Alnylam Pharmaceuticals, Inc. · NASDAQ:ALNY
Alnylam Pharmaceuticals — Annual Biologics Symposium 2018
34 slides from Alnylam Pharmaceuticals's conference presentation, May 9, 2018
Slide 1: Nonclinical Safety Evaluation of GalNAc-siRNA Conjugates
Nonclinical Safety Evaluation of GalNAc-siRNA Conjugates Joe Cichocki, Ph.D. Early Development, Alnylam Pharmaceuticals Annual Biologics Symposium May 9th, 2018 © 2018 Alnylam Pharmaceuticals, Inc. 1
Slide 2: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 3: Alnylam Clinical Development Pipeline
Alnylam Clinical Development Pipeline Focused in 3 Strategic Therapeutic Areas (STArs): Genetic Medicines Cardio-Metabolic Diseases Hepatic Infectious Diseases 3
Slide 4: Challenges In Delivery Of siRNA Therapeutics
Challenges In Delivery Of siRNA Therapeutics • Large (15-17 kDa), net negative charge, unstable in serum OH O • Can be modified to promote active hepatocellular uptake • Lipid nanoparticles OH • Require i.v. dosing • Conjugation with ligand • Can be given s.c. F OCH3 • Can be chemically-modified to
Slide 5: GalNAc-siRNA Conjugates Preclinical considerations
GalNAc-siRNA Conjugates Preclinical considerations N-acetylgalactosamine (GalNAc) • Fate of sense and anti-sense strands • Off-target toxicity • Limited tissue distribution, targeted delivery to liver • Short plasma half-life; long tissue half-life; long PD effects • Metabolized by serum and tissue
Slide 6: Regulatory Challenges For RNAi Therapeutics
Regulatory Challenges For RNAi Therapeutics Preclinical considerations Not a biologic….but not a small molecule! Treated (more) like a small molecule by regulators Separate regulatory guidance not in place for RNAi Paving the way for future RNAi therapeutics to ensure timely delivery of these medica
Slide 7: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 8: Liver: Distribution of ASGPR Expression Across Species
Liver: Distribution of ASGPR Expression Across Species Rat Mouse NHP Human 8
Slide 9: Consistent Plasma and Liver Exposure Profiles Across Programs (Rat Data)
Consistent Plasma and Liver Exposure Profiles Across Programs (Rat Data) Dose vs Plasma AUC Dose vs Liver AUC Compound PBS siRNA1 siRNA2 siRNA3 siRNA4 siRNA5 siRNA6 9
Slide 10: Long PD Effect and Long Tissue Half-life
Long PD Effect and Long Tissue Half-life • Maximum pharmacologic effect is not immediate! Typically 2-3 weeks after dosing • Toxicological considerations • Test article typically still present in liver after a 13 week recovery period • Durable pharmacology • Toxicological considerations 10
Slide 11: Lack Of Cross Reactivity Between Rodents And Humans
Lack Of Cross Reactivity Between Rodents And Humans • In some instances, we can get target knockdown in nonclinical species and humans • We always get cross reactivity to cynomolgus monkey • Often, we do not have cross reactivity to rodent • Can still evaluate chemical toxicity • Cannot investigate
Slide 12: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Early tox screening ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposur
Slide 13: Candidate Selection: Strategies to Mitigate Off-Target Toxicity
Candidate Selection: Strategies to Mitigate “Off-Target” Toxicity • Bioinformatics • BLAST sequence against human and tox species genomes (1,000s of sequences) • Predict potency • Exclude sequences with high homology to “off-target” transcripts • Chemistry • Modify sense and antisense strands to pro
Slide 14: siRNA-GalNAc Conjugates Platform-wide Toxicological Responses
siRNA-GalNAc Conjugates Platform-wide Toxicological Responses Rodent (typically Rat) ◦ Liver – Hepatocellular vacuolation. Increased number and size of normal rat hepatocellular vacuoles. Contain neutral lipid. – Increased single cell necrosis at high doses and tissue exposures – Increased mitosis a
Slide 15: Hepatocellular Vacuolation, Rodents Only
Hepatocellular Vacuolation, Rodents Only • Typically seen at all doses (including controls) • Usually minimal to moderate increase above background control levels • Dose-dependent incidence and severity • Not associated with changes in serum liver enzyme activity • Partially-to-fully reversible Vehi
Slide 16: Hepatocellular Vacuolation, Rodents Only (ORO)
Hepatocellular Vacuolation, Rodents Only (ORO) Control, Female 100mg/kg, Female 16
Slide 17: Hepatocellular Vacuolation in SD Rats
Hepatocellular Vacuolation in SD Rats 140 mg/kg cumulative dose associated with minimal vacuolation (score = 10) Most pharmacological doses ≤ 2.5 mg/kg Minimal vacuolation observed at doses > 50X pharmacological dose Black line = Model fit Grey line highlights concentration of drug needed to produce
Slide 18: Hepatocellular Single Cell Necrosis, Rats Only
Hepatocellular Single Cell Necrosis +/- Increased mitoses & regeneration, Rats Only • Most severe lesion observed, occasionally with minimal increase in serum liver enzyme activity (typically <2- fold over control) • Occurs at drug liver levels approaching tissue saturation generally at tissue expos
Slide 19: siRNA Class-Wide Toxicities: Test Article Accumulation, Rats
siRNA Class-Wide Toxicities: Test Article Accumulation, Rats Basophilic granules Basophilic granules in renal tubule epithelium • Rat proximal renal tubules ◦ Rare in NHP or other species, only at extremely high doses -- not seen in 13 week NHP studies • Severity: Minimal to moderate, dose dependent
Slide 20: siRNA Class-Wide Toxicities: Test Article Accumulation, NHPs
siRNA Class-Wide Toxicities: Test Article Accumulation, NHPs Basophilic granules in macrophages +/- vacuolation • Kupffer cells, lymph nodes • Severity: Minimal to moderate, dose dependent ◦ e.g. ≥ 30 mg/kg qW ◦ Partially reversible in recovery Liver NHP (siRNA) • Not related to setting NOAEL ◦ No d
Slide 21: Comparable toxicological profile following sub-chronic or chronic dosing in rats
Comparable toxicological profile following sub-chronic or chronic dosing in rats Comparative Toxicological Profile – Typical ALNY conjugate Dose qW*3 qM*7 (mg/kg) 10-15 No findings Liver: min.-mod. hepatocellular vacuolation 30-50 Liver: min.-mod. hepatocellular vacuolation Liver: min.-mod. hepatoce
Slide 22: Chronic Tox Study in Sprague-Dawley Rats
Compound X Ultra-rare disease indication, no approved therapy Chronic Tox Study in Sprague-Dawley Rats • 0, 20, 50, 200 mg/kg (qM*7) • No changes in target organ toxicity between 8-wk repeat dose and chronic studies ◦ Liver (minimal-to-marked hepatocellular vacuolation, minimal hepatocellular hypert
Slide 23: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 24: The Standard Test Battery For Genotoxicity ICH S2(R1)
The Standard Test Battery For Genotoxicity ICH S2(R1) Small Molecules Guidance* GalNAc-siRNA Conjugate Result Standard Battery Systems Bacterial reverse Salmonella typhimurium and mutation (Ames) test: Negative (all programs) Escherichia coli point mutations In vitro mammalian test: Metaphase chromo
Slide 25: Revusiran: 2-Year Carcinogenicity Assessment in Sprague-Dawley Rats
Revusiran: 2-Year Carcinogenicity Assessment in Sprague- Dawley Rats • 0, 10, 30, or 100 mg/kg (qW dosing) • No Revusiran-related effects on survival • Revusiran-related non-adverse decreased bodyweight gain and terminal bodyweight observed in both sexes • No evidence of neoplasia or hyperplasia • T
Slide 26: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 27: Fitusiran: 26-Week Rat Chronic Toxicity, Juvenile and Male Reproductive Outcome
Fitusiran: 26-Week Rat Chronic Toxicity Juvenile and Male Reproductive Functional Outcome Results (naive females mated with Fitusiran-treated males) • Juvenile tox: no effects on growth (crown-rump measurement), bone measurement (tibia and femur), and sexual maturation (physical development) • Male
Slide 28: siRNA-GalNAc Placental Exposures
siRNA- GalNAc Placental Exposures Inhibits Maternal Target, But no effect on Fetus Day: 0 3 6 9 12 15 18 21 Conception siRNA siRNA siRNA siRNA siRNA 24-hour Blood, organ, Urine fetus collection • Target Silencing at 10 mg/kg • ~95% knockdown of mRNA in maternal liver; no significant silencing in pla
Slide 29: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 30: Specialty Toxicity Evaluations of GalNAc-siRNA Conjugates
Specialty Toxicity Evaluations of GalNAc-siRNA Conjugates • Cardiovascular safety ◦ Single dose and multi-dose CV Safety Pharmacology studies in NHP at doses up to 300 mg/kg negative for GalNAc-siRNA conjugates. No effects on conduction or hemodynamics. ◦ No expected effects on hERG or ion channel a
Slide 31: Presentation Topics
Presentation Topics • Investigational RNAi Therapeutics ◦ Alnylam Pipeline ◦ Summary of points to consider in toxicology assessments • Summary of Key Toxicology Profiles ◦ PK/PD considerations ◦ Repeat-dose toxicity: target organs & pathologic effects; dose response and exposure relationships ◦ Chro
Slide 32: Clinical Perspective: Givosiran (ALN-AS1) Phase 1C Study
Clinical Perspective: Givosiran (ALN-AS1) Phase 1C Study in Patients with Acute Hepatic Porphyria 2.5 mg/kg qM 13-week (qW) toxicity data Species HED HED Adverse Findings NOAEL NOAEL (mg/kg) (cumul. mg/kg) Rat 5 20 None 32 Monkey 50 200 None
Slide 33: Summary: Emerging safety profiles for GalNAc-siRNA Conjugates
Summary: Emerging safety profiles for GalNAc-siRNA Conjugates The safety profiles of GalNAc-siRNA conjugates are generally consistent between programs ◦ Rat more sensitive than NHP – NOAEL in NHP always highest dose tested (up to 300 mg/kg) ◦ Target organs – 1º - Liver, 2º - Kidneys, Reticuloendothe
Slide 34: Thank You!
34 Thank You! LAlnylan,
Alnylam Pharmaceuticals — Annual Biologics Symposium 2018 on 6ix