UNIVERSITA’ DEGLI STUDI DI PALERMO

FACOLTA’ DI FARMACIA

Design, synthesis and biological evaluation of new inhibitors of

carcinogenic processes

DIPARTIMENTO FARMACOCHIMICO TOSSICOLOGICO E BIOLOGICO

Dott. Ilenia Abbate

2

Cancer cells are less sensitive to the therapy and repopulate the tumor, facilitating tumor progression

Migratory growth factorMigratory growth factor

PROTO-ONCOGENEPROTO-ONCOGENE

STAT3c-MYC

growth factor-growth factor-induced induced

mitogenesismitogenesis

RasERK

MERKCyclin/cdks

promotes G1/S phasepromotes G1/S phase

PI 3 kinaseAkt

survival survival

SIGNAL-TRANSDUCTION PATHWAYS CORRELATED TO TUMOR PROGRESSION

Without fuctional HSP90, these proteins undergo proteasome-mediated degradation, leading to cell cycle arrest and apoptosis.

4

HSP90 chaperone stabilizes

oncoproteins

Inhibiting HSP90 degradesoncoproteins, stopping

tumor growth

5

ability to evade apoptosis ability to be self-sufficient for growth ability to invade surrounding tissue and to metastasize to distant sites

Multiple HSP90 client proteins mediate acquisition and maintenance of the six properties necessary for transformation of a normal cell into a cancer cell:

ability to undergo limitless replicationability to promote neoangiogenesis ability not to respond to antigrowth signals.

•Folding

•Biological fuction

•Stabilization/Degradation

•Activation

•Folding

•Biological fuction

•Stabilization/Degradation

•Activation

CLIENTS

PROTEINS

•Protein kinases (Her-2/ErbB2 e Akt)•Cdk4/ciclin D complex•c-Raf-1•HIF-1α•p53•Steroid hormone receptors

HSP90 stress condition: heat, irradiation, ROS (reactive oxygen radicals), toxins, lack of

nutrients, hypoxia, bacterial and viral infections.

-C-terminal domain : 10kDa, omodimerization and co-chaperones recruitment super-chaperone complex;

-middle region: 35kDa, binding site for client proteins and ATPase activity;

-N-terminal domain: 25kDa, binding site for ATP/ADP and inhibitors, such as geldanamycin and radicicol.

OH

HO

Cl

O

O

O

O

Me

O

O

NH

O

Me

MeO

OCONH2

MeMeO

Me

Me

OH

R

Geldanamicina R= OMe17 DMAD R= NHCH2CH2NMe2

17 AAG R= NHCH=CH2

N

N

NH2

RX

Y

OMe

OMeOMe

O

O

OO

HO

OH

O

O

HO

OH

OH

Derrubone

Apigenina

PU3 X=Y=H R= C4H9

PU24FCl X=F Y=Cl R= (CH2)3C CH

Radicicolo

N N

H

R4OH

HOR1

R2

R3

G3129 Et - O- CH2- O- COOH

CCT018159 Et - O- (CH2)2- O- Me

VER- 49009 Cl OMe H CONHEt

R1 R2 R3 R4

BINDING PROTEIN…

geldanamycingeldanamycin

HYDROPHOBIC INTERACTION:Leu48, Lys58,

Met98, Thr109, Val136, Phe138, Val150, Val186

H-BOND ACCEPTORS/DONORS: Asp51, Asp54, Lys58, Asp93,

Gly97, Lys112, Phe139,Thr184

N S

NN

NN

O

N

NH

NH

O

PURINES12

PYRAZOLES28

SULFONAMIDES9

HSP90 INHIBITORS

PU1 PU2 PI1 PI2 SU1 SU2

HYPO1 HYPO2

ZINC DATABASE5,627,809 compounds

Lipinsky’s Rule

Molecular DockingHTVS Glide level

MolecolareDockingSP Glide level

MATCHING HITS

MOLECULAR DOCKING/

PHARMACOPHORE APPROACH

MOLECULAR DOCKING/

PHARMACOPHORE APPROACH

N

N

NHAr

NHO2S N

N

RR

O

Ar

N

N

NH

SO2 R

Ar

O

N

CN

NH2

12

3 4

N N

O

NHSO2Me

R1 R2

R3

9a- k

N

NH2NC

NHSO2MeR1

R2

R3

10a- k

X S

NN

NN

O

R'

R

11

N N

O

NHSO2Me

R1 R2

R3

9a- k

N

NH2NC

NHSO2MeR1

R2

R3

10a- k

O

NHO2S

6

O

NH2

CH3SO2Cl

Pyridine

5

R1

R2

R3

O

NaOH/H2O/EtOH

O

R1

R2

R3

NHO2S

7a- k

8a- k

N2H4/H2O

AcOH

NCCH2CNNH4Aca: R1=Cl, R2=R3=H

b: R1=R2=H, R3=NMe2 c: R1=R3=H, R2=F d: R1=OCH3, R2=R3=H e: R1=H, R2=R3=OCH3 f: R1=R2=H, R3=OCH3 g: R1=R2=H, R3=OH h: R1=OH, R2=OCH3, R3=H i: R1=R3=H, R2=OCH3

j: R1=R2=H, R3=NEt2 k: R1=Br, R2=R3=H

R=CN, PhX= CH, N

- CH2CH2CH2COOH - CH2CH2CH2COOCH2CH3

- CH2COOCH2CH3

- CH2CH2CH2Cl - CH2CH2CH2N(CH2CH2CH3)2

- CH2CH2CH2 N N CH3- CH2CH2CH2 N

- CH2CH2CH2CONHCH2COOCH2CH3- CH2CONHCH2COOCH2CH3

- CH2CH2CH2CONHCH2COOH - CH2CONHCH2COOH

- CH2COOH

- CH2CH2CH2CONHCH2CH2

NH

N - CH2CONHCH2CH2NH

N

- CH2CH2CH2CONHCHCH2

NH

NCOOCH3

- CH2CONHCHCH2

NH

NCOOCH3

- CH2CONHCH2CONHCH2CH2

NH

N

- CH2CONHCH2CONHCHCH2NH

NCOOCH3

- CH2CH2CH2CONHCH2CONHCH2CH2

NH

N

- CH2CH2CH2CONHCH2CONHCHCH2NH

NCOOCH3

- C

CHCOOCH2CH3

CH2COOCH2CH3 - C

CHCOOH

CH2COOH

X S

NN

NN

O

H

R

15

12

X

CN

F

HSCH2COOEt

NaH DMSO X

CN

SO

OX S

O

O

NH2

X S

O

O

NN

N

R

N

13

14

EtONa/EtOH

X S

NN

NN

O

R'

R

11

X S

NN

N

N

O

R

R'

16

NATIONAL CANCER INSTITUTE, Bethesda

DEVELOPMENTAL THERAPEUTIC PROGRAM

Leukemia

Non Small Cell Lung Cancer

Colon Cancer

SNC Cancer

Melanoma

Ovarian cancer

Renal Cancer

Prostate Cancer

Sub Panels of Human Tumor Cell

Lines

Breast Cancer

15

S

N

N

O

NN

Ph

HN

O

NH

N

K-562:pGI50=7.64

LEUKEMIACNS CANCER

SF-539:pGI50=7.69

RENAL CANCER

SN-12C:pGI50=7.96

16 side chain O

16

N

NH2NC

NHSO2MeN

10 b

60 cell lines single dose of 10 uM

TUMOR

• GIST• NSCLC• PROSTATE (PTEN-/-)• BREAST (HER2 +)• MELANOMA

HSP90 CLIENT PROTEIN

• c-Kit• EGFR / C-met• AR / p-Akt• HER2 / ER• b-Raf (V800E)

DIPARTIMENTO FARMACOCHIMICO, TOSSICOLOGICO E BIOLOGICO

• Prof. Anna Maria Almerico• Prof. Antonino Lauria• Prof. Gaetano Dattolo

• Prof. Maria A. Livrea• Prof. Luisa Tesoriere• Dott. Carla Gentile