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Neglected Diseases -‐ Drugs for Neglected Diseases Ini3a3ve (DNDi) and Medicines for Malaria Venture (MMV)
Luiz Carlos Dias
Ins:tute of Chemistry – UNICAMP Campinas – SP, BRAZIL
www.dndi.org
q 100 milhoes de pessoas sob risco na America La4na (AL)
q Endêmica em 21 países da América La4na 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 la4no 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 Cons4tui a maior causa de incapacidade provocada por doenças tropicais em adultos jovens e uma causa comum de insuficiência cardíaca em muitos países da América La4na
q Numero de pacientes crescendo em regioes nao-‐endemicas
q Atualmente DNDi es4ma que menos de 1% das pessoas infectadas recebem tratamento
Chagas Disease
Partnership between DNDi and: LAFEPE – Brazil Fundacion Mundo Sano And Ministerio Saude – Argentina ELEA produces ABARAX
Lead Optimization Latin America (LOLA)
The Lead Optimization Latin America (LOLA) consortium: collaborative drug discovery for
Neglected Tropical Diseases (NTDs)
DNDi
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
Origins of leads against T. cruzi Early leads for new drugs for Chagas disease
¨ Monocyclic series
¤ TDR screening campaign ¤ TDR op4misa4on project
¨ Bicyclic series
¤ NIH funded screen of the Broad Ins4tute compound collec4on
N
CN
S
HN
O
S
N
F
LOLA4IC50 = 0.03 µM (in vitro)
H3C N
CH3CN
S
TDR30139IC50 = 0.34 µM (in vitro)
O
S
Design and Analysis of new targets Collabora4ve effort by UNICAMP, AbbVie, Simon Campbell & DNDi
Synthesis UNICAMP, Campinas
Primary Parasitology USP São Carlos and LMPH, Antwerp
in vitro ADME Abbvie, Chicago
Secondary Parasitology Swiss Tropical Ins4tute Formula:on – in vivo PK
Wuxi AppTech, Shanghai
Mouse model of Chagas Disease LSHTM, London
Early screening cascade & partners
General Synthesis
NH2
SNC+Me Me
O OMe N
H
MeCN
S
Et3Nethanol
reflux, 30 min
Schmidt, U.; Kubitzek, H. Chem. Ber. 1960, 93, 1559-1565. TDR30139 analogues
thiopyridone
monocyclic cyanopyridines
Me N
MeCN
S R3
bicyclic cyanopyridines
NH2
SNC+
H Ar
O
N O
Et3N, ethanolreflux, 30 min
then piperidinereflux, 18 h
Boc thiopyridone
NH
N
S
CNAr
Boc
N
ArCN
S R3
NR
NIH lead analogues Abdel-Wadood, F. K.; Abdel-Monem, M. I.; Fahmy, A. M.; Geies, A. A. J. Chem. Res. 2008, 89-94.
MAD328IC50 > 100 µM
N
N
CH3
H3C SO
S
Synthesis of TDR30139 derivatives
TDR91228IC50 = 1.2 µM
H3C N
CH3CN
S
S
OH
TDR100524IC50 = 26 µM
H3C N
CH3CN
S
OS
TDR100612IC50 = 70 µM
H3C N
CH3CN
OO
S
TDR30139IC50 = 0.34 µM
H3C N
CH3CN
SO
S
LOLA67IC50 = 0.58 µM
SN
CNCH3
H3CO
F
N
HN
S
S
CNHN
O
F
LOLA3IC50 = 0.31 µM
N
N
S
S
CNHN
O
F
LOLA4IC50 = 0.03 µM
N
HN
S
CNHN
OLOLA48
IC50 = 7.9 µMF
HCl
TDR95696IC50 = 2.0 µM
N
CH3
HO
CN
SO
S
monocyclic
bicyclic
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:
• Experiment kindly carried out by collaborators at GSK, Tres Cantos, and Dundee Drug Discovery Unit
TDR30139T. cruzi IC50 = 0.34 µMCYP51 IC50 > 10 µM
N SO
SCN
CH3
CH3
N SO
CH3
CH3
CN
TDR91219T. cruzi IC50 = 0.7 µMCYP51 IC50 > 10µM
Kine:c Solubility Results
N
N
CN
SO
S
HN F
LOLA4
HN
N
CN
SO
S
HN F
LOLA3
N
N
CN
SO
S
HN F
Boc
LOLA2
N SO
CH3
CH3
CN F
LOLA67
K.S. (pH 2.0) < 1 µM K.S. (pH 7.4) < 1 µM
K.S. (pH 2.0) < 1 µM K.S. (pH 7.4) < 1 µM
K.S. (pH 2.0) > 200 µM K.S. (pH 7.4) = 2.65 µM
K.S. (pH 2.0) > 200 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
Shanghai, China
10 mg/mL
10% DMSO, 10% Cremophor EL,
40% PEG400, 40% Water; step by step
Poor plasma solubility
In vivo (mouse) PK studies
Acute mouse model of Chagas Disease
SN
CN
O
FH3C
H3C
IC50 = 0.58 µM
LOLA67(MAD431)
cLogP = 3.74 ± 0.53
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
Erythrocyte
Industry Academia
P. berghei liver stage assay GNF Novar4s/ UCSD, USA
In vitro blood stage ac4vity Swiss TPH, Switzerland
P. cynomolgi hypnozoite assay BPRC, Netherlands
Parasite Reduc4on Rate in vivo hu-‐SCID model GSK Tres Cantos, Spain
Gamete forma4on assays Imperial College UK
Resistance risk assessment Columbia University, USA
In vitro DMPK In silico modelling
In vitro DMPK In vivo DMPK Phys Chem measurements
Molecular Weight 380.40 In vitro human/ rodent mics (Clint µl/min/mg)
17.35 (human mics) 70.42 (mouse mics)
24.33 (rat heps)
LogD for pH 7.4 3.0 Rodent oral bioavailability 61% at 5mg/kg (close derivative)
Whole cell potency (IC50 nM) (NF54) 23 Rodent iv clearance estimated
(Cl ml/min/kg at dose) 59 at 2mg/kg
(close derivative) Cross resistance (IC50 nM) (K1, HB3, 7G8, TM80C2B, D6, V1/ S, Db2, FCB)
19-25 Rodent Vd, t1/2 (L/Kg, h)
Cytotoxicity THP1 (µM) >50 (close derivative)
In vivo efficacy Peters 4 day test (Pb/ Pf ED90 - mg/kg)
In vitro PRR (Log PRR) 3.1 (ATQ like) AUC at ED90 (nM.h)
Exo-erythrocytic stages (Y/N)* Pberghei liver: Y Pcynomolgi liver: N
In vivo PRR (comparable with which known antimalarial)
Solubility (µM) 14 Cyp inhibition (3A4, 2C9, 2D6, 1A2, 2C19) IC50 (µM)
Permeability: Human Caco2 AB pH6.5 (1E-6 cm/s) 34.3 hERG IC50 (µM) >33.3
(close derivative)
Protein binding (human %) >96.3 Additional data
Frontrunner profile MMV085400 Confidential
Project Highlights – MMV085400
31
– 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 IC50 (nM) MMV085497 KDU691
(Novartis) BQR695 (Novartis) MMV390048
P.vivax PI4K (isolated enzyme) 6.4 1.5 3.5
P.falciparum 24 (NF54) 118 (field isolate) 71 28
P.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
Brian Brown, Mira Hinman, Yvonne C. Martin, and Dale Kempf
Prof. Adriano Andricopulo, Marco Dessoy and Brian Slafer
James Mills
Manu De Rycker Marcel Kaiser
Prof. Louis Maes, An Matheeussen, Margot Desmet
Charlie Mowbray, Eric Chatelain Leandro Christmann and Simon Campbell
Wen Hua
Alan Brown
Acknowledgements Susann Krake, Pablo Martinez and Maitia Labora
Sue Charman
Mark Wenlock and Stefan Kavanagh
Sergio Wittlin
Paul Willis, Coline Legrand and Simon Campbell