1. UNIVERSITA’ DEGLI STUDI DI PALERMO
FACOLTA’ DI FARMACIA
DIPARTIMENTO FARMACOCHIMICO TOSSICOLOGICO E BIOLOGICO
Design, synthesis and biological evaluation of new inhibitors of carcinogenic processes
Dott. Ilenia Abbate

2. TUMOR PROGRESSION
Cancer cells are less sensitive to the therapy and repopulate the tumor, facilitating tumor progression
Migratory growth factor
growth factor-
induced
mitogenesis
STAT3
c-MYC
PROTO-ONCOGENE
PI 3 kinase
Akt
survival
Ras
ERK
MERK
Cyclin/cdks
promotes G1/S phase

3. SIGNAL-TRANSDUCTION PATHWAYS CORRELATED TO TUMOR PROGRESSION
Without fuctional HSP90, these proteins undergo proteasome-mediated degradation, leading to cell cycle arrest and apoptosis.

4. …and Signaling Pathways
HSP90 chaperone
stabilizes oncoproteins
Inhibiting HSP90 degrades
oncoproteins, stopping tumor growth

5. 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 evade apoptosis
ability to be self-sufficient for growth
ability to invade surrounding tissue and to metastasize to distant sites
ability to undergo limitless replication
ability to promote neoangiogenesis
ability not to respond to antigrowth signals.

6. HSP90: molecular chaperone
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

7. 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.

8. INHIBITORS
HSP90

9. HYDROPHOBIC INTERACTION:
BINDING PROTEIN…
geldanamycin
HYDROPHOBIC INTERACTION:
Leu48, Lys58,
Met98, Thr109,
Val136, Phe138,
Val150, Val186
H-BOND ACCEPTORS/DONORS:
Asp51, Asp54,
Lys58, Asp93,
Gly97, Lys112,
Phe139,Thr184

10. PHARMACOPHORE APPROACH
HSP90 INHIBITORS
ZINC DATABASE
5,627,809 compounds
PURINES 12
PYRAZOLES 28
SULFONAMIDES 9
Lipinsky’s Rule
PU1
PU2
PI1
PI2
SU1
SU2
Molecular Docking
HTVS Glide level
MolecolareDocking
SP Glide level
HYPO1
HYPO2
MOLECULAR DOCKING/
PHARMACOPHORE APPROACH
MATCHING
HITS
112 COMPOUNDS

11. 112 DERIVATIVES

12. dihydropyrazole 9 and 2-amino-3-cyanopyridine 10
SYNTHESIS
dihydropyrazole 9 and 2-amino-3-cyanopyridine 10
a: 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

13. pyridinothieno/benzothieno-triazolO-pyrimidines
R=CN, Ph
X= CH, N R’ A B C D E F G H I J K L M N O P Q R S T U V

14. NATIONAL CANCER INSTITUTE, Bethesda DEVELOPMENTAL THERAPEUTIC PROGRAM
BIOLOGICAL SCREENING
NATIONAL CANCER INSTITUTE, Bethesda
DEVELOPMENTAL THERAPEUTIC PROGRAM

15. NATIONAL CANCER INSTITUTE DEVELOPMENTAL THERAPEUTICS PROGRAM
MEAN GRAPH AND DOSE RESPONSE CURVES
16 side chain O
LEUKEMIA
RENAL CANCER
CNS CANCER
SN-12C:pGI50=7.96
SF-539:pGI50=7.69
K-562:pGI50=7.64

16. One DOSE RESPONSE CURVES
MEAN GRAPH
One DOSE RESPONSE CURVES
60 cell lines
single dose of 10 uM

17. HSP90 CLIENT PROTEIN
c-Kit
EGFR / C-met
AR / p-Akt
HER2 / ER
b-Raf (V800E)
TUMOR
GIST
NSCLC
PROSTATE (PTEN-/-)
BREAST (HER2 +)
MELANOMA

18. …THANKS FOR YOUR ATTENTION
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
…THANKS FOR YOUR ATTENTION