Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011

PHARMACOLOGICAL INFLUENCE OF SIGNALING

Tímea Berki and Ferenc BoldizsárSignal transduction

Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011

TÁMOP-4.1.2-08/1/A-2009-0011

Potential drug targets in signaling pathways

PIP2 PIP3 PIP2PIP3

PI3K AKTPDK PTEN

Ribosome protein rRNA↑ Translation

S

M

G2G1

↑ Cell cycle

Cyclin D Cyclin A,ECyclin D

Cyclin B

GRB2SOSRAS

RAF

MEK1/2

MAPK1,2,3

GTP GDP

RAS

Specific gene products

Receptor and nonreceptortyrosine kinases

G-proteincoupled receptors

PLC PKC

AC PKA

G protein

ATFc-Jun c-Myc NF-IL6

Nucleus

4-EBP p70S6

mTOR

B

A

C

D

E

F

CDK4, 6 CDK2

CDK1

TÁMOP-4.1.2-08/1/A-2009-0011

Various levels of intervention I

Protein

RNADNA

ProteolysisB ACD

E

F

G

P

Substrate

ATP

TÁMOP-4.1.2-08/1/A-2009-0011

Various levels of intervention II• Blockade of cell surface receptors• Inhibition of signal transmission (eg. kinase

inhibitors)• Interference with the turnover of signaling

proteins (eg. proteosomal degradation, siRNS)

TÁMOP-4.1.2-08/1/A-2009-0011

Blockade of surface receptors with monoclonal antibodiesAntibody Brand name Approval

dateType Target Indication

Adalimumab Humira 2002 Human Inhibition of TNF- signalingα Several auto-immune disorders

Bevacizumab Avastin 2004 HumanizedVascular endothelial growth factor (VEGF)

Colorectal cancerAge related macular degeneration

Cetuximab Erbitux 2004 ChimericEpidermal growth factor receptor

Colorectal cancerHead and neck cancer

Infliximab Remicade 1998 Chimeric Inhibition of TNF- signalingα Several autoimmune disorders

Panitumumab

Vectibix 2006 HumanEpidermal growth factor

receptorColorectal cancer

Ranibizumab Lucentis 2006 HumanizedVascular endothelial growth factor A (VEGF-A)

Macular degeneration

Rituximab Rituxan 1997 Chimeric CD20 Non-Hodgkin lymphoma

Tositumomab Bexxar 2003 Murine CD20 Non-Hodgkin lymphoma

Trastuzumab Herceptin 1998 Humanized ErbB2 Breast cancer

TÁMOP-4.1.2-08/1/A-2009-0011

Selected kinase inhibitors in clinical development

Target Agent Structure Develpoment stage

Growth-factor-receptor inhibitors

EGFR

IMC-C225 cetuximab (Erbitux, Imclone)ABX-EGF (Abgenix)EMD 72000 (Merck KgaA Darmstadt)RH3 (York Medical Bioscience)ZD1839 gefitinib (Iress; AstraZeneca)MDX-447 (Medarex/Merc KgaA)OSI-774 eriotinib (Tarceva; OSI-Pharmaceuticals)CI-1033/PD183805 (Pfizer)EKB-569 (Wyeth Ayerst)GW2016/572016 (GlaxoSmithKLine)

Monoclonal antibodyMonoclonal antibodyMonoclonal antibodyMonoclonal antibodyMonoclonal antibody bivalentSmall-molecule kinase inhibitorSmall-molecule kinase inhibitor

Small-molecule kinase inhibitorSmall-molecule kinase inhibitorSmall-molecule kinase inhibitor

Phase IIIPhase IIPhase IPhase IIPhase IPhase IIIPhase III

Phase IIPhase IPhase I

HER2/neu

Trastuzumab (Herceptin; Genentech)MDX-210 (Medarex/Novartis)2C4 (Genentech)17-AAG (Kosan)

Monoclonal antibodyMonoclonal antibodyMonoclonal antibodyeldananycin derivate inhibits HSP90

RegisteredPhase IPhase IPhase I

PDGFR/c-Kit/BCR-ABL Imatinib (STI571/Gleevec; Novartis) Small-molecule kinase inhibitor Registered

Ras inhibitors Ras

ISIS 2503 (Isis Pharmaceuticals)R115777 (Johnson and Johnson)SCH66336 (Schering-Plough)BMS214662 (Bristol-Myers-Squibb)

Antisense oligonucleotideFarnesyl transferase inhibitorFarnesyl transferase inhibitorFarnesyl transferase inhibitor

Phase IIPhase II/Phase IIIPhase IIPhase II

Raf inhibitors RafISIS 5132/CGP698-46AL-779, 450 (Merck)BAY 43-9006 (Onyx/Bayer)

Antisense oligonucleotideSmall-molecule kinase inhibitorSmall-molecule kinase inhibitor

Phase II

Phase II

MEK inhibitors MEKPD 184352/CI-1040 (Pfizer)U-0126 (Promega)

Small-molecule kinase inhibitorSmall-molecule kinase inhibitor

Phase IIPhase I

mTOR inhibitors mTOR

CCI-779 (Wyeth)RAD001 (Novartis)

Rapamacyn/sirolimus (Wyeth)

Inhibits mTOR kinase by binding to FKBP12Inhibits mTOR kinase by binding to FKBP12

Inhibits mTOR kinase by binding to FKBP12

Phase IIPhase I as a cancer therapeuticPhase II/III as an immunosuppressantRegistered as an immunosuppressant

Cyclin-dependent-kinase inhibitors

CDK

Flavopirodol/HMR-1275 (Aventis)E7070 (EISAI)CYC202 (Cyclacel)BMS-387032 (Bristol-Myers-Squibb)

Small-molecule kinase inhibitorSmall-molecule kinase inhibitorSmall-molecule kinase inhibitorSmall-molecule kinase inhibitor

Phase IIPhase IPhase IPhase I

Other targets and agents

PKC

ISIS 3521/LY900003 Affinitak (Isis Pharmaceuticals)CGP41251PKC412 (Novartis)Bryostatin-1 (GPC Biotek)UCN-01 (Kyowa Hakko Kogyo)

Antisense oligonucleotide

Staurosporine analogueSmall-molecule kinase inhibitorStaurosporine analogue

Phase III

Phase IIPhase IIPhase I/II

PKC-b LY333531 (BI Lilly) Small-molecule kinase inhibitorPhase I oncologyPhase II/III diabetic neuropathy

PDK1 UCN-01 (Kyowa Hakko Kogyo) Staurosporine analogue Phase I/II

TÁMOP-4.1.2-08/1/A-2009-0011

Calcineurin and rapamycin

R

FKBP12

FK506

FKBP12

Cell-cycleTranslation

mTOR

FRB

CyPA

CsA

Cation stressT-cell activation

Calcineurin A

B

CaM

TÁMOP-4.1.2-08/1/A-2009-0011

Rapamycin

Apoptosis

PI3K

AKT

Bcl-2

p70S6KPHASI

P27kip1

G1 S phase

mTOR

?

a gb

IL2

IL2R

R FKBP12

TÁMOP-4.1.2-08/1/A-2009-0011

Toll-like receptor inhibitors

MyD88 TRIF

TLR3TLR7

TLR2

PKA

SP600125SB203580

U0126

PD98059

H-89

Celastrol

Bay11-7082

2-Aminopurine

PepinhTRIFPepinhMYD

TAK1 PKR

p38 JNK

MKKs lkBp50

p65

MyD88

Polymixin B

CLI095

LPS

TLR4

MyD88

MD2LBP

dsRNA

BX795TBK1IKKe

MDA-5RIG-1

IPS1

TLR9

Chloroquine

JAK2

Wortmannin

Rapamycin

LY294002

AG490

mTOR

PI3K

CD14

OxPAPC

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ERB signaling intervention

Tyrosine kinase inhibitors(e. g. tryphostins)

Unfolded, inactiveErbB dimer

ErbB dimerLigand

Hsp90 inhibitors(e. g. geldanamycin)

Hsp90Phosphoproteins and

downstream signalling events

Anti-ErbB antibody(e. g. Herceptin)

Nucleus

ErbB gene

Immature ErbB

ER/Golgi

Translation

Triplex-forming oligos,antisense oligos

Ribozymes

Transcription

scFvs

TÁMOP-4.1.2-08/1/A-2009-0011

Proteosome inhibitors-Bortezomib

Bortezomib

Cytokines activatecell-surface receptors

↓ Cytokines

P105 precursorprocessed

Proteasome

Degraded IκB

Activated NF-kB translocatesto the nucleus

↓ Anti-apoptoticfactors

Activation of NF-kBby degradation of IkB

↓ Inflammatorymolecules

lkBp50

p65

p105

↓ Cell adhesionmolecules

p50

lkBp65

p50

p65

Nucleus

NF-kB activatestranscription

p50

p65NF-kB

O H OH

N

N

O

N BNH

OH

TÁMOP-4.1.2-08/1/A-2009-0011

Apoptosis signaling intervention

Chemotherapy

DNA damage

Bcl-2 inhibitors

IAP inhibitors

TRAIL

DR4/DR5

HDAC inhibitors

Bcl-2Bcl-XL

Bfl-1/A1c-FLIPDcR1

Proteasome

c-FLIP

Caspase-9Cytc

Caspase-8

APOPTOSIS

Bcl-2 Bcl-xL

BakBax

c-IAP1c-IAP2XIAP

p53

tBid Pro-caspase-3

NF-kB

IAPs

lkBP

P

Caspase-3

Bid

Proteasomeinhibitors

lkB

NF-kB

FADD

Smac/Diablo

TÁMOP-4.1.2-08/1/A-2009-0011

HSP-90 inhibitors

Ubiquitin ligase CHIP

Proteasome

Ubiquitin proteasome-dependent degradation

Early complex

HSP40HSP70

CLIENT

Intermediate complex

HSP90

HSP90

HSP40HSP70

HOPCLIENT

CLIENTUBUB UBUBUB

HSP90

HSP90

Geldanamycin binds to theATP-binding site of HSP90

ATPADP

Mature complex

HSP90

HSP90

CLIENTCDC37

p23Immunophilin

HARCAH1

+