Corporate PresentationNovember 2014

Non-Confidential

EnBiotix Summary

• Formed in 2012 by Boston University (BU) & Apeiron Partners; extensive patent portfolio licensed

• Focused on anti-bacterial applications of systems & synthetic biology platforms of Prof. Jim Collins’ lab at BU

• Product concept: ABX – potentiator combinations with significant potential to revitalize $30B antibiotic market

• First corporate deal finalized in 4Q2013 with multinational healthcare company; terms not disclosed

• EnBiotix team comprised of proven biotech and anti-bacterial product development executives

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EnBiotix Anti-infective Platforms & Products

• Systems biology platforms:– Novel computational methods & algorithms to separate signal

relevance from noise in large, complex data sets– Applicable to wide variety of datasets & disease states– Foundation for internal product programs

• Anti-persister platform:– Targeting eradication of bacterial persisters– EBX-001: tobramycin + potentiator for Rx of P. aeruginosa infections in

CF patients; ↑tobramycin killing 10,000 times against persisters– EBX-002: gentamycin + potentiator for treatment of chronic, catheter-

associated urinary tract infections

• Synthetic biology platform:– Engineered bacteriophage for delivery of wide variety of payloads;

EPP-001 for prosthetic joint infections lead program– Inducible protease system for tunable target degradation; allows

precise functional assessments of targets, pathways, MOAs

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Pre

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Preclinical

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Phase 1

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Phase 1

Phase 2

Phase 2b/3

Phase 2

Phase 3

Phase 3

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2014 2015 2016 2017 2018 2019 2020 2021 2022 2023 2024 2025

EnBiotix Product Pipeline 4

Persisters Platform (EBX)

Engineered Phage Platform (EPP)

EPPfor PJI

EBXfor caUTI

EBXfor CF

James J. Collins, Ph.D. EnBiotix Co-Founder and Chair, SAB

• Focused on synthetic biology and systems biology; particular focus on network biology approaches to antibiotic action and bacterial defense mechanisms

• Technologies licensed by > 25 biotech, pharma and medical devices companies

• Has helped to launched a number of companies, including Sample6 Technologies (formerly Novophage) and Joule Unlimited.

• Investigator, Howard Hughes Medical Institute

• William F. Warren Distinguished Professor, University Professor and Professor of Biomedical Engineering, Co-Director, Center for BioDynamics, Boston University

• Co-founder, Wyss Institute for Biologically Inspired Engineering, Harvard University; Visiting Professor in the Department of Systems Biology at Harvard Medical School

• Awards: 2012 Sanofi Pasteur Award; NIH Director's Pioneer Award, the Ellison Medical Foundation Senior Scholar Award in Aging, the inaugural Anthony J. Drexel Exceptional Achievement Award, the Lagrange Prize from the CRT Foundation in Italy; the Scientific American 50 - the top 50 outstanding leaders in science and technology; MacArthur Foundation "Genius Award", becoming the first bioengineer to receive this honor

• Member of the National Academy of Sciences, National Academy of Engineering, the Institute of Medicine and the American Academy of Arts & Sciences; Fellow of the American Physical Society, the Institute of Physics, the American Institute for Medical and Biological Engineering. 1990, Ph.D., Medical Engineering, University of Oxford

• 1987-1990, Rhodes Scholar

• 1987, B.S., Physics, summa cum laude; class valedictorian, College of the Holy Cross

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Founders, Board, Management

• Founders: – Apeiron Partners LLC (Boston, MA): Life sciences transaction advisory &

investment firm focused on corporate/academic spin-outs; $2 billion worth of companies formed/deals closed since inception

– Naeja Pharmaceutical, Inc. (Edmonton, AB, Canada): Med/syn-chem firm focused on ABXs; responsible for discovery of tazobactam: annual sales> $1 BB

– Great Lakes Drug Development, Inc. (Ann Arbor, MI): Devises integrated development strategies focused on key decision points and value creation

• Board: – Jeff Wager, MD (Chair): Apeiron founder, Bank of Tokyo, Harvard VC Fund, Z-Cube

(Zambon Group (Italy) corporate VC fund, CEO/Artisan Pharma (sold to Asahi 2011)– Denny Ausiello: 1996 – 2013; Physician-in-Chief, MGH; member, Board of Directors

& Chair, Science & Technology Committee, Pfizer; Member, Board of Directors: TARIS, Alnylam; member, Institute of Medicine (1999) & American Academy of Arts & Sciences (2003)

– CEOs of NAEJA (Chris Micetich) and GLDD (Dan Hartman, MD)

• Key Management: – Jeff Wager, MD: CEO– Jeff Radding, Ph.D.: 18 yrs Lilly as Anti-infective Drug Discovery Principal Research

Scientist and Group Leader; VP, Epitome Biosciences, Indiana University Ph.D., Yale post-doc

– Diane Joseph-McCarthy, Ph.D.: AstraZeneca as Director of Anti-infective Chemistry 4 yrs , Wyeth 10 yrs, MIT PhD., Harvard post-doc

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Anti-Persister Platform: EBX-001 for P. aeruginosa Infections

in Cystic Fibrosis Patients

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EBX-001 for P. aeruginosa Infections in Cystic Fibrosis Patients

• Up to 85% of CF patients develop chronic Pseudomonas aeruginosa infections by age 18-25

• Inhaled antibiotics approved to treat such patients include tobramycin as cornerstone of therapy

• Tobramycin 2013 sales ~$350M; nebulized tobramycin coming off-patent October 2014

• EBX-001 combines tobramycin with potentiatoras an inhaled treatment

• Our data/patents cover the product combination and the indication broadly

• In the clinic within 12-18 months from funding

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EBX-001 Program Collaborators, Advisors & Consultants

• Sam Moskowitz, MD, Massachusetts General Hospital– Leading basic and translational research lab in CF with a focus on Pseudomonas aeruginosa,

the major pathogen in CF– Associate Professor of Pediatrics and Director of CF Basic Science Program and Clinical

Program at MGH– Prominent investigator with the CF Foundation; participated in the TDN since 1999, Chair or

Vice Chair of its National Resource Center Committee from 2006-2012

• Chet Leach, Ph.D. – Ph.D. Pulmonary Toxicology– Lilly, 3M, Nektar, Lovelace Respiratory Research Institute– Developed most of non-clinical development for Novartis’ dry powder formulation of inhaled

Tobramycin

• “Dutch” VanDevanter, Ph.D.– Ph.D. Medicinal Chemistry– Ex-Pathogenesis, Chiron; co-project leader (with Bruce Montgomery) for aztreonam

development– Senior consultant to CF Foundation on variety of programs

• Bonnie Ramsey, M.D.– Program Director and Director of the Clinical Core at the Therapeutic Development Network

Coordinating Center (TDN-CC) at the Seattle Children’s Research Institute

• Lovelace Respiratory Research Institute (www.lrri.com) – Leading non-profit CRO specializing in inhaled ROA drug development

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Bacterial Persisters: Chronic and Recurrent Infections

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Metabolite-Enabled Eradication of Bacterial Persisters is Proton Motive Force Mediated

KR Allison et al., Nature, 2011

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Tobramycin-Metabolite Combination:Enhanced Efficacy Towards P. aeruginosaIn Vitro and In Vivo

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Carbon sources

Tobramycin 40mg/l

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In vivo bronchitis mouse modelIn vitro with persisters*

*Persisters selected by pretreatment with ofloxacin

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13EBX-001 for the CF Market

• ~70,000 patients worldwide; mostly in North America & Europe; 85% of patients overall receive TOBI

• Inhaled Tobramycin (TOBI/Novartis) 2012 sales of $311M; ~$350M sales in 2013

• Average annual cost of TOBI therapy = $20-25K

• EBX-001 pricing expected to be equal or superior to TOBI Podhaler if exacerbation and therapy intensity profile equal or superior (per 28 day course pricing of TOBI and Cayston both priced at ~$3500)

Anti-Persister Platform: EBX-002: gentamycin + potentiator for treatment of chronic, catheter-associated urinary tract infections

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Executive Summary: EBX-002

• Leverages gentamicin + mannitol mouse work cited in 2011 Nature paper• Product concept: development of proprietary gentamicin + potentiator i.v.

piggyback for in-hospital Rx of catheter-associated UTIs• Significant unmet need:

– 450,000 cases annually in U.S. alone– 4th most common hospital-acquired infection (HAI) in the U.S– 68% of UTIs in acute care hospital settings are estimated to be catheter-associated– 13,000 deaths (mortality rate 2.3%) attributed to UTIs annually in the U.S. – CAUTIs result in longer hospital stay; leading cause of 2° bloodstream infection– Using antibiotics to treat UTIs without symptoms contributes to antibiotic

resistance and can lead to the development of Clostridium difficile infection

• Patents: – We hold patents covering the combination of gentamicin (or any other

aminoglycoside) with a metabolite like mannitol

• Clinical/Regulatory: – FDA/EMA know gentamicin and mannitol well– Pursuing investigator-sponsored clinical trial of gent & mannitol – Since both products already approved by FDA, clinical path very straightforward

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In Vivo Potentiation Of Gentamicin Eradication of E. coli Biofilms

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In vivo studies in a mouse chronic, urinary-tract-infection model show treatment with a mannitol-gentamicin combination reduces the bacterial load on the catheters by about an order of magnitude, compared to gentamycin alone

Engineered Bacteriophage Platform

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Bacteriophage as Antibacterials

• Bacteriophage (phage) are viruses that only infect bacteria– Self-replicating inside the bacteria

– Capture bacterial machinery for DNA and protein synthesis

– Can ultimately kill bacteria by lysis of the cell

• Bacteriophage as antibacterial agents– Discovered in 1915 by Frederick Twort and further characterized

as bacterial viruses by Félix d’Hérelle in 1917

– First documented successful use of phage therapy in humans: children with dysentery by d’Hérelle in 1919

– Discovery of small molecule antibiotics (penicillin, 1928); become predominant anti-bacterial therapy

– Phage therapy development continues in other parts of the world

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Why Engineered Bacteriophage?

• Natural phage limitations

– Limited host range for infection

– Killing limited to infected bacteria only

– For sufficient pathogen coverage, phage cocktails required

– Use of phage cocktails present barriers to regulatory approval

• Engineered phage advantages

– Can generate anti-bacterial effect to uninfected bacteria:

• Mitigates need for phage cocktails with

• Reduced barriers to regulatory approval

– Adds functionality to phage therapy beyond lysis of the cell

– Can combine multiple modalities to tailor anti-bacterial mechanism-of-action & efficacy

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Multiple Modalities for Engineered Phage

• Engineered enzymatic phage– Dispersin B or other enzymes for degrading biofilms

• Engineered antimicrobial peptide (AMP) phage– Secretion/release of AMPs

• Engineered antibiotic potentiating phage– Lex3A repression of RecA

– Targeting other mechanisms of bacterial resistance

• Combinations of above to optimally address bacterial resistance mechanisms and targets

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Wide Range of Engineered Phage Product Candidates Possible

• Rx Indications:– Wound infections– Pneumonia/CF/VAP/bronchiectasis– Prevention/salvage of medical device/prosthetic infections– Preventing/treating catheter-related biofilms/infections– Eye & ear infections

• Rx/Consumer products:– Oral health: toothpastes, mouthwashes– Skin care: acne, soaps, hand-sanitizers, lotions

• Agricultural/Animal Health Indications: – Eradicating bacterial pests for bio-ag applications– Eradicating bacterial pathogens in aquaculture– Eradicating bacterial pathogens in livestock

• Industrial Applications:– Biofilms found in oil pipelines, industrial settings: microbially induced

corrosion– Biofilms found in HVAC systems, where they impede heat transfer– Current treatments are chemical: toxic and ineffective

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Engineered Bacteriophage Platform:EPP-001 for Prosthetic Joint Infections

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The PJI Problem: Pete the Ironworker

Patient: Pete is a 60 year-old ironworker, a robust man with years of experience buildingbridges across the nation. Three years ago, Pete had both knees replaced. He recently felt sharp pain and discomfort in one knee and was unable to walk with out assistance.

Diagnosis: Infection of the artificial knee joint through aspiration of pus from the kneeand subsequent microbiological characterization as a Staph infection.

Treatment: Surgery, debridement, and removal of the prosthetic knee joint, replacingit with a temporary articulating joint with an antibiotic spacer, antibiotic cement and antibioticcalcium pellets, along with concurrent IV antibiotics (picc line) for 6 -12 weeks, followed by a second surgery to remove the temp knee joint and replace it with a new permanent prosthetic joint.

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• Staphylococcal infections in prosthetic joints– 70% failure rate for salvage therapy: antibiotic Rx unable to clear infection,

requiring surgical replacement of prosthesis, similar to the patient profiled

– ~2.5 M cases of knee/hip replacments annually: 720K & 330K in US; 550K & 750K in EU; 70K & 40K in Japan; PJI incidence is ~ 2-2.5%

– Total cost to U.S. hospitals alone for Rx of PJIs projected at $1.62B by 2020

– S. aureus and CoNS predominant isolates from PJIs

Unmet Medical Need: Prosthetic Joint Infections

SM Kurtz et al., J. Arthroplasty 27:61 (2012); TKA (Total knee arthroplasty), THA (Total hip arthroplasty)

Projected number of PJIs in US Estimated cost PJIs in the US

Why Salvage Therapy Doesn’t Work 25

The Natural Shelter for Bacteria

Why Salvage Therapy Doesn’t Work (2) 26

• Bacteria protect themselves from antibiotics by:

- secreting biofilm in response to antibiotics and other stresses

- creating a physical barrier - the biofilm - thereby reducing antibiotic access to bacteria

- altering their metabolism within the biofilm to escape antibiotic action (persister state)

• Failure of antibiotics in PJI salvage therapy is NOT due to antibiotic resistance, but due to bacterial tolerance of antibiotics

• Antibiotic-tolerant bacteria lead to chronic device colonization and a nidus for infection, resulting in the high failure rate of salvage therapy

Disperse the biofilm!

EnBiotix’s Solution: Engineered Phage to Dipserse Biofilms

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Proof-of-Concept: Engineered Phage with a Biofilm Degrading Enzyme

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Bacterial Recovery from Biofilm (E. coli TG1)

Time course Inoculum Ratio

Engineered phage with biofilm degrading enzyme significantly decreases recovery of viable bacteria

• 10,000-fold over untreated • 100-fold over natural phage• with very low inoculum

Advantages of Engineered Phage Over Natural Phage for Biofilm Dispersal

• Natural phages have been reported to disperse biofilms, BUT…….

– Natural phage degrade peptidoglycan only…..

– This inhibits new biofilm formation, but does not affect existing mature biofilms in established infections

• Engineered phage adds capability to natural phage by……

– Adding functionality to specifically degrade mature biofilms

– Combining multiple payloads into a single phage(i.e. depolymerase + endonuclease)

– Secreting engineered payloads to affect bacteria not directly infected by phage

– Improving host range to mitigate the need for phage cocktails

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Advantages of Engineered Phage Over Enzymes Alone for Biofilm Dispersal

• Enzymes are currently manufactured or in development for biofilm removal….

– Pulmozyme for cystic fibrosis (P. aeruginosa)

– Dispersin B in combination with antimicrobials for wound care

• Engineered phage has advantages over enzyme therapies….

– Engineered phage replicate continuously secreting enzyme until there are no further bacterial to infect & kill

– This contrasts with exogenous enzyme administration, which is rapidly degraded

– Biofilms have multiple components that can be targeted by engineered phage (e.g, depolymerase + endonuclease)

– Engineered phage also kill bacteria, whereas enzymes do not

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Biofilm-Degrading Engineered Phage Products Applicable to Broad Range of Biofilms

del Pozo and Patel Clin Pharm Ther , 2007

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32Other EnBiotix Programs in Development Targeting Biofilms: Infections in CF Patients

• Pseudomonas infections in cystic fibrosis

– P. aeruginosa biofilms are highly resistant to antibiotics

– Antibiotics promote biofilm formation in CF infection (Hoffman et al, Nature 2005; 436:1171)

– Biofilm promotes antibiotic resistance in CF infection (Mah et al, Nature 2003; 426:306)

– Persistent chronic infections in CF result in recurrent exacerbations

– Total annual CF-related health care cost in US estimated at $1.47 B (2009) (O’Sullivan et al, Manag Care 2011; 20:37-44)

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Other EnBiotix Programs in Development Targeting Biofilms: Central Venous Catheter-Associated Infections

• Catheters are a nidus for infection with pathogens S. aureus, CoNS & Pseudomonas

• Catheters are readily colonized by bacterial biofilms

• Central venous catheter (CVC)-associated bloodstream infections (BSIs) in US

– Estimated 250,000 total and 80,000 ICU cases of CVC-associated BSIs annually

– Non-ICU cost estimated at $25,000, ICU cost $34,508--$56,000/episode

– Attributable mortality is 12-25%

– Attributable cost is estimated at $7.00B – $8.73B annually in the US

Engineered Bacteriophage Platform:Anti-Microbial Peptide & Genetic Regulator

Delivery Modalities

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Engineered Antimicrobial Peptide PhageCombining AMP and Bacteriophage

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• Strains infected with engineered AMP phage generate free AMP– Infected strain killed by phage lysis and AMP activity– Non-infected strains killed by free AMP

• Overcomes limited host range of phage• Targets localized non-infected bacterial strains• Two-pronged strategy for phage therapy (lysis and AMP activity)

36Engineered Antimicrobial Peptide PhageTreatment of E. coli with T7-AMP

T7.Indol-OmpA: Indolicidin-OmpA signal sequence fusionT7. CecP1-OmpA: Cecropin P1-OmpA signal sequence fusion

E. Coli Kill Curve for T7. AMP-OmpA Phage

• E. coli infected with AMP-OmpA fusions secrete active AMP

• Exploring secreted AMP inhibition of growth with other bacterial strains

Antibiotic Potentiating Phage Can Target Diverse Resistance Mechanisms

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Kohanski et al. Cell 130:797, 2007. Lu & Collins PNAS 106:4629 2009

Potentiating Phage Enhances Killing of Major Antibiotic Classes & Overcomes Resistance

Thank You

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