bet protacs are more broadly effective than bet inhibitors · bet inhibitors selectively disrupt...
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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
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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
0 5 1 0 1 5
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0IV V e h ic le B IW
A R C C -2 9 1 2 M P K b iw IV
A R C C -2 9 6 M P K b iw IV
A R C C -2 9 3 M P K b iw IV
T re a tm e n t D a ySU
-DH
L-6
Tu
mo
r V
olu
me
(m
m3)
O T X 0 1 5 5 0 M P K q d P O
P O V e h ic le Q D
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
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
0 5 1 0 1 5 2 0 2 5
0
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5 0 0 0
7 5 0 0
1 0 0 0 0
1 2 5 0 0
1 5 0 0 0
1 7 5 0 0
2 0 0 0 0
0 . 0
0 . 5
1 . 0
1 . 5
A R V - 4 5 4 P K / P D
T i m e ( h )
Pl
as
ma
C
on
c.
(
nM
)
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la
ti
ve
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RD
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L
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el
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P l a s m a C o n c . ( n M )
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la
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le
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Relative BRD4 levels
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tive B
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ls
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sm
a C
on
c.
(nM
)
DC90
Plasma
Level
Time Post-Dose (h)
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
13
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