Fully automated fast-flow synthesis of antisense phosphorodiamidate morpholino oligomers

Abstract

Rapid development of antisense therapies can enable on-demand responses to new viral pathogens and make personalized medicine for genetic diseases practical. Antisense phosphorodiamidate morpholino oligomers (PMOs) are promising candidates to fill such a role, but their challenging synthesis limits their widespread application. To rapidly prototype potential PMO drug candidates, we report a fully automated flow-based oligonucleotide synthesizer. Our optimized synthesis platform reduces coupling times by up to 22-fold compared to previously reported methods. We demonstrate the power of our automated technology with the synthesis of milligram quantities of three candidate therapeutic PMO sequences for an unserved class of Duchenne muscular dystrophy (DMD). To further test our platform, we synthesize a PMO that targets the genomic mRNA of SARS-CoV-2 and demonstrate its antiviral effects. This platform could find broad application not only in designing new SARS-CoV-2 and DMD antisense therapeutics, but also for rapid development of PMO candidates to treat new and emerging diseases.

Recyclable cooperative catalyst for accelerated hydroaminomethylation of hindered amines in a continuous segmented flow reactor

“…Carbon–nitrogen (C–N) bonds are widely common in bioactive molecules and thus, rapid C–N bond formation reactions are essential tools for the generation of large molecular libraries for structure-activity-relationship mapping (SAR), and the on-demand synthesis of isotopically labeled tracers with relatively short half-life such as the 11 C labeled PET tracers 6 – 8 . The emerging trend towards decentralized manufacturing of pharmaceuticals and fine chemicals promotes continuous flow production as the preferred method of choice 9 – 11 . Four conventional processes are typically used in the large scale synthesis of aliphatic amines: (1) amination of alcohols, (2) amination of alkyl halides, (3) reduction of nitrile compounds, and (4) reductive amination of carbonyl compounds 3 , which all require multistep processing and purification.…”

Inhibition of SARS-CoV-2 by Targeting Conserved Viral RNA Structures and Sequences

“…Spike protein-targeting antibodies (e.g., bamlanivimab) can effectively neutralize the virus and prevent viral entry ( Gottlieb et al, 2021 ). RNA-targeting antisense oligonucleotides (ASO) or small molecules will degrade the viral RNA genome or hinder RNA translation ( Li et al, 2021b , Li et al, 2021a. ; Lulla et al, 2021 ; Rosenke et al, 2021 ; Sun et al, 2021a ; Zhang et al, 2021 ; Zhu et al, 2021 ).…”

Automated Flow Synthesis of Peptide–PNA Conjugates

“…The automated flow PNA synthesizer consists of seven modules including a central control computer, solution storage system, three HPLC pumps, three multiposition valves, heating elements, reaction zone, and a UV–vis detector. A modular script in the Mechwolf programming environment 31 controls the instrument ( Figure 2 a). During a coupling reaction, three HPLC pumps draw reagents stored under nitrogen atmosphere from the storage module, and the desired PNA monomer, activator, and base solutions are merged using a valve.…”