Drug screening against Yellow fever Virus Lucio Freitas Junior - PowerPoint PPT Presentation

“Drug screening against Yellow fever Virus” Lucio Freitas Junior Departamento de Micorbiologia Instituto de Ciências Biomédicas Universidade de São Paulo

The Phenotypic Screening Platform at ICB-USP Chemical and Genetic screening Cellular models of disease Chagas disease Leishmaniasis Dengue Yellow Fever Worms Chikungunya Zika Secondary assays for hit MoA and target deconvolution Phenotypic assays

Phenotypic Screening Platform

Screening methodology Reading Fixation and labeling of with DAPI and with Anti-Flavivirus Huh7 cels + ZIKV or YFV antibody + compound 72 h incubation at 37 o C and 5% C0 2 Data analysis

From ideas to Preclinical Candidates DRUG METABOLISM & MEDICINAL CHEMISTRY PHARMACOKINETICS BIOLOGY SCREENING LAB Ensaio Pré-clínico Fase I Hits Lead • Structure- • Eficácia in vivo Activity • PK/PD Relationship • Toxicologia (SAR) • In vitro ADME • Eficácia in vivo • PK/PD • Toxicologia (preliminar) • Target ID

Why is Repurposing import ?

Input Image High Content Screening for Yellow fever YFV

Nuclei Selection YFV

Cytoplasm Selection YFV

Select Population I YFV

Alexa Intensity YFV

Infected cells YFV

FDA for Yellow fever

Screening of a FDA-approved drug library for inhibitors of Y fever virus infection in human cells

Yellow fever assay SOFOSBUVIR 150 1.5 Normalized Activity % 100 1.0 Cell Ratio 50 0.5 0 0.0 -4 -2 0 -50 -0.5 log [Cpd], mg/mL Compound Max activity (%) EC50 (mg/mL) CC50 (mg/mL) S.I. Sofosbuvir 100.07 0.0013 ND 307.665

Yellow fever assay HARVONI 150 1.5 Normalized Activity % 100 1.0 Cell Ratio 50 0.5 0 0.0 -4 -2 0 -50 -0.5 log [Cpd], mg/mL Compound Max activity (%) EC50 (mg/mL) CC50 (mg/mL) S.I. Harvoni 100.07 0.0002 0.1089 659.04

Acknowledgements Phenotypic Screening Platform Carolina Borsoi Moraes Adalberto Araujo-Junior Caio H. Franco Department of Microbiology Luisa Naves Luís Carlos de Souza Ferreira Laura Alcantara Denise Pilger Marcia Mayer Gabriel Padilla Rafaela Bonotto Paolo Zanotto Giovana Cintra Edison Durigon Bruna Wey Joao Renato Rebello luciofreitasjunior@gmail.com Sergio Schenkman