Non-invasive small animal imaging in cancer drug research and developmentCathy Zhang, Translational Research Group, Oncology Research Unit, Pfizer

Global Research and Development, La Jolla Laboratories

Modalities Light Output Application Characteristics Clinical Imaging

BLIBioluminescence

luciferase labeled cells /luciferin injection

anatomical, functional, molecular

high throughput, low cost high sensitivity/low resolution

No

Fluorescence fluorescence labeled

molecules or cells (exogenous/intrinsic)

anatomical, functional, molecular

high sensitivity (low cost)

Limited

18FLT-PET injection of 3’-Fluoro-3’deoxythymidine

anatomical, functional

high resolution(high cost)

Yes

Ultrasound interaction of sound waves with living tissues

anatomical, functional

high resolution (vasculature)

Yes

CT n/a anatomical, functional

high resolution (bone, lung)

Yes

Preclinical imaging modalities overview

Target – P-cadherin, CDK4 and Chk1

Cell 1 Cell 2

PlasmaMembrane

Intracellular Space

Cytoplasm

CadherinDimers

actincytoskeleton

AdherensJunction

cell Proliferation

Target gene transcription

Wnt

Β-catenin/Tcf

Cell surface glycoprotein /involved in Ca++-dependent cell-cell adhesion.

Signalling is mediated by β-catenin

In the cytoplasm, β-catenin is stabilized by Wnt signaling and translocates to the nucleus

In the nucleus, β-catenin/partners w/ Tcf factor to regulate expression of oncogenes such as, c-myc, survivn, and cyclin D1 etc.

nucleus

cell invasion

tissue integrity

P-Cadherin target key profile

Bioluminescence

day 33 ProSenseProSenseInjectionInjection

Fluorescence

Normal Lung

SRC Model

Days Post Tumor Implant36 552814

0 25 50 75 100

0

25

50

75

100

0

25

50

75

100

MouseSurvival

Perc

ent s

urvi

val %

of Survival

0 25 50 75 100

Primary TumorGrowth

Days Post Tumor Implant

Tumor Burdenin Lungs

4

8

12

16

0BLI

(pho

tons

/sec

x 1

08 )

Disease progression in MDA-MB-435-HAL SRC model

Primary tumor and lung metastasis can be longitudinally tracked via BLI and FLI

FMT (VisEn)

IVIS200 (Caliper)

PF-03732010 displays antitumor and antimetastatic property

Vehicle PF-2010(20 mg/kg)

MDA-MB-435HAL-CDH3-SRC

Dorsal

Ventral

Vehicle

PF-0373201020 mg/kg

Lymph Node Metastasis

Primary Tumor

PC3M-CDH3

PF-03732010 induces antitumor and antimetastasis property in multiple models

0 50 100 150 200

1

5

20 CTC (hAlu)

PF-0

3732

010

(mg/

kg)

% of Control

H&E Staining

Tumor growth inhibition of PF-03732010 in SC and SRC Model

MDA-MB-435HAL-CDH3

10 20 30 40 50 60

VehiclePF-2010, 40 mg/kgPF-2010, 20 mg/kg

Days Post Tumor Implant

PF-2010, 10 mg/kg

4

8

12

16

BLI

x 1

08 (pho

tons

/sec

)

10 20 30 40 50 600

200

400

600

800

1000 ControlPF2010, 20mg/kg

Days post tumor implant

Tuno

r vo

lum

e (m

m3 )

SRC SC0

30

60

90

120

Vehicle PF3732010

Rel

ativ

e Tu

mor

Siz

e(%

of C

ontr

ol)

SRC implant

SC implant

PF-03732010 displays better efficacy in the MDA-MB-435HAL-CDH3 SRC model than SC model

72 hr

SRC tumorsc tumor

Tumor implanted in sc (right flank) and subrenal capsule (left side)SC injection with P-Cad IgGDrug level reached Cmax in 24 hr IgG distributes to sc and SRC location

24 hr

1 hr

Fluorescence imaging detect the IgG (PF-03732010) distribution

M1 M2 M3

0 168 336 504 672

10

100

1000

10 mg/kg IV10 mg/kg s.c.

T1/2 = ~ 7 daysCL = 0.01ml/min/kgVss = 0.18 L/kg

Time, hr

Seru

m c

once

ntra

tion,

nM

IgG Distribution

LungLive

rKidneySm. In

tSkin

SRC TumorSC Tumor

0

2

4

6

8

10

24 hr72 hr

Inte

nsity

(x 1

09 pho

tons

/sec

)

IVIS200 (Caliper)

PF-201020 mg/kg

Vehicle

β-Catenin Merged Caspase 3

IHC –β-Catenin change - target modulationIncreased apoptosis & decreased proliferation

Ki67

PD modulation with the treatment of P-Cadherin IgG

[18F]FLT-PET ImagingSuppression of [18F]FLT uptake

kidney

Vehicle

PF-2010

CT Imaging[18F]FLT-PET/CT 18FLT-PET

Day 42

tumor

Vehicle

PF-2010

Vehicle

PF-2010 Tumor in SRC Tumor in Lung

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