Dunbar, C. E. et al. Gene therapy comes of age. Science 359, 175–185 (2018).
Haque, U. & Yokota, T. Enhancing antisense oligonucleotide-based therapeutic delivery with DG9, a versatile cell-penetrating peptide. Cells 12, 2395–2419 (2023).
Wan, W. B. & Seth, P. P. The medicinal chemistry of therapeutic oligonucleotides. J. Med. Chem. 59, 9645–9667 (2016).
Mullard, A. 2023 FDA approvals. Nat. Rev. Drug Discov. 23, 88–95 (2024).
Roberts, T. C., Langer, R. & Wood, M. J. A. Advances in oligonucleotide drug delivery. Nat. Rev. Drug Discov. 19, 673–694 (2020).
Campbella, M. A. & Wengel, J. Locked vs. unlocked nucleic acids (LNAvs.UNA): contrasting structures work towards common therapeutic goals. Chem. Soc. Rev. 40, 5680–5689 (2011).
Anand, P., Zhang, Y., Patil, S. & Kaur, K. Metabolic stability and targeted delivery of oligonucleotides: advancing RNA therapeutics beyond the liver. J. Med. Chem. 68, 6870–6896 (2025).
Moody, E. R., Obexer, R., Nickl, F., Spiess, R. & Lovelock, S. L. An enzyme cascade enables production oftherapeutic oligonucleotides in a single operation. Science 380, 1150–1154 (2023).
Wolfgang, K.-D. et al. Synthesis of oligodeoxynucleoside phosphorodithioates via thioamidites. J. Am. Chem. Soc. 111, 2321–2322 (1989).
Stec, W. J. & Wilk, A. Stereocontrolled synthesis of oligo(nucleoside phosphorothioate)s. Angew. Chem. Int. Ed. 33, 709–722 (1994).
Khvorova, A. & Watts, J. K. The chemical evolution of oligonucleotide therapies of clinical utility. Nat. Biotechnol. 35, 238–248 (2017).
Huang, P.-J. J. & Liu, J. W. Rational evolution of Cd2+-specific DNAzymes with phosphorothioate modified cleavage junction and Cd2+ sensing. Nucleic Acids Res. 43, 6125–6133 (2015).
McKenzie, L. K. et al. Recent progress in non-native nucleic acid modifications. Chem. Soc. Rev. 50, 5126–5164 (2021).
Laurent, Q. et al. Oligonucleotide phosphorothioates enter cells by thiol-mediated uptake. Angew. Chem. Int. Ed. 60, 19102–19106 (2021).
Crooke, S. T. et al. Cellular uptake and trafficking of antisense oligonucleotides. Nat. Biotechnol. 35, 230–237 (2017).
Purcell, J. & Hengge, A. C. The thermodynamics of phosphate versus phosphorothioate ester hydrolysis. J. Org. Chem. 70, 8437–8442 (2005).
Hoy, S. M. Nusinersen: first global approval. Drugs 77, 473–479 (2017).
Lamb, Y. N. Inclisiran: first approval. Drugs 81, 389–395 (2021).
Duschmale, J. et al. In vitro and in vivo properties of therapeutic oligonucleotides containing non-chiral 3′ and 5′ thiophosphate linkages. Nucleic Acids Res. 48, 63–74 (2020).
Jahns, H. et al. Chirality matters: stereo-defined phosphorothioate linkages at the termini of small interfering RNAs improve pharmacology in vivo. Nucleic Acids Res. 50, 1221–1240 (2022).
Poredoš, T. et al. Why and how to control P-chirality in phosphorothioated therapeutic oligonucleotides: analytical challenges associated with determination of stereochemical composition. Org. Process Res. Dev. 28, 4194–4214 (2024).
Iwamoto, N. et al. Control of phosphorothioate stereochemistry substantially increases the efficacy of antisense oligonucleotides. Nat. Biotechnol. 35, 845–851 (2017).
McIntosh, J. A. et al. A kinase-cGAS cascade to synthesize a therapeutic STING activator. Nature 603, 439–443 (2022).
Okaa, N. & Wada, T. Stereocontrolled synthesis of oligonucleotide analogs containing chiral internucleotidic phosphorus atoms. Chem. Soc. Rev. 40, 5829–5843 (2011).
Eckstein, F. Phosphorothioates, essential components of therapeutic oligonucleotides. Nucleic Acid Ther. 24, 374–387 (2014).
Sakamuri, S. et al. Identification of a tricyclic PIII chiral auxiliary for solid-supported synthesis of stereopure phosphorothioate-containing oligonucleotides. ChemBioChem 21, 1298–1303 (2020).
Iyer, R. P., Yu, D., Ho, N.-H., Tan, W. & Agrawal, S. A novel nucleoside phosphoramidite synthon derived from 1R, 2S-ephedrine. Tetrahedron Asymmetry 6, 1051–1054 (1995).
Wilk, A., Grajkowski, A., Phillips, L. R. & Beaucage, S. L. Deoxyribonucleoside cyclic N-acylphosphoramidites as a new class of monomers for the stereocontrolled synthesis of oligothymidylyl- and oligodeoxycytidylyl- phosphorothioates. J. Am. Chem. Soc. 122, 2149–2156 (2000).
Lu, Y. & Just, G. Stereoselective synthesis of R(P)- and S(P)-dithymidine phosphorothioates via chiral indolooxazaphosphorine intermediates derived from tryptophan. Angew. Chem. Int. Ed. 39, 4521–4524 (2000).
Oka, N., Yamamoto, M., Sato, T. & Wada, T. Solid-phase synthesis of stereoregular oligodeoxyribonucleoside phosphorothioates using bicyclic oxazaphospholidine derivatives as monomer units. J. Am. Chem. Soc. 130, 16031–16037 (2008).
Iwamoto, N., Oka, N., Sato, T. & Wada, T. Stereocontrolled solid-phase synthesis of oligonucleoside H-phosphonates by an oxazaphospholidine approach. Angew. Chem. Int. Ed. 48, 496–499 (2009).
Kandasamy, P. et al. Impact of guanidine-containing backbone linkages on stereopure antisense oligonucleotides in the CNS. Nucleic Acids Res. 50, 5401–5423 (2022).
Kandasamy, P. et al. Control of backbone chemistry and chirality boost oligonucleotide splice switching activity. Nucleic Acids Res. 50, 5443–5466 (2022).
Stec, W. J. et al. Deoxyribonucleoside 3′-O-(2-thio- and 2-oxo-‘spiro’-4,4-pentamethylene-1,3,2-oxathiaphospholane)s: monomers for stereocontrolled synthesis of oligo(deoxyribonucleoside phosphorothioate)s and chimeric PS/PO oligonucleotides. J. Am. Chem. Soc. 120, 7156–7167 (1998).
Nawrot, B. et al. 1,3,2-Oxathiaphospholane approach to the synthesis of P-chiral stereodefined analogs of oligonucleotides and biologically relevant nucleoside polyphosphates. Pure Appl. Chem. 80, 1859–1871 (2008).
Misiura, K., Szymanowicz, D., Olesiak, M. & Stec, W. J. DBU-assisted 1,3,2-oxathiaphospholane ring-opening condensation with selected O-, S-, N- and C-nucleophiles. Tetrahedron Lett. 45, 4301–4305 (2004).
Knouse, K. W. et al. Unlocking P(V): reagents for chiral phosphorothioate synthesis. Science 361, 1234–1238 (2018).
Huang, Y. et al. A P(V) platform for oligonucleotide synthesis. Science 373, 1265–1270 (2021).
Zheng, B. et al. P(III) vs P(V): a P(V) reagent for thiophosphoramidate linkages and application to an asymmetric synthesis of a cyclic dinucleotide STING agonist. J. Org. Chem. 87, 1934–1940 (2022).
Oka, N. & Wada, T. Advances in DNA-based nanotechnology themed issue. Chem. Soc. Rev. 40, 5730–5744 (2011).
Guga, P. & Stec, W. J. Synthesis of phosphorothioate oligonucleotides with stereodefined phosphorothioate linkages. Curr. Protoc. Nucleic Acid Chem. 14, 4.17.1–4.17.28 (2003).
Featherston, A. L. et al. Catalytic asymmetric and stereodivergent oligonucleotide synthesis. Science 371, 702–707 (2021).
DiRocco, D. A. et al. A multifunctional catalyst that stereoselectively assembles prodrugs. Science 356, 426–430 (2017).
Marzijarani, N. S. et al. New mechanism for cinchona alkaloid-catalysis allows for an efficient thiophosphorylation reaction. J. Am. Chem. Soc. 142, 20021–20029 (2020).
Lovinger, G. J., Sak, M. H. & Jacobsen, E. N. Catalysis of an SN2 pathway by geometric preorganization. Nature 632, 1052–1059 (2024).
Wang, M. et al. Catalytic asymmetric synthesis of the anti-COVID-19 drug remdesivir. Angew. Chem. Int. Ed. 59, 20814–20819 (2020).
Formica, M. et al. Catalytic enantioselective nucleophilic desymmetrisation of phosphonate esters. Nat. Chem. 15, 714–721 (2023).
Nugent, B. M., Yoder, R. A. & Johnston, J. N. Chiral proton catalysis: a catalytic enantioselective direct aza-Henry reaction. J. Am. Chem. Soc. 126, 3418–3419 (2004).
Struble, T. J., Lankswert, H. M., Pink, M. & Johnston, J. N. Enantioselective organocatalytic amine-isocyanate capture-cyclization: regioselective alkene Iodoamination for the synthesis of chiral cyclic ureas. ACS Catal. 8, 11926–11931 (2018).
Crocker, M. S., Deng, Z. & Johnston, J. N. Preparation of N‑aryl amides by epimerization-free umpolung amide synthesis. J. Am. Chem. Soc. 144, 16708–16714 (2022).
Nassir, M. et al. Stereocontrolled radical thiophosphorylation. J. Am. Chem. Soc. 145, 15088–15093 (2023).
Kanatsu, K. et al. Discovery and characterization of stereodefined PMO-gapmers targeting tau. Mol. Ther. Nucleic Acids 36, 102404–102417 (2025).
