ChemRxiv
These are preliminary reports that have not been peer-reviewed. They should not be regarded as conclusive, guide clinical practice/health-related behavior, or be reported in news media as established information. For more information, please see our FAQs.
1/1
2 files

Automated Fast-Flow Synthesis of Peptide Nucleic Acid Conjugates

preprint
submitted on 23.02.2021, 21:13 and posted on 24.02.2021, 11:59 by Chengxi Li, Alexander J. Callahan, Kruttika-Suhas Phadke, Bryan Bellaire, Charlotte E. Farquhar, Genwei Zhang, Carly K. Schissel, Alexander J. Mijalis, Nina Hartrampf, Andrei Loas, David E. Verhoeven, Bradley Pentelute
Peptide nucleic acids (PNAs) are charge-neutral oligonucleotides with emerging potential for treatment of genetic, acquired, and viral diseases, including COVID-19. Their challenging synthesis, however, limits their use for rapid therapeutic intervention and widespread application. Here, we report a highly efficient technology that utilizes a fully automated fast-flow instrument to manufacture cell-penetrating peptide-conjugated PNAs (PPNAs) in a single shot. The machine is rapid: each amide bond is formed in 10 seconds and the synthesis of an 18-mer bioactive PNA is complete in one hour. Anti-IVS2-654 PPNA synthesized in a single shot with this instrument presented over 16-fold activity compared to unmodified PNA in a splice-correction assay. We demonstrated the utility of this approach by chemically synthesizing an eight-member anti-SARS-CoV-2 PPNA library within one day. A designer PPNA targeting the 5’ untranslated region of SARS-CoV-2 genomic RNA reduced the viral titer by over 95% in live virus infection assays (IC50: 0.8 mM). Our technology can rapidly yield on-demand PPNA candidates to tackle newly-emerging viral pathogens.

Funding

Novo Nordisk A/S

History

Email Address of Submitting Author

blp@mit.edu

Institution

Massachusetts Institute of Technology

Country

USA

ORCID For Submitting Author

0000-0002-7242-801X

Declaration of Conflict of Interest

B.L.P. is a co-founder of Amide Technologies and Resolute Bio. Both companies focus on developing protein and pep-tide therapeutics.

Exports