DDD107498: A Novel

Clinical Candidate for

Malaria

Dr. Kevin ReadNHS Research Scotland, 2016

Defeating Malaria Together

The Drug Discovery Unit • A “Biotech company” within a university

– State of art equipment and facilities

– Current funding streams - ~£33 million

– 85 scientists, attracted from across the biopharma industry and

academia

• Translational Research Engine

– Combines excellence in discovery science with drug discovery

industry expertise

– Delivers preclinical candidates and advances novel targets via

collaboration and partnership

• Complements the pharmaceutical industry

– Neglected Tropical Diseases (NTDs)

– Innovative Targets Portfolio (ITP)

UNMET NEED

Disease New Cases(per annum)

Health burden(million DALYsa)

HIV / AIDS

1.5 millionTuberculosis

0.6 millionMalaria

1.2 million

Sleeping sickness

Chagas' disease

Leishmaniasis (VL)

Schistosomiasis

2 million

9.6 million

214 million

8,000

7 million (total)

400,000

42 million (Rx)

92

44

55

1.2

0.5

3.3

4.0

18,000

48,000

43,000

22,000

Deaths(per annum)

Cryptosporidiosis 64 million 27,0002.1

Current Areas of Focus - NTDs

a Disability-adjusted life years lost to the community, c.f. War = 20 million DALYs

Target

screen

Target hits Cell hits LeadsPreclinical

candidate

Cell

screen

Fully characterised &

genetically validated target

Existing drugs

Animal

screen

Fragment or lead-like

compounds

4

Hit

Discovery

Hits to

Leads

Lead

Optimization

Target

Selection

Assay

Development

Hit

Validation

Pre-clinical

Development

Three Approaches to Drug Discovery:Target-based, Phenotypic and Drug Repurposing

Drug-like compounds

Clinical

Development

Product

registration

10+ years and as much as £800 Million

• Phase II clinical candidate for visceral leishmaniasis

• Preclinical development for Malaria 2015

• Preclinical candidates

• Visceral leishmaniasis: Two selected in 2016; differing MoAs

• Animal trypanosomiasis: candidate in field trials

• In vivo proof of concept

• Chagas’ disease: 3 series with animal efficacy, 1 lead optimisation

• Tuberculosis: 2 series in late lead optimisation in collaboration

with TBDA

Track Record of Delivery – NTDs

Malaria

About Malaria

Malaria Report 2014: www.who.int

Half of the world’s population, 3.2

billion people, is at risk of malaria

198 millions cases in 2013

584 000 deaths, mostly among

children under five (437 000)

90% of all malaria deaths occur in

Africa

$12 billion lost GDP

Consumes 40% of public health

spending

Malaria is caused by parasites of the

species Plasmodium that are

transmitted by the bites of the

female Anopheles mosquitoes

There are four types of human Plasmodium:

Plasmodium falciparumPlasmodium vivaxPlasmodium malariaePlasmodium ovale

Plasmodium falciparum and Plasmodium vivax are the most commonPlasmodium falciparum is the most deadly

Quinoline and amino-alcohols

Artemesinin analogues

Halofantrine

Quinolines, etc.

Chloroquine

Amodiaquine

Quinine

Mefloquine

Halofantrine

Resistance (20¢)

Safety / resistance

Compliance / safety / resistance

Resistance / safety / cost

Safety / resistance / cost

Artemesinins

Artemether

Arteether

Artesunate

Emerging resistance /

Compliance / safety / GMP

standard / availability / cost of raw

material

ACTs

Lumefantrine /

artemether (Coartem)

AS/AQ (Coarsucam)

Resistance potential /

compliance / ($2.40) / availability

Antifolates

Sulphadoxine /

pyrimethamine

Resistance (25¢)

Naphthoquinone

Atovaquone /

proguanil

Resistance potential / cost ($73)

Chloroquine

NCl

HNN

Quinine

N

H3CO

HON

H

HH2C

CH3

O

O

CH3

RO

H3C OO

Dihydroartemsinin R=H Artemether R=Methyl Arteether R=Ethyl Artesunate R=Succinyl

S

H2NOCH3

OCH3

NN

N

OO

H

Sulphadoxine

N

N

NH2Cl

NH2H3C

Pyrimethamine

NCF3

N

CF3

HO

H

Mefloquine

OH

O

O

Cl

N N N CH3

NH NH CH3Cl

H H H

ProguanilAtovaquone

N

HNN

OH

Cl

Amodiaquine

N CH3

CH3

Cl Cl

F3COH

N

H3C

H3C

Cl Cl

Cl

OH

Lumefantrine

Why we need new drugs for malaria

Other antimalarials

Drug resistance is a major challenge…

• Signs of resistance to artemisinin have emerged in SE Asia

• Why? People often do not take complete doses, or take single agent

therapies e.g. chloroquine, or substandard drugs

Dondorp et al., N. Engl. J. Med., 2009; 361(5): 455–467

Menard et al., Nature, 2014; 505, 50–55

CHILDREN& PREGNANT

WOMEN

SINGLE DOSECURES

PREVENTIONOF RELAPSE

TRANSMISSIONBLOCKING

CHEMOPREVENTION

1x

Malaria R&D challenges

Malaria R&D challenges

• Better medicines for uncomplicated malaria • Tackling resistance to first-line anti-malarials

• Aiming for a single-dose cure

• Medicines for vulnerable populations• Formulations for children

• Protecting pregnant women

• Medicines for malaria elimination/eradication• Stopping relapse (P. vivax)

• Blocking transmission and chemoprevention

DDU Malaria Project Starting Point:

Hit Discovery

• In late 2009 the DDU’s focused kinase compound library was screened

phenotypically against P. falciparum

• 3 chemical series taken forward with

MMV funding starting April 2010

• Candidate selected October 2013

• Partnered with Merck Serono for Clinical

Development in late 2014

POTENCY SELECTIVITY

KINETICS SAFETY

Hit Discovery Workflow

MMV Progression Criteria used to drive project

• SYBRgreen 384-well assay validated versus [3H]-hypoxanthine method

Assay development

• Screen at 3 μM against P. falciparum• >70% inhibition in single point assay (Z’ = 0.84)

4731 compounds

• Purity and identity of hits confirmed

• Potency confirmed in 10 point assay 120 hits

• Chemical clustering

• Human cell counter screen11 series

• Overlap with MMV portfolio4 series

• Hit and analogues confirmed

• Mammalian kinase panel screen3 series

Hits to lead

Kinase Library - PI Frequency Distribution

0

50

100

150

200

250

300

350

400

-50

-43

-36

-29

-22

-15 -8 -1 6

13

20

27

34

41

48

55

62

69

76

83

90

97

10

4

>1

10

PI

Nu

mb

er

of

co

mp

ou

nd

s

Mefloquine

Concentration (uM)

0.0001 0.01 1

Pe

rce

nt

Inh

ibiti

on

0

20

40

60

80

100

Hit Series

EARLY LEADS

Series ID MMV02 MMV03 MMV04

EC50 vs. P f (M) 0.2 0.3 0.1 <0.1

EC50 vs. MRC5 (M) 39 > 50 20 > 10 fold

logD pH 7.4 4.0 3.2 4.3 <5

MW 342 346 418 <500

No. of examples <1 mol 3 4 5

• MMV03 series dropped due to flat SAR

• MMV02 series stopped for developability issues

MMV04 Series: Hits to Lead - focus on

properties

MMV04 Series

Pf EC50 (nM) 120 350 700 50

MW 418 357 377 430

LogP 4.3 3.7 3.7 2.1

Solubility (μM)

39 36 180 >230

MLM Cli (ml/min/g)

5.3 8.6 3.4 0.8

PPB (%) >99 97 96 59

Meets MMV progression criteria

Lead Optimization – Getting the right balance

Potency: Pf EC50 1 nMBioavailability: high (74%)Efficacy (rodent malaria):ED90: 0.6mg/kg, Oral route

Potency: Pf EC50 50 nMBioavailability: lowEfficacy (rodent malaria):15mg/kg bid for 4 daysIntraperitoneal injection

POTENCY SELECTIVITY

PK SAFETY

DDD107498: Active against drug-resistant

parasites

0.1

1

10

100

1000

NF54(sensitive)

D6 (MFQ)K1

(CQ,SUL,PYR, CYC)

W2 (CQ,SUL, PYR,

CYC))

7G8 (CQ,PYR, CYC))

TM90C2A(CQ, PYR,

MFQ, CYC)

V1/S (CQ,SUL, PYR,

CYC)

EC5

0(n

M)

DDD107498

Artesunate

Chloroquine

No cross-resistance to current antimalarial drugs

Cellular Selectivity: > 20,000 fold (MRC-5) and > 59,000 (HEPG-2)

CQ: Chloroquine

MFQ: Mefloquine

SUL:Sulphadoxine

PYR: Pyrimethamine

CYC:cycloguanil

DDD107498: Active against recent clinical

isolates

Active against Both P. falciparum and P. vivax clinical isolates

More potent than all current drugs

Drug n Median EC 50 [range] nM n Median EC 50 [range] nM

Chloroquine 28 106.0 [28.5-218.8] 22 165.9 [40.2-383.4]

Amodiaquine 27 16.9 [7.8-28.3] 21 26.4 [5.3-49.5]

Piperaquine 28 26.4 [7.4-58.1] 22 24.3 [3.4-81.4]

Mefloquine 28 10.6 [2.8-31.8] 22 21.3 [2.4-81.4]

Artesunate 27 6.6 [1.7-25.9] 22 3.1[0.3-19.7]

DDD107498 28 0.8 [0.5-3.3] 21 0.5 [0.3-1.4]

P. falciparum P. vivax

Fresh clinical isolates from southern Papua, Indonesia

Studies carried out by Jutta Marfurt and Ric Price, Menzies School of Health Research

DDD107498: Highly potent against multiple life

cycle stages

DDD107498: Good in vitro Developability

19

DDD107498

clogP 3.3

logD (experimental) 2.5

MWt 468

in vitro Cli (mouse, rat, dog, cyno.,human) (ml/min/mg microsomal protein)

≤0.017, ≤0.013, 0.036, 0.038,

0.021

PPB % (mouse, rat, dog, human) 63, 68, 67, 77

Solubility (SGF, FeSSIF, FaSSIF) (M) >6410, >6410, 3970

Permeability PAMPA (nm/s) 76

pKa (experimental) 8.71 and 6.82

SGF Stability >4h

Plasma and blood stability ( mouse/human) >8h

CYP450 Inhibition (1A2, 2C9, 2C19, 2D6,

3A4) (M)

≥ 77

Bioactivation risk (GSH trapping, MDI) No adducts, No MDI

Good oral bioavailability and long half life. Vd high

0.0

20.0

40.0

60.0

80.0

100.0

120.0

0 500 1000 1500 2000 2500

Wh

ole

blo

od

co

nc

en

trat

ion

(ng

/mL

)

Time-point (min)

The mean concentration-time profile of DDD00107498 following a single Oral administration at 3 mg free base/kg to the female NMRI mouse (n=3)

DDD107498: Pharmacokinetics in Mouse

Mouse IV 1 mg/kg (n=3) Mouse PO 3 mg/kg (n=3)

Clb 12 ml/min/kg Cmax 90 ng/ml

Vdss 15.0 L/kg Tmax 1 h

T½ 16.0 h T½ 19.3 h

F% 74

P.Berghei mouse model ED90 = 0.6mg/kg PO

0 1 2 3 4 5 6 7

0.01

0.1

1

10

DDD00107498(mg/Kg)

Limit of detection

Treatment

ED90

0

0.6

0.1

0.05

0.3

1

3

Day after infection

% p

aras

item

ia

Blood Stage Efficacy in P. falciparum

SCID mouse model

-2

-1

0

1

0.01 0.1 1 10 100Vehicle

Dose (mg/Kg)

log

10 (

% p

aras

item

ia)

ED90= 0.62 mg/Kg

ED90 0.6 mg/kg

MPC = 10-12 ng/mL

Targets Multiple Life-Cycle Stages:

Transmission

Transmission

blocking stages

EC50 ~ 1-10nM

P. berghei

ookinetes.

EC50 ~ 5nM

P. falciparum &

P. vivax blood

stage form

EC50 ~ 1nM

P. yoelli & P.

berghei liver

schizonts

EC50 ~ 1nM

Transmission blocking potential: In VitroKoen Dechering, TropIQ

• Standard Membrane Feeding Assay (SMFA)

DDD107498 demonstrates very potent in vitro

transmission blocking potential

Gamete Formation:

• Male EC50 1.8 nM

• Female EC50 1.2 nM

Transmission Blocking Potential: In VivoBob Sinden/Michael Delves, Imperial College, London

• Mice infected with P. berghei and then used to feed mosquitos.

DDD107498 dosed at 3 mg/kg orally

• Significant reduction in live oocysts found in mosquitos that had

fed on treated mice

• Intensity (oocysts per midgut): reduction by 98%

• Prevalence (infected mosquitos): reduction by 90%

• Subsequent transmission to naïve mice inhibited by 90%.

• Could be due to killing gametocytes, preventing exflagellation

or oocyst development

DDD107498 demonstrates very strong in vivo

transmission blocking potential

Targets Multiple Life-Cycle Stages:

Chemoprotection

Transmission

blocking stages.

EC50 ~ 1-10nM

P. berghei

ookinetes.

EC50 ~ 5nM

P. falciparum &

P. vivax blood

stage form.

EC50 ~ 1nM

P. yoelli & P.

berghei liver

schizonts.

EC50 ~ 1nM

DDD107498: Potent Chemoprotection In VitroElizabeth Winzeler, Stephen Meister, UCSD

DDD107498

EC50 (nM)

Atovaquone

EC50 (nM)

0.66 106

0.52 1.5

0.55 1.3

0.44 0.59

0.45 1.1

0.11 0.30

• DDD107498 incubated with P. berghei infected liver cells

• Potent across liver stage life cycle

• Potent when given for brief period during invasion (protocol 1)

• Potent when given 24 hours after infection (protocol 5)

Suggests DDD107498 may not require constant drug

pressure to have chemoprotective effect

Drug

Treatment

Protocol

1

2

3

4

5

6

Invasion Liver Schizont development

Liver cells infected with sporozoites

DDD107498: Potent Chemoprotection in VivoDennis Kyle, USF

• Mice dosed with

DDD107498 two hours

prior to infection with

sporozoites

• No sign of infection in

mice treated at 3 mg/kg

after day 30 and limited

sign in mice treated at 1

mg/kg (4/5 mice cured)

• Exposure consistent with

other PK

Minimum fully protective

dose set at 3mg/kg

100 100

80

25

100

00

20

40

60

80

100

120

DDD, 10mg/kg

DDD, 3mg/kg

DDD, 1mg/kg

DDD, 0.3mg/kg

ATQ, 3mg/kg

Notreatement

Pe

rce

nt

Percentage Mice Parasite Free, Day 30

DDD107498 Mode of Action: Inhibition of Translation Elongation Factor 2

3’5’

3’5’

ribosome

3’5’

pep debondforma on

GTP:PfeEF2

GDP:PfeEF2

mRNA

tRNA

BlockedbyDDD107498

• Responsible for GTP-dependent translocation of the ribosome along mRNA

• Essential for protein synthesis

Asexual blood-stage P. falciparum cultured in the presence of DDD107498 at 5 x

EC50 until parasites became resistant

• Determined by sequence analysis of DNA from ten resistant mutant lines

• Confirmed by molecular and cellular biology experiments

DDD107498

DDD107498: Good Pharmacokinetic

profile in preclinical species

Sprague Dawley Rat – Oral Dose (5mg/kg)

0

5

10

15

20

25

30

0 6 12 18 24 30 36 42 48

Whole

Blo

od C

oncentr

atio

n (

ng/m

L)

Time after dose (hours)

Beagle Dog - Oral dose (3 mg/kg)

Long predicted human T½

Safety/Toxicology Profile of DDD107498

30

Phototox: No phototoxicity flag: UV scan 290-700nm

Potencies in Off-Target Assays (IC50 values)

• Ion Channels• hERG (IKr) 25 µM*

• hNaV1.5 16 µM

• hKV1.5 > 100 µM

• CaV1.2 > 100 µM

• Dundee Kinase Panel (125 human kinases) > 10 µM

• Broad Screening (receptors, enzymes, ion channels)• Ricerca (68 targets) > 10 µM

• CEREP (100 targets) > 10 µM

Rat 7 day safety study – no major issues, satisfactory safety margins

Genotox: Ames, in vitro and in vivo micronucleus studies negative

_________________________________________________________________________________________________________________________________________________

*hERG IC50/free Cmax at predicted human dose: 184 fold

DDD107498: Summary

Possible single dose blood-stage treatment with

transmission blocking (TCP2; TCP3b) and chemoprotection

(TCP4) potential

Excellent agreement in potency between blood and transmission assays

Novel mode of action - active against resistant strains

Good pharmacokinetic properties

Low clinical drug-drug Interaction risk

Acceptable safety margins

No safety (toxicological) concerns have been identified to

date

Cost of goods estimated to deliver at <$1/dose

Under clinical development by MMV and Merck

DDD107498: Phase 0/Phase I project plan

CMC

Non-clinical Evaluation

Clinical Development

2016 2017 2018

M A M J J A S O N M A MD J F JFJ N DJ A S O M A MJ F J N DJ A S O

Human bio-analytical method Validation

Embryo-foetal studies (DRF and main: GLP)

Pivotal toxicology studies (GLP)

Safety pharmacology

Animal bio-analytical method Validation

Clinical trial supplies for Phase I

Study planning Single Ascending Dose Phase I

Human Malaria Challenge Phase I

Multiple Ascending Dose Phase I

Juvenile Tox (DRF: GLP)

Food effect Phase I

Drug Substance development for Phase II

Drug Product development for Phase II

Clinical trial supplies for Phase IIa

Drug Substance & Drug Product for Phase I already available

Final report

Nature, 2015, 522,

315-320

The Dundee Team:Beatriz BaraganaIrene Hallyburton

Neil NorcrossCaroline Wilson

Raffaella GrimaldiLaste Stojanovski

Frederick SimeonsMaria Osuna-Cabello

Suzanne NorvalKevin ReadIan Gilbert

Alan Fairlamb

Acknowledgements

Dundee

Achim Porzelle

Andrew Woodland

Jennifer Riley

Fabio Zuccotto

John Thomas

Julie Frearson

Paul Wyatt

David Gray

Columbia

David Fidock

Marcus Lee

Tara Abraham

Mariana Almedia

Rajshekhar Basak

MMV

Paul Willis

Sir Simon Campbell

Lidiya Bebrevska

Pete Siegl

Carol Sibley

Didier Leroy

Brice Campo

Sanger

Julian Rayner

Thomas Otto

William Proto

GSK

Francisco Javier Gamo

Inigo Angulo-Barturen

Santiago Ferrer-Bazaga

Maria Santos Martinez

Laura Maria Sanz

Maria Belen Jimenez-DiazUCSD

Elizabeth Winzeler

Stephan Meister

Imperial College

Robert Sinden

Michael Delves

Andrew Blagborough

Andrea Ruecker

Leanna Upton

Paddy Brock

Tom Churcher

Katarzyna Sala

Sara Zakutansky

Monash

Susan Charman

Michael Campbell

TropIQ

Koen Dechering

Robert Sauerwein

Judith Bolscher

SwissTPH

Sergio Wittlin

USF

Dennis Kyle

Anupam Pradhan

Menzies

Ric Price

Jutta Marfurt

Grennady Wirjanata

Eskitis Institute

Vicky Avery

Funding

MMV

Wellcome Trust

B&MGF

NIH

European Union

“Science is the most communal

of human endeavours”