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 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