BET PROTACs Are More Broadly Effective Than BET Inhibitors. Dr. Kevin Coleman Arvinas, LLC New Haven, CT

Transcription

1 BET PROTACs Are More Broadly Effective Than BET Inhibitors Dr. Kevin Coleman Arvinas, LLC New Haven, CT 1

2 Disclosures I am an employee of and have an equity stake in Arvinas 2

3 Arvinas: The Protein Degradation Company Private company, founded July 2013 Founder: Dr. Craig Crews Licensed Technologies for Targeted Degradation of Proteins from Yale University 3

4 PROTACs Induce The Rapid Degradation Of Target Proteins Lys Ubiquiti Lys Degraded protei Target protein E3 ligase Target protein E3 Ligase 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 4

5 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 H441 NSCLC Histone Valent and Zuber (2014) Cell Cycle 13:689 Shimamura et al. (2013) Clin Cancer Res 19:

6 ARV-825 Potently And Rapidly Degrades BRD4 In A Cereblon- And Proteasome-Dependent Manner ARV-825 ARV-825 ARV-825 BRD4 Actin Namalwa 0.1 mm ARV mm ARV-825 Namalwa Ramos ARV-825 is a potent degrader of all BET family members Lu et al. (2015) Chem. Biol 22:755 Namalwa 6

7 BET PROTACs Cause Prolonged BRD4 Degradation, MYC Suppression and Inhibition of Cell Proliferation Burkitt's Lymphoma Namalwa cells Compound Incubation 12 h Time after compound washout Compound Washout Immunoblot Cell Proliferation Lu et al. (2015) Chem. Biol 22:755 7

8 C a o v -3 -P R O T A C C a o v -3 -In h ib ito r C O V P R O T A C C O V In h ib ito r C O V P R O T A C C O V In h ib ito r C O V P R O T A C C O V In h ib ito r E S -2 -P R O T A C E S -2 -In h ib ito r K U R A M O C H I-P R O T A C K U R A M O C H I-In h ib ito r O V C A R -3 -P R O T A C O V C A R -3 -In h ib ito r O V P R O T A C O V In h ib ito r O V S A H O -P R O T A C O V S A H O -In h ib ito r R M U G -S -P R O T A C R M U G -S -In h ib ito r S N U P R O T A C S N U In h ib ito r T Y K -n u -P R O T A C T Y K -n u -In h ib ito r A P R O T A C A In h ib ito r C O V P R O T A C C O V In h ib ito r H O P R O T A C H O In h ib ito r K G N -P R O T A C K G N -In h ib ito r M C A S -P R O T A C M C A S -In h ib ito r O A W 4 2 -P R O T A C O A W 4 2 -In h ib ito r O V 7 -P R O T A C O V 7 -In h ib ito r O V IS E -P R O T A C O V IS E -In h ib ito r O V M A N A -P R O T A C O V M A N A -In h ib ito r O V T O K O -P R O T A C O V T O K O -In h ib ito r P A -1 -P R O T A C P A -1 -In h ib to r R K N -P R O T A C R K N -In h ib ito r R M G -I-P R O T A C R M G -I-In h ib ito r S K -O V -3 -P R O T A C S K -O V -3 -In h ib ito r T O V D -P R O T A C T O V D -In h ib ito r T O V -2 1 G -P R O T A C T O V -2 1 G -In h ib ito r A 2 0 -P R O T A C A 2 0 -In h ib ito r B C P -1 -P R O T A C B C P -1 -In h ib ito r D B -P R O T A C D B -In h ib ito r D O H H -2 -P R O T A C D O H H -2 -In h ib ito r F a ra g e -P R O T A C F a ra g e -In h ib ito r G R A N T A P R O T A C G R A N T A In h ib ito r H 9 -P R O T A C H 9 -In h ib ito r H H -P R O T A C H H -In h ib ito r H T -P R O T A C H T -In h ib ito r H u T P R O T A C H u T In h ib ito r H u T 7 8 -P R O T A C H u T 7 8 -In h ib ito r J e K o -1 -P R O T A C J e K o -1 -In h ib ito r J M 1 -P R O T A C J M 1 -In h ib ito r M a v e r-1 -P R O T A C M A V E R -1 -In h ib ito r M C P R O T A C M C In h ib ito r M in a m i-1 -P R O T A C M in a m i-1 -In h ib ito r M J -P R O T A C M J -In h ib ito r M L M A -P R O T A C M L M A -in h ib ito r N A L M -1 -P R O T A C N A M L -1 -in h ib ito r P fe iffe r-p R O T A C P fe iffe r-in h ib ito r O C I-L Y P R O T A C O C I-L Y In h ib ito r R E C -1 -P R O T A C R E C -1 -In h ib ito r R L -P R O T A C R L -In h ib ito r S R -P R O T A C S R -In h ib ito r S U -D H L -2 -P R O T A C S U -D H L -2 -In h ib ito r S U -D H L -4 -P R O T A C S U -D H L -4 -In h ib ito r S U -D H L -6 -P R O T A C S U -D H L -6 -In h ib ito r T o le d o -P R O T A C T o le d o -In h ib ito r T U R -P R O T A C T U R -In h ib ito r U P R O T A C U In h ib ito r W S U -D L C L 2 -P R O T A C W S U -D L C L 2 -In h ib ito r Z P R O T A C Z In h ib ito r C e ll P ro life ra tio n E C 5 0 (m M ) C e ll P ro life ra tio n E C 5 0 (m M ) Tumor Cell Lines Are Consistently More Sensitive To BET Degradation Than To BET Inhibition Log scale: mm ARCC-29 (BET PROTAC) OTX015 (BET Inhibitor) Ovarian Cancer Cell Lines 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 200 nm 20 nm 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 8

9 BET PROTACs Have Superior Anti-Proliferative Activity in DLBCL Cells Compared to BETi s BRD4-binding moiety VHL-binding moiety ARV-771 (Active BET PROTAC) U2932 Raina et al. (2016) PNAS 113:7124 ARV-766 (Inactive diastereomer) RI-1 SU-DHL-6 SU-DHL-5 SU-DHL-10 9

10 BET PROTACs Have Superior Apoptotic Activity In DLBCL Cells Compared to BET Inhibitors Caspase 3/7 BET PROTACs have also been shown to have superior apoptotic activity than BET inhibitors in ovarian and prostate cancer cell lines 10

11 S U -D H L -6 T u m o r V o lu m e (m m 3 ) Intermittent Dosing Of The BET PROTAC ARCC-29 Causes Tumor Regression IV V e h ic le B IW P O V e h ic le Q D A R C C M P K b iw IV A R C C M P K b iw IV A R C C M P K b iw IV O T X M P K q d P O T r e a tm e n t D a y 27% TGI (3 MPK ARCC-29) 44% TGI (50 MPK OTX015) 68% TGI (6 MPK ARCC-29) Tumor regression (12 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 11

12 Plasma Conc. (nm) P l a s m a C o n c. ( n M ) R e l a t i v e C l e a v e d C a s p 3 L e v e l s R e l a t i v e B R D 4 L e v e l s The Bet PROTAC ARCC-29 Causes Rapid BRD4 Degradation And Caspase Activation SU-DHL-6 DLBCL Xenograft Model BRD4 cl.casp3 hmito BRD4 cl.casp3 hmito BRD4 cl.casp3 hmito BRD4 cl.casp3 hmito Veh 1h 3h Veh 6h 9h Veh 12h 15h Veh 18h 24h DC 90 Plasma Level P r e d i c t e d D C A R V P K / P D ARCC-29 P l a s m a C o n plasma c. ( n M ) conc.(nm) Relative R e l a t i v e B RBRD4 D 4 L e v elevels l s Relative cleaved caspase3 levels T i m e ( h ) Time Post-Dose (h) Relative BRD4 Levels Relative Cleaved Caspace3 Levels 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

13 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 13

14 Acknowledgements Arvinas Biology Kanak Raina Jing Lu Martha Altieri Ann Marie Rossi Deborah Gordon Ryan Willard Jim Winkler Arvinas Chemistry Yimin Qian Xin Chen Jing Wang Hanqing Dong Andy Crew Yale University John Hines Craig Crews 14