Lassa Virus Therapeutics and Target Product Profile

Sina Bavari, Ph.D. US Army Medical Research Institute of Infectious Diseases (USAMRIID) Frederick, MD [email protected]

Objectives:

1) How to identify and evaluate potential therapeutics

2) Current bench research and clinical grade “therapeutics"

4) Ideal therapeutics for emerging pathogens under difficult situation

4) Target Product Profile for therapeutic use of drugs agisnt Lassa Fever Virus

Lassa Fever Virus Target Product Profile

This presentation will not cover IPEP, PEP, or other prophylaxis types of use. Therapeutic Targets for high consequence Viruses

Fusion/ Bunyaviruses (RVFV) Viral proteins: Arenaviruses (GP2) Coronaviruses (S protein) entry Paramyxoviruses (F protein) Arenaviruses (GP1, GP2) inhibitors Coronaviruses (S protein) Paramyxoviruses (G, F proteins) Bunyaviruses (Gn, Gc proteins) Abs Kinase Bunyaviruses (RVFV) Cellular receptors: Arenaviruses (LASV) Coronaviruses (DPP4) inhibitors Paramyxoviruses (ephrin-B2/-B3) Arenaviruses (multiple) Proteasome Bunyaviruses (RVFV) inhibitors Viral proteases: Coronaviruses (3CLpro, PLpro) Nucleoside RNA polymerase Cellular proteases: analogs (Bunya, Arena, Paramyxo) Coronaviruses (cathepsins B, L, TMPRSS2) Protease Arenaviruses (SKI-1) inhibitors

Paramyxoviruses (M protein) Host modulators/ Bunyaviruses (SKI-1) Assembly inhibitors immunomodulators Bunyaviruses Arenaviruses (Z protein) Coronaviruses Paramyxoviruses (M protein) Bunyaviruses (RVFV) Egress inhibitors

From Campbell Biology: Concepts & Connections (5th ed) High Content Imaging Assay for Viral Infection

1. Cell Plating PE Janus MDT 20 hrs

2. Compound Treatment 2 hrs

3. Virus Dispensing 48 hrs BSL2,3,4 +24hrs DRAQ5 anti-Virus IgG-Alexa488 4. Immuno-fluorescence Staining

PE Opera 5. Confocal High Throughput Imaging

Nuclei Cytoplasm Pathogen Pos. Cells 6. Image Analysis

7. Multi-parametric Data Analysis

GeneData Screener and PE SpotFire

Veronica Soloveva, , 8 April 5 2018 Therapeutic evaluation of compounds against viral infection Viral infection of HeLa Cells

DMSO [Inhibitor ] No infection (+ control) (- control)

EC50, uM

HCI HCI 0.215 1 0 0

P laque Plaque assay 0.145

7 5 RT-PCR RT-PCR 0.172

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% % % Inhibition 0 N=2 -9 -8 -7 -6 -5 -2 5 Loglo g M Concentration [M] Veronica Soloveva, , 8 April 2018 Potential Lassa Virus Therapeutics

Antivirals Host-response modifiers Potential Lassa Virus Therapeutics

Antivirals

Synthetic Biologics IFNab Small Molecule Nucs mAbs NA-based (T-705) Blood product Small Molecule non-Nucs (LHF-535) SiRNA PMO Ideal therapeutics for emerging infections

1) Easy to access, easy to store, and widely available 2) Inexpensive 3) Safety/safety/safety 4) Easy to administer 5) Long-shelf life 6) Clear uncomplicated regulatory path for approval 7) Broad activity against multiple families of viruses 8) Treat/intervene at any stages of infection 9) Can be added to other treatment options 10)Well distributed to multiple tissues based on the infection pattern

Therapeutics: Lassa Virus Clinical disease: • Ranges from mild flu-like illness to severe hemorrhagic fever • Deafness is a sequelae in some survivors Therapeutic strategies: • Difficult to target entry – different arenaviruses use different cellular receptors; • Target cell processes critical for entry and replication (e.g. kinase inhibitors) Compound Mechanism Stage Notes Some effect if administered early; Tx Ribavirin Nucleoside analog Approved in humans efficacy needs to be formally investigated. In vivo efficacy –small Synergistic effect in combination with Favipiravir Nucleobase animals Ribavirin? ST-193 Entry inhibitor In vivo efficacy – guinea pigs Inhibits pH-induced membrane fusion Genistein , many Entry inhibitor/host In vitro data EC50 nM-uM Isoflavone; kinase inhibitor/Nuc others modulator/replication Kineta/Wellcome Trust LHF-535 Entry inhibitor Finishing Pre-clinical Finishing pre-IND studies In vivo efficacy – guinea May not be consistent with TPP (cost, huMAbs Antiviral pigs/NHPs manufacturing, etc) IFN consensus; combination with IFN-alfacon-1 Immune modulator Approved in humans Ribavirin or other antivirals LASV Therapeutic (Reactive/Emergency Use) TPP (Preliminary Draft Apr 2018) Category Threshold Optimal Adults (18-62 years), excluding pregnant Children, Adults, Geriatrics (2-62+ years), Patient Population & lactating women suitable for pregnant & lactating women 42d - 3mo: Regulated NHP safety or Human safety evaluated: NHV/patients Safety other appropriate models (<50 subjects) Well tolerated, transient, manageable No critical or severe adverse events; AEs Tolerability tolerable AEs acceptable (eg: rash, GI) - No drug-drug interactions - Acceptable for use in pregnant women Effective vs various LASV (Broad Effective vs ≥ 2 arenaviruses (current Spectrum/outbreak strain), 2-log outbreak strain and or other AVs) , 3-log Efficacy reduction viremia in serum , Decrease reduction viremia in serum, Decrease clinical signs by 50%, Survival benefit clinical signs by 80%, Survival benefit >50% >80% Route of Administration Parenteral (IV/IM/subcutaneous) Oral and parenteral Frequency of Dosing Continuous infusion or TID QD or BID Duration of Treatment 21 days 10-14 days Onset of Action Efficacy <24hr after confirmed infection Efficacy >24hr after confirmed infection Storage Condition 2-8oC 20oC – 35oC Shelf Life 2 years (cold chain acceptable) 5 years (room temperature) Manufacturing/Stock- Efficient, high yield synthesis (1,000-Ms Synthesis (100-1,000) treatments) pilling treatments) Affordable in limited healthcare areas Cost of Goods Global deployment opportunity $1-5 $10-20 Patient Population Safety Tolerability

Efficacy

Route of Administration Frequency of Dosing Duration of Treatment Onset of Action Storage Condition Shelf Life Manufacturing/Stock-pilling Cost of Goods Lassa Virus Therapeutics: Target Product Profile

Category Threshold Optimal Children, Adults, Geriatrics Adults (18-62 years), Patient (2-62+ years), suitable for excluding pregnant & Population pregnant & lactating lactating women women 42d - 3mo: Regulated NHP Human safety evaluated: Safety safety or other NHV/patients (<50 appropriate models subjects) Well tolerated, transient, No critical or severe manageable AEs adverse events; tolerable Tolerability - No drug-drug interactions AEs acceptable (eg: rash, - Acceptable for use in GI) pregnant women Lassa Virus Therapeutics: Target Product Profile

Category Threshold Optimal Effective vs ≥ 2 Effective vs various LASV arenaviruses (current Broad Spectrum/outbreak outbreak strain and or strain), 2-log reduction other AVs) , 3-log Efficacy viremia in serum , reduction viremia in Decrease clinical signs by serum, Decrease clinical 50%, Survival benefit signs by 80%, Survival >50% benefit >80% Route of Parenteral Oral and parenteral Administration (IV/IM/subcutaneous) Frequency of Continuous infusion or QD or BID Dosing TID Duration of 21 days 10-14 days Treatment Lassa Virus Therapeutics: Target Product Profile

Category Threshold Optimal Efficacy <24hr after Efficacy >24hr after Onset of Action confirmed infection confirmed infection Storage 2-8oC 20oC – 35oC Condition 2 years (cold chain 2-8oC 5 years (room Shelf Life acceptable) temperature) Efficient, high yield Manufacturing/ Synthesis (100s-1,000) synthesis (more than Stock-pilling treatments) 1,000-treatments) Affordable in limited Global deployment Cost of Goods healthcare areas $10-20 opportunity $1-5