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