Translational Platform for the Development of Targeted

Therapeutics

Ondřej Kalous, MD Associate Project Scientist

UCLA Translational Oncology Research Laboratories (TORL)

Jonsson Comprehensive Cancer Center

Los Angeles, CA, USA

UCLA/TORL Translational Cancer Research Program

• Aims to rapidly translate discoveries from the laboratory to the development of clinical studies (bench to bedside)

• Not just a one-way path • Clinical observations, ideas, and samples from

clinics are transferred to the laboratory (bedside to bench)

• Requires frequent communications between the laboratory and clinical sites

• Relies on ‘translational thinking’ from both laboratory scientists and physicians

UCLA/TORL Components

• Proliferation assays conducted on a panel of ~ 600 cell lines

• Calculating outcome measures (i.e., IC50, IC100) • Identification of sensitive and resistant histologies and

histological subtypes • Analysis of in vitro biomarkers using genotype-

response and protein expression-response association screens

• In vivo testing using s.c. xenograft models of human cancer (selective)

• Preclinical data used to support clinical trials conducted by TRIO network

UCLA/TORL Panel of Cell Lines

• ~600 human cancer cell lines from 15 different cancer types

• Obtained from commercial vendors (ATCC, DSMZ, JCRB, ECACC) or established internally from patient samples

• Verified as unique via mitochondrial DNA sequencing

• STR profiling to match with online data sources

UCLA/TORL Cell Line Panel

UCLATORL Proliferation Assay Protocol

Summary of Outcome Measures

• Primary outcome measure: % generational inhibition

𝐷𝑜𝑢𝑏𝑙𝑖𝑛𝑔𝑠 𝑓𝑟𝑜𝑚 𝐵𝑎𝑠𝑒𝑙𝑖𝑛𝑒 =

log 𝐴𝑣𝑔 𝐷𝑎𝑦5 𝐶𝑜𝑢𝑛𝑡 − log 𝐴𝑣𝑔 𝐷𝑎𝑦0 𝐶𝑜𝑢𝑛𝑡

log 2

% 𝐼𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 = 1 − 𝑑𝑜𝑢𝑏𝑙𝑖𝑛𝑔𝑠 𝑢𝑛𝑑𝑒𝑟 𝑡𝑟𝑒𝑎𝑡𝑚𝑒𝑛𝑡

𝑑𝑜𝑢𝑏𝑙𝑖𝑛𝑔𝑠 𝑤𝑖𝑡ℎ𝑜𝑢𝑡 𝑡𝑟𝑒𝑎𝑡𝑚𝑒𝑛𝑡× 100%

• IC50 = dose of drug necessary to prevent 50% of cell population growth

• IC100 = dose of drug where cell population does not grow from baseline

• Dose response data condensed to single-value summary measures used to rank order cell lines by response

Genomic and Proteomic Characterization

• Four main data sources: – Whole-exome point mutation data from Cancer

Cell Line Encyclopedia

– Copy number alteration data from comparative genomic hybridization arrays (UCLA/TORL)

– mRNA expression by Agilent 44K microarrays (UCLA/TORL)

– Reverse phase protein array for total and phosphorylated protein (n = 225) levels (from MD Anderson)

• Cell line and patient derived xenograft models representing each of the known cancer sub-types

• Our own dedicated vivarium with a capacity for carrying 1,400 animals managed by a team of experienced technologists and veterinarians

• Inform the design of patient clinical trials by determining the most active and safest single agent and drug combination strategies

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Assessment of Drug Activity Assessment of Drug Safety

In vivo Program at UCLA-TORL

TRIO network • TRIO is a not-for-profit CRO that is dedicated to advancing

translational cancer research by bringing forward innovative and targeted therapeutic concepts into the clinical trial setting

• TRIO offices are based in Canada, France and Uruguay

• International network of 2000 investigators and 450 cancer centers in over 45 different countries

• TRIO comes from the merger of CIRG (Cancer International Research Group) and TORI (Translational Oncology Research International), and the wide group of preclinical researchers at UCLA JCCC

• TRIO is developing from proof of concept up to registrational phase III trials in breast, ovary, GI and other cancers

PALBOCICLIB (PD-0332991)

CDK-4/6 INHIBITOR

UCLA TORL/TRIO Translational Platform

Rb as Master-Regulator of the R-point

14

palbociclib

Richard Finn, AACR 2014

Breast Cancer Research, 2009

16

• RB and cyclin D1 expression were higher and CDKN2A (p16) was lower in sensitive cell lines.

Differentially Expressed Genes Between Sensitive and Resistance Cell Lines

Finn et al., Breast Cancer Research 2009

Palbociclib Synergizes the Effects of Tamoxifen in ER+ Breast Cancer in vitro

Finn et al., Breast Cancer Research 2009

Palbociclib + Letrozole Phase Ib Trial

• To test the safety of palbociclib in combination with letrozole (aromatase inhibitor)

• Oral palbociclib 125 mg once daily for 3 weeks followed by 1 week off treatment in a 28-day cycle, combined with letrozole 2.5 mg once daily

• The most common treatment-related adverse events were neutropenia, leucopenia, and fatigue

PALOMA-1/TRIO-18

Phase 2 Design

N = 66

1:1

Part 1

ER+, HER2–

BC

R

A

N

D

O

M

I

Z

A

T

I

O

N

PD 0332991

125 mg QDa +

Letrozole

2.5 mg QD

Letrozole

2.5 mg QD

Part 2

N = 99

1:1

ER+, HER2–

BC with

CCND1 amp

and/or

loss of p16

R

A

N

D

O

M

I

Z

A

T

I

O

N

PD 0332991

125 mg QDa +

Letrozole

2.5 mg QD

Letrozole

2.5 mg QD

Stratification Factors

• Disease Site (Visceral vs Bone only vs Other)

• Disease-Free Interval (>12 vs ≤12 mo from end of

adjuvant to recurrence or de novo advanced disease) a Schedule 3/1.

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ree S

urv

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rob

ab

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%)

84 67 60 47 36 28 21 13 8 5 1PAL+LET81 48 36 28 19 14 6 3 3 1LET

Number of patients at risk

Progression-Free Survival (ITT)

PAL + LET (N=84)

LET (N=81)

Number of Events (%) 41 (49) 59 (73)

Median PFS, months (95% CI)

20.2 (13.8, 27.5)

10.2 (5.7, 12.6)

Hazard Ratio (95% CI)

0.488 (0.319, 0.748)

p-value 0.0004

Finn et al, Lancet Oncol 2015

Palbociclib Timeline

• Late 1990s: Parke Davis / Onyx /Pfizer develop a novel small molecule (PD 0332991) that is potent and selective against Cyclin D-CDK4/6, no clear indication for use

• 2007: Pfizer contracts with UCLA/TORL for preclinical development

• 2009: UCLA/TORL publishes observation that ER+ breast cancers are uniquely sensitive to CDK4/6 inhibition

• 2010: Phase 1 trial shows hints of efficacy, with minimal toxicity

• 2011: TRIO begins biomarker-informed Phase 2

• 2013: Interim analysis shows major improvement in PFS

– FDA grants “Breakthrough Therapy” status in April, 2013

– TRIO begins enrollment in large Phase 3

• 2014: Final analysis from Phase 2 confirms strong efficacy signal

– Phase 3 study is fully enrolled and ongoing

Dacomitinib (pan-HER inhibitor) in Breast Cancer

• Dacomitinib has a strong antiproliferative activity in HER2-amplified breast cancer cell lines

• Dacomitinib maintains high activity in HER2- amplified cell lines with de novo and acquired resistance to trastuzumab and acquired resistance to lapatinib.

Kalous et al., Mol Cancer Ther 2012

AMG 900 pan-Aurora Kinase Inhibitor in Breast Cancer

• AMG 900 has potent anti-proliferative effects in breast cancer cell lines regardless of subtype

• Neither gene copy number changes nor expression of Aurora kinases A/B/C predicted for response to AMG 900

• AMG 900 preferentially inhibits the proliferation of breast cancer cell lines with dysfunctional p53 and low protein expression of p21, a downstream target of p53

Kalous et al., Breast Cancer Res Treat, 2013

Translational component in cancer research

• Can identify sensitive subtypes within histologies and novel predictive markers of response

• Can help select the right patients for clinical trials

• Can speed up the development of targeted therapeutics

• Can save financial resources

• All to benefit our patients

Thank you

• Dennis Slamon

• Richard Finn

• Dylan Conklin

• Erika von Euw

• Neil O’Brien

• UCLA/TORL staff