Discovery Research 09/09/2004

Synthesis and Aromatase Inhibitory Activity of

Novel Pyridine-Containing Isoflavones
Young –Woo Kim, John C Hackett and Robertt W. Brueggemeier
College of Pharmacy, The Ohio State University
J. Med. Chem. 2004, 47, 4032 - 4040

Dr. Venugopal Rao Veeramaneni

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 Highlights of the presentation

Introduction of Cancer.
Details of Breast Cancer.
Aromatase enzyme and Steroid biosynthesis.
Aromatase Inhibitors.
Design and Synthesis of Isoflavones.
Results and Discussions.
Conclusions.

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Cancers are a group of diseases that cause cells in
the body to change and grow out of control.
control
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 Cell Structure & Cell Division

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5
 Cancers (Tumors)

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 Estimated Cancer Deaths in 2004

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 Breast Cancer

Normal Breast Structure

Lobules ⇒ Produce Milk

Ducts ⇒ Connect Lobules to Nipple
Fatty Tissue
Ligaments
Blood Vessels
Lymphatic Vessels

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 Normal Breast
A = Ducts
B = Lobules
C = Dilated Section of duct to hold milk
D = Nipple
E = Fat
F = Pectorails major muscle
G = Chest wall

A = Normal Duct cell

B = Basement membrane
C = Lumen (center of duct)

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 Cancer in the Breast

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 > 2 cm to < 5cm < 5cm, spread to
=< 2cm Advanced Cancer
Lymph node
98 % 88 – 76 %
49 – 46 % 16 %

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 Risk Factors

Family History Reproductive History

Genetic Risks Oral Contraceptives
Personal History Hormone Replacement
Certain types of Alcohol
breast cancer Weight
Menstrual History

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 Early Detection Methods

Breast Self Examination.

Clinical Breast Examination.

Screening Mammography.

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 Mammography

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 Aromatase

Cytochrome P450 enzyme complex.

First reported in human placental tissues by
K J Rayan in 1959.
Main expression sites: ovarian granulosa cells,
placental syncytiotrophoblast, adipose tissue,
skin fibroblasts and the brain. .
Role in estrogen biosynthesis.

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 Hormone Dependent Breast Cancer

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 Biosynthesis of Steroid Hormones
O
O
OH

HO HO
Cholestrol 17 hydroxypregnenlone
O
Progesterone
Side Chain Cleavage 17, 20 - Lyase
17 α-hydroxylase
4,5-Isomerase 17 α-hydroxylase O
O O
OH

HO
O dehydroepiandrosterone
17 hydroxyprogesterone HO
Pregnenolone
dehydrogenase
17, 20 - Lyase
OH
O

O HO
androstenedione androstenediol
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 Biosynthesis of Steroid HormonesContd…
Androstenedione Androstenediol
Aromatase 4,5-Isomerase
OH
OH
O
Aromatase
dehydrogenase
HO O
HO estradiol testosterone
estrone
O
O O
HO OH
O2 O2 HO - H2O

O O
O
androstenedione 19-hydroxy androstenedione 19,19I - dihydroxyandrostenedione
N
N
N
N

O Fe
Fe
O
N
O
O2
N
N
HCO2H
N

O OH
O O
HO O HO OH
O H
19-oxoandrostenedione H HO estrone
O O

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Classes of Drugs Used Treat Breast Cancer

Class Action Examples

SERMs (Selective Bind estrogen receptors in Tamoxifen (Nolvadex)

Estrogen Receptor breast cancer cells Evista (Raloxifene)
Modulators) Fareston (Toremifen)
Aromatase Inhibitors Prevent Production of Aromasin (Exemestane)
estrogen in adrenal glands Femara (Letrozole)
Arimdex (anastrozole)
Megace (Megestrol)
Biologic Response Bind with certain proteins Herceptine (Trastuzumab)
modulators on breast cancer cells,
preventing their growth

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Classification of aromatase Inhibitors

Generation Type 1 Type 2

(Steroidal) (Nonsteroidal)
First None Aminoglutethimide

Second Formestane Fadrozole

Rogletimide
Third Exmestane Anastrozole
Letrozole
Vorozole
Irreversibly inactivates the reversibly bind the enzyme
enzyme

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 Aromatase Inhibitors
OH O OH O O
OH
Ph OH
O N

H Ph H H
O
O O OH O O CH3
O
Megestrol Tamoxifen O
Ellence OH Formestane
1973, Astrazeneca OH
1963, BMS 1992, Novartis
1984, Pfizer NH2 O OH O
N OH
N
OH
N N N
N N N
N
N Fadrozole
Anastrozole O O OH O O CH3

1995, Astrazeneca Letrozole N Adriamycin OH

1995, Novartis 1996, Novartis 1996, BMS NH2
H3C O
N
O O O OH
N N
HO H N P OH
4 N
N N O
H H OH
HO N
N O O O P OH N
Xeloda H Aromasin Zometa
F OH Cl N
Vorozole
1998, Roche 1999, Pfizer 2000, Novartis
Pre-Registered, Janssen
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 Designing of Novel Inhibitor
O OH O OH
OH O OH O

HO O HO O
HO O
Biochanin A Chrysin Genistein
Biological Testing, Lion Phase II, Yamanouchi Phase II, Nihon University
R1
O

R3
R2 O S
R1 = H, Me, OMe, OH
R2 = OH, OMe, OBn
R3 = Allyl, Benzyl, Pyridylmethyl
2,4,7-tri substituted isoflavone

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 Synthesis of Isoflavones

R1 R1
O O

R2OH
Ph3P, DIAD
R2
HO OH
THF, O °C, 0.5 h O OH
(2)
(1) R1 = H, Me, OMe
R1 = H, Me, OMe R2 = Me, Bn

R1 R1
O O

n-BuN.HSO4, aq. NaOH

BBr3, CHCl3, r.t or
CS2, R3X, THF, r.t R2 R3
BF3.OEt2, Me2S, CH2Cl2 R3
overnight O O S HO O S
(3) r.t, overnight
(4)
R1 = H, Me, OMe, OH
R2 = OH, OMe, OBn R2 = OH, OMe, OBn
R3 = Allyl, Benzyl, Pyridylmethyl R3 = Allyl, Benzyl, Pyridylmethyl

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 Results and Discussion
O
O O
O
O

Bn Bn
O O S Bn Bn
O O S O O S
IC 50 100 IC 50 100
IC 50 100
O O
O

O O S O O S
O O S
IC 50 = 1.6 N
IC 50 100 N
IC 50 = 9.2
N
O O
O O

Bn
O O S
O O S O O S
IC 50 = 0.21 N
IC 50 = 3.0 N IC 50 = 3.1 N

O
OH O
O O

HO O S
HO O S HO O S
IC 50 = 0.61 IC 50 = 0.22 N
N IC 50 = 0.28 N

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 Results and Discussion
O
O
OH O O
O
O

O O S O O S
HO O
N
Biochanin A N

IC 50 = 34 µM IC 50 = 3.1 µM IC 50 = 0.53 µM
NH2

O
O O

HO O S O O S
O N O
H N N
Aminoglutithimide
IC 50 = 2.8 µM IC 50 = 0.21 µM IC 50 = 0.22 µM

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 Conclusions
Aromatase inhibitory activity can be achieved with
Isoflavone Scaffold.

The SAR studies indicates binding modes of 7-

prortected analogues might be different from 7-
hydroxy analogues.

Based on kinetic studies these compounds compete at

active site of aromatase with the natural substrate,
androstenedione.

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 Thanks To All

Arise awake, and stop not

Till the goal is reached
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