Slide 1: Resurrecting Dr. Edward Jenner's 1796 Horsepox Vaccine to Protect Against Mpox and Smallpox
© 2025 Tonix Pharmaceuticals Holding Corp. Resurrecting Dr. Edward Jenner’s 1796 Horsepox Vaccine to Protect Against Mpox and Smallpox UT Southwestern Microbiology Department Seminar Seth Leder man, M.D. March 11, 2025 Version P06050 March 11, 2025 (Doc 1566)
Slide 2: Cautionary Note on Forward-Looking Statements
© 2025 Tonix Pharmaceuticals Holding Corp. Cautionary Note on Forward - Looking Statements Certain statements in this presentation regarding strategic plans, expectations and objectives for future operations or results are “forward - looking statements” as defined by the Private Securities Litigatio
Slide 3: Key Contributors
© 2025 Tonix Pharmaceuticals Holding Corp. Key Contributors 3 Univ. of Alberta Ryan Noyce David Evans Tonix Sina Bavari Farooq Nasar Scott Goebel Zeil Rosenberg
Slide 4: In 1798, Edward Jenner Described Vaccination with the Virus that Causes Cow Pox
© 2025 Tonix Pharmaceuticals Holding Corp. 4 In 1798, Edward Jenner Described Vaccination with the “Virus” that Causes Cow Pox for Preventing Smallpox Jenner, E. (1798) “ The Inquiry ” • Full title: “An Inquiry Into the Causes and Effects of the Variolae Vaccinae , a Disease Discovered in Some of th
Slide 5: Two Vaccines FDA-Approved for Smallpox and Mpox
© 2025 Tonix Pharmaceuticals Holding Corp. Two Vaccines FDA - Approved for Smallpox and Mpox Both are derived from Jenner’s Vaccine from >200 years ago ACAM2000 (live - virus vaccine) - Emergent • Replicating - based on a clone of live - virus vaccinia ( Dryvax ®) • Single - dose • Provides durable
Slide 6: Live Virus Smallpox Vaccines in the National Stockpile Are Derived From the NYCBH Strain
© 2025 Tonix Pharmaceuticals Holding Corp. Live Virus Smallpox Vaccines in the National Stockpile Are Derived From the NYCBH Strain 6 Propagation of Jenner’s vaccinia in cows rapidly increases genetic diversity and results in many strains with varying properties 1,2 1800s WHO selects the NYCBH strai
Slide 7: Passage or Zoonosis Leads to Genomic Deletions and New Host Range
© 2025 Tonix Pharmaceuticals Holding Corp. Passage or Zoonosis Leads to Genomic Deletions and New Host Range 1,2 7 Passage Species jumping Modern vaccinia (ACAM2000 ® , approved) Passage MVA (Jynneos ® , approved) ITR deletions Extensive deletions Host range Passage is a primitive tool to induce del
Slide 8: U.S. Recognizes Smallpox Preparedness as a Priority
© 2025 Tonix Pharmaceuticals Holding Corp. U.S. Recognizes Smallpox Preparedness as a Priority National Stockpile Expansion is Recommended by Experts 8
Slide 9: Mpox and Smallpox Reports by U.S. Agencies & Institutions
© 2025 Tonix Pharmaceuticals Holding Corp. Mpox and Smallpox Reports by U.S. Agencies & Institutions Multiple recent statements by U.S. Agencies warning about smallpox and monkeypox 1 - 6 U.S. National Academy of Sciences Consensus Report (March, 2024) 6 ‒ “Additionally, safer, single - dose vaccine
Slide 10: Mpox Outbreak 2022-23: Clade IIb: WHO Public Health Emergency
© 2025 Tonix Pharmaceuticals Holding Corp. Mpox Outbreak 2022 - 23: Clade IIb: WHO Public Health Emergency Global Health Concern (PHEIC) 10 Risk of Spread and Lethality of Clade IIb • Case Fatality Rate (CFR): 0.1% to 3.6% Lower compared to Clade I • Primarily spread through sexual contact among g
Slide 11: New Clade Ib Mpox Declared PHEIC by WHO in August 2024
© 2025 Tonix Pharmaceuticals Holding Corp. New Clade Ib Mpox Declared PHEIC* by WHO** in August 2024 and Reaffirmed in February 2025 Clade Ib - first wave in Democratic Republic of Congo (DRC) • Spreads in households • Affects children Additional emerging mutation • Potentially lower mortality • H e
Slide 12: Non-replicating MVA Requires Two Doses: Drop-off in Protection
© 2025 Tonix Pharmaceuticals Holding Corp. Non - replicating MVA Requires Two Doses: Drop - off in Protection from Mpox With Only One Dose 12 ACIP Oct 25, 2023 U.S. Mpox Vaccine Coverage in High - Risk Groups (CDC) 1 - dose: 38.8% 2 - dose: 24.3% 37% Drop Out
Slide 13: Jenner was not Just a Country Doctor
© 2025 Tonix Pharmaceuticals Holding Corp. Jenner was not Just “a Country Doctor” Linnean Society member (1788) and Royal Society 1 fellow (1789) - PRIOR to Vaccination Jenner was the first and favorite student of John Hunter (1728 - 1793) − Hunter ran a private “medical school” in London • Surgeon
Slide 14: Edward Jenner Successfully Used Vaccination to Protect Against Smallpox
© 2025 Tonix Pharmaceuticals Holding Corp. • Jenner "vaccinated” healthy individuals with material from the lesions, which he called “vaccine” (from vacca , Latin for “cow”) • The p ustule matter from “cow pox” sores on a milkmaid’s hands; conferred protection against future challenges with smallpox
Slide 15: Edward Jenner Successfully Used Vaccination: Virus came from Horses
© 2025 Tonix Pharmaceuticals Holding Corp. Jenner "vaccinated” healthy individuals with “vaccine” • F rom vacca , Latin for “cow” The material from “cow pox” sores conferred protection against smallpox virus • Originally came from a milkmaid’s hands; Jenner suspected that the virus originated in hor
Slide 16: First Live Virus Vaccine: Edward Jenner's Inquiry (1798) – 1/2
© 2025 Tonix Pharmaceuticals Holding Corp. First Live Virus Vaccine: Edward Jenner’s Inquiry 1 (1798) – 1/2 “There is a disease to which the Horse from his state of domestication is frequently subject. The Farriers have termed it the Grease . It is an inflammation and swelling in the heel, from whic
Slide 17: First Live Virus Vaccine: Edward Jenner's Inquiry (1798) – 2/2
© 2025 Tonix Pharmaceuticals Holding Corp. First Live Virus Vaccine: Edward Jenner’s Inquiry 1 (1798) – 2/2 1 Jenner, E. “An Inquiry Into the Causes and Effects of the Variolae Vaccinae , a Disease Discovered in Some of the Western Counties of England, Particularly Gloucestershire, and Known by the
Slide 18: Loy's Account of Some Experiments (1801)
© 2025 Tonix Pharmaceuticals Holding Corp. Loy’s “ Account of some experiments 1 (1801) “This fact induces me to suspect, that two kinds of Grease exist, differing from each other in the power of giving disease to the human or brute animal: and there is another circumstance which renders this suppos
Slide 19: Equination: Use of Smallpox Vaccines Directly from Horse Lesions
© 2025 Tonix Pharmaceuticals Holding Corp. Equination 1 : Use of Smallpox Vaccines Directly from Horse Lesions (Without Passage Through Cows) Both Jenner and Loy used vaccine from horses; subsequently “ Equination ” was used in Europe in parallel with “vaccination” − Jenner believed that his “cowpox
Slide 20: 2006 Sequence and Analysis of the Horsepox Genome
© 2025 Tonix Pharmaceuticals Holding Corp. 2006 Sequence and Analysis of the Horsepox Genome 1 20 “ It is likely that a once naturally circulating but now rare VACV - like virus(s) from which current strains are derived was introduced as a vaccine virus, and the agent of horsepox has been surmised a
Slide 21: 2015 Genetic Analysis of Vaccinia Vaccines: Horsepox-like Virus Ancestor?
© 2025 Tonix Pharmaceuticals Holding Corp. 2015 Genetic Analysis of Vaccinia Vaccines: Horsepox - like Virus Ancestor? 1 21 “The biological origin of VACV is uncertain, although it has been suggested that a horsepox - like virus was an ancestor, even though a surviving horsepox virus (HPXV) genome h
Slide 22: David Evans: Speciation and Gene Loss in Vaccinia
© 2025 Tonix Pharmaceuticals Holding Corp. David Evans: Speciation and Gene Loss in Vaccinia Evans, “…the process of speciation appears to be associated with gene loss.” 1 − “ Relationship between DPP25 and horsepox virus. An important aspect of poxvirus evolutionary modeling concerns the hypothesis
Slide 23: Proposed Evolution of Vaccinia Vaccines
23 Relationship to Smallpox Incidence and Eradication Proposed Evolution of Vaccinia Vaccines TIME *Rough approximation (not data derived) 1977 Eradication SMALLPOX CASES IN US/EUROPE* ACAM2000 1798 1902 1931 2007 Dryvax ® swarm Mulford Vaccine Jenner’s Vaccine (Horsepox - like) Clonal Diversity 23
Slide 24: Synthesis of Horsepox (HPXV, TNX-801)
© 2025 Tonix Pharmaceuticals Holding Corp. Synthesis of Horsepox (HPXV, TNX - 801) 2018 1 24 1 Noyce RS, Lederman S, Evans DH. PLoS One . 2018 Jan 19;13(1):e0188453. doi : 10.1371/journal.pone.0188453. PMID: 29351298; PMCID: PMC5774680.
Slide 25: Genome Assembly (212 kbp): TNX-801 Core Genome Based on HPXV Strain MNR-76
© 2025 Tonix Pharmaceuticals Holding Corp. Genome Assembly (212 kbp ): TNX - 801 Core Genome i s Based on HPXV Strain MNR - 76 1,2 25 Sequence: GenBank entry DQ792504; DNA: GeneArt 1 Noyce, RS, Lederman S, Evans DH. PLoS ONE. 2018; 13(1): e0188453 https://doi.org/10.1371/journal.pone.0188453 2 Tulma
Slide 26: TNX-801 (Live HPXV for Percutaneous Administration)
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 (Live HPXV for Percutaneous Administration) 26 Vaccine based on sequence of isolated horsepox (HPXV) clone 1 − Synthesized 2 since 1976 isolate was not available outside of the U.S. Centers for Disease Control and Prevention (CDC) − No new gene el
Slide 27: Vaccination with TNX-801: Immunity with Low Reactogenicity
© 2025 Tonix Pharmaceuticals Holding Corp. Vaccination with TNX - 801 ( r HPXV ): Immunity with Low Reactogenicity ( i.e. , Better Tolerability) 27 100 80 60 40 20 0 0 3 6 9 12 15 Days post - infection Survival (%) TNX - 801 + LD of VACV Dryvax + LD of VACV LD of VACV 0 5 10 15 20 25 30 15 10 5 0 -
Slide 28: Horsepox Compared to Cowpox and Vaccinia Strains
© 2025 Tonix Pharmaceuticals Holding Corp. Horsepox Compared to Cowpox and Vaccinia Strains 1 Consistent with Near “Primordial” Strain Status 28 1 Evans, D. U. of Alberta (2018) with permission
Slide 29: Evolution of the Vaccinia Genome
© 2025 Tonix Pharmaceuticals Holding Corp. Evolution of the Vaccinia Genome 29 Recent studies (particularly from José Esparza & colleagues ) demonstrate that horsepox and horsepox - like viruses were used as smallpox vaccines in the 1800s 1 - 3 HPXV or HPXV - like Jenner’s vaccinia (cause of “ gease
Slide 30: Horsepox: Relationship to Jenner's Vaccinia
© 2025 Tonix Pharmaceuticals Holding Corp. Horsepox: Relationship to Jenner’s Vaccinia Horsepox environmental isolate sequenced in 2006 shares a common ancestor with vaccinia and could be considered a strain of vaccinia − Similar to cowpox with “intact” inverted terminal repeats (ITRs) – could be co
Slide 31: Deduced Relationship of Horsepox with Jenner's Vaccinia and circa 1960 Vaccinia
© 2025 Tonix Pharmaceuticals Holding Corp. Deduced Relationship of Horsepox with “Jenner’s Vaccinia” and “circa 1960 Vaccinia” Vaccines 1976 Mongolian Field Isolate 2 Arm - to - arm Molecular Biology “Vaccine Farms” Cow production started ~1875: Selection for Growth, Reactivity or Increased Virulenc
Slide 32: TNX-801 (Live HPXV for Percutaneous Administration)
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 (Live HPXV for Percutaneous Administration) 32 Vaccine based on sequence of isolated horsepox (HPXV) clone 1 − Synthesized 2 since 1976 isolate was not available outside of the U.S. Centers for Disease Control and Prevention (CDC) − No new gene el
Slide 33: TNX-801 Immunogenicity and Efficacy in NHPs - 2023
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 Immunogenicity and Efficacy in NHPs - 2023 33 Noyce RS, et al. Viruses . 2023 Jan 26;15(2):356. doi : 10.3390/v15020356. PMID: 36851570; PMCID: PMC9965234.
Slide 34: Survival: 100% of TNX-801 Vaccinated NHPs Survived Lethal MPXV Challenge
© 2025 Tonix Pharmaceuticals Holding Corp. Survival: 100% of TNX - 801 Vaccinated NHPs Survived Lethal MPXV Clade 1 Intratracheal Challenge 34 No deaths in TNX - 801 vaccinated groups Noyce et al . Viruses. 2023 Jan 26;15(2):356.
Slide 35: No Lesions Were Observed After TNX-801 Vaccination
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 Vaccination/MPXV Clade 1 Challenge: No Lesions Were Observed After TNX - 801 Vaccination 35 TNX - 801 (High Dose) TNX - 801 (Low Dose) rVACV Mock Noyce et al . Viruses. 2023 Jan 26;15(2):356.
Slide 36: TNX-801 Vaccination: Minimal MPXV Virus Shedding
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 Vaccination: Minimal MPXV Virus Shedding 36 TNX - 801 (High Dose) TNX - 801 (Low Dose) rVACV Mock Potential to Reduce Forward Transmission Noyce et al . Viruses. 2023 Jan 26;15(2):356.
Slide 37: Conclusions from NHP MPXV Challenge Study
© 2025 Tonix Pharmaceuticals Holding Corp. Conclusions from NHP MPXV Challenge Study 37 A single dose of TNX - 801 (horsepox) vaccination was well tolerated ‒ No severe adverse events ‒ Tolerability compares favorably to ACAM2000 – recently approved by US FDA for mpox 1 TNX - 801 vaccination via tra
Slide 38: TNX-801 in Primary Cell Lines and Immunocompromised Mice – 2024
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 in Primary Cell Lines and Immunocompromised Mice – 2024 ( mSphere ) 38 Trefry SV, et al. mSphere . 2024 Dec 19;9(12):e0026524. doi : 10.1128/msphere.00265 - 24. Epub 2024 Nov 13. PMID: 39535212; PMCID: PMC11656774.
Slide 39: TNX-801 has Reduced Virulence Relative to circa 1960 Vaccinia
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 has Reduced Virulence Relative to “circa 1960 Vaccinia” 39 Comparisons in vitro: 1) Plaque phenotype: VACV ( 3 - 4 mm) vs. TNX - 801 ( 1 - 2 mm) 2) Multi - step growth kinetics: ▪ Immortalized cell lines: TNX - 801 10 - to 1 00 - fold less v
Slide 40: Conclusion: TNX-801 is Less Virulent than 20th Century Vaccinia
© 2025 Tonix Pharmaceuticals Holding Corp. Conclusion: TNX - 801 is 10 - to - 1000 - fold Less Virulent than 20 th Century Vaccinia (VACV) 40
Slide 41: TNX-801 Lacks Lethality Associated With Older Smallpox Vaccine Strains (1 of 2)
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 Lacks Lethality Associated With Older Smallpox Vaccine Strains (Lister, NYC Board of health) in Double KO IFN - α R - / - and IFN - γ R - / - Mice 41 IND strain was deposited by the US Army in 1963 Farooq Nasar et al, Tonix unpublished data (1 of
Slide 42: TNX-801 Lacks Lethality Associated With Older Smallpox Vaccine Strains (2 of 2)
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 Lacks Lethality Associated With Older Smallpox Vaccine Strains (Lister, NYC Board of health) in Double KO IFN - α R - / - and IFN - γ R - / - Mice 42 IND strain was deposited by the US Army in 1963 Farooq Nasar et al, Tonix unpublished data *p <0.
Slide 43: High Dose TNX-801 is Unable to Cause Disseminated Infection
© 2025 Tonix Pharmaceuticals Holding Corp. High Dose TNX - 801 is Unable to Cause Disseminated Infection in Double KO IFN - α R - / - and IFN - γ R - / - Mice 43 IND strain was deposited by the US Army in 1963
Slide 44: Single Vaccination of TNX-801 Provides Durable Protection Against RPXV (1 of 2)
© 2025 Tonix Pharmaceuticals Holding Corp. Single Vaccination of TNX - 801 Provides Durable Protection Against Rabbitpox Virus (RPXV) Challenge 44 Farooq Nasar et al, Tonix unpublished data (1 of 2)
Slide 45: Single Vaccination of TNX-801 Provides Durable Protection Against RPXV (2 of 2)
© 2025 Tonix Pharmaceuticals Holding Corp. Single Vaccination of TNX - 801 Provides Durable Protection Against Rabbitpox Virus (RPXV) Challenge 45 Farooq Nasar et al, Tonix unpublished data (2 of 2)
Slide 46: Modern Cowpox and Vaccinia Are Endemic in the Environment Worldwide
© 2025 Tonix Pharmaceuticals Holding Corp. 16 Modern Cowpox and Vaccinia Are Endemic in the Environment Worldwide 46 Modern cowpox and vaccinia are environmentally endemic and are able to infect many animal hosts They are found widely distributed in important agricultural species and companion anima
Slide 47: Commercial Applications of Licensed Recombinant Poxvirus-Based Vaccines
© 2025 Tonix Pharmaceuticals Holding Corp. Commercial Applications of Licensed Recombinant Poxvirus - Based Vaccines 47 1 Taylor J, et al. Avian Dis. 1996;40(1):173 - 180. 2 The Poultry Site. March 14, 2005. Accessed July 9, 2021. https://www.thepoultrysite.com/news/2005/03/merial - launches - new -
Slide 48: Approved Recombinant Poxvirus-Based Commercial Products
© 2025 Tonix Pharmaceuticals Holding Corp. Approved Recombinant Poxvirus - Based Commercial Products 1 - 3 48 Doses released to environment Host restricted? Poxvirus vector Location Application / disease Product 2 billion (as of 2006) No Replication competent TROVAC - AIV H5N1 Mexico, Central Americ
Slide 49: Environmental Distribution: Vaccinia Released Aerially as Rabies Vaccine
© 2025 Tonix Pharmaceuticals Holding Corp. Environmental Distribution: Vaccinia Released Aerially as Rabies Vaccine 49 RABORAL V - RG ® is an oral vaccine based on vaccinia – rabies - glycoprotein recombinant virus used to prevent the spread of rabies among wildlife populations 1 - 3,* − RABORAL has
Slide 50: Illustrative Safety Spectrum Of Pox-based Vaccine Vectors
© 2025 Tonix Pharmaceuticals Holding Corp. Illustrative Safety Spectrum Of Pox - based Vaccine Vectors Optimizing Live Virus Vaccines 50 ~1 death per million ~10 deaths per million 20 th Century smallpox vaccines Dryvax Lister TianTan Copenhagen Tashkent MVA Canarypox Fowlpox Minimally Replicating L
Slide 51: Horsepox Protection and Tolerability in Animals Potentially Decouples Immunity from Reactogenicity
© 2025 Tonix Pharmaceuticals Holding Corp. Horsepox Protection and Tolerability in Animals Potentially Decouples Protective Immunity from Reactogenicity Conventional view holds that reactogenicity correlates with protection Protective immunity is not necessarily related to reactogenicity − Reactogen
Slide 52: Horsepox: More (Regulatory) Genes Confer Tolerability
© 2025 Tonix Pharmaceuticals Holding Corp. Horsepox: More (Regulatory) Genes Confer Tolerability For “20 th Century vaccinia vaccines”, the process of “Passage” through cows or birds was a primitive form of genetic engineering − “Passage” through cows resulted in gene deletions that may have increas
Slide 53: Live Virus Vaccine Platform: Recombinant Pox Vaccine (RPV) Platform
© 2025 Tonix Pharmaceuticals Holding Corp. Live Virus Vaccine Platform: Recombinant Pox Vaccine (RPV) Platform 53 Monkeypox and Smallpox Future Pandemics & New Infectious Diseases (“Hard Targets”) COVID - 19 Biodefense Using Proven Science To Address Challenging Disease States, We Have Created A Pro
Slide 54: TNX-1800 Immunogenicity in Hamsters and Rabbits - 2023
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 1800 (SARS - CoV - 2 spike – Expressing HPXV) Immunogenicity in Hamsters and Rabbits - 2023 54 Awasthi M, et al. Viruses . 2023 Oct 21;15(10):2131. doi : 10.3390/v15102131. PMID: 37896908; PMCID: PMC10612059.
Slide 55: Recombinant SARS-CoV-2 Vaccine Generation (TNX-1800 Expresses Spike)
© 2025 Tonix Pharmaceuticals Holding Corp. Recombinant SARS - CoV - 2 Vaccine Generation (TNX - 1800* Expresses Spike) 55 Development of HPXV as a recombinant Delivery Vector Platform Central General Region 36Kp - 146Kbp Left Terminal Region 60Kbp Right Terminal Region 147 - 212 kbp Essential Viral
Slide 56: TNX-1800 Immunogenicity and Efficacy in NHPs - 2023
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 1800 Immunogenicity and Efficacy in NHPs - 2023 56 Awasthi M, et al. Viruses . 2023 Oct 21;15(10):2131. doi : 10.3390/v15102131. PMID: 37896908; PMCID: PMC10612059.
Slide 57: Immunogenicity: All NHPs in TNX-1800 Group Had Neutralizing Antibody Response
© 2025 Tonix Pharmaceuticals Holding Corp. T N X - 1 8 0 0 D i l u e n t T N X - 1 8 0 0 D i l u e n t T N X - 1 8 0 0 D i l u e n t T N X - 1 8 0 0 D i l u e n t T N X - 1 8 0 0 D i l u e n t T N X - 1 8 0 0 D i l u e n t 0 100 200 300 400 M i c r o n e u t r a l i z a t i o n T i t e r -1 Days Pos
Slide 58: Vaccination with TNX-1800 Inhibits SARS-CoV-2 Replication in NHPs
© 2025 Tonix Pharmaceuticals Holding Corp. Vaccination with TNX - 1800 results in the inhibition of SARS - CoV - 2 Replication in Vaccinated NHPs 58 NHPs were vaccinated day 0 and challenged day 41; “Day 47” is 6 days after challenge
Slide 59: TNX-801 is Potential Vaccine for Mpox and Smallpox
© 2025 Tonix Pharmaceuticals Holding Corp. TNX - 801 is Potential Vaccine for Mpox and Smallpox Platform to express other viral antigens Animal studies show TNX - 801 protects against mpox − Appears to provide mucosal immunity after percutaneous vaccination (May prevent forward transmission) Single
Slide 60: Tonix Platform Selected by NIH/NIAID: Project NextGen COVID
© 2025 Tonix Pharmaceuticals Holding Corp. Tonix Platform Selected by NIH/NIAID : Project NextGen COVID 60
Slide 61: Tonix has Capacity and Technology to Develop and Produce TNX-801
© 2025 Tonix Pharmaceuticals Holding Corp. Tonix has Capacity and Technology to Develop and Produce TNX - 801 and Other HPXV Vaccines 61 Potential to manufacture at scale − Low dose because replication amplifies dose in vivo Believed will be thermo - stable in ultimate lyophilized formulation R&D Ce
Slide 62: Classical Technologies for Attenuating Pox Vaccines
© 2025 Tonix Pharmaceuticals Holding Corp. “Classical” Technologies for Attenuating Pox Vaccines Deletions by Serial Passage vs Targeted Genomic Modification 1 62 Targeted genomic manipulation ( 1990’s) Systematic deletion of known virulence and host range factors ( e.g., Enzo Paoletti 2 and Virogen
Slide 63: Interpretation of Evolution and Real-World Evidence to Recreate an Extinct Virus Vaccine
© 2025 Tonix Pharmaceuticals Holding Corp. Interpretation of Evolution and Real - World Evidence to Recreate an Extinct Virus Vaccine: Larger “Model” *This is a conceptual view to illustrate whether these genes are active or not and does not indicate the actual number, size, or location of the genes
Slide 64: Recreating an Extinct Vaccine Virus
© 2025 Tonix Pharmaceuticals Holding Corp. Recreating an Extinct Vaccine Virus “Evolutionary Intelligence”: Unknown evolutionary pressures resulted in Horsepox − Harnessing the results of large numbers of genetic events: both deletions and reactivations • In the parlance of “AI/Large Language Models
Slide 65: Contributors
© 2025 Tonix Pharmaceuticals Holding Corp. Contributors 65 Tonix Christy Raney Amy Cregger Chase A. Gonzales Brittany Layton Robert Enamorado Nelson Martinez Deborah Gohegan Massoudeh Masoud - Bahamiri Jennifer Cho Dawn Myscofski Tinoush Moulaei Nastasza Ziółkowska Current Addresses 1 University of
Slide 66: Selected Publications
© 2025 Tonix Pharmaceuticals Holding Corp. Selected Publications 1. Noyce RS, Lederman S, Evans DH. Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragment s. PLoS One. 2018 Jan 19;13(1):e0188453. doi : 10.1371/journal.pone.0188453. PMID: 29351298; PMCID: PMC577
Slide 67: Thank You
© 2025 Tonix Pharmaceuticals Holding Corp. Thank You
Slide 68: MPXV clade IIb is more attenuated than clade IIa
© 2025 Tonix Pharmaceuticals Holding Corp. MPXV clade IIb (MA - 2022) is 10,000 - to 100,000 - fold more attenuated than clade IIa (WR 7 - 61 and US - 2003) 68 *p <0.0001 *p <0.01 Farooq Nasar et al, Tonix unpublished data Double KO IFN - α R - / - and IFN - γ R - / - mice