Tonix Pharmaceuticals Holding Corp. · NASDAQ:TNXP
Tonix Pharmaceuticals — Wnt/β-Catenin Targeted Drug Development Summit (2023)
36 slides from Tonix Pharmaceuticals's conference presentation, 2023
Slide 1: Targeting the Wnt/b-catenin pathway as a broad-spectrum antiviral strategy
Targeting the Wnt/b-catenin pathway as a broad-spectrum antiviral strategy 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: Research approaches
Researchapproaches Experimental Focus § •Identification Flaviviruses of key host factors/pathways that •are Alphaviruses utilized or affected by multiple RNA viruses • HIV § •Pharmacological Coronaviruses PR I S CR targeting of these host Restriction & factors/pathways should Dependency Factors resu
Slide 4: Host-targeted antivirals to be part of this arsenal?
“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.” White et al, 2021 Host-targeted
Slide 5: Peroxisomes are targeted during RNA virus infection
Peroxisomes are targeted during RNA virus infection • Abundant metabolic organelles in the cytoplasm • Catabolize very long chain fatty acids • Regulate reactive oxygen species • Produce specialized phospholipids (e.g. plasmalogens)
Slide 6: Flavivirus infection results in loss of peroxisomes
Flavivirus infection results in loss of peroxisomes Mock WNV Mock WNV Peroxisome aNS2B-3 Peroxisome You, Hou et al. 2015
Slide 7: Why would a virus want to deplete the peroxisome pool?
Why would a virus want to deplete the Trends in Cell Biology peroxisome pool? Ferreira et al, 2022 Trends in Cell Biology Figure 1. Mitochondrial antiviral signaling (MAVS) protein-dependent antiviral signaling. Upon virus entry, viral RNA is released into the cytosol where it is recognized by retin
Slide 8: SARS-CoV-2 is highly sensitive to Interferon (IFN)
SARS-CoV-2 is highly sensitive to Interferon (IFN) SARS-CoV-2 Is Sensitive to Type I IFN Pretreatment Lokugamage et al. FIG 1 SARS-CoV-2 is sensitive to type I IFN pretreatment. (A) Vero E6 cells were treated with 1,000 U recombinant type I (hashed line) IFN or mock (solid line) for 18 h prior to in
Slide 9: SARS-CoV-2 depletes functional peroxisomes
SARS-CoV-2 depletes functional peroxisomes SARS-CoV-2 Mock 24-hr 48-hr 72-hr a-PTS1 Flavivirus Knoblach et al, 2021 X Fujiki et al, 2014 Mock 24 48 72 hpi SARS-CoV-2
Slide 10: Respiratory Syncytial Virus (RSV) also reduces peroxisome pool
Respiratory Syncytial Virus (RSV) also reduces peroxisome Respiratory pool syncytial virus infection depletes peroxisomes too RSV F Peroxisomes Merge Mock RSV
Slide 11: As does HIV....
< T-eq1/73H asejolul sewosixoled Jo Jequinu eajejay Peroxisome
Slide 12: Canonical Wnt pathway
Canonical Wnt pathway
Slide 13: HIV-1 Vpu suppresses peroxisome biogenesis
HIV-1 Vpu suppresses peroxisome biogenesis Vpu reduces metabolic activity of peroxisomes RESEARCH ARTICLE Host-Microbe Biology crossm RESEARCH ARTICLE Host-Microbe Biology The HIV-1 Accessory Protein Vpu Downregulates Peroxisome crossm Biogenesis The HIV-1 Zaikun Xu, Accessory Robert Lodge, a Christ
Slide 14: Wnt/b pathway inhibits peroxisome biogenesis
§ Wnt/b pathway inhibits peroxisome biogenesis Hypothesis: Inhibiting Wnt/b pathway will induce peroxisomes and reduce virus replication via enhanced IFN response Test effects of Wnt/b pathway inhibitors on peroxisome density
Slide 15: Wnt/β-catenin inhibitors increase peroxisome density
Wnt/β-catenin inhibitors increase peroxisome density DMSO Pyrvinium KYA1797K E7449 Xu, Elaish, Wong et al, in preparation But do they inhibit virus replication?
Slide 16: Wnt/β-catenin inhibitors reduce SARS-CoV-2 replication in multiple cell types
Wnt/β-catenin inhibitors reduce SARS-CoV-2 replication in multiple cell types Calu-3 NHBE Calu3 NHBE Calu3-human lung adenocarcinoma NHBE-normal human bronchial epithelial cells Xu, Elaish, Wong et al, in preparation
Slide 17: Reducing β-catenin levels induces peroxisome proliferation and enhances IFN response
Reducing β-catenin levels induces peroxisome proliferation and enhances IFN response X Suggests 38that antiviral effects of Wnt/b catenin inhibitors is Sawa et al, 2013 not due to off target effects Xu, Elaish, Wong et al, in preparation
Slide 18: Wnt/β-catenin inhibitors increase peroxisome density in Vero cells but do not reduce virus replication
Wnt/β-catenin inhibitors increase peroxisome density in Vero cells but do not reduce virus replication V e ro E 6 , 2 4 h p i 6 S A R S - C o V - 2 v ir u s tit e r ( P F U /m l, L o g ) 5 4 M M µM µM µM µM µM µM µM µM µM µM µM µM O µM n n S M 1 0 1 0 0 0 1 0 0 1 1 1 0 0 1 1 1 1 1 0 1 1 1 D 1 4 4 K
Slide 19: Some Wnt/β-catenin inhibitors decrease virus replication when added post-infection
S A R S - C o V - 2 v ir u s t it e r ( P F U /m l, L o g ) 3 4 5 6 D M S O IW P -O L 1 * G K -9 7 W 4 n t- C N 5 C 9 B -0 K 8 Y 4 A 6 E 1 T 7 C 9 7 K * -1 9 2 2 1 P 5 y 9 rv in iu m * iC R T -1 S 4 M 0 4 6 9 0 E 7 4 4 9 1 2 h p o s t- in fe c tio n tr e a tm e n t, 2 4 h p i S A R S - C o V - 2 v i
Slide 20: Drugs with high SIs chosen for testing against Variants of Concern and small animal studies
Drugs with high SIs chosen for testing against Variants of Concern and small animal studies E7449 KYA1797K IWP-01 Yamaguchi et al, 2020
Slide 21: Peroxisome-modulating drugs are effective against SARS-CoV-2 Variants of Concern
Peroxisome-modulating drugs are effective against SARS-CoV-2 Variants of Concern S A R S - C o V - 2 v ir u s t it e r ( P F U /m l, L o g ) S A R S - C o V - 2 v i r u s t it e r ( P F U / m l , L o g ) In d ia B .1 .6 1 7 .2 U K B .1 .1 .7 ( D e lt a ) 6 (A lp h a ) 6 * 5 * * 5 * * * * * * 4 * * 4
Slide 22: Some Wnt/b-catenin inhibitors reduce replication of other human coronaviruses
V ir u s t it e r ( P F U /m l, L o g ) V ir u s t it e r ( P F U /m l, L o g ) 0 1 2 3 4 5 0 1 2 3 4 5 DMS O DMS O IW P - O1 IW P - O1 LGK- LGK- 974 974 W n t- W n t- C59 C59 NCB NCB -0 8 4 6 -0 8 4 6 KYA1 KYA1 * * 797K 797K ETC- ETC- 1922 1922 159 159 P y rv P y rv * in iu m * in iu m iC R T iC R
Slide 23: Wnt/b-catenin inhibitors reduce replication of other RNA viruses
Wnt/b-catenin inhibitors reduce replication of other RNA viruses 7 Flavivirus 8 Alphavirus M A Y V t itr e ( P F U /m L , L o g ) Z IK V titr e (P F U /m L , L o g ) * * 7 * * * * 6 6 ** 5 * 5 4 K K 9 9 1 1 m m O O 4 4 7 7 -O -O iu iu S S 4 4 9 9 M M 7 7 7 P 7 P in in E E 1 1 D IW D IW rv rv A A Y Y
Slide 24: In vivo testing of Wnt inhibitors
In vivo testing of Wnt inhibitors Drugs administered intranasally to female BALB/c mice (5 in each group) Intranasal challenge with 5 x 103 pfu of mouse-adapted SARS-CoV-2
Slide 25: KYA1797K
KYA1797K § Destabilizes b-catenin by activating Axin-GSK3b complex § Tested via IP administration in mice (20 mg/kg/day) ú Here limited to 3 mg/kg due to solubility
Slide 26: E7449
E7449 § Other names: Stenoparib § Dual inhibitor of PARP1/2 & tankyrase1/2 § Orally bioavailable § Phase1/2 study for cancer indications ú Well tolerated (50-800 mg dosing) in humans ú 0% cytotoxicity in human cells at 10 µM in vitro
Slide 27: Wnt inhibitors have modest effect on virus-induced weight loss
Wnt inhibitors have modest effect on virus- induced weight loss 110 N o r m a liz e d w e ig h t ( % ) D M S O /S F M 100 D M S O /S A R S - C o V -2 E 7 4 4 9 /S A R S - C o V -2 90 6 mg/kg 80 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 T im e a f t e r c h a ll e n g e ( d ) 110 N o r m a liz e d w e ig h
Slide 28: Anesthesia/IN drug administration causes mild transient weight loss
Anesthesia/IN drug administration causes mild transient weight loss 110 D M S O /S F M N o r m a liz e d w e ig h t ( % ) K Y A 1 7 9 7 K /S F M 105 E 7 4 4 9 /S F M 100 95 90 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 T im e a f t e r c h a ll e n g e ( d )
Slide 29: Wnt inhibitors reduce viral load in lungs
Wnt inhibitors reduce viral load in lungs 4 DPI, Lung, N=5 PMP70 Nucleocapsid SARS-CoV-2 titer (PFU/g tissue, Log) ** 6 *** 5 4 Uninfected DMSO 3 SO K 9 44 97 M E7 17 D YA K Xu, Elaish, Wong et al, in preparation E7449 KYA1797K
Slide 30: Wnt inhibitors reduce proinflammatory markers in lungs
Wnt inhibitors reduce proinflammatory markers in lungs IL-6 IL-1β 40 8 Host Gene RNA (RFC) Host Gene RNA (RFC) 30 6 20 4 10 2 0 0 49 K O O 9 K O O 97 S S 74 44 97 S S M M 17 M M E E7 D D 17 D D A A Y Y K K SARS-CoV-2 SARS-CoV-2 4-days post-infection Xu, Elaish, Wong et al, in preparation
Slide 31: Wnt inhibitors reduce expression of miRNAs that suppress peroxisome biogenesis
Wnt inhibitors reduce expression of miRNAs Wntthat suppress inhibitors peroxisome suppress expression biogenesis of PEX-targeting miRNAs (No HIV-1 Vpu suppresses infection)biogenesis peroxisome Flavivirus 4 Relative miRNA level (Fold) 3 2 Fujiki et al, 2014 1 * * * * * * * * * * * * * * 0 O1 m O1 m
Slide 32: Summary
Summary § Inhibition of Wnt/b catenin pathway induces peroxisomes and enhances IFN response during viral infection § Significantly reduces SARS-CoV-2 replication in vitro and in vivo § Broad-spectrum activity against other RNA viruses
Slide 33: Potential benefits of targeting peroxisomes for antiviral therapy
Potential benefits of targeting peroxisomes for antiviral therapy § Drug candidates with good safety profiles ú Wnt inhibitors ú Peroxisome proliferator-activated receptor agonists § Reduce inflammation? § Prophylactic and early therapeutic use? § Do not induce IFN in absence of viral infection
Slide 34: Inducing peroxisome biogenesis may have multiple health benefits
Inducing peroxisome biogenesis may have multiple health benefits Zalckvar and Schuldiner, 2022
Slide 35: Ongoing/Future studies
Ongoing/Future studies § In vivo efficacy of post-infection administration of Wnt/b catenin inhibitors § Increase bioavailability of drugs? Oral? Nebulizer? Derivatives? § Testing other Wnt/b catenin inhibitors and peroxisome proliferators alone and in combination § High throughput screening for nov
Slide 36: Acknowledgements
Acknowledgements Dr. Zaikun Xu Dr. Mohamed Elaish Dr. Jason Wong Wil Branton Dr. Natacha Ogando Dr. David Evans Dr. Chris Power Ray Ishida Dr. Rick Rachubinski Dr. Barbara Knoblach Dr. Joaquin Lopez Orozco Dr. Lara Mahal Dr. Irv Mayers Dr. Ryan Noyce
Tonix Pharmaceuticals Holding — Wnt/β-Catenin Targeted Drug Development Summit (2023) on 6ix