Neglected Diseases - Drugs for Neglected Diseases Ini a ve (DNDi) and Medicines for Malaria Venture (MMV)
Luiz Carlos Dias Ins tute of Chemistry – UNICAMP Campinas – SP, BRAZIL www.dndi.org
Chagas Disease
q 100 milhoes de pessoas sob risco na America La na (AL) q Endêmica em 21 países da América La na 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 la no 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 Cons tui a maior causa de incapacidade provocada por doenças tropicais em adultos jovens e uma causa comum de insuficiência cardíaca em muitos países da América La na q Numero de pacientes crescendo em regioes nao-endemicas q Atualmente DNDi es ma que menos de 1% das pessoas infectadas recebem tratamento Partnership between DNDi and:
LAFEPE – Brazil
Fundacion Mundo Sano And Ministerio Saude – Argentina
ELEA produces ABARAX
The Lead Optimization Latin America (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 Chagas disease ¨ Monocyclic series
CH3 CN S
H3C N S O TDR30139
IC50 = 0.34 µM (in vitro)
¤ TDR screening campaign ¤ TDR op misa on 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 Ins tute compound collec on Early screening cascade & partners
Design and Analysis of new targets Collabora ve 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 Ins tute Formula on – 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
Kine c 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
Theore cal concentra on: 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 Es ma-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, es ma-se em 1000 o número de óbitos devido à malária por ano. q Tem um custo mundial anual es mado em 34 milhões de DALYs, cons tuindo a quarta causa de DALYs nos países em desenvolvimento. q Em 2011, es ma-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, es ma-se em 1 milhão o número de casos e que 20% da população está sob algum risco de contrair a infecção. ACT = Artemisinin- based Combina on 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 Novar s/ UCSD, USA
In vitro DMPK In vivo DMPK In vitro blood stage ac vity Phys Chem Swiss TPH, Switzerland measurements
Gamete forma on assays Erythrocyte Parasite Reduc on Rate Imperial College UK in vivo hu-SCID model GSK Tres Cantos, Spain
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