1

http://www.utsouthwestern.edu/kidneycancer

COI: Patent HIF-2 biomarkerResearch funding – Peloton Therapeutics

Scheuermann et al. PNAS 2009

B

A

B

A

HRE (DNA)

HIF-2 HIF-1

HRE (DNA)

HIF-1HIF-2

Scheuermann et al. Nat Chem Biol 2013

High-Throughput Screen

HIF2-I

Development of a HIF-2 inhibitor (HIF2-I)

PT2399

Validating HIF-2 as a target for ccRCC

Pavia-Jimenez et al., Nat Protoc 2014

>100 lines

2

Validating HIF-2 as a target

Human tumors in mice - do they reproduce: Histological appearance? Gene expression? Mutations? DNA copy number alterations? Treatment responsiveness?

Sivanand et al., Sci Transl Med 2012Adapted from Sivanand et al., Sci Transl Med 2012

Unsupervised hierarchical clustering of gene expression shows similarities between tumors and corresponding tumorgrafts

HIF2-I (PT2399) is active against human ccRCC transplants in mice

Days

Tum

or v

olum

e (m

m3 )

Vehicle

Sunitinib

HIF2-I

HIF2-I is active in 50% of ccRCC

Inte

rmed

.Se

nsiti

veR

esis

tant

VehicleSunitinibHIF2-I

267 mice from 22 independently derived TG lines (18 ccRCC)

Chen et al., Nature 2016

HIF2-I (PT2399) has greater activity than sunitinib and is better tolerated

Chen et al., Nature 2016

3

HIF2-I (PT2399) inhibits proliferation & angiogenesis in sensitive ccRCC

HIF2-Iveh

XP37

3XP

144

CD

31H

&E

XP37

3Ki67

veh HIF2-I

XP16

4XP

490

XP16

9XP

373

vehicle HIF2‐I

V4237

V5239 V5229 P5231 P5240

V4236 V4241V4234

V3290 V3294 V3287 V3281

V3212 V3224 V3210 V3214

Chen et al., Nature 2016

HIF2-I (PT2399) suppresses VEGF only in the tumor

Circulating Human VEGF (tumor) Circulating Mouse VEGF (host)

Chen et al., Nature 2016

HIF2-I (PT2399) dissociates HIF-2 in both sensitive & resistant tumors

RES

ISTA

NT

SEN

SITI

VE

Chen et al., Nature 2016

HIF2-I (PT2399) is a highly specific inhibitor

Veh HIF2-I Veh HIF2-I

Sensitive Resistant

VehicleHIF2-ISunitinib

Up 195 0

Down 297 0

Total 492 0

Differential Gene Expression

Sensitive ResistantVeh v HIF2-I Veh v HIF2-I

Rel

ativ

em

RN

Aex

pres

sion

Rel

ativ

em

RN

Aex

pres

sion

CCND1 TGFA

hVEGF SERPINE1 IGFBP3

SLC2A1

Chen et al., Nature 2016

4

Higher HIF-2in sensitive tumors

HIF-2α HIF-1αHIF-2αHIF-1α

XP46

9XP

374

XP16

4

XP49

0XP

530

XP46

2HIF2-

Perc

enta

ge p

ositi

ve c

ells

Chen et al., Nature 2016

Identification of biomarkers of HIF-2-dependency

RNAseq

Chen et al., Nature 2016

Acquired resistance?P3283V3286

Forw

ard

Rev

erse

G323E(HIF-2)

V1849 P5123

Forw

ard

Rev

erse

F446L(HIF-1)

HIF-1 HIF-2

0500

100015002000250030003500400045005000

‐20 0 20 40 60 80 100 120 140 160 180 200

Tum

or V

olum

e (m

m3 )

XP164

3283

3288

3296

3295

3299

3286 Veh

Sunitinib

HIF2-I

Input

IP HIF-1

V3290

V3294

V3298

P3283

P3288

P3297

V3290

V3294

V3298

P3283

P3297

Tubulin

post-resistance

HIF‐2

HIF‐1

P3288

pre-resistance

vehicle HIF2-I

HIF-2and HIF-1mutants preserve HIF complexes despite HIF2-I (PT2399)

FLAG-HIF-1

Input IP FLAG

Vehicle Vehicle PT2399PT2399

+-+

+-

--

--+

+ +

++

- -

Tubulin

HA

FLAG

FLAG-HIF-1(F466L)HA-HIF-2

HA-HIF-2 (G323E)

+-+

+-

--

--+

+ +

++

- - +-+

+-

--

--+

+ +

++

- - +-+

+-

--

--+

+ +

++

- -

Chen et al., Nature 2016

5

Baseline Characteristics

Pharmacokinetics & Pharmacodynamics

• Targeted Cmin of 280 ng/mL exceeded at ≥ 400 mg b.i.d.

• Increased exposure up to 800 mg b.i.d. (no further increase beyond 800 mg b.i.d.)

Toxicity

6

Duration of Treatment Prolonged stable disease in heavily pretreated patient (7 prior lines)

Kidney

-12 37 100 225 350 Days

Pelvis

Liver

PT2385

Chen et al., Nature 2016

Higher exposure correlates with anti-tumor activity

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

7

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

Second generation inhibitor, PT2977 –greater potency in preclinical models

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

PT2977 Phase I

Slide 13

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

PT2977 Phase I Pharmacokinetics

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

PT2977 - Pharmacodynamics

PT2385

8

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

Response Duration

Presented By Kyriakos Papadopoulos at 2018 ASCO Annual Meeting

Responses in ccRCC

PT2977 – VHL pts RCC

Inclusion Criteria:• von Hippel Lindau disease with a germline VHL

mut.• At least 1 measurable solid RCC tumor

(diagnosis of RCC can be radiologic). • VHL disease-associated tumors in other organ

systems.Exclusion Criteria:• Prior systemic anti-cancer therapy (includes anti-

VEGF therapy and investigational agents).• Immediate need for surgical intervention for tumor

treatment.• Metastatic disease.

University of Colorado Cancer Center Not yet recruitingAurora, Colorado, United States, 80045Yale Cancer Center Not yet recruitingNew Haven, Connecticut, United States, 06519National Institutes of Health Clinical Center RecruitingBethesda, Maryland, United States, 20892Massachusetts General Hospital RecruitingBoston, Massachusetts, United States, 02114University of Michigan RecruitingAnn Arbor, Michigan, United States, 48109Memorial Sloan Kettering Cancer Center Not yet recruitingNew York, New York, United States, 10065Wake Forest Not yet recruitingWinston‐Salem, North Carolina, United States, 27103University of Pennsylvania Medical Center Not yet recruitingPhiladelphia, Pennsylvania, United States, 19104University of Pittsburgh Medical Center Not yet recruitingPittsburgh, Pennsylvania, United States, 15232Vanderbilt Medical Center RecruitingNashville, Tennessee, United States, 37232UT Southwestern Medical Center Not yet recruitingDallas, Texas, United States, 75239MD Anderson Cancer Center RecruitingHouston, Texas, United States, 77030Huntsman Cancer Institute RecruitingSalt Lake City, Utah, United States, 84112Aarhus University Hospital Not yet recruitingAarhus, DenmarkHospital Georges Pompidou Not yet recruitingParis, FranceUniversity Medical Center Utrecht Not yet recruitingUtrecht, NetherlandsCambridge University Hospitals Not yet recruitingCambridge, United KingdomGuy's and St. Thomas' Hospitals Not yet recruitingLondon, United KingdomChristie NHS Foundation Trust Not yet recruitingManchester, United Kingdom

June 6, 2018

Photo, courtesy of Brian Coats

Conclusions HIF-2 is a valid target in ccRCC. Inhibition of the HIF-2 transcription factor abrogates tumor growth in >50% of human

ccRCC tumors implanted in mice, including tumors resistant to sunitinib. HIF2-I effectively (and specifically) dissociates HIF-2 from HIF-1 in human ccRCC

implanted in mice. HIF-2 inhibition results in the downregulation of HIF-2 target genes and decreased

circulating levels of tumor-produced VEGF. HIF-2 inhibitors are more effective and better tolerated than sunitinib in TG models. Primary resistance occurs despite dissociation of the HIF-2 complex in tumors. Sensitive and resistant tumors can be distinguished by HIF-2 levels and gene

expression. Resistance develops in sensitive tumors due to binding site and “second-site

suppressor mutations.” A Phase I clinical trial of PT2385 showed that HIF-2 inhibitors are safe and may benefit

a subset of patients, but resistance develops. Second generation inhibitors (PT2799) have greater potency. HIF-2 inhibitors are well tolerated and associated with anemia. Close monitoring for

hypoxia is needed (which can be severe).

9

Brugarolas LabAlana Christie

Roy EliasAdrien Jump

Eric MaRenee McKay

Tiffani McKenzieOreoluwa OnaboluGopinath Prakasam

Nirmish Singla Christina Stevens Vanina Toffessi Layton Woolford

Juan YangHui Ye

Xiaolin Zhu

Wenfang ChenShannon Cohn

Yifeng GuHaley Hill

Allison JoyceBlanka Kucejova

Andrea Pavia-JimenezSamuel Peña-LlopisSharanya Sivanand

Tram Anh TranSilvia Vega-Rubin de Celis

Shanshan WangNick Wolff

Toshinari YamasakiAnum Yousuf

Avery Anderson Megan Dougherty

Debbie HarveyMarieshia HicksChey Reynolds

FundingVirginia Murchison Linthicum Endowment

Cancer Prevention and Research Institute of TexasNIH, National Cancer Institute

Peloton Therapeutics, Inc

Kidney Cancer ProgramUrology

Aditya Bagrodia Jeff CadedduJeff GahanYair Lotan

Vitaly MargulisGanesh Raj

Arthur Sagalowsky

Medical OncologyYull Arriaga

Alex BowmanKevin CourtneyEugene FrenkelHans Hammers

Radiation OncologyNeil Desai

Michael FolkertRaquibul Hannan

Robert TimmermanZabi Wardak

PathologyPayal Kapur

Dinesh Rakheja

RadiologyAlberto Diaz de Leon

Ananth Madhuranthakam Ivan PedrosaXiankai Sun

Lori Watumull

Illumina Inc. Mark Ross

David Bentley

Mayo ClinicRichard W. Joseph

Daniel J. SerieJeanette Eckel-Passow

Thai HoJohn C. ChevilleAlexander Parker

GenentechAnwesha Dey

Steffen DurinckEric W. StawiskiZora ModrusanSekar Seshagiri

NCILaura Schmidt

Marston Linehan

Other CollaboratorsKevin Jones

Brian Rini

UTSW CollaboratorsLaura Banaszynski

Tom CarrollRalph DeBerardinis

Bob HammerLisa KinchSri Malladi

Josh MendellBruce Posner

Noelle WilliamsJin Ye

Yonghao Yu

Phase 1 Trial (PT2385)Kevin CourtneyJeffrey Infante

Elaine LamRobert Figlin

Brian RiniNaseem Zojwalla

Keshi WangEli WallaceJohn Josey

Toni Choueiri

BioinformaticsMin Kim

Satwik Rajaram Phillip Reeder

Tao WangJason Xie

Guanghua Xiao Yang XieHe Zhang

Clinical CoordinatorsJenny Chang

Dendra Von Merveldt