Automated Flow Synthesis of Peptide-PNA Conjugates.
Chengxi LiAlex J CallahanKruttika S PhadkeBryan H BellaireCharlotte E FarquharGenwei ZhangCarly K SchisselAlexander J MijalisNina HartrampfAndrei LoasDavid E VerhoevenBradley L PentelutePublished in: ACS central science (2021)
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 (IC 50 = 0.8 μM). Our technology can deliver PPNA candidates to further investigate their potential as antiviral agents.
Keyphrases
- sars cov
- high throughput
- nucleic acid
- deep learning
- single cell
- respiratory syndrome coronavirus
- induced apoptosis
- machine learning
- cancer therapy
- loop mediated isothermal amplification
- patient reported outcomes
- cell cycle arrest
- cell therapy
- photodynamic therapy
- endoplasmic reticulum stress
- copy number
- stem cells
- drug delivery
- human health
- signaling pathway
- dna methylation
- risk assessment
- cell death
- bone marrow
- amino acid
- quantum dots