Tonix Pharmaceuticals Holding Corp. · NASDAQ:TNXP
Tonix Pharmaceuticals — Technical Presentation (June 2022)
47 slides from Tonix Pharmaceuticals's technical presentation, Jun 8, 2022
Slide 1: The virus-host interface: A treasure trove of novel antiviral targets
The virus - host interface: A treasure trove of novel antiviral targets Tom C. Hobman Department of Cell Biology, Li Ka Shing Institute of Virology University of Alberta
Slide 2: Disclosures
Disclosures • Received research funding from Tonix Pharmaceuticals • Licensing agreement with Tonix Pharmaceuticals
Slide 3: Burden of epidemic/pandemic viruses
Burden of epidemic/pandemic viruses ■ Smallpox virus , influenza virus, Dengue virus, HIV, chikungunya virus, Ebola virus, HCV, HBV, Zika virus, SARS - CoV - 2 …. ■ Billions of people infected since 19 th century ■ Hundreds of millions of deaths ■ Enormous economic and social impact 0 >$82 trillion
Slide 4: Paucity of antivirals for emerging viruses
Paucity of antivirals for emerging viruses ■ Conventional antivirals target viral proteins □ Take years to develop □ Often highly specific ■ Vaccines are most effective in preventing viral diseases □ Highly specific ■ Need broad spectrum antivirals as first line of defense against emerging viruses
Slide 5: Taking advantage of the matchup
T ak i n g advantage of the matchup Youtube.com ~10 genes >20,000 genes
Slide 6: Targeting host-dependency factors is an exciting new frontier for antivirals
T a r get i n g host - dependency factors is an exciting new frontier for antivirals • Viruses are wholly reliant on host cellular pathways for replication & assembly • Vulnerabilities • Conserved pathways that can be targeted • Multiple viruses affected • Less chance of developing resistance
Slide 7: The calcium pump SPCA1 is required for infectivity of multiple RNA viruses
The calcium pump SPCA1 is required for infectivity of multiple RNA viruses including measles, Dengue, West Nile, Zika and chikungunya viruses Hoffmann et al, 2017 Cell Host Microbe Proof of principle
Slide 8: >50,000-fold reduction in SARS-CoV-2 replication
>50,000 - fold reduction in SARS - CoV - 2 replication
Slide 9: Quote on pandemic response and self-administered drugs
“For the present pandemic response, and for future pandemics …. the scientific community must be ready with an arsenal of easily self - administered drugs that can be tested in rapid, efficient clinical trials immediately after the causative viral agent is identified.”
Slide 10: Experimental approaches
Experimental approaches • Determine which cellular pathways are perturbed or activated • Test effects of pathway agonists/inhibitors on virus replication • Flaviviruses • Alphaviruses • HIV • Coronaviruses
Slide 11: Zika virus (ZIKV) distribution
Zika virus (ZIKV) distribution ResearchGate
Slide 12: ZIKV transmission cycles
ZIKV transmission cycles Sharma et al, 2019
Slide 13: ZIKV causes persistent infection and antagonizes the interferon response
ZIKV causes persistent infection and uses multiple strategies to antagonize the interferon response Kumar, Hou et al, 2016; Kumar et al, 2018; Limonta et al, 2019; Airo et al, 2022 Capsid
Slide 14: FGF2 mRNA is upregulated during ZIKV infection
Kumar et al, 2018 (Sertoli cells) Limonta et al, 2019 (Fetal astrocytes) Human fetal astrocytes Fibroblast growth factor - 2 (FGF2) mRNA is upregulated 270 - 8600X during ZIKV infection • FGF2 signaling suppresses IFN!
Slide 15: Ras/Raf/MEK pathway and antiviral response publications
Vol. 80, No. 9 J OURNAL OF V IROLOGY , May 2006, p. 4422 – 4430 0022 - 538X/06/$08.00 + 0 doi:10.1128/JVI.80.9.4422 – 4430.2006 Copyright © 2006, American Society for Microbiology. All Rights Reserved. Negative Regulation of the Alpha Interferon - Induced Antiviral Response by the Ras/Raf/MEK Pathwa
Slide 16: Flavivirus capsids downregulate the IFN response
DENV, JEV, MVEV, SLEV, YFV, ZIKV vs Control 12,966 genes with information 3621 Differentially expressed 3546 Downregulated 75 upregulated APOL1 HERC5 TRANK1 BORCS7 THEMIS2 IFI16 TNFSF10 SERPING1 CD274 SP110 SAMD9L PARP10 DDX58 ATF3 HLA - B UBA7 IFIH1 IRF7 HSPA6 IFI6 PLEKHA4 ISG 15 BST 2 IFIT 1 IFIT
Slide 17: ZIKV capsid inhibits RIG-I ubiquitination by TRIM25
ZIKV capsid inhibits RIG - I ubiquitination by TRIM25 Yang et al, 2021 Airo et al, 2022
Slide 18: Flaviviruses use many strategies to inhibit IFN response
Flaviviruses use many strategies to inhibit IFN response • Non - structural proteins block IFN induction and signaling by targeting NF B, IRF3 and STAT2 • Mechanisms known in many cases • Upregulated FGF signaling blocks IFN induction and ISG expression • MEK - dependent pathway • Capsid proteins
Slide 19: Conserved interactions between flavivirus capsid proteins and host proteins
Ma et al, 2011 Conserved interactions between flavivirus capsid proteins and host proteins You, Hou et al, 2015
Slide 20: Peroxisomes are membranous organelles
Peroxisomes are membranous organelles that: • Catabolize very long chain fatty acids • Regulate reactive oxygen species • Are sites of biosynthesis for specialized phospholipids • Control inflammation
Slide 21: Knowledge gap regarding viruses and peroxisomes
Knowledge gap regarding viruses and peroxisomes ViralZone Mohan et, 2002
Slide 22: Peroxisomes are antiviral signaling platforms
Ferreira et al, 2022 Peroxisomes are Antiviral Signaling Platforms
Slide 23: What happens to peroxisomes during viral infection?
What happens to peroxisomes during viral infection?
Slide 24: Flavivirus infection results in loss of peroxisomes
Flavivirus infection results in loss of peroxisomes You, Hou et al . 2015 Peroxisome Mock WNV
Slide 25: HIV infection also depletes peroxisomes
HIV - 1 infection also depletes peroxisomes Xu et al , 2017 HIVE - HIV with encephalitis HAND - HIV - Associated Neurocognitive Disorder
Slide 26: Different mechanisms, same result
Different mechanisms, same result…. Wong et al , 2018
Slide 27: SARS-CoV-2 infection depletes functional peroxisomes
Mock 24 hpi 48 hpi 72 hpi SARS - CoV - 2 SARS - CoV - 2 infection depletes functional peroxisomes Knoblach et al, 2021
Slide 28: SARS-CoV-2 infection impairs import of matrix proteins
SARS - CoV - 2 infection impairs import of matrix proteins PTS1 cargo X X X X X
Slide 29: Some viruses upregulate peroxisomes
Ferreira et al, 2022 Some viruses upregulate peroxisomes
Slide 30: Peroxisome pool is maintained by main pathways
Peroxisomes pool is maintained by 2 main pathways Ma et al, 2011
Slide 31: Genetic induction of peroxisomes by Pex11 inhibits ZIKV replication
Genetic induction of peroxisomes by Pex11 inhibits ZIKV replication by enhancing IFN response Wong et al, 2019
Slide 32: The ISG Viperin associates with peroxisomes
The ISG Viperin associates with peroxisomes Khantisitthiporn et al, 2021 • Potentiates the innate antiviral response • Positive feedback mechanism? • Positioning peroxisomes near the mitochondrial/MAM MAVS signaling synapse?
Slide 33: The power and serendipity of collaboration
Hopfensperger et al, 2020 The power and serendipity of collaboration…… RESEARCH ARTICLE Host - Microbe Biology The HIV - 1 Accessory Protein Vpu Downregulates Peroxisome Biogenesis Zaikun Xu, a Robert Lodge, b Christopher Power, c,d,f,g Eric A. Cohen, b,h Tom C. Hobman a,d,e,f Sawa et al, 2013 So wh
Slide 34: Drugs that inhibit Wnt signaling pathway
Pai et al, 2017 Lots of drugs that inhibit Wnt signaling pathway Do these types of drugs induce peroxisomes??
Slide 35: Wnt/β-catenin inhibitors increase peroxisome density and potentiate IFN response
Wnt/β - catenin inhibitors increase peroxisome density and potentiate the IFN response during viral infection Xu, Wong et al, in preparation
Slide 36: Wnt/β-catenin inhibitors inhibit SARS-CoV-2 replication in multiple cell types
0 50000 100000 150000 200000 250000 * * * * * * * * * * SARS - CoV - 2 virus titer (PFU/ml) 0 20000 40000 60000 80000 * * * * * * * * * * SARS - CoV - 2 virus titer (PFU/ml) Wnt/β - catenin inhibitors inhibit SARS - CoV - 2 replication in multiple cell types Calu - 3 NHBE • No effect in Vero cells •
Slide 37: Reducing β-catenin levels induces peroxisome proliferation and enhances IFN response
Reducing β - catenin levels induces peroxisome proliferation and enhances IFN response Sawa et al, 2013 X Xu, Wong et al, in preparation
Slide 38: Drugs with high SIs chosen for testing against variants
Yamaguchi et al, 2020 IWP - 01 KYA1797K E7449 Drugs with high SIs chosen for testing against Variants of Concern and small animal studies
Slide 39: Peroxisome-modulating drugs are effective against SARS-CoV-2 Variants of Concern
Peroxisome - modulating drugs are effective against SARS - CoV - 2 Variants of Concern DMSO 0 200000 400000 600000 800000 1000000 D614G * * * * * * * * IWP - O1 Pyrvinium KYA1797K SARS - CoV - 2 virus titer (PFU/ml) 0 100000 200000 300000 400000 UK B.1.1.7 (Alpha) * * * * * * * DMSO SARS - CoV - 2 v
Slide 40: Peroxisome-inducing drugs reduce replication of other RNA viruses
Peroxisome - inducing drugs reduce replication of other RNA viruses Xu, Wong et al, in preparation
Slide 41: SARS-CoV-2 is highly sensitive to Interferon
SARS - CoV - 2 is highly sensitive to Interferon IFN added pre - infection IFN added post - infection Lokugamage et al , 2020
Slide 42: In vivo testing of Wnt inhibitors
In vivo testing of Wnt inhibitors
Slide 43: Mouse SARS-CoV-2 lung titers: promising trend
4 DPI, Lung, N=5 60000 40000 20000 0 SARS - CoV - 2 titer (PFU/ml) Promising trend…… • Mice infected with 10x 3 pfu of mouse - adapted SARS - CoV - 2 • Drugs administered IP pre - and post - infection
Slide 44: Summary
Summary ■ RNA viruses employ a highly diverse array of tactics to block innate immune signaling ■ This includes novel mechanisms to disrupt biogenesis and/or function of peroxisomes during infection ■ Wnt signaling plays a role in peroxisome homeostasis ■ Genetic or pharmacological upregulation of p
Slide 45: Potential benefits of targeting peroxisomes for antiviral therapy
Potential benefits of targeting peroxisomes for antiviral therapy ■ Drugs ……..lots of them! □ Wnt inhibitors □ Peroxisome proliferator - activated receptor agonists □ Good safety profiles □ Bench to bedside route shorter ■ Broad - spectrum antiviral activity ■ Reduce inflammation? ■ Prophylactic and
Slide 46: Ongoing studies and future directions
Ongoing studies and future directions ■ Determine how SARS - CoV - 2 depletes peroxisomes □ Candidate viral proteins and interactome studies ■ In vivo studies □ Intranasal and oral delivery routes ■ Testing antiviral activity of Wnt inhibitors in combination with other drugs ■ Screening libraries fo
Slide 47: Acknowledgements
Acknowledgements Hospital Foundation we University ©° CIHR IRS¢ Li Ka Shing Institute of Virology
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