Circio Holding Asa · OSL:CRNA
Circio Holding Asa — Corporate Update (October 2023)
35 slides from Circio Holding Asa's corporate update, Oct 11, 2023
Slide 1: Building next generation RNA therapeutics
circio Building next generation RNA therapeutics Investor webcast 11 October 2023
Slide 2: Forward-looking statements
This report contains certain forward-looking statements based on uncertainty, since they relate to events and depend on circumstances that will occur in the future and which, by their nature, will have an impact on the results of operations and the financial condition of Circio Holding ASA and the C
Slide 3: Agenda
2. circVec Data 3. Rare disease data 4. Summary & Next steps 3
Slide 4: Citations of circRNA research
6,373 citations 30 September 2011 922 citations 2,291 citations 4
Slide 5: circRNA provides a toolbox to create a novel class of medicines
circRNA provides a toolbox to create a novel class of medicines Extended RNA durability Zi Enhanced protein expression circRNAisresistantto “2 exonuclease degradation, improving durability vs. mRNA mRNA circRNA ) X > microRNA sponging _ Regulatory functionality 5 circio
Slide 6: circVec: DNA or viral vector producing circRNA
` circVec DNA or viral vector DNA Inject circRNA biogenesis circRNA Intra-cellular protein expression 6
Slide 7: circRNA vs. mRNA intra-cellular half-life and expression
Shorter intra-cellular half-life Longer intracellular half-life Exponential decay High steady-state expression Less protein yield Higher protein yield → → (1) Based on published mRNA data and information from circRNA companies (2) Based on published synthetic circRNA data 7 (3) Based on Circio’s in
Slide 8: Temporal vector-based RNA expression dynamics; circRNA vs. mRNA
Temporal vector-based RNA expression dynamics; circRNA vs. mRNA Input assumptions for simulation: circRNA reaches Non-dividing target cells substantially higher Vector half-life: 100 days and more durable steady-state levels mRNA production: 10 molecules / hr mRNA half-life: 9 hrs * circRNA producti
Slide 9: circVec application areas
Gene therapy Vaccines Gene therapy Gene therapy Oncology Vaccines Improved expression Single-dose vaccine Enable repeat-dosing Improved uptake and reduced dosing Therapeutic protein for gene therapy Reduced vs. mRNA AAV delivery to tumors Enhanced nuclear uptake immunogenicity Advantage: efficient d
Slide 10: Collaboration with Neoregen announced 10 October
Collaboration announced 10 October Gene therapy Gene therapy Test delivery of circVec DNA vectors Vaccines using Neoregen´s proprietary peptide chemistry Enable repeat-dosing Improved uptake for gene therapy Reduced Enhanced nuclear uptake immunogenicity Advantage: repeat dosing and manufacturing Ch
Slide 11: Current focus areas
Current focus areas Designed for intra-cellular circRNA supply, durable protein expression and targeted regulatory functionality
Slide 12: circVec for AAV gene therapy in rare disease
Rare Disease vector 15x durability Shorter durability RNA 3-5x translation rate Lower steady-state Protein 10-15x maximum protein levels protein levels circVec could enable improved safety, lower dosing and reduced cost for AAV gene therapy 12
Slide 13: Rare disease indication prioritization
Rare Disease Initial Screening n Ipsum Lorem = 364 • Therapeutic Focus: Rare genetic disorders • Development Stage: Clinical or Market validation achieved 1st Prioritization Criteria n= Lorem 128 Ipsum • Etiology: monogenic disorders only (exclude polygenic disorders) • In vivo validation: availabil
Slide 14: AATD is a major unmet medical need
Rare Disease Current treatment options AATD is a major unmet medical need manifested in liver and lung Lung-associated AATD Replacement therapy with an alpha-1 proteinase inhibitors Weekly IV infusions Bronchodilators and inhaled steroids used for mild symptoms Lack of functional AAT protein Toxic a
Slide 15: Vaccines: development plan and target indication
Vaccines Development plan & target indication circVac non-replicating AdV vector Major infectious diseases, incl. influenza, shingles, malaria Establish single dose vaccine concept circVec inserts Out-license technical concept for clinical development following pre-clinical PoC 1-2 circRNAs Upcoming
Slide 16: circVac collaboration with Curiel laboratory
Vaccines circVac non-replicating AdV vector circVec inserts 1-2 circRNAs Collaboration to be initiated in October To be performed in the laboratory of professor David Curiel Test novel concept for circVac Flu Durable antigen Immune response vaccination expression booster
Slide 17: circVec data
3. Rare disease data 4. Summary & Next steps 17
Slide 18: Screening of natural circRNA-producing sequences
` mRNA AAAAAAAA Screening of most effective circRNA-producing sequences invented by nature Linear splicing circRNA protein Backsplicing 5 expression expression circRNA 4 / GAPDH L2 L1 L3 L4 Expression of endogenous circRNA 3 NGS analysis of 300+ RNAseq datasets 2 ⍺-ORF 1 ⍺-Ctrl 0 IR1 IR2 IR3 IR4 Top
Slide 19: Design optimization for circVec v1.0
´ Design optimization for circRNA biogenesis circVec v1.0 design - outperforming mRNA circRNA-specific RT-PCR; L1 = top wild-type Western blot, protein expression 400 5-10x improvement 300 on top natural locus 300 200 / GAPDH 200 100 Payload 100 00 1 2 3 4 5 6 7 8 IR 9 IR 0 11 1_ 1_ 1_ 1_ 1_ 1_ 1_ 1
Slide 20: circRNA outperforms mRNA in vitro
Accumulation of circVec v1.0 circRNA and protein payload over time, RT-PCR and Western blot circVec v1.0 RNA stability RT-qPCR, nascent vs. total RNA RNA expression protein expression circRNA outperforms mRNA in vitro – comparative in vivo experiments ongoing with circVec v1.0 20
Slide 21: circVec v1.0 circRNA vs. mRNA luciferase reporter expression
circVec v1.0 circRNA vs. mRNA luciferase reporter expression; time course1 + 200% more circRNA Doubled reporter protein + 100% expression from circRNA vs. mRNA after six days Equal more mRNA - 100% - 200% mRNA only better at 24h time - 300% point, due to faster biogenesis Hours 24 48 72 96 120 144 2
Slide 22: SPIKE expression from circVac
SPIKE expression from circVac, RNA and protein level RNA expression Protein expression circRNA > mRNA SPIKE/GAPDH mRNA circRNA SPIKE protein 0.8 mRNA circRNA expression @ 96hr 48hr 48hr 96hr 96hr / GAPDH 0.6 /GAPDH 0.4 ⍺-ORF SPIKE-encoding 0.2 circRNA accumu- lates over time 0.0 ⍺-Hexon 8 6 8 6 _4 _
Slide 23: circVec v2.0 sequence optimization
circVec sequence optimization, protein expression level 15 Relative expression 10 circVec v2.0 achieves 3-10x1 higher protein 5 10x yield than v1.0 design 0 VE H iE K C C P1 R lo 3 C _ f C er r PC CV _F V V R BV ru ve PV i H V_ C VB SV iHR H B H V_ v B tM 3_ EM lo VB EM C 23 1 Level of improvement i
Slide 24: In vivo confirmation of circVec v1.0
circVec v1.0 circRNA biogenesis and protein expression confirmed ✓ for DNA vector in immunodeficient mice ✓ circVec v1.0 circRNA biogenesis and protein expression confirmed in solid tumors from replicating Adenovirus (AdV) circVec vector ✓ circVac v1.0 immunogenicity confirmed for non-replicating Ad
Slide 25: Rare disease data
Rare disease data 4. Summary & Next steps 25 eircio
Slide 26: Depleting mutant form and replenishing functional protein by circVec
Rare Disease Depleting mutant form and replenishing functional protein by circVec - reverses toxic protein accumulation in liver and restores normal function in lung DNA or viral vector circVec 26
Slide 27: Depleting mutant form and replenishing functional protein by circVec
Rare Disease Depleting mutant form and replenishing functional protein by circVec - reverses toxic protein accumulation in liver and restores normal function in lung circRNA encoding functional AAT protein ✓ Functional AAT protein DNA or viral vector circVec 27
Slide 28: Depleting mutant form and replenishing functional protein by circVec
Rare Disease Depleting mutant form and replenishing functional protein by circVec - reverses toxic protein accumulation in liver and restores normal function in lung circRNA encoding functional AAT protein ✓ Functional AAT protein DNA or viral vector circVec Removal of mutant mRNA Abnormal AAT p
Slide 29: circVec AAT expression in liver cells
Rare Disease circVec AAT expression in liver cells Depleting mutant form and replenishing functional HepG2 AAT1 Protein Expression protein by circVec 250 > durability - reverses toxic protein accumulation in liver and restores normal function in lung 24 hr w/ circRNA 200 48 hr 72 hr 150 ng/mL 96 hr
Slide 30: mutAAT knock-down with circVec
Rare Disease circVec AAT expression in liver cells Depleting mutant form and replenishing functional HepG2 AAT1 Protein Expression protein by circVec 250 > durability - reverses toxic protein accumulation in liver and restores normal function in lung 24 hr w/ circRNA 200 48 hr 72 hr 150 ng/mL 96 hr
Slide 31: Summary & Next steps
ZA Summary & Next steps 31 eircio
Slide 32: Summary of circVec achievements and next steps
10x biogenesis rate vs. DNA and virus-based 3-5x enhanced protein miRNA sponging activity “nature´s best design” circRNA biogenesis expression vs. mRNA confirmed 15x extended half-life Optimized circVec 2.0 Validated for multiple Additional KD functi- vs. mRNA in vitro design established protein typ
Slide 33: Upcoming AAV and DNA vector experiments
2023 Oct-Dec Evaluation of circRNA in vitro expression profile from AAV virus compared to 1 AAV conventional mRNA-based AAVs 2 DNA v1 Oct-Dec Characterization of DNA format 1 as vector for circRNA expression 3 DNA v2 Nov-Jan Characterization of DNA format 2 as vector for circRNA expression
Slide 34: Upcoming vaccine, reporter and AAT experiments
2023 Vaccine Oct-Dec Evaluation of immunogenicity and T-cell responses in mice immunized 1 efficacy with circVac v1.0 expressing COVID Spike protein Reporter Nov-Dec Characterization of circVec v2.0 Firefly luciferase reporter expression level 2 expression and durability durability AAT 3 expression
Slide 35: Circio is the only significant player in DNA-format circRNA
Circio is the only significant player in the DNA-format circRNA space Enhanced durability and protein expression from circRNA is expected to translate into lower dosing of DNA-format applications, which may solve both potency, toxicity and cost challenges facing current ”gold-standard” gene therapy