Silence Therapeutics PLC · NASDAQ:SLN
Silence Therapeutics — Oligo Oxford 2018
36 slides from Silence Therapeutics's conference presentation, Apr 10, 2018
Slide 1: Development of innovative hepatocyte-targeted siRNA conjugates for liver-related disorders
Oligo Oxford 2018 Development of innovative hepatocyte-targeted siRNA conjugates for the treatment of liver-related disorders. Adrien Weingärtner, PhD, Group Leader Drug Delivery 10 APRIL 2018
Slide 2: Disclaimer
Disclaimer The information contained in this presentation is being supplied and communicated to you on a Securities in the Company have not been, and will not be, registered under the United States confidential basis solely for your information and may not be reproduced, further distributed to Secur
Slide 3: About Silence Therapeutics
About Silence Therapeutics Only quoted European RNA Our interference player facilities Key Performance Indicators: HQ in > Net cash position of £43M at 2nd of Jan 2018 London Headcount: > 45 FTE (30 in Berlin, R&D and 15 London, Corporate and R&D) R&D in Listing: Berlin > AIM, London Stock Exchange
Slide 4: Pipeline
Pipeline Building a proprietary portfolio, two projects advanced into preclinical development Our Programs 4Q 2018 Mid Mid 2019 2019 Mid 2019 Out-Licensed Programs © Silence Therapeutics 2018 4
Slide 5: GalNAc conjugated siRNA technology
GalNAc conjugated siRNA technology © Silence Therapeutics 2018 5
Slide 6: GalNAc-conjugated siRNA
GalNAc-conjugated siRNA Linker siRNA GalNAc gene silencing hepatocyte targeting Advantages of GalNAc-siRNA technology > GalNAc targets therapeutic siRNA molecules specifically to hepatocytes > Highly specific by targeting a single transcript > Patient-friendly via infrequent subcutaneous administrat
Slide 7: Gene silencing with GalNAc-conjugated siRNA in hepatocytes
Gene silencing with GalNAc-conjugated siRNA in hepatocytes extracellular intracellular Endosomal Escape Recognition & binding GalNAc-siRNA Recycling ASGPR Receptor (ASGPR) RNAi RISC AAA Target mRNA silencing > Intracellular delivery mediated through receptor endocytosis © Silence Therapeutics 2018 7
Slide 8: SLN124 for the treatment of iron overload disorders
SLN124 for the treatment of iron overload disorders © Silence Therapeutics 2018 8
Slide 9: Treatment of iron overload disorders
Treatment of iron overload disorders GOAL > Provide an effective and safe novel treatment option for patients with iron overload conditions, such as β-Thalassemia RATIONALE > Target a key modulator in iron regulation with a GalNAc-siRNA molecule providing a highly specific, effective & safe option t
Slide 10: Iron Overload Disorders
Iron Overload Disorders Affected organs by iron overload Diseases with iron overload > β-Thalassemia > Hereditary Haemochromatosis > Myelodysplastic Syndrome > Aplastic Anaemia > Sideroblastic Anaemia If untreated, iron accumulation in organs leads to severe damage, e.g. heart, liver & endocrine org
Slide 11: SLN124 mechanism of action
SLN124 mechanism of action HJV BMPR Macrophages > TMPRSS6 (Transmembrane BMP Protease, Serine 6) is a negative regulator of the Ferroportin BMP/SMAD signaling TMPRSS6 P Fe pathway Iron recycling > Inhibition of TMPRSS6 in P SMAD hepatocytes induces Hepcidin Hepcidin expression > Hepcidin reduces Iro
Slide 12: Silencing TMPRSS6 lowers serum iron levels in mice
Silencing TMPRSS6 lowers serum iron levels in mice Study design d1 d4 SC, n=4-6 mice TMPRSS6 mRNA (liver) Hepcidin mRNA (liver) Iron (serum) 4 * 40 N o r m a lis e d T M P R S S 6 m R N A N o r m a lis e d H e p c id in m R N A 1 .0 * S e r u m ir o n [µ m o l/L ] 3 30 ** * 2 20 0 .5 1 10 * 0 .0 0 0
Slide 13: SLN124 lowers iron levels for at least 6 weeks after single administration in mice
SLN124 lowers iron levels for at least 6 weeks after single administration in mice Study design d1 wk 1 wk 3 wk 6 SC, n=4 mice, 3 mg/kg TMPRSS6 mRNA (liver) Iron (serum) 40 N o r m a lis e d T M P R S S 6 m R N A 1 .0 S e ru m iro n [µ m o l/L ] 30 ** 20 0 .5 *** 10 *** *** *** 0 .0 0 1 1 3 6 w eeks
Slide 14: SLN124 - Summary
SLN124 - Summary > Highly potent, selective and long acting GalNAc-siRNA > Efficacious in lowering blood iron and well tolerated in healthy mice and non-human primates after single subcutaneous administration > Demonstrated therapeutic efficacy in clinically relevant animal disease models > Currentl
Slide 15: Technology Innovation: making a good thing better
Technology Innovation: making a good thing better © Silence Therapeutics 2018 15
Slide 16: R&D with focus on portfolio and innovation
R&D with focus on portfolio and innovation Technology Innovation Drug Discovery & Development > Improve performance of our GalNAc- > Build a proprietary therapeutic siRNA molecules portfolio by applying validated siRNA technologies > Strengthen and broaden IP portfolio > Partner programs in a strate
Slide 17: Unmodified GalNAc-conjugated siRNA is quickly degraded in biological fluids
Unmodified GalNAc-conjugated siRNA is quickly degraded in biological fluids Non-stabilised Serum exonucleases incubation 0h 4h 3d intact endonucleases misc enzymes degraded Stabilised 0h 4h 3d intact degraded end protection internal protection linker stabilisation >Incorporation of chemically modifi
Slide 18: Examples of stabilising chemically modified nucleotides
Examples of stabilising chemically modified nucleotides Modified 2‘-OH Modified phosphate O Base O Base O Base O Base O O O O O O O O O R 2N O OH O F O OMe O O P P P O O Base S O Base O O Base O O O RNA - 2-'OH 2'-F 2'-OMe LNA O Base O Base O Base O Base O OH O OH O OH O F O O O PO PS Amidates O O O
Slide 19: Internal siRNA modifications for endonuclease protection
Internal siRNA modifications for endonuclease protection © Silence Therapeutics 2018 19
Slide 20: Minimising the use of non-natural nucleotides
Minimising the use of non-natural nucleotides endonucleases Hundreds of chemical modification patterns ≥50% non-natural In vitro activity: without GalNAc, with various modifications, patterns, sequences Internal siRNA modifications Serum stability: with or without GalNAc Primary hepatocyte activity:
Slide 21: Serum stability and activity of molecules with reduced non-natural nucleotide content
Serum stability and activity of molecules with reduced non-natural nucleotide content Serum stability A x1 B x2 x3 x4 x5 C x6 x7 x8 x9 intact 50% serum 3d at 37oC degraded Silencing activity in primary hepatocytes % non-natural 50% 34% 18% nucleotides >Molecules with dramatically reduced non-natural
Slide 22: In vivo activity of GalNAc-siRNA molecules with reduced non-natural nucleotide content
In vivo activity of GalNAc-siRNA molecules with reduced non-natural nucleotide content Study design d1 d15 SC, n=6 mice Silencing activity in mice (target mRNA levels in liver tissue) % non-natural 50% 34% 18% 2 .0 nucleotides N o r m a lis e d t a r g e t m R N A 1 .5 1 .0 0 .5 0 .0 PBS C TR L 1 0
Slide 23: Summary – Internal GalNAc-siRNA modifications for endonuclease protection
Summary – Internal GalNAc-siRNA modifications for endonuclease protection endonucleases Identified chemical modification patterns that > Reduce dramatically non-natural nucleotide content from ≥50% to less than 15% > Retain outstanding nuclease stability, as well as robust silencing activity in vitr
Slide 24: Stabilisation of the linker
Stabilisation of the linker © Silence Therapeutics 2018 24
Slide 25: Performance of GalNAc-siRNA molecules with stabilised linker
Performance of GalNAc-siRNA molecules with stabilised linker Serum stability Silencing activity in primary hepatocytes 50% FBS unstable stable control at 37°C 0h 4h 3d 0h 4h 3d 0h 4h 3d intact degraded nM GalNAc- siRNA >GalNAc-siRNA with stabilised linker produces robust silencing activity in primar
Slide 26: In vivo activity of GalNAc-siRNA molecules with stabilised linker
In vivo activity of GalNAc-siRNA molecules with stabilised linker Study design d0 d7 d14 d21 d28 2 mg/kg SC, n=4 mice Silencing activity in mice (target mRNA in liver) 1.5 Normalised target mR NA 1.0 0.5 0.0 P B S C TR L d7 d14 d2 1 d28 d7 d14 d21 d2 8 u n s ta b le sta b le >GalNAc-siRNA with stabi
Slide 27: Antisense GalNAc-siRNA Conjugates
Antisense GalNAc-siRNA Conjugates Background: Affinity of the ligand to the ASGPR was reported to increase dramatically from monomeric to dimeric and trimeric but not much add on was shown by tetrameric clusters. Hence, triantenary GalNAc clusters with distances of 15-25Å between the carbohydrates a
Slide 28: Serum stability of siRNA-GalNAc conjugates
Serum stability of siRNA-GalNAc conjugates Serum stability 0h 4h 72h siRNA in 50% FCS RNA isolation & gel @ 37°C electrophoresis serum stability over three days monomer dimer trimer tetramer monomer dimer trimer tetramer control 0h 4h 3d 0h 4h 3d 0h 4h 3d 0h 4h 3d 0h 4h 3d 0h 4h 3d 0h 4h 3d 0h 4h 3d
Slide 29: In vitro activity of 3' anti-sense conjugates
In vitro activity of 3‘ anti-sense conjugates Study Design 0h 24h seeding primary murine hepatocytes RNA isolation & & co-treatment with siRNA TaqMan analysis 3’ anti-sense GalNAc-conjugates Normalised target mRNA © Silence Therapeutics 2018 29
Slide 30: In vitro activity of 5' sense conjugates
In vitro activity of 5‘ sense conjugates Study Design 0h 24h seeding primary murine hepatocytes RNA isolation & & co-treatment with siRNA TaqMan analysis 5’ sense GalNAc-conjugates Normalised target mRNA © Silence Therapeutics 2018 30
Slide 31: Duration of target knock down in vivo following single dose of GalNAc conjugates
Duration of target knock down in vivo following single dose of GalNAc conjugates d7 Study Design d0 d14 d28 s.c. 1 mg/kg n=4 ELISA of serum protein 3’ anti-sense GalNAc-conjugates 125 PBS s e r u m p r o t e in [ % o f P B S ] 100 3 '- a n ti- s e n s e - m o n o m e r 75 3 '- a n ti- s e n s e - d
Slide 32: Duration of target knock down in vivo following single dose of GalNAc conjugates
Duration of target knock down in vivo following single dose of GalNAc conjugates d7 Study Design d0 d14 d28 s.c. 1 mg/kg n=4 ELISA of serum protein 5’ sense GalNAc-conjugates 125 PBS s e r u m p r o t e in [ % o f P B S ] 100 5 '- s e n s e - m o n o m e r 75 tr im e r ic r e fe r e n c e 50 5 '- s
Slide 33: Duration of target knock down in vivo following single dose of GalNAc conjugates
Duration of target knock down in vivo following single dose of GalNAc conjugates d7 Study Design d0 d14 d28 s.c. 1 mg/kg n=4 ELISA of serum protein dimeric GalNAc-conjugates 125 PBS s e r u m p r o t e in [ % o f P B S ] 100 3 '- a n ti- s e n s e - d im e r 75 tr im e r ic r e fe r e n c e 5 '- s e
Slide 34: Summary – valency of GalNAc-cluster and position
Summary – valency of GalNAc-cluster and postion GalNAc-clusters > with two carbohydrates are as good clusters of as tree and four > conjugation to the sense strand results in better in vivo performance © Silence Therapeutics 2018 34
Slide 35: Technology Innovation: Summary
Technology Innovation: Summary • highly stable and potent GalNAc molecules (in vitro and in vivo) • reduced internal non-natural nucleotides by up to 80% • internal patterns are universally applicable • reduced COG of building blocks: by >25% • improved GalNAc-linker designs for simpler synthesis •
Slide 36: Acknowledgements
Acknowledgements Silence Technology Innovation Team: Marie Lindholm, Judith Hauptmann, Lucas Bethge Silence Drug Development Team: Manuela Aleku, Christian Frauendorf & Jens-Uwe Künstler Silence Preclinical Drug Discovery Team: Ute Schaeper, Steffen Schubert & Ulrich Zügel © Silence Therapeutics 201