Preclinical Models for Developing Therapy for Pediatric Solid Tumors Preclinical Models for Developing Therapy for Pediatric Solid Tumors uses and limitations

Preclinical Models for Developing Therapy for Pediatric Solid Tumors uses and limitationsPeter J. Houghton, Ph.D.Solid Malignancies ProgramSt. Jude Childrens Research Hospital

Problems in using Preclinical DataEarly drug discovery is conducted under defined/standardized environment.Pediatric cancer models are not part of this process either in industry or NCI.Preclinical data using pediatric models are generated in an uncontrolled or regulated environment.Such data are derived from experimental systems that are not validated using experimental design and interpretation that lacks consistency or rigor.

Informative Non-Clinical DataPharmacology and pharmacokineticsSafetyEfficacyBehaviourLong term effectsDevelopmental aspectsOther? Pharmacodynamics

Uses:Where Do Animal Models Fit in Drug Development for Childhood Cancer?Drug acquired NCI/Industry/AcademiaPhase IPhase IIIdentification of active agents/analogsOptimization of administration schedules and drug combinations.Prioritization of agents for phase IRational decisions to advance/stop developmentPotential to focus phase II trialsPotential to relate target inhibition to biological response

Models Identify Clinically Active Agents Prospectively

RHABDOMYOSARCOMACOLON CARCINOMA

DRUG

XENOGRAFTS (%)

CLINICAL (%)

VINCRISTINE

78

59

CYTOXAN

44

54

DACTINOMYCIN

11

24

ADRIAMYCIN

19

31

MELPHALAN

83

81

DRUG

XENOGRAFTS (%)

CLINICAL (%)

VINCRISTINE

0

SENSITIVITY OF WILMS TUMOR XENOGRAFTS

TUMOR

VINCRISTINE

CYTOXAN

TOPOTECAN

IRINOTECAN

WT1

++++++

++

++++++

++++++

WT7

++++++

++++++

++++++

++++++

WT8

++++++

++++

++++++

++++++

WT9

++++++

+++

++++++

++++++

WT10

++++++

++++++

++++++

++++++

SKNEP

++

+++++

++

++

++ PROGRESSIVE DISEASE

+++ STABLE DISEASE

++++ PARTIAL REGRESSION

+++++ COMPLETE REGRESSION WITH REGROWTH

++++++ COMPLETE REGRESSION W/O REGROWTH

Topotecan Lactone AUC Associated with Response in Neuroblastoma Xenografts0255075100125150AUC (ng-hr/mL)NB1382.2NB1643NBEBNB1771NB1691PRCR290

Targeted Topotecan Neuroblastoma Protocol NB97113 Courses; 92% Targeting Success Results 28 evaluable patients TPT median dose 3.0 mg/m2 17 partial responses (60%) 11 stable disease no tumor progressionsPK StudiesDose Adjustment Schema (100 20 ng*hr/ml) Day 1 2 3 4 5 6 7 8 9 10 11 12TPT TPT TPT TPT TPT X X TPT TPT TPT TPT TPTDoseAdjustSantana et al.(submitted)

Retrospective Analysis of Response-Exposure Relationships for Pediatric Tumors XenograftsDrugs that failedDrugs that worked

Drug

AUC@Mouse MTD

AUC@Human MTD

Effective dose range (fold from MTD)

DMP840

15-20

~2-3

Carzelesin

~80

10

Irinotecan (SN38)

~16

>100

Evaluation of MGI-114(Phase II in COG?)Systemic exposure is still > 10-fold higher than in children @MTD

Tumor

Dose level (mg/kg)

7.0

4.6

3.0

2.0

1.3

DAOY

++++++

++++++

++++++

+++++

++

D283

++++++

++++++

++++++

+++

+

SJ-BT27

ND

++++++

++++++

+++

+

SJ-BT29

++++++

+++

+++

ND

ND

SJ-BT33

-

-

-

-

-

SJ-BT34

++++++

++++++

++++++

ND

ND

SJ-BT36

++++++

++++++

++++

+++

+++

SJ-BT37

++++++

+++++

+++

+

-

SJ-BT40

++++

++++

++

+

-

SJ-BT12

ND

++++

+/-

-

-

SJ-BT16

+++

++++

+++

++

+

SJ-GBM2

++++++

++++++

+

ND

ND

NB-1771

ND

ND

+

-

-

NB-1382

++++

++++

+++

ND

ND

NB-1643

++++++

++++++

++++

ND

ND

NB-1691

+++++

+++

++

+

ND

Rh18

++++++

++++

++++

+++

++

Rh28

++++++

++++++

++++++

+++++

+++

Rh30

ND

ND

ND

++++++

++++++

Rh36

++++

+++

++

+

-

CR+PR/N

14/16

14 /18

8/18

3 /15

1/14

Schedule-dependent antitumor activity of topotecanControlTPT (daily x 5) every 21 days Total dose = 22.5 mg/kgTPT (daily x 5) x 2 every 21 days Total dose = 22.5 mg/kgTumor volume (cm3)1010.1WeekWeek024681012Week468021210024681012

Discriminating Between Analogs: Osteosarcoma Models

The Challenge of Molecularly-Targeted DrugsTumor models must accurately recapitulate activity of signal transduction pathways.-orthotopic or subcutaneous?- expression profiling- proteomics profiling

WT6 vs parentWT8 vs parentXENOGRAFTXENOGRAFTPRIMARY TUMORPRIMARY TUMOR

P. Hedge et al. ASCO #535AGSK/Cytokinetics

Evaluation of SB713489 Study #Tumor15 mg/kg10 mg/kg5 mg/kgCR/PR@ 15mg/kgToxdeaths1998SKNEP+--0/50/51999SKNEP+++--2/51/52005SJ-WT7+++++++++5/50/52009SJ-WT7++++++: CR with regrowth ++++: all tumors have partial regression (PR > 50% volume reduction) +++: stable disease ++: growth delay ~ 2 tumor volume doubling times +: growth delay ~ 1 tumor volume doubling time -: no growth inhibition

Study 2026SJ-WT10

A

Study #:2026

Tumor/Passage:SJ-WT10p9

Transplant Date:3/27/03

Treatment:Control

Technician:JDC

Data Set #1Day0# of mice =5% Survival

Date5/11/99wt of mice=118100

MouseDiameterVolume% ChangeRTV>4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X4X0.000.000.000.000.000.000.000.000.000.00

% Survival

WeeksABCDEFGHIJ

0.00100.00100.00100.00100.00100.00100.00100.00100.00100.00100.00

1.00100.00100.00100.00100.000.000.000.000.000.000.00

2.00100.00100.00100.00100.000.000.000.000.000.000.00

3.000.00100.00100.00100.000.000.000.000.000.000.00

4.000.00100.00100.000.000.000.000.000.000.000.00

5.000.00100.00100.000.000.000.000.000.000.000.00

6.000.00100.00100.000.000.000.000.000.000.000.00

7.000.00100.000.000.000.000.000.000.000.000.00

8.000.00100.000.000.000.000.000.000.000.000.00

9.000.00100.000.000.000.000.000.000.000.000.00

10.000.00100.000.000.000.000.000.000.000.000.00

11.000.00100.000.000.000.000.000.000.000.000.00

12.000.00100.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

0.000.000.000.000.000.000.000.000.000.000.00

Tumor Volume Group A

Week#1#2#3#4#5

0.000.980.160.221.040.57

1.002.210.390.472.111.44

2.004.721.031.294.632.81

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group B

Week#1#2#3#4#5

0.000.530.230.240.221.07

1.000.230.050.060.090.39

2.000.000.000.000.000.12

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.050.000.000.000.00

10.000.130.000.000.070.00

11.000.380.000.000.220.06

12.000.590.000.000.310.03

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group C

Week#1#2#3#4#5

0.000.360.130.610.261.17

1.000.300.150.670.290.83

2.000.240.110.450.250.89

3.000.570.261.080.662.47

4.000.900.141.751.082.54

5.001.380.201.900.002.60

6.002.425.653.170.005.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group D

Week#1#2#3#4#5

0.000.350.230.210.230.45

1.000.410.460.140.320.65

2.000.891.000.370.911.89

3.002.870.001.080.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group E

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group F

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group G

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group H

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group I

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

Tumor Volume Group J

Week#1#2#3#4#5

0.000.010.010.010.010.01

1.000.000.000.000.000.00

2.000.000.000.000.000.00

3.000.000.000.000.000.00

4.000.000.000.000.000.00

5.000.000.000.000.000.00

6.000.000.000.000.000.00

7.000.000.000.000.000.00

8.000.000.000.000.000.00

9.000.000.000.000.000.00

10.000.000.000.000.000.00

11.000.000.000.000.000.00

12.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

0.000.000.000.000.000.00

&A

Page &P

graph1

&F

graph1

100100100100100100100100100100

100100100100111111

100100100100222222

3100100100333333

41001004444444

51001005555555

61001006666666

710077777777

810088888888

910099999999

101001010101010101010

111001111111111111111

121001212121212121212

0000000000

0000000000

0000000000

0000000000

0000000000

0000000000

0000000000

weeks

% Survival

% Survival

graph 2

0.98269127330.15712720380.21737826611.04241718210.5746551919

2.20760070930.39262182080.47272134542.1086775621.4413815018

4.71862950011.03003706061.28968427174.63410866772.80616875

33333

44444

55555

66666

77777

88888

99999

1010101010

1111111111

1212121212

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor volume

Control

0.53070045640.23121146260.24278537380.2165052621.0687375106

0.22713668150.05349885450.05808711590.09493993530.3939197432

0.00000052360.00000052360.00000052360.00000052360.1188472516

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.00000052360.00000052360.00000052360.00000052360.0000005236

0.04707969930.00000052360.00000052360.00000052360.0000005236

0.12813907360.00000052360.00000052360.07404791050.0000005236

0.38361610460.00000052360.00000052360.21606963790.0579783838

0.58982868750.00000052360.00000052360.31367677320.0325151076

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

SB713489 15 mg/kg (q4dx3)2 repeat cycles at 21 day intervals

0.35742729860.12999165830.61047228410.25668750631.1703741756

0.29561618740.14545919710.67063947570.29400958120.8332403748

0.23997321620.10699057520.45034068980.25039168170.8856900748

0.57215185720.26308799061.08015193710.65501534512.4738622149

0.89801189450.1428004511.74953733961.08397472852.536300539

1.3775547570.19706578631.895610476352.5974955315

2.42334692465.64783051533.169583875664.9959909549

77777

88888

99999

1010101010

1111111111

1212121212

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume


0.35257815370.23395487930.2100321180.22713668150.4475053947

0.4091186320.45819944730.13854942320.31535418560.6450360599

0.88903931941.00072553680.36725856610.91499780221.8882129804

2.869171939431.082699464233

44444

55555

66666

77777

88888

99999

1010101010

1111111111

1212121212

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume


A

B

C

D

E

F

G

H

I

J

weeks

Relative Tumor Volume

RTV

0.00001

0.00001

0.00001

0.00001

0.00001

0.00001

0.00001

0.00001

0.00001

0.00001

1

1

1

1

1

1

1

1

1

1

2

2

2

2

2

2

2

2

2

2

3

3

3

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5

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5

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11

11

11

11

11

11

11

11

11

11

12

12

12

12

12

12

12

12

12

12

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

1

1

1

1

1

1

1

1

1

2.2902024114

0.3411454407

0.9803912412

1.3216619338

5.3237684847

0.0222422465

0.7928741305

3.3590744101

0.0000016632

2.0119083446

6.6463069813

0.0000016632

2.5733726842

0.0000016632

2.6736494104

0.0000016632

14.9220853668

0.0000016632

0.0000016632

0.0177439414

0.1166944018

0.3550180813

0.5181323927

&F

0.010.010.010.010.01

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

Tumor Group F

0.010.010.010.010.01

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

Tumor Group G

0.010.010.010.010.01

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

Tumor Group H

0.010.010.010.010.01

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

Tumor Group I

0.010.010.010.010.01

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

00000

weeks

Tumor Volume

Tumor Group J

weeks

Tumor Volume

Tumor Group E

0.00001

0.00001

0.00001

0.00001

0.00001

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0.01

0.01

0.01

0.01

0.01

Informative Non-Clinical DataPharmacology and pharmacokineticsSafetyEfficacyBehaviourLong term effectsDevelopmental aspectsOther? Pharmacodynamics

PI-3KTargets of Cap Dependent TranslationCyclin D1ODC1MYCHIF1a

Aktp70S6KmTORIRS-1PDK1/2PTENGTPGDPmLST8RaptorIGF-IITSC1TSC2AMPKAMPLow Energy (High AMP)Low Amino Acid PoolsTOP Dependent Translation?Increased translationGTPLKB1Rapamycin (CCI-779)TARGET INHIBITIONIGF-IIBIOLOGICALREADOUTPHARMACOKINETICS

Developing Initiatives for a National Consortium to Identify and Prioritize New Agents

RFA to systematically characterize models through genomic/proteomic screens to identify potential molecular targets (POPP-TAP).

RFP to establish preclinical screening program that will identify agents having high priority for pediatric Phase I testing through the Childrens Oncology Group.

Effective use of Non-clinical DataStandardization of experimental procedures: difficult, but a realistic goal.Standardization of acceptable criteria for assessing drug activity.GLP compliance: very expensive, most academic centers would be excluded.

Documents

Preclinical Models for Developing Therapy for Pediatric Solid Tumors Preclinical Models for Developing Therapy for Pediatric Solid Tumors uses and limitations