Tyrosine Kinase Inhibitors

Introduction
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Cancer:
2
nd
leading cause of early death in Canada

1 in 3 Canadians will develop cancer

New Emerging Class of Anticancer Drugs:
Tyrosine Kinase Inhibitors

Canadian Cancer Society, 2001

Overview
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

What are tyrosine kinases?

What is their role in cancer?

Can their activity be inhibited?

Are the inhibitors effective?

How will they affect cancer treatment?


Tyrosine Kinases
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

What are Tyrosine Kinases?
*Not Really
Photo Courtesy of NBC.com
Tyrosine Kinases- Introduction
Tyrosine Kinases (TK)
Enzymes
Phosphorylate Tyrosine (Y) residues
Adenosine-PPP
(ATP)
Adenosine-PP
(ADP)
Y Y Y
P
Substrate
TK
Phosphorylated
Substrate
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Tyrosine Kinases- Introduction
Transduce signals that direct
Growth
Division
Migration
Synthesis
Apoptosis

Major classification as:
1- Receptor Tyrosine Kinases (RTKs)
2- Protein Tyrosine Kinases (PTKs)
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Receptor Tyrosine Kinases (RTKs)
Cell surface receptors


Extracellular to Intracellular signalling


Families:
Insulin receptors
Growth factor receptors

1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure
Structurally:
Extracellular
Cell Membrane
Intracellular
Ligand Binding Domain
Transmembrane Domain
Juxtamembrane Domain
Kinase Domain
Carboxy-Terminal Tail
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure & Function

Extracellular
Cell Membrane
Intracellular
Ligand Binding Domain
Typically glycosylated
Basis for RTK categorization


1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure & Function

Extracellular
Cell Membrane
Intracellular
Transmembrane Domain
Single transmembrane pass
Alpha helix
Membrane anchor

1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure & Function

Extracellular
Cell Membrane
Intracellular
Juxtamembrane Domain
Tyrosine residues:
autophosphorylated
Substrate recruitment
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure & Function

Extracellular
Cell Membrane
Intracellular
Kinase Domain
Highly conserved catalytic
region
ATP & Substrate binding
sites
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Structure & Function

Extracellular
Cell Membrane
Intracellular
Carboxy-Terminal Tail
Tyrosine residues:
autophosphorylated
Substrate recruitment
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Adapted from Hubbard,1999
Epidermal Growth Factor Receptor & Family
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Insulin Receptor & Family
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Platelet-Derived Growth Factor Receptor& Family
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Vascular Endothelial Growth Factor Receptor & Family
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Fibroblast Growth Factor Receptor & Family
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Hepatocyte Growth Factor Receptor & Family
Adapted from Hubbard,1999
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Some Major RTK Families
Ret Receptor (Orphan)
RTK Activation & Signal Transduction
Ligand
1.
Inactive Receptors
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
Ligand
1.
Inactive Receptors
2.
Receptor
Dimerization
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
Ligand
P
P
P
P
P
P
1.
Inactive Receptors
2.
Receptor
Dimerization
3.
Receptor
Autophosphorylation
and Activation
Extracellular
Intracellular
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
P
P
P
P
P
P
Extracellular
Intracellular
Ligand
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
P
P
P
P
P
P
Extracellular
Intracellular
4.
Intracellular
signalling
molecule
Ligand
Y
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
P
P
P
P
P
P
Extracellular
Intracellular
4.
Intracellular
signalling
molecule
5.
Recruitment &
phosphorylation of
signalling molecule
Ligand
Y
Y
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
P
P
P
P
P
P
Extracellular
Intracellular
4.
Intracellular
signalling
molecule
5.
Recruitment &
phosphorylation of
signalling molecule
Ligand
ATP ADP
Y
Y
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
P
P
P
P
P
P
Extracellular
Intracellular
4.
Intracellular
signalling
molecule
5.
Recruitment &
phosphorylation of
signalling molecule
6.
Activation of
signalling molecule
& downstream
signalling pathways
P
Ligand
ATP ADP
Y
Y
Y
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK Activation & Signal Transduction
Extracellular
Intracellular
Nucleus
Transcription
Factors
Proliferation, Differentiation,
Migration, Metabolism
JAK
STAT
Homo/Heterodimer
(MAP Kinases)
Ras
Raf
MEK
ERK
PI-3K
PKB
FKHR1 BAD
Apoptosis
PLC
IP
3

DAG
Ca
++
PKC
Adapted from Schlessinger, 2000
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTK: Key Concept
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Transduce extracellular signals to
intracellular signalling pathways

Achieved by:
Extracellular ligand binding

Phosphorylation of signalling molecules

Activation of intracellular signalling
pathways
Cytoplasmic proteins

Transduce signals to downstream
signalling pathways





9 Families,
Structural and functional characteristics

Protein Tyrosine Kinases (PTKs)
ATP ADP
Y Y Y
P
Substrate
TK
Phosphorylated
Substrate
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Src Family
Mitogenesis, T- and B- cell activation
PDGFR & EGFR

JAK (Janus Kinase) Family
Association with all Cytokine receptors
Activates STATs (Signal transducers and activators of
transcription)


Other Families: FAK, Abl, Csk,
Syk/ZAP70, Ack
Some Notable PTK Families
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Basic structure:
PTK Structure
N-terminal region
Association Domain(s)
Kinase Domain
C-Terminal region
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Basic structure:
PTK Structure
Association Domain(s)
PTK-Protein, PTK-Lipid, PTK-DNA
SH2 (Src Homology 2), SH3 (Src Homology 3),
PTB (Phosphotyrosine Binding), PH (Pleckstrin
Homology) domains
Recruited to Y- P
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Basic structure:
PTK Structure
Kinase Domain
Catalytic region
ATP & Substrate binding
sites
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Similar to RTKs, generally:

1- Recruited to upstream signalling molecule
RTK, Protein Kinases, GPCR (G-Protein Coupled
Receptor)

2- Activated by
phosphorylation/dephosphorylation

3- Phosphorylate downstream signalling
molecule at Tyrosine residue
PTK Activation
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

PTK Signal Transduction
Nucleus
Transcription
Factors
Proliferation, Differentiation,
Migration, Metabolism
STAT
Homo/Heterodimer
(MAP Kinases)
Ras
Raf
MEK
ERK
PI-3K
PKB
FKHR1 BAD
Apoptosis
Adapted from Schlessinger, 2000
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

RTKs, GPCRs,
Protein Kinases,
Ca
++

PTK: Key Concept
1. Introduction
2. Overview
3. Tyrosine Kinases
- Introduction
- Receptor TKs
- Protein TKs
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Transduce upstream signals to
intracellular signalling pathways

Achieved by:
Recruitment & activation

PTK phosphorylation of downstream
substrate

Transmission of intracellular signalling
pathway
What is their role to they play in cancer?
Tyrosine Kinases & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

*Not Really
Photo Courtesy of NBC.com
Cancer:
Disease of uncontrolled cellular growth and
proliferation
Inherited abnormalities and/or mutations


Growth & proliferation signals:
Potentially oncogenic


Tyrosine Kinases & Cancer- Introduction
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Genetic TK mutations: constitutive activity

Diseases caused by TK mutation:

Chronic myelogenous leukemia (CML)
Abl-BCR PTK fusion protein

Anaplastic large cell lymphoma
ALK-NPM RTK fusion protein

Glioblastoma multiforme (human brain tumor)
EGFR RTK mutation
Mutation & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Genetic TK mutations: constitutive activity

Diseases caused by TK mutation:

Chronic myelogenous leukemia (CML)
Abl-BCR PTK fusion protein

Anaplastic large cell lymphoma
ALK-NPM RTK fusion protein

Glioblastoma multiforme (human brain tumor)
EGFR RTK mutation
Mutation & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

CML: 20% Adult leukemia
95% of CML: Chromosomal Translocation
Chromosome 22: Breakpoint cluster region (BCR)
Chromosome 9: Abl PTK






BCR-Abl fusion protein: constitutively active PTK
Chronic Myelogenous Leukemia (CML)
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from Whittaker &
Skeffington, 2000
Auto-stimulation of cell

Autocrine Loops & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Nucleus
GF synthesis
Proliferation
Receptor and/or GF over-
expression

Cancer
EGF-EGFR loop: Breast,
Bladder, Lung, Brain
PDGF-PDGFR loop:
Brain, Vasculature
IGF1-IGF1R loop: Breast,
Lung, Prostate

Auto-stimulation of cell

Autocrine Loops & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Nucleus
GF synthesis
Proliferation
Receptor and/or GF over-
expression

Cancer
EGF-EGFR loop: Breast,
Bladder, Lung, Brain
PDGF-PDGFR loop:
Brain, Vasculature
IGF1-IGF1R loop: Breast,
Lung, Prostate

erbB-2 or HER-2/NEU
EGFR family members

Low expression in normal breast tissue

Over-expression >65% ductal carcinomas
Confers anti-estrogen (Tamoxifen) resistance
not expressed in benign breast tumors

Constitutive activity:
Single point mutation in transmembrane
domain
EGF-EGFR in Breast Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Angiogenesis:
Generation & growth of new blood vessels

Tumor growth limited to 1-2mm
3
In absence of blood supply

Tumor: secretes growth factors
Response to hypoxia
Promote angiogenesis

VEGFs, FGFs, PDGFs
Angiogenesis & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
- Introduction
- TK Mutation
- Autocrine Loops
- Angiogenesis
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Cancer is disease of uncontrolled
proliferation

TKs transduce cellular proliferation signals

Increased TK activity may be oncogenic

TKs required for angiogenesis


Key Concept: Tyrosine Kinases & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Can their activity be inhibited?
Tyrosine Kinases & Cancer
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

*Not Really
Photo Courtesy of NBC.com
Tyrosine Kinase Inhibitors
Normal kinase activity
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

N-Terminal Lobe
C-Terminal Lobe
Lys
Adenosine-PPP
Substrate
Substrate
P
Kinase Domain
Tyrosine Kinase Inhibitors
Compete for ATP binding site
Inhibit P transfer to substrate tyrosine







No signal transduction
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

N-Terminal Lobe
C-Terminal Lobe
Lys
TK Inhibitor
Substrate
Kinase Domain
Adenosine-PPP
Tyrosine Kinase Inhibitors
Promising Tyrosine Kinase Inhibitors:
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from Morin, 2000
Key Concept: Tyrosine Kinase Inhibitors
Synthetic molecules

Compete for ATP binding site of kinase
domain

Inhibit phosphorylation of downstream
signalling molecule
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Preclinical Results: TK Inhibitors
Are the inhibitors effective?
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

*Not Really
Photo Courtesy of NBC.com
Preclinical Results: STI571 (Gleevec
TM
)
PDGFR & BCR-Abl selective inhibitor
CML & GI Stromal tumor treatment

in vitro apoptosis results: BCR-Abl inhibition
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
-STI571
-ZD1839
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from: Mow et al., 2002
Preclinical Results: STI571 (Gleevec
TM
)
In vivo (mice) tumor growth results:
PDGFR inhibition
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
-STI571
-ZD1839
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from: Sjblom, et al., 2001
Preclinical Results: ZD1839 (Iressa
TM
)
EGFR selective inhibitor
Affinity for erbB2

in vitro growth results:
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
-STI571
-ZD1839
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from: Moulder, et al., 2001
Preclinical Results: ZD1839 (Iressa
TM
)
In vivo (mice) tumor growth:
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
-STI571
-ZD1839
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Modified from: Moulder, et al., 2001
Clinical Results: TK Inhibitors
How will they affect cancer treatment?
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

*Not Really
Photo Courtesy of NBC.com
Clinical Results: Imatinib mesylate (Gleevec
TM
)
Pharmacokinetics:
400mg or 600mg once/day

98% orally bioavailable


Elimination :

P450 CYP3A4 major metabolizing enzyme


1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
- Imatinib
mesylate
-ZD1839
8. Other Notables
9. Summary &
Conclusion

Clinical Results: Imatinib mesylate (Gleevec
TM
)
Myeloid Leukemia Study- Phase III Trial
532 patients with phase CML
400mg/day

60% Major Hematologic response
No cells with Philadelphia Chromosome

95% Complete Response
White blood cell count < 10
10
cells/L


1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
- Imatinib
mesylate
-ZD1839
8. Other Notables
9. Summary &
Conclusion

Clinical Results: ZD1839 (Iressa
TM
)
Pharmacokinetics- Phase I trail
300mg, 400mg, & 525mg once/day
Antitumor effects below dose-limiting
toxicity (>700mg/day)


Orally bioavailable

No observed renal or cardiac toxicity

Phase II & III trials ongoing
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
- Iminitab
mesylate
-ZD1839
8. Other Notables
9. Summary &
Conclusion

Clinical Results: ZD1839 (Iressa
TM
)
Antitumor effects:
Phase I - difficult to assess, however:







Promising...

1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
- Iminitab
mesylate
-ZD1839
8. Other Notables
9. Summary &
Conclusion

Ranson et al., 2002
Other Notables
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Anything Else?
*Not Really
Photo Courtesy of NBC.com
Other Notables
1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Resistance



Herceptin
TM
(Trastuzumat):
FDA approved
Humanized monoclonal antibody
Selective for erbB-2/HER-2/NEU
Treatment erbB-2 metastatic breast cancer



Serine/Threonine Kinases
Summary
TKs transduce signalling events
Phosphorylating substrates
Signals direct: growth, proliferation,
migration, synthesis

Increases in TK activity can be oncogenic

TK inhibitors compete for ATP site
Impede Phosphate transfer to substrate

Promising clinical data

1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

Conclusion
Promising Anticancer Drugs


1 already on market (Gleevec
TM
)


Many in development


A cure for cancer in 10 years?


1. Introduction
2. Overview
3. Tyrosine Kinases
4. Tyrosine Kinases
& Cancer
5. Inhibitors
6. Preclinical
7. Clinical
8. Other Notables
9. Summary &
Conclusion

References
Tyrosine Kinase Structure, Activation & Function

Carrera, A.C., Alexandrov, K., & Roberts, T.M. (1993) The conserved lysine
of the catalytic domain of protein kinases is actively involved in the
phosphotransfer reaction and not required for anchoring ATP. Proc. Natl.
Acad. Sci. USA 90: 442-46
Hubbard, S.R., & Hill, J.H. (2000) Protein tyrosine kinase structure. Ann Rev
Biochem 69:373-98
Hubbard, S.R. (1999) Structural analysis of receptor tyrosine kinases. Prog
Biophys Mol Biol 71: 343-58
Hubbard, S.R., Mohammadi, M., & Schlessinger, J. (1998) Autoregulatory
mechanisms in protein-tyrosine kinases. J. Biol. Chem. 273(20): 11987-90
Ihle, J.N. (1995) Cytokine receptor signalling. Nature 377: 591-94
Mayer, B.J., & Baltimore, D. (1993) Signalling through SH2 and SH3
domains. TiCB 3: 8-13
References (2)
Tyrosine Kinase Structure, Activation & Function

Mohammadi, M., McMahon, G., Sun, L., Tang, C., Hirth, P., Yen, B.K.,
Hubbard, R., & Schlessinger, J. (1997) Structures of the tyrosine kinase
domain of fibroblast growth factor receptor in complex with inhibitors.
Science 276: 955-960
Schenk, P, & Snaar-Jagalska, B.E. (1999) Signal perception and transduction:
the role of protein kinases. Biochim Biophys Acta: 1-24
Schlessinger, J. (2000) Cell signalling by receptor tyrosine kinases. Cell 103:
211-225
Ullrich, A., & Schlessinger, J. (1990) Signal transduction by receptors with
tyrosine kinase activity. Cell 61: 203-212
Weiss, A., & Schlessinger, J. (1998) Switching signals on or off by receptor
dimerization. Cell 94: 277-80
References (3)
Tyrosine Kinases & Cancer

Canadian Cancer Society (2001) General Facts. www.cancer.ca
Kolibaba, K.S. & Druker, B.J. (1997) Protein tyrosine kinases and cancer.
Biochim Biophys Acta 1333: F217-48
Kumar, R., & Yarmad-Bagheri, R. (2001) The role of HER2 in angiogenesis.
Sem Oncol 25(5 suppl 16): 27-32
Morin, M.J. (2000) From oncogene to drug: development of small molecule
tyrosine kinase inhibitors as anti-tumor and anti-angiogenic agents. Oncogene
19: 6574-83
Noonberg, S.B., & Benz, C.C. (2000) EGFR tyrosine kinase inhibitors as
anticancer agents. Drugs 59(4): 753-764
Whittaker, E., Skeffington, B.S., & McCann, S. (2000) CLM, a paradigm of
malignancy: new developments in the treatments of haematological
malignancies. TSMJ 1: 22-24
References (4)
Tyrosine Kinases Inhibitors

Moulder, S.L., Yakes, F.M., Muthuswamy, S.K., Bianco, R., Simpson, J.F., &
Arteaga, C.L. (2001) Epidermal growth factor receptor (HER1) tyrosine
kinase inhibitor ZD 1839 (Iressa) inhibits HER2/neu (erbB2)-overexpressing
breast cancer cells in vitro and in vivo. Canc Res 61: 8887-95
Mow, B.M.F., Chandra, J., Svingen, P.A., Hallgren, C.G., Weisberg, E.,
Kottke, T.J., Narayanan, V.L., Litzow, M.R., Griffin, J.D., Sausville, E.A.,
Tefferi, A., & Kaufmann, S.A. (2002) Effects of the Bcr/abl kinase inhibitors
STI571 and adaphostin NSC 680410 on chronic myelogenous leukemia cells
in vitro. Blood 99(22): 664-71
Novartis (2001) Gleevec
TM
.

www.novartis.com
Ranson, M., Hammond, L.A., Ferry, D., Kris, M., Tullo, A., Murray, P.I.,
Miller, V., Averbuch, S., Ochs, J., Morris, C., Feyereislova, A., Swaisland, H.,
& Rowinski, E.K. (2002) ZD1839, a selective oral epidermal growth factor
receptor-tyrosine kinase inhibitor, is well tolerated and active in patients with
solid, malignant tumors: Results of a phase I trial. J Clin Oncol 20(9): May 1
References (5)
Tyrosine Kinases Inhibitors

Sjoblom, R., Shimizu, A., OBrien, K.P., Pictras, K., Cin, P.D., Buchdunger,
E., Dumanski, J.P., Ostman, A., & Heldin, C.-H. (2001) Growth inhibition of
Dermatofibrosarcoma Protuberans tumors by the platelet-derived growth
factro receptor antagonist STI571 through induction of apoptosis. Canc Res
61: 5778-83
Sun, L., & McMahon, G. (2000) Inhibition o tumor angiogenesis by synthetic
receptor tyrosine kinase inhibitors. Drug Dis. Today 5(8): 344-53
Thiesing, J.T., Ohno-Jones, S., Kolibaba, K.S., & Druker, B.J. (2000) Efficacy
of STI571, and Abl tyrosine kinase inhibitor, in conjunction with other
antileukemic agents against Bcr-Abl-positive cells. Blood 96(9): 3195-99