense oligonucleotide (ASO) in muscle will be beneficial for developing ASO therapeutics targeting genes expressed muscle. We hypothesized that improving albumin binding facilitate traversal from blood compartment to interstitium tissues enhance uptake. synthesized structurally diverse saturated and unsaturated fatty acid conjugated ASOs with a range hydrophobicity. The affinity conjugates plasma proteins improved chain length highest was...
Mutant huntingtin suppression with antisense oligonucleotides reverses cognitive impairments in a mouse model of Huntington’s disease.
Huntington disease (HD) is an inherited, fatal neurodegenerative disorder caused by a CAG repeat expansion in the huntingtin gene. The mutant protein causes neuronal dysfunction and degeneration resulting motor dysfunction, cognitive decline, psychiatric disturbances. Currently, there no altering treatment, symptomatic therapy has limited benefit. pathogenesis of HD complicated multiple pathways are compromised. Addressing problem at its genetic root suppressing expression promising...
Phosphorothioate-modified antisense oligonucleotides (PS-ASOs) interact with a host of plasma, cell-surface and intracellular proteins which govern their therapeutic properties. Given the importance PS backbone for interaction proteins, we systematically replaced anionic PS-linkages in toxic ASOs charge-neutral alkylphosphonate linkages. Site-specific incorporation alkyl phosphonates altered RNaseH1 cleavage patterns but overall rates activity versus on-target gene cells mice were only...
The PS modification enhances the nuclease stability and protein binding properties of gapmer antisense oligonucleotides (ASOs) is one very few modifications that support RNaseH1 activity. We evaluated effect introducing stereorandom chiral mesyl-phosphoramidate (MsPA) linkages in DNA gap flanks ASOs characterized these on RNA-binding, stability, binding, pro-inflammatory profile, activity toxicity cells mice. show all a ASO can be replaced with MsPA without compromising chemical RNA affinity...
Abstract Therapeutic oligonucleotides are often modified using the phosphorothioate (PS) backbone modification which enhances stability from nuclease mediated degradation. However, substituting oxygen in phosphodiester with sulfur introduce chirality into such that a full PS 16-mer oligonucleotide is comprised of 215 distinct stereoisomers. As result, role on performance antisense (ASOs) has been subject debate for over two decades. We carried out systematic analysis to determine if...