uniQure N.V. · NASDAQ:QURE
uniQure — ASGCT 2019
20 slides from uniQure's conference presentation, Aug 13, 2019
Slide 1: Towards AAV5-mediated Gene Therapy for Hemophilia A with a Factor IX Variant
Towards AAV5-mediated Gene Therapy for Hemophilia A with a Factor IX Variant that functions independently of FVIII Ying Poi Liu, PhD uniQure Biopharma B.V., Amsterdam, The Netherlands
Slide 2: Forward-looking statements
This presentation contains forward-looking statements. All statements other than statements of historical fact are forward-looking statements, which are often indicated by terms such as “anticipate,” “believe,” “could,” “estimate,” “expect,” “goal,” “intend,” “look forward to,” “may,” “plan,” “poten
Slide 3: Hemophilia A; a bleeding disorder due to lack of FVIII
Hemophilia A; a bleeding disorder due to lack of FVIII - X-linked bleeding disorder - Deficiency in coagulation factor FVIII that serves as a cofactor for factor IX for activation of the coagulation cascade - Spontaneous bleeds - Classified into severe, moderate and mild (<1%, 1-5% and >5-40% of FVI
Slide 4: Why not express FVIII in the liver?
Why not express FVIII in the liver? • Endogenous FVIII synthesis in endothelial cells and not hepatocytes • Production site and protein load may activate the unfolded protein response in vitro and in vivo (Dorner et al. 1989, Malhotra et al 2008, Brown et al 2011, Zolothukhin et al 2016, uniQure unp
Slide 5: uniQure's approach: FIX variant
uniQure’s approach: FIX variant Novel Approach • Expression of a FIX variant with FVIII-independent FX activity using AAV5 vector Non thrombogenic Correction of Normal Hepatocyte friendly hemophilia activation Non immunogenic phenotype Low predicted immunogenicity risk Efficacious in patients Long-t
Slide 6: FIX-FIAV activates FX in the absence of FVIII
FIX-FIAV activates FX in the absence of FVIII FIX-FIAV: FIX-FIAV wt FIX FIX FX L6F, V181I, K265A and I383V Not hyperactive K265A and normally FVIII activated I383V V181I FIX-FIAV FVIII-independent activation of FX L6F Adapted from Kristensen L. H. et al Biochem J. 2016 2019| 6 CONFIDENTIAL
Slide 7: AMT-180: AAV5-Q1-FIX-FIAV is physiologically activated
AMT-180: AAV5-Q1-FIX-FIAV is physiologically activated AAV5-FIX-FIAV AAV5 FIX - The inactive FIX-FIAV zymogen is expressed - Activation is required FIX-FIAV Q1 hFIXco-FIAV gene Promoter FX activation in the absence of FVIII ss AAV vector genome L6F, K265A, V181I, I383V 2019| 7 CONFIDENTIAL
Slide 8: Studies to show proof of concept of FIX-FIAV in vitro and in vivo
Studies to show proof of concept of FIX-FIAV in vitro and in vivo In vitro, cells Wt mice HemA mice Cynomolgus Macaques FIX protein FIX protein FIX protein FIX protein FVIII-independent FVIII-independent Safety / tolerability of activity activity the AAV product 2019| 8 CONFIDENTIAL
Slide 9: FIX-FIAV shows 32% of FVIII-independent activity in APTT and thrombin generation assay
FIX-FIAV shows 32% of FVIII-independent activity in APTT and thrombin generation assay Catalytic K265A Residues Stable FIX Protein Functional expression in Purification Evaluation human cell line I383V One stage clotting assay (APTT) F V III- d egeneration Thrombin p le t e d p la s massay a 0 .5 p
Slide 10: FIX-FIAV is not hyperactive and requires physiological activation
FIX-FIAV is not hyperactive and requires physiological activation (same as FIX) Western blot FIXa FIX-WT FIX-FIAV t=0 t=60 t=0 t=60 188 98 62 55kDa = FIX 49 45kDa = FIXa 38 28 2 0 1 9 | 10 CONFIDENTIAL
Slide 11: FVIII-independent activity upon AAV injection in hemophilic mice
FVIII-independent activity upon AAV injection in hemophilic mice n=10, male Clotting (APTT) assay FVIII KO mice + Calcium IV dose 5e13 gc/kg generation FXa Activator + phospholipids FVIII-independent activity vs FIX protein W e e k 5 F V III in d e p e n d e n t a c tiv ity v s F IX a n tig e n F V
Slide 12: FIX-FIAV shows a therapeutic meaningful FVIII-independent activity in hemophilic plasma
FIX-FIAV shows a therapeutic meaningful FVIII-independent activity in hemophilic plasma • Normalisation of the FVIII-independent Summary efficacy AMT-180 activity to 100% of FIX protein • Recombinant FIX-FIAV • ~24% of FVIII-independent activity in ✓ 29% FVIII-like activity in thrombin hemophilic mi
Slide 13: FIX-FIAV expression in NHPs expected to translate to therapeutically relevant activity
FIX-FIAV expression in NHPs expected to translate to therapeutically relevant FVIII independent activity in humans h F IX p r o te in (% ) in N H P s male Cynomolgus macaque n=2 250 v e h ic le IV, 9e13 gc/kg adapted delivery 200 A A V 5 -L P 1 -F IA V h F IX p r o te in ( % ) 1 vehicle treated NHP
Slide 14: Safety assessments: non thrombogenic & low predicted immunogenicity risk
Safety assessments: non thrombogenic & low predicted immunogenicity risk Thrombogenicity • No elevation of coagulation activation markers: TAT + D-dimer levels in AAV-injected mice and NHPs • Histopathological examination of the NHP organs did not show signs of thrombus formation Immunogenicity • In
Slide 15: Conclusions and future plans
Conclusions and future plans Conclusions Ongoing & Future plans • AMT-180 is expected to prevent bleeds; sufficient • Full biochemical characterization of recombinant thrombin generation & clot formation FIX-FIAV protein • Hepatocyte friendly • Thrombin generation & clotting activity of AAV- injecte
Slide 16: Acknowledgements
Acknowledgements • Mettine H.A. Bos • Keiran Sinclair • Viola J.F. Strijbis • Caroline Brennan • Pieter Reitsma • Joachim Schwäble • Emily Mallet • Karin Huber • H. Fogg • Erhard Seifried • T. Jones • Juan Manuel Iglesias • Linda Tan • Michael Roberts Research Immunology Vector and process developme
Slide 17: uniQure
uniQure
Slide 18: FVIII-independent activity in AAV-FIX-FIAV transduced cells
FVIII-independent activity in AAV-FIX-FIAV transduced cells • AAV transduction of Huh-7 cells • Dose-dependent FIX-FIAV protein expression (ELISA) and FVIII-independent activity as measured by APTT clotting assay relative to a FVIII standardd F IX p r o te in le v e ls in H u h -7 F V III in d e p e
Slide 19: FIX-FIAV protein expression in hemophilic mice upon AAV injection
FIX-FIAV protein expression in hemophilic mice upon AAV injection n=10, male FVIII KO mice IV dose 5e13 gc/kg F IX -w t e x p r e s s io n in h e m o p h ilic m ic e F IX -F IA V e x p r e s s io n in h e m o p h ilic m ic e 2400 2400 A 12L B 14L 2000 2000 12R 15L F IX p r o te in (% ) F IX p r o t
Slide 20: Dose-dependent increase in FVIII independent activity in hemophilic mice
Dose-dependent increase in FVIII independent activity upon increased FIX-FIAV expression in hemophilic mice Day 0 Weeks 1, 2, 5 Week 8 Phenotypic correction IV tail vein and 8 Sacrifice assay injection Collect blood FVIII-/- mice Week 8 FVIII mimetic activity n=11 Dosis= high:1,44e14, mid: 5e13, low