Slide 1: Synthesis and application of stereopure guanidine-containing backbone to multiple oligonucleotide modalities in preclinical studies
Synthesis and application of stereopure guanidine-containing backbone to multiple oligonucleotide modalities in preclinical studies Pachamuthu Kandasamy Vice President , Medicinal Chemistry Wave Life Sciences Aug 28-31, 2022 IRT 3NA 2022
Slide 2: Forward looking statements
Forward looking statements This document contains forward-looking statements. All statements other than statements of historical facts contained in this document, including statements regarding possible or assumed future results of operations, preclinical and clinical studies, business strategies, r
Slide 3: PRISM platform enables rational drug design
PRISM platform enables rational drug design Sequence Chemistry 5’ B B: bases R: 2’ modifications A, T, C, mC, G, U, OMe, MOE, F, 3’ 2’ other modified bases other modifications X R 5’ B X: backbone chemistry PO, PS, PN 3’ 2’ Stereochemistry R Chiral control of any stereocenter 5’ modifications, backb
Slide 4: PRISM backbone modifications
PRISM backbone modifications Stereorandom PN variations (eg, targeting ligands) Rp PS PS Sp PS PO PN Stereorandom Rp PN 4 Sp PN
Slide 5: Advances in stereopure monomer synthesis & manufacturing
Advances in stereopure monomer synthesis & manufacturing Chiral auxiliary Amidites • High yield (up to 95%) • High selectivity L-Prolinol • Multi-kg scale Sp Monomer • Various 2’-modifications • Standard protecting groups ➢ Bz, iBu, Ac D-Prolinol Rp Monomer R'= Ph, R''= CH3 R'= CH2SiCH3Ph2, R''= H R
Slide 6: Efficient scalable process for synthesis
Efficient scalable process for synthesis ❑ Std CNE amidite for PO ❑ Chiral amidite for both PS and PN ❑ High diastereoselectivity ❑ High coupling efficiency Standard C&D process ❖ On column DEA wash ❖ Ammonium hydroxide DEA wash Scalable process ❖ 96-well plate to GMP manufacturing ❖ Comparable yiel
Slide 7: Characterization of stereopure dimers
Characterization of stereopure dimers Yield (Rp:Sp) Stereorandom 93.26% (47.27: 52.73) Stereopure Rp 84.97% Stereopure Sp (99.25:0.75) Rp-PN-fCfC 83.91% (0.22:99.78) RP-UPLC 31P NMR Dimer Amidite Rp:Sp 2’-MOE-G + 2’OMe-U L-PSM 99.8:0.2 D-PSM 0.9:99.1 2’-MOE-5MeC + 2’OMe-U L-PSM 99.9:0.1 D-PSM 0.4:99
Slide 8: Potency is enhanced with addition of PN modifications across modalities
Potency is enhanced with addition of PN modifications across modalities Silencing Splicing Editing Target knockdown (% remaining) % Skipping % Editing 100 Improved knockdown 80 Improved skipping Improved editing % Editing 60 40 20 0 10 -8 10 -6 10 -4 10 -2 10 0 10 2 Ranked by potency of reference Co
Slide 9: WVE-004 treatment resulted in durable reduction of poly-GP biomarker in mouse spinal cord & cortex
WVE-004 treatment resulted in durable reduction Silencing of poly-GP biomarker in mouse spinal cord & cortex1,2 Silencing – RNase H • WVE-004 leads to variant-selective silencing of C9orf72 transcripts Endogenous ➢ Contains PN chemistry RNase H ➢ Lowers expansion-containing transcripts ➢ Preserves h
Slide 10: PN modification improves silencing for GalNAc-siRNAs by increasing Ago2 loading in mice
PN modification improves silencing for GalNAc- Silencing siRNAs by increasing Ago2 loading in mice Silencing – RNAi • ~80% silencing HSD17B13 mRNA 14-weeks Endogenous AGO2 mRNA (liver) Human HSD17B13 post-single dose with PN-containing siRNA RISC transgenic mice 3 mg/kg SC dose • Significantly great
Slide 11: PN modification increases exon skipping, restores dystrophin expression and prolongs survival in dKO mice
PN modification increases exon skipping, restores dystrophin Splicing expression and prolongs survival in dKO mice Increased survival in dKO mice Splicing R2: 75 mg/kg, bi-weekly SC PBS PS/PO utr/mdx23 dKO mice PN/PS/PO PS/PO R1 Dosing regimens PN/PS/PO R2 R1: 150 mg/kg weekly Restored Reading Frame
Slide 12: WVE-006 directs RNA base editing to increase serum AAT protein above potential therapeutic threshold in mice
WVE-006 directs RNA base editing to increase serum AAT protein above potential therapeutic threshold in mice Editing Editing • WVE-006: PN-modified AIMer for SERPINA1 RNA editing • ~50% SERPINA1 mRNA editing in liver of AATD mouse model AIMers engage endogenous • WVE-006 increases serum AAT protein
Slide 13: Impact of PN chemistry highlighted in three high-impact publications this year
Impact of PN chemistry highlighted in three high- impact publications this year Silencing Splicing RNA Editing • Thanks to all colleagues and contributors from Wave Life Sciences 13