Automated Flow Synthesis of Peptide-PNA Conjugates

• Main Authors: Li, Chengxi (Author), Callahan, Alex J (Author), Phadke, Kruttika S (Author), Bellaire, Bryan (Author), Farquhar, Charlotte E (Author), Zhang, Genwei (Author), Schissel, Carly K (Author), Mijalis, Alexander J (Author), Hartrampf, Nina (Author), Loas, Andrei (Author), Verhoeven, David E (Author), Pentelute, Bradley L (Author)
• Format: Article
• Language: English
• Published: American Chemical Society (ACS), 2022-03-15T19:07:21Z.
• Subjects: Article
• Online Access: Get fulltext

 Antisense peptide nucleic acids (PNAs) have yet to translate to the clinic because of poor cellular uptake, limited solubility, and rapid elimination. Cell-penetrating peptides (CPPs) covalently attached to PNAs may facilitate clinical development by improving uptake into cells. We report an efficient technology that utilizes a fully automated fast-flow instrument to manufacture CPP-conjugated PNAs (PPNAs) in a single shot. The machine is rapid, with each amide bond being formed in 10 s. Anti-IVS2-654 PPNA synthesized with this instrument presented threefold activity compared to transfected PNA in a splice-correction assay. We demonstrated the utility of this approach by chemically synthesizing eight anti-SARS-CoV-2 PPNAs in 1 day. A PPNA targeting the 5' untranslated region of SARS-CoV-2 genomic RNA reduced the viral titer by over 95% in a live virus infection assay (IC50 = 0.8 μM). Our technology can deliver PPNA candidates to further investigate their potential as antiviral agents.