Bicycles® - An entirely new class of therapeuticsPaul BeswickBicycle Therapeutics

The challenges in treating cancer

ELRIG April 20192

Tumours can be “silent”

Are difficult to differentiate from normal tissue

Can be hard to access

Actively suppress the immune system

Heterogeneous and evolvingDiverse set of

diseases

Overview

• Bicyclic peptides: A completely new, disruptive therapeutic modality

• Sir Greg Winter technology, platform derisked, industrialized, reduced to practice and validated

• Internal oncology pipeline, multiple therapeutic themes, BT1718 in Ph1: funded by CRUK. Partnered outside oncology

• UK /US presence, world class team & strong clinical / scientific collaborations

• >£65M Series B funded

Jan-193

Highly constrained: high affinity, exquisite selectivity, excellent stability

Large binding footprint: disrupt protein-protein interactions

Fully synthetic: NCE classification and synthetic control

Highly flexible modality: modular building blocks retain pharmacology

Adjustable PK: excellent tissue penetration, renal elimination, tuneable T1/2

ELRIG April 20194

COOH

X

NH2

Linear peptide

NH2COOH

Bicycle

Chemical modification with scaffold

Bicycles®: a new therapeutic modality

Loop 1

Loop 2

Scaffold

ELRIG April 20195

Comparison of therapeutic modalities

Antibody ScFv (fragment) Bicycle Small molecule

Mw (kDa) 150 28 1.5-2 <0.8

Volume of distribution

Low (vascular) Intermediate Whole body Typically whole body

t 1/2

Days to weeks Minutes to daysMin to hours

(tunable). Days possible2

Hours (tunable)

Clearance hepatic Renal, hepatic Renal Renal, hepatic

Tumour penetranceLow (outer rim only) Low (poor exposure) High High

Target classes Many, small pockets restricted

Many, small pockets restricted

All tested successful, PPI trivial

Small pockets, PPI rare

Selectivity Highly Highly Highly Poor

Modularity Low (bi-specifics) Possible, difficult Trivial (“Lego like”) Low

Synthesis Complex biologic Complex biologic Chemical, trivial Chemical, trivial

Immunogenicity Possible Frequent None detected None

(to scale)

Peptide imaging agents in the clinic based on:

Human• RGD (fibronectin)• Vasoactive intestinal peptide• Somatastatin-14Other species• Exendin-4 (GLP-1 homologue)• Bombesin (GRP homologue)• Venoms & toxins

Current generation of peptidic imaging agents & approved drugs all inspired by nature

ELRIG April 20196

64Cu-DOTA-TATE111In-DTPA-Octreotide

DOTA-TATEDTPA-octreotide

The Bicycle platform can deliver novel tumour targeting peptides

ELRIG April 20197

Protein III

Linear peptide

Bicycle DNA Sequence

Gene III

Phage particle

Bicycle

Chemical modification with scaffold

Diverse Bicycle phage libraries (>1015)

Evolution driven, informed selection

3Amplify

2Select

1Cyclise

POC in 6 wk Optimised

lead in 9mnth

Extremely large and diverse chemical library Low synthetic burden

ELRIG April 20198

Tractable target classes

Enzymes

Serine proteases

Other proteases

Metalloenzymes

Matrix metalloproteinases

Coagulation factors

Other enzymes

Immune checkpoint

TNFR superfamily members

IG domain receptors

Signalling

Receptor Tyrosine kinases

Interleukin receptors

Interleukins

Growth Factors

Cytokines

AdhesionIntegrins

Other cell adhesion proteins

GPCRsChemokine receptors

Adrenergic receptors

OtherHeat shock proteins

Serum proteins

Bicycles®: many shapes to drug many targets

Receptor ANTAGONIST

Receptor AGONIST

Neutral BINDER

Enzyme INHIBITOR

Natural ligand

Bicycle

>90 diverse targets screened80% success rate

Bicycle® – large molecular footprint drives affinity and selectivity between close homologues

ELRIG April 20199

CA IX Ki = 25 nMCA XII Ki = 6 nM

CA IX Ki = 7.5 nMCA XII Ki > 2000 nM

Bicycleinhibitors

Human Kallikrein

Ki (nM)

Rat Kallikrein

Ki (nM)

Thrombin

Ki (nM)

Plasmin

Ki (nM)

FactorXla

Ki (nM)

FactorXlla

Ki (nM)

Exemplar 1 0.8 17.6 >10,000 >15,000 >50,000 >10,000

Exemplar 2 0.2 3.7 >10,000 >35,000 15,000 >10,000

Homologue active site sequence identity 85% 92% 100% 85%

Acetazolamide Bicycle

Tolerance to conjugation is built-in

ELRIG April 201910

1 nm

Bicycle

Tag to specific target

• Small molecule drugs

• Other Bicycles (tandems)

• Chelated radionuclides

• Fluorescent dyes

• Affinity tags

• PK extenders

Bacteriophage

900 nm x 7 nm

In vitro tools

Fluorescent probe

Phage bulk readily replaced without

compromising binding

DOTA

68Ga

In vivo tools/diagnostics

Case Study: MT1-MMP Targeting BTC – BT1718

11

Rosse et al., 2014, PNAS 111, pp1872–1879

Proven tumour delivery with Bicycle Toxin Conjugates: targeting MT1-MMP

• Membrane type 1 matrix metalloproteinase

• Low expression in normal adult

• Strong correlation with invasiveness in cancer cells

12 ELRIG April 2019

HumanMT1-MMPKd (nM)

MouseMT1-MMPKd (nM)

MT2-MMPKd (nM)

MT3-MMPKd (nM)

MT5-MMPKd (nM)

MMP1Kd (nM)

MMP2Kd (nM)

2.6 1.8 >10000 >10000 >2000 >1000 >1000

Bicycle binder to MT1-MMP:

DOTA

68Ga

Bicycles® are retained in tumours and rapidly cleared from systemic circulation

ELRIG April 201913

Bicycle show superior retention in tumours and lower background vs antibodies

Ph

oto

aco

usti

c s

ign

al in

ten

sit

y

(ch

an

ge f

rom

baselin

e)

15m

in 2h 24

0

40

80

120

160

Antibody

Bicycle

High tumour retentionIdeal distribution for imaging

40-60 min

68Ga MT1-MMP Bicycle 68Ga MT1-MMP Antibody

Tumour

40-60 min

Heart

Liver

Bicycle® radio conjugate - kinetics of distribution and clearance

14

Figure 22. Whole-body coronal slices (0.8 mm) from µPET imaging 0-20 min p.i. (A), 20-40 min p.i. (B), and 40-60 min p.i.

(C).

A B C

Bladder

L.Kidney

R.Kidney

Liver

Bladder

L.Kidney

R.Kidney

Sk.Muscle

Heart

Tumour

Tumour

0-20min 20-40min 40-60min

Tumour

Muscle

Expanded scale

68Ga conjugated MT1-MMP targeting Bicycle

ELRIG April 2019

Bicycle® toxin conjugates show profound efficacy

ELRIG April 201915

Days after start of dosing

Tum

our

Vol

ume

(mm

3 )

0 10 20 300

1000

2000

3000 Vehicle

BT171810mg/kg biw

^ ^ ^^ ^^ ^

Large 1000mm3 CDX (EBC1)

Antigen mediated cell killing Clears heterogenous PDXs

Cell-derived xenografts Patient-derived xenografts

Days after start of dosing

Tu

mo

ur

Vo

lum

e (

mm

3)

20 40 600

500

1000

1500Vehicle, iv, biw

BT1718, 1 mpk, biw

BT1718, 3 mpk, biw

BT1718, 10 mpk, biw

 ^

 ^ ^^ ^^

Days after start of dosing

Tum

ou

r V

olu

me

(m

m3 )

0 20 40 600

1000

2000

3000Vehicle, iv, biw

Docetaxel 20mg/kg, qw

^ ^ ^ ^ ^ ^ ^ ^

BT1718 3mg/kg, biw

BT1718 10mg/kg, biw

DM1Toxin

Cleavable linker

O

O

O

N

O

NHOH

O

ClO

H

H

ON

O

O

S S

O

SpacerTargeting Bicycle

BT1718: MT1-MMP targeting Bicycle Drug Conjugate Clears large tumours as quickly as small

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

0

1 0 0 0

2 0 0 0

3 0 0 0

^ ^ ^

V e h ic le i . v b iw

B T 1 7 1 8 1 0 m g / k g b iw

D o c e t a x e l 2 0 m g / k g q w

^ ^^ ^ ^ ^

D a y s a f t e r s t a r t o f d o s i n g

Tu

mo

ur

Vo

lum

e (

mm

3)

Large 1000mm3 PDX (Lu-01-0046)

Vehicle day 14

BT1718day 28

Vehicle day 7

BT1718day 28

Case Study: EphA2 Targeting BTC - BT5528

16

EphA2: Biological rationale

• Erythropoietin-producing hepatocellular A2 receptor

• Member of Eph subfamily of receptor tyrosine kinases

• Regulates cell migration, adhesion proliferation and differentiation

• Overexpression in human cancers, correlates with tumourprogression

• Key area for pharma companies, multiple programs in discovery, and clinical stages

17

Benign Breast

Invasive ductal carcinoma

% s

am

ple

s /

cate

go

ry

Benign Breast Breast carcinoma0

20

40

60

80

100negative

weak

moderate

strong

Zelinski et al Cancer Res 61: 2301-2306 (2001)

ELRIG April 2019

Biodistribution of 68Ga labelled Bicycle® shows excellent tumour targeting

18

BCY6099

DOTA

68Ga

Physicochemical properties of Bicycles have profound effect on distribution

PET imaging of HT-1080 xenograft at 60 minutes

ELRIG April 2019

Prototype Bicycle(hydrophobic)

BT5528: Rapid discovery of EphA2 targeted Bicycle®

Toxin Conjugate

19

huEphA2Kd (nM)

moEphA2Kd (nM)

ratEphA2Kd (nM)

huEphA1Kd (nM)

huEphA3Kd (nM)

huEphA4Kd (nM)

huEphA5Kd (nM)

1.2 2.5 3 >5000 >5000 >5000 >25000

• Matrix of ~70 conjugates synthesized and screened

• Identify optimal toxin, cleavable linker, molecular spacer

• BT5528 identified as candidate BTC

Val-Citcleavable linker

Molecular spacer

Targeting Bicycle

BCY6099

MMAE

ELRIG April 2019

Phage hit → Candidate

selection: 1 year

Extensive tumour penetration maintains efficacy even in very large PDX model

• BT5528 maintains efficacy seen in CDX models even in large PDX

• Patient-derived xenograft

• Lung adenocarcinoma

• Heterogeneous tumour

• 1000mm3 at dosing start

• Significant regression of tumour after 21d dosing 3mg/kg qw

• ADC shows no efficacy

• Dosed 3mg/kg qw

• PET imaging shows rapid penetration of Bicycle conjugate into tumour

• ADC data shows largely vascular distribution

• BT5528 in pre-clinical development

20

0 7 1 4 2 1

0

1 0 0 0

2 0 0 0

3 0 0 0

4 0 0 0

^ ^ ^^

V e h i c l e i . v q w

B T 5 5 2 8 3 m g / k g q w

A D C 3 m g / k g q w

D a y s a f t e r s t a r t o f d o s i n g

Tu

mo

ur

Vo

lum

e (

mm

3)

Bicycle distribution at 60 min

ADC distribution at 60 min

Cai W et al, Quantitative radioimmunoPET

imaging of EphA2 in tumor-bearing mice. Eur

J Nucl Med Mol Imaging. 2007

ELRIG April 2019

Bicycles® can meet many of the challenges in oncology

ELRIG April 201921

Tumours can be “silent”

• Large toolkit of novel probes

Are difficult to differentiate from normal tissue

• Highly selective to tumour target• Combine in bispecifics tandem etc.

Can be hard to access• Size and PK accesses

tumours efficiently

Actively suppress the immune system

• Multimeric immune receptor agonists• Targeted systemic delivery of innate

immune activators

Heterogeneous and evolving• Superior penetration & bystander effect

kills whole tumour• Extensive arson of different anti-cancer

targeting agents

Diverse set of diseases

• Companion diagnostics to stratify patients

• Prof. Matthias Eder and group at DKFZ, Heidelberg

Deutschen Krebsforschungszentrum/German Cancer Research Centre

• Bioprobe Ltd

• Team at Bicycle UK & US

Acknowledgements

ELRIG April 201922

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