BET PROTACs Are More Broadly Effective Than BET Inhibitors

1

Dr. Kevin Coleman

Arvinas, LLC

New Haven, CT

Disclosures

I am an employee of and have an equity stake in Arvinas

2

3

Private company, founded July 2013

– Founder: Dr. Craig Crews

– Licensed Technologies for Targeted Degradation of Proteins from Yale University

Arvinas: The Protein Degradation Company

PROTACs Induce The Rapid Degradation Of Target Proteins

PROTACs (Proteolysis-Targeting Chimera) are bifunctional chimeric molecules that

recruit an E3 ligase to a target protein to induce its degradation

– Have successfully degraded proteins using ligands that bind to several E3 ligases, including

cereblon, IAP2, MDM2, VHL

PROTAC technology is robust:

– Have degraded >85% of proteins tested & potentially can degrade any unwanted protein

– Degradation of unwanted proteins is fast (hours), durable (days), and potent (pM)

PROTAC technology is broadly applicable across target classes & therapeutic areas

Lys

E3

Ligase

Target

protein

Lys

Target

protein

Ubiquitin

E3

ligase

Degraded

protein

4

BRD4 Is An Important Cancer Epigenetic Regulator

BRD4 is a member of the Bromodomain and Extra-Terminal motif (BET) family of

proteins

BRD4 is an epigenetic reader of acetylated histones and regulates gene

transcription through the recruitment of other proteins to super-enhancer regions

BET inhibitors selectively disrupt transcription of a number of important cell- and

lineage-specific genes including oncogenes, i.e., MYC

Valent and Zuber (2014) Cell Cycle 13:689

Histone

5

H441 NSCLC

Shimamura et al. (2013) Clin Cancer Res 19: 6183

ARV-825 Potently And Rapidly Degrades BRD4 In A Cereblon- And Proteasome-Dependent Manner

6

ARV-825

Namalwa

Namalwa

Namalwa Ramos

0.1 mM ARV-825 0.1 mM ARV-825

ARV-825 ARV-825BRD4

Actin

ARV-825 is a potent degrader of all

BET family membersLu et al. (2015) Chem. Biol 22:755

BET PROTACs Cause Prolonged BRD4 Degradation, MYC Suppression and Inhibition of Cell Proliferation

7

Time after compound washout

Burkitt's Lymphoma

Namalwa cells

Compound Incubation

12 h

Compound

Washout

Immunoblot

Cell

Proliferation

Lu et al. (2015) Chem. Biol 22:755

Ovarian Cancer Cell Lines

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Tumor Cell Lines Are Consistently More Sensitive To BET Degradation Than To BET Inhibition

Ovarian cancer cell lines are consistently more

sensitive to BET protein degradation than they

are to BET protein inhibition

54% of ovarian cancer cell lines are resistant to

10 mM OTX015

Breast and NSCLC cell lines are similarly more

sensitive to BET PROTACs compared to BET

inhibitors

ARCC-29 (BET PROTAC)

OTX015 (BET Inhibitor)

Lymphoma cell lines are more sensitive to

BET inhibition compared to ovarian,

breast, and NSCLC cell lines

Nevertheless, lymphoma cell lines are

largely more sensitive to BET protein

degradation than they are to BET protein

inhibition

Log scale: 0.0001-10 mM

8

20 nM

200 nM

BET PROTACs Have Superior Anti-Proliferative

Activity in DLBCL Cells Compared to BETi’s

9

BRD4-binding moiety VHL-binding moiety

ARV-771

(Active BET PROTAC)

ARV-766

(Inactive diastereomer)

U2932 RI-1

SU-DHL-6 SU-DHL-5 SU-DHL-10

Raina et al. (2016) PNAS 113:7124

BET PROTACs Have Superior Apoptotic Activity

In DLBCL Cells Compared to BET Inhibitors

10

Caspase 3/7

BET PROTACs have also been shown to have superior apoptotic activity

than BET inhibitors in ovarian and prostate cancer cell lines

Intermittent Dosing Of The BET PROTAC ARCC-29 Causes Tumor Regression

11

Tumor regression (12 MPK ARCC-29)

68% TGI (6 MPK ARCC-29)

44% TGI (50 MPK OTX015)

27% TGI (3 MPK ARCC-29)

Once-weekly dosing of the BET PROTAC ARCC-29 also results in tumor

stasis or regression in human prostate and ovarian cancer xenograft models

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The Bet PROTAC ARCC-29 Causes Rapid BRD4 Degradation And Caspase Activation

• Free fraction of ARCC-29 stays above the predicted plasma concentration required to

achieve 90% BRD4 knockdown for ~ 6 hours

• Maximal BRD4 knockdown occurs at 3 hours followed by robust caspase activation at

6 hours

• BRD4 protein levels return to untreated levels 24 h post-dose

BRD4

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BRD4

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BRD4

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SU-DHL-6 DLBCL Xenograft Model

BET PROTACs Are More Broadly Effective Than BET Inhibitors

BET PROTACs cause robust BRD4 degradation and more prolonged c-Myc

suppression in vitro and have greater efficacy in vivo compared to BET

inhibitors

Tumor cell lines representing different tumor types are consistently more

sensitive to BET protein degradation than BET protein inhibition

BET PROTACs have superior anti-proliferative and apoptotic activity in

DLBCL, prostate cancer, and ovarian cancer cells compared to BET inhibitors

Intermittent dosing of BET PROTACs caused tumor regression or stasis in

DLBCL, prostate cancer, and ovarian cancer xenograft models

Arvinas’ BET PROTAC is planned to enter the clinic in 2H17

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Acknowledgements

Kanak Raina

Jing Lu

Martha Altieri

Ann Marie Rossi

Deborah Gordon

Ryan Willard

Jim Winkler

14

Arvinas Chemistry Yale UniversityArvinas Biology

Yimin Qian

Xin Chen

Jing Wang

Hanqing Dong

Andy Crew

John Hines

Craig Crews