Download Neglected Diseases

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 Chagas Disease 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 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
DNDi
consultant
Lead Optimization Latin America (LOLA)
Origins of leads against T. cruzi
Early leads for new drugs for Chagas disease
¨  Monocyclic series CH3
CN
H3C
N
S
S
O
TDR30139
IC50 = 0.34 µM (in vitro)
¤  TDR screening campaign ¤  TDR op4misa4on project ¨  Bicyclic series S
CN
N
N
H
N
S
F
O
IC50
LOLA4
= 0.03 µM (in vitr o)
¤  NIH funded screen of the Broad Ins4tute compound collec4on Early screening
cascade & partners
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 Secondary Parasitology Swiss Tropical Ins4tute in vitro ADME Abbvie, Chicago Formula:on – in vivo PK Wuxi AppTech, Shanghai Mouse model of Chagas Disease LSHTM, London General Synthesis
monocyclic cyanopyridines
Me
O
S
O
Me
Et3N
ethanol
reflux, 30 min
Me
+
NC
Me
CN
Me
NH2
N
H
S
CN
Me
N
R3
S
thiopyridone
TDR30139
analogues
Schmidt, U.; Kubitzek, H. Chem. Ber. 1960, 93, 1559-1565.
bicyclic cyanopyridines
Ar
O
H
+
Ar
Et 3N, ethanol
reflux, 30 min
S
NC
NH2
Boc
then piperidine
reflux, 18 h
Boc N
O
Ar
CN
N
N
H
S
R
CN
N
N
S
R3
thiopyridone
Abdel-Wadood, F. K.; Abdel-Monem, M. I.; Fahmy, A. M.; Geies, A. A. J. Chem. Res. 2008, 89-94.
NIH lead
analogues
Synthesis of TDR30139 derivatives
CH3
CN
S
CN
N
N
CH 3
S
LOLA4
IC 50 = 0.03 µM
H3C
H
N
F
O
N
CN
S
S
TDR91228
IC50 = 1.2 µM
CN
OH
H 3C
N
H 3C
O
N
S
TDR100612
IC50 = 70 µM
TDR100524
monocyclic
IC50 = 26 µM
CH3
CN
CH 3
CN
N
O
O
S
S
HN
S
CH 3
H
N
S
HO
CN
F
H3C
O
LOLA3
IC 50 = 0.31 µM
N
N
S
S
S
O
TDR95696
IC50 = 2.0 µM
S
O
TDR30139
IC50 = 0.34 µM
CH 3
N
HN
HCl
N
CH3
bicyclic
CN
CN
H
N
S
O
LOLA48
IC50 = 7.9 µM
H3C
F
N
F
S
O
LOLA67
IC50 = 0.58 µM
H3C
N
S
S
O
MAD328
IC50 > 100 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
CH3
N
CN
S
S
O
TDR30139
T. cruzi IC50 = 0.34 µM
CYP51 IC50 > 10 µM
CH3
N
S
O
TDR91219
T. cruzi IC50 = 0.7 µM
CYP51 IC50 > 10µM
•  Experiment kindly carried out by collaborators at GSK, Tres Cantos, and
Dundee Drug Discovery Unit
Kine:c Solubility Results CH3
CN
S
F
Boc
CH 3
N
S
CN
N
N
O
F
LOLA2
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
S
S
CN
N
S
O
LOLA67
HN
H
N
H
N
S
F
N
O
H
N
S
O
LOLA3
K.S. (pH 2.0) > 200 µM
K.S. (pH 7.4) = 2.65 µM
CN
N
LOLA4
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 F
Formulation studies on LOLA67
In vivo (mouse) PK studies H3 C
H3C
N
CN
S
F
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 BPRC, Netherlands In vitro DMPK In silico modelling P. berghei liver stage assay GNF Novar4s/ UCSD, USA In vitro DMPK In vivo DMPK Phys Chem measurements Gamete forma4on assays Imperial College UK In vitro blood stage ac4vity Swiss TPH, Switzerland Erythrocyte
Resistance risk assessment Columbia University, USA Parasite Reduc4on Rate in vivo hu-­‐SCID model GSK Tres Cantos, Spain Confidential
Frontrunner profile MMV085400
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)
Molecular Weight
Cross resistance (IC50 nM)
(K1, HB3, 7G8, TM80C2B, D6, V1/
S, Db2, FCB)
19-25
Cytotoxicity THP1 (µM)
>50
(close derivative)
In vitro PRR (Log PRR)
3.1 (ATQ like)
Exo-erythrocytic stages (Y/N)*
Solubility (µM)
Rodent Vd, t1/2 (L/Kg, h)
In vivo efficacy Peters 4 day test
(Pb/ Pf ED90 - mg/kg)
AUC at ED90 (nM.h)
Pberghei liver: Y In vivo PRR (comparable with which
Pcynomolgi liver: N known antimalarial)
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)
Protein binding (human %)
>96.3
Additional data
>33.3
(close derivative)
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
IC50 (nM)
MMV085497
KDU691
(Novartis)
BQR695
(Novartis)
P.vivax PI4K (isolated enzyme)
6.4
1.5
3.5
P.falciparum
24 (NF54)
118 (field isolate)
71
31P.falciparum PI4K resistant
29
MMV390048
28
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,
Yvonne C. Martin, and Dale Kempf
Marcel Kaiser
Alan Brown
Manu De Rycker
James Mills
Wen Hua
Charlie Mowbray, Eric Chatelain
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