Neglected Diseases - Drugs for Neglected Diseases Iniave (DNDi) and Medicines for Venture (MMV)

Luiz Carlos Dias Instute of Chemistry – UNICAMP Campinas – SP, BRAZIL www.dndi.org

Chagas Disease

q 100 milhoes de pessoas sob risco na America Lana (AL) q Endêmica em 21 países da América Lana e Central q Aproximadamente 8 milhões de indivíduos infectados na AL q Aproximadamente 55.000 novos casos a cada ano q Mata mais na região lano americana do que qualquer outra doença parasitária, incluindo a malária q Causa de 14.000 a 21.000 mortes por ano na região, sendo cerca de 5.000 por ano no Brasil q Custo mundial anual de 430.000 anos de vida perdidos ajustados por incapacidade (DALYs) q Constui a maior causa de incapacidade provocada por doenças tropicais em adultos jovens e uma causa comum de insuficiência cardíaca em muitos países da América Lana q Numero de pacientes crescendo em regioes nao-endemicas q Atualmente DNDi esma que menos de 1% das pessoas infectadas recebem tratamento Partnership between DNDi and:

LAFEPE – Brazil

Fundacion Mundo Sano And Ministerio Saude –

ELEA produces ABARAX

The Lead Optimization (LOLA) consortium: collaborative drug discovery for Neglected Tropical Diseases (NTDs)

Luiz Carlos Dias1, Marco A. Dessoy1, Brian W. Slafer1, Adriano Andricopulo2, Dale Kempf3, Brian Brown3, Mira Hinman3, Yvonne C. Martin3, Charles E. Mowbray4, Simon F. Campbell5

1Instituto de Química – UNICAMP, Campinas, Brazil 2Laboratorio de Química Medicinal e Computacional, Centro de Biotecnologia Molecular Estrutural– USP, São Paulo, Brazil 3AbbVie Inc., Chicago, USA 4Drugs for Neglected Diseases initiative (DNDi), Geneva, Switzerland 5Independent consultant

DNDi Lead Optimization Latin America (LOLA) Origins of leads against T. cruzi Early leads for new drugs for ¨ Monocyclic series

CH3 CN S

H3C N S O TDR30139

IC50 = 0.34 µM (in vitro)

¤ TDR screening campaign ¤ TDR opmisaon project ¨ Bicyclic series

S

CN N H N F N S O LOLA4

IC50 = 0.03 µM (in vitro) ¤ NIH funded screen of the Broad Instute compound collecon Early screening cascade & partners

Design and Analysis of new targets Collaborave effort by UNICAMP, AbbVie, Simon Campbell & DNDi

Synthesis UNICAMP, Campinas

Primary Parasitology in vitro ADME USP São Carlos and LMPH, Antwerp Abbvie, Chicago

Secondary Parasitology Swiss Tropical Instute Formulaon – in vivo PK Wuxi AppTech, Shanghai

Mouse model of Chagas Disease LSHTM, London General Synthesis monocyclic cyanopyridines Me Me Et3N CN ethanol CN O O S reflux, 30 min NC Me N S 3 Me Me + NH2 H Me N S R thiopyridone

Schmidt, U.; Kubitzek, H. Chem. Ber. 1960, 93, 1559-1565. TDR30139 analogues

bicyclic cyanopyridines

Ar Ar Et3N, ethanol Boc CN R CN O S reflux, 30 min N N + NC H Ar NH2 then piperidine N S N S R3 reflux, 18 h H

Boc N O thiopyridone NIH lead Abdel-Wadood, F. K.; Abdel-Monem, M. I.; Fahmy, A. M.; Geies, A. A. J. Chem. Res. 2008, 89-94. analogues Synthesis of TDR30139 derivatives

CH3 CH3 S CN CN S S CH3 CN N H3C N S CN H3C N O H O N F OH O N S S TDR91228 H3C N S O TDR100612 LOLA4 IC = 1.2 µM 50 IC = 70 µM IC = 0.03 µM TDR100524 50 50 monocyclic IC50 = 26 µM CH3 S CN S CN CH3 HN H CN HO N S N F S O N S O H3C N S TDR95696 LOLA3 O IC50 = 2.0 µM IC50 = 0.31 µM TDR30139 IC50 = 0.34 µM CH3 N

CH3 H C N S S CN bicyclic 3 HN CN F O H MAD328 HCl N N S H3C N S IC50 > 100 µM O O LOLA48 F LOLA67 IC50 = 7.9 µM IC50 = 0.58 µM MOA is not CYP51 inhibition

• TDR30139 & TDR91219 have promising in vitro activity against T. cruzi • Hit to lead chemistry in progress at University of Campinas • Check for CYP51 inhibition before investing too much effort:

CH3 CH3 CN CN S

CH3 N S CH3 N S O O

TDR30139 TDR91219 T. cruzi IC50 = 0.34 µM T. cruzi IC50 = 0.7 µM CYP51 IC50 > 10 µM CYP51 IC50 > 10µM • Experiment kindly carried out by collaborators at GSK, Tres Cantos, and Dundee Drug Discovery Unit

Kinec Solubility Results

CH 3 S CN F Boc CN N CH N S H 3 N F O N S LOLA67 O K.S. (pH 2.0) < 1 µM LOLA2 K.S. (pH 2.0) < 1 µM K.S. (pH 7.4) < 1 µM K.S. (pH 7.4) < 1 µM

S S

CN CN HN N H H N F N F N S N S O O LOLA3 LOLA4 K.S. (pH 2.0) > 200 µM K.S. (pH 2.0) > 200 µM K.S. (pH 7.4) = 2.65 µM K.S. (pH 7.4) < 1 µM

Theorecal concentraon: 200 µM K.S. Buffer: 50 µM phosphate buffer, pH 2.0 and 7.4 Formulation studies on LOLA67 H3C CN F In vivo (mouse) PK studies H3C N S O LOLA67 (MAD431)

IC50 = 0.58 µM Acute mouse model of Chagas Disease cLogP = 3.74 ± 0.53

Poor plasma solubility

10 mg/mL

10% DMSO, 10% Cremophor EL, 40% PEG400, 40% Water; step by step

Shanghai, China Summary

• Cyanopyridine series – Synthetic chemistry is the key to progress – Encouraging in vitro profiles of lead compounds – Leads scaled up for formulation and in vivo studies – Mouse pk carried out – Applying metabolite ID to guide design – Aim to test leads in a mouse model of Chagas disease soon

• Apply medicinal chemistry & drug discovery principles to other new chemical series from Pfizer and AbbVie

• Extend the LOLA consortium – DMPK, in vivo models, more chemistry, safety/toxicology,… – Maintain the excellent, close teamwork www.mmv.org

Combating malaria with the power of research

Malaria is caused by protozoan parasites of the genus Plasmodium – single-celled organisms that cannot survive outside of their host(s). Malaria is the leading parasitic cause of morbidity and mortality worldwide, especially in developing countries where it has serious economic and social costs.

u Endemic q Esma-se em 655.000 óbitos anuais causados pela malária no mundo, sendo 91% somente na África e 86% em crianças com menos de 5 anos de idade. q Nas Américas, esma-se em 1000 o número de óbitos devido à malária por ano. q Tem um custo mundial anual esmado em 34 milhões de DALYs, constuindo a quarta causa de DALYs nos países em desenvolvimento. q Em 2011, esma-se que houve 216 milhões de casos de malária ao redor do mundo, com aproximadamente 81% (ou 174 milhões de casos) na África e 91% dos casos causados pelo P. falciparum. q Nas Américas em 2011, esma-se em 1 milhão o número de casos e que 20% da população está sob algum risco de contrair a infecção. ACT = Artemisinin- based Combinaon Therapy:

Unicamp/MMV Anti-malarial drug discovery Project

BRAZIL HETEROCYCLES

Defeating Malaria Together Key Partners for screening

Academia Industry

P. cynomolgi hypnozoite assay In vitro DMPK BPRC, Netherlands In silico modelling P. berghei liver stage assay GNF Novars/ UCSD, USA

In vitro DMPK In vivo DMPK In vitro blood stage acvity Phys Chem Swiss TPH, Switzerland measurements

Gamete formaon assays Erythrocyte Parasite Reducon Rate Imperial College UK in vivo hu-SCID model GSK Tres Cantos,

Resistance risk assessment Columbia University, USA Confidential Frontrunner profile MMV085400

17.35 (human mics) In vitro human/ rodent mics Molecular Weight 380.40 70.42 (mouse mics) (Clint µl/min/mg) 24.33 (rat heps) 61% at 5mg/kg LogD for pH 7.4 3.0 Rodent oral bioavailability (close derivative) Whole cell potency (IC nM) Rodent iv clearance estimated 59 at 2mg/kg 50 23 (NF54) (Cl ml/min/kg at dose) (close derivative)

Cross resistance (IC50 nM) (K1, HB3, 7G8, TM80C2B, D6, V1/ 19-25 Rodent Vd, t1/2 (L/Kg, h) S, Db2, FCB) >50 In vivo efficacy Peters 4 day test Cytotoxicity THP1 (µM) (close derivative) (Pb/ Pf ED90 - mg/kg)

In vitro PRR (Log PRR) 3.1 (ATQ like) AUC at ED90 (nM.h)

Pberghei liver: Y In vivo PRR (comparable with which Exo-erythrocytic stages (Y/N)* Pcynomolgi liver: N known antimalarial) Cyp inhibition Solubility (µM) 14 (3A4, 2C9, 2D6, 1A2, 2C19) IC50 (µM) Permeability: Human Caco2 AB >33.3 34.3 hERG IC (µM) pH6.5 (1E-6 cm/s) 50 (close derivative)

Protein binding (human %) >96.3 Additional data Project Highlights – MMV085400

– PI4K inhibitor with apparent different resistance profile than other PI4K inhibitors in the MMV portfolio – No potency loss for: • PI4K resistant strain • 8 P.falciparum drug resistant field isolates – Transmission blocking and liver stage activity – PI4K inhibitor / PRR à slow killer (Atovaquone like) – Target Candidate Profile: • 2 (long duration) • 3b (transmission blocking) or 4 (chemoprotection)

PI4K-study KDU691 BQR695 MMV085497 MMV390048 IC50 (nM) (Novartis) (Novartis)

P.vivax PI4K (isolated enzyme) 6.4 1.5 3.5

P.falciparum 24 (NF54) 118 (field isolate) 71 28

31P.falciparum PI4K resistant 29 PI4K Inhibitors

Targeting Plasmodium PI(4)K to eliminate malaria: Nature 2013, 504, 248-253 (Novartis) Kinase Activity

• 80 Human Kinases assay at 10µM (AbbVie) • Hits followed up for IC50, no major issues for leads • Good kinase selectivity • Only one kinase had <100 fold selectivity (PDGFRA with 85 fold)

• Human Lipid Kinase assay at 10µM (University of Dundee) • Possible selectivity issue human vs. plasmodium PI3Kα, PI3Kβ, PI4Kβ • Good selectivity needed, ideal >100x

• Use of dockings planned: human vs. plasmodium to design more selective compounds Acknowledgements

Prof. Adriano Andricopulo, Marco Dessoy and Brian Slafer

Prof. Louis Maes, An Matheeussen, Margot Desmet

Brian Brown, Mira Hinman, Alan Brown Yvonne C. Martin, and Dale Kempf

Marcel Kaiser Manu De Rycker

James Mills

Charlie Mowbray, Eric Chatelain Wen Hua Leandro Christmann and Simon Campbell Acknowledgements

Susann Krake, Pablo Martinez and Maitia Labora

Sergio Wittlin Mark Wenlock and Stefan Kavanagh

Sue Charman

Paul Willis, Coline Legrand and Simon Campbell