Oestrone

It is known for a long time that the uterine cycle has been controlled by hormones. But the first evidence for this claim was given by Butenandt & Doisy in 1929 after they isolated the active substance oestrone from the urine of pregnant women. Oestrone is the first known member of sex hormone. Its m.p. is 259oC, [ ]D is +170o. Molecular formula is C18H22O2. It forms an oxime. Hence one O atom may be ketonic in nature.

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Oestrone forms a monoacetate & monomethyl ether. So the 2nd O atom may be a hydroxyl group. Since oestrone couples with diazonium salts in alkaline solution the –OH group may be phenolic. When distilled with Zn dust it forms chrysene, which reveals that oestrone is structurally related to steroids. The X-ray studies showed that it contains steroid nucleus with keto and –OH groups are present at opposite ends of the molecule. On catalytic hydrogenation it takes up 4 moles of hydrogen to give octahydrooestrone, C18H30O2. Here 1 mole of hydrogen is used to reduce keto group and the remaining 3 moles are used to reduce 3 = bonds, hence we can expect a benzenoid ring in oestrone if these 3 = bonds are seen in one ring.

The presence of phenolic –OH group accounted the benzenoid ring in it. When methyl ether of oestrone is subjected to Wolff-Kishner reduction followed by Se distillation, 7-methoxy-1,2cyclopentenophenanthrene is formed. The structure of this compound was established by the following synthesis. OH
CH2MgBr O H3C H3C

- H2O

+
H3CO H3CO

H3C

AlCl3
H3CO

Se
CH3

320 °C
H3CO

H3CO

7-Methoxy-1,2-cyclopenteno phenanthrene

Thus the benzene ring in oestrone is ring A, and the phenolic –OH group is at position 3. Hence we can sketch the skeleton of oestrone as follows.
C A
HO

D

B

Now the job is to fix the –C=O group. The above skeleton accounts only for 17 C atoms. Later in 1935 Cook et al. showed the presence of an angular methyl group at position 13. When methyl ether of oestrone is treated with methyl magnesium bromide and other reagents as given below the final product is 7-methoxy-3’,3’-dimethyl-1,2cyclopentenophenanthrene, V.

The formation of V can be explained only if there is a keto group at position 17 & an angular methyl group at position 13. It should be noted in the given reactions below, the dehydration is accompanied by the migration of angular methyl group. Several known examples are there for this kind of migration. O CH OH
CH3
3

CH3

CH3MgI
H H3CO H3CO H

KHSO4 (-H2O)

I
CH3 CH3

II
CH3 CH3 CH3 CH3

H2 - Pt
H3CO H3CO

Se
H3CO

III

IV

V

Thus the structure of oestrone is given below
CH3 H H HO H O

Oestrone

Synthesis:
The above structure of oestrone has been confirmed by its synthesis by Anner and Miescher in 1948. They started the synthesis using phenanthrene derivative, VI, which was synthesized by Bachmann et al. in 1942.

CH3 H O H H3CO

CH3 COOMe H

COOMe CH2CO2Me

+

BrCH2CO2Me

+ Zn
H3CO

H

OH

POCl3 C5H5N

VI
CH3 CH3 CH3

COOMe

H2 Pd/C (sepn.)
H

-

COOMe

(i) aq. MeOH - KOH (ii) H+
H

COOMe

CHCO 2Me

CH2CO2Me

CH2CO2H

VII
CH3 COOMe CH3 COOMe

(COCl)2
H CH2COCl

(i) CH2N2 (ii) AgOH/MeOH
H

CH2CH2CO2Me

(i) KOH; 180 °C (ii) PbCO3; 320 °C

CH3 H H H3CO H

O

CH3

O

C5H5N.HCl
H HO

H H

(+)(-) - Oestrone
D. Ilangeswaran 8

Johnson et al. in 1958,1962 have also carried out total synthesis of oestrone. In this synthesis each step was stereospecific.
CH CH2Br MeO NEt 2

C2HNa DMF
MeO

Et2NH CH2O
O CH3 MeO

H2SO4 Hg2+
CH3 O

CH2 H3C O O MeO

O

O

O

TsOH
MeO CH3 OH CH3 H H HO H O

OHO

MeO

CH3

H2 - Ni
H MeO

K/NH2 NH4Cl
H MeO

H H

(i) CrO3 (ii) HBr/AcOH

(+ or - ) Oestrone

Torgov et al. synthesized oestrone in 1960-1962 as follows.
O CH2 H3C OH O O CH3 O

CH2=CHMgBr
MeO MeO

OHMeO

O

CH3

O CH3

O

TsOH H2 - Ni
MeO MeO H

(i) K / NH2 (ii) CrO3

CH3 H H MeO H

O

CH3 H H HO H

O

C5H5N.HCl

(+ or -) Oestrone
D. Ilangeswaran 10

OESTRADIOL
 There are two stereoisomers α- and -. Of these two - form is more active.  The - isomer was isolated from the ovaries of sows by Doisy et.al. in 1935. The α- isomer was isolated from the pregnant urine of mares by Wintersteiner et. al. in 1938.  The m.p. of α- isomer is 178oC and that of isomer is 222oC. The [α]D for α = +81o, for = +54o.

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Constitution
1. Molecular formula: C18H24O2. 2. Presence of two –OH groups: Easy formation of diacetyl derivative confirm this. 3. Nature of –OH groups: One is phenolic while the other one is secondary alcoholic since on oxidation oestradiol yields oestrone. 4. When oestrone on reduction yields oestradiol. Hence oestradiol can have the same C skeletal framework as oestrone.
D. Ilangeswaran 12

The phenolic methyl ester of estradiol on heating with ZnCl2 undergoes a rearrangement with the migration of angular methyl group to cyclopentane ring. The rearranged product on Se dehydrogenation yields 7-methoxy-3’methyl-1,2-cyclopentenophenanthrene. On these facts the structure of estradiol would be assigned as I.
D. Ilangeswaran 13

OH H
CH2N2

OH H
ZnCl2

H HO
Oetradiol - 17

H H3CO CH3 H
Se

H

H

CH3

H H3CO

H

Dehydrogenation

H3CO
7-Methoxy-3'-methyl-1,2-cyclopenteno phenanthrene

OH H H HO
Oetradiol - 17
D. Ilangeswaran

OH H

H HO

H

H

Oetradiol - 17
14

Conversion of Estrone into Estradiol
O H
Al(iPrO)3

OH H

H HO
Estrone

H

OR LiAlH4

H HO
Estradiol

H

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Conversion of Estrone into Estriol
O H
Isopropyl

OAc H
PhCO3H

H HO
Estrone

H

Acetate

H AcO

H

OAc O H H AcO H
LiAlH4

OH H H HO
Estriol

OH H

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Equilenin
This has been isolated from pregnant mares by Girard et al. in 1932. It has m.p. 258-259oC and [ ]D +87o. Molecular formula is C18H18O2. The reactions of equilenin show that a phenolic hydroxyl group and a ketonic group present. It also contain five double bonds.

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• When methyl ether of equilenin is treated with methyl magnesium bromide, then the alcohol dehydrated, catalytically reduced finally dehydrogenated with se. The product is 7-methoxy-3’,3’-dimethyl-1,2cyclopentenophenanthrene, the same product as in oestrone. • Hence oestrone & equilenin are having the same structural unit except the later has two more double bonds. • The absorbtion spectrum of equilenin shows that it is a naphthalene derivative. • As ring A in oestrone is benzenoid, it appears probable that ring B in equilenin is also benzenoid. • Hence rings A & B form naphthalene nucleus in equilenin.
D. Ilangeswaran 18

All the foregoing reactions of equilenin can be readily explained by assuming that I is structure of equilenin. Further evidence to this structure is given by Marker et al. as equilenin may be readily reduced to oestrone by Na/ethanol mixture.

H3C

CH3 CH3

O CH3

O

Na
H HO HO

H H HO H

C2H5OH

II

I Equilenin

III Oestrone

Synthesis
Bachmann synthesized compound IV. Using IV, Johnson synthesized equilenin as follows.
NH2 NH2 NHCOCH 3

KOH
HO 3S HO

(CH3CO)2O
HO

(i) (CH3)2SO4 - NaOH (ii) Hydrolysis
CH2OH

Cleve's acid
NH2 I

(i) NaNO2-H2SO4
H3CO CH2Br

(i) Mg (ii) H
H O H H

PBr3
H3CO

(ii) KI

H3CO

Malonic ester
H3CO

COOH

(i) SOCl2 (ii) SnCl4 H CO 3 IV

O

synthesis H3CO

Johnson's synthesis starting from (IV)
CHO O H3CO N O NH2OH.HCl H3CO OH O

HCO2C2H5 CH3ONa

CH3CO2H

IV

N

Tautomn.
OH

OH

- H2O
O

N

CN

(CH3)3COK
OK
- +

Isoxazole
CH3 CN CH3 CN

CH3I
O

Methyl succinate (CH3)3COK

Thorpe
CH2CO2CH3 CO2CH3

reaction

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CH3 NH CO2K CO2CH3

CH3 O

CH3 O

(i) Ba(OH)2 (ii) HCl
COOH

boil with C5H5N.HCl - HCl

H2 Pd - C V

CH3 O

CH3 O

HCl
H H3CO

CH3CO2H
HO

H

VI

VII

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Progesterone
This was first isolated in a pure form by Butenandt et al. in 1934 from the corpora lutea of pregnant sows. Molecular formula is C21H30O2. Its m.p. 128oC and [ ]D +192o. The chemical reactions of progesterone show that there are 2 keto groups present. On catalytic reduction it adds on 3 moles of hydrogen to form a dialcohol, C21H36O2. So there may be one = bond.

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The parent hydrocarbon of progesterone is C21H36 which corresponds to the general formula CnH2n-6 of a tetracyclic compound. X-ray studies reveals that progesterone contains steroid nucleus. This is further supported as progesterone is obtained from stigmasterol & cholesterol. The absorption spectrum shows that it is an , -unsaturated ketone. Hence the double bond may be present between position 4 an d 5. The various synthesis of progesterone given below confirm its structure.

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Butenandt et al. in 1943
Progesterone from Stigmasterol
CH3 H3C CH3 CH3 H AcO H H AcO Br Br COOH H3C CH3 COOH CH3 CH3 H3C CH3 CH3 H H H CH3 CH3 CH3

Br2

O3

Stigmasteryl acetate
H3C CH3 CH3 H AcO Br Br H H

Zn CH3CO2H
AcO

CH3 H

H H

(i) C2H5OH - HCl (ii) C6H5MgBr (iii) - H2O

Acetate of 3 -hydroxybisnorchol-5-enic acid

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H3C CH3 CH3 H AcO H H

CPh2

H3C CH3

H3C O CH3 CH3 H H H

O

(i) Br2 (ii) CrO3
AcO

CH3 H Br Br H3C CH3 CH3 H H H

H H

(i) Zn/AcOH (ii) Hydrol.
AcO

Pregnenolone Oppenauer oxidation

O

O

Progesterone

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Butenandt et al. 1939
H3C CH3 CH3 H HO H H HO

Progesterone from Cholesterol
CH3 CH3 CH3 H H H CH3 O

(i) Ac2O (ii) HCN
H3C O CH3 CH3 H H

Cholesterol
CH3 CH3 H AcO H OH CN

Dehydroepiandrosterone
CH3 CH3 H AcO H3C O CH3 H H CN

POCl3
H

CH3MgBr
H AcO H3C O CH3

(i) H2/Raney Ni (ii) Hydrol.

CH3 H

H H

Oppenauer oxidation
O

CH3 H

H H

HO

Pregnenolone
D. Ilangeswaran

Progesterone

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Diosegnin occurs as a glycoside in the root of Trillium erectum
Progesterone from Diosgenin (Marker et al. 1940,1941)
H3C CH3 CH3 H HO H H O CH3 O CH3 H AcO H H H3C CH3 O CH3 O Ac

Ac2O 200 °C

CrO3

Diosgenin
H3C CH3 CH3 H H H HO O

H3C O CH3 CH3 H H H O

H3C O CH3

(i) H2 - Pd (ii) Hydrol.

Oppenauer oxidation

CH3 H

H H

AcO

Pregnenolone

Progesterone

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Butenandt et al.in 1934
Progesterone from Pregnanediol
CH3 H CH3 CH3 H HO H H H O CH3 H H H H O Br H OH H3C CH3 O H3C CH3 CH3 H H H O

CrO3

Br2

Pregnanediol

Pregnanedione
H3C CH3 CH3 H O H H O

C5H5N (-HBr)

Progesterone

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Shepherd et al. 1955, more practical synthesis (note the enamine step)
Progesterone from Ergosterol
H3C CH3 CH3 H HO H CH3 CH3 CH3 H3C CH3 CH3 H O H CH3 CH3 CH3

Oppenauer oxidation

HCl in CH3OH

Ergosterol
CH3 CH3 CH3

Ergosterone
H3C CH3 CH3 CH3 CH3

H3C CH3 CH3

HCl

CH3 H

H H

H2 Pd - C

H MeO

H

O

Isoergosterone

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H3C CH3 CH3 H O H H

CH3 CH3 CH3 H3C CH3 CH3 H O H H CHO

O3

C5H10NH

H3C CH3 CH3 H O H H

H3C N CH3 CH3 H O H H

O

Na2Cr2O7 AcOH

Progesterone

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Testosterone
Among the 5 derivatives of androgens that function as steroid hormones the most potent is testosterone. Testosterone was isolated by E. Laquer et. al. in 1935 from testes. About 10 g of testosterone is obtained from 100 Kg of testes. Its m.p. is 155oC, [α]D = +109o and λmax = 240nm.
D. Ilangeswaran 32

Physiological Action
I. Testosterone is a real male sex hormone while other androgens are metabolic products of testosterone. II. It stimulates the development of secondary male sex characteristics. III. It assists in bringing about the descent of the testes in cryptorchidism. IV. It inhibits the secretion of anterior pituitary gonadotropins. V. Testosterone and its derivatives have been found useful in the treatment of advanced metastatic carcinoma of breast. VI. Rarely, it may produce jaundice. VII. It is used in the treatment of the menopausal syndrome combined with estrogens.

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Constitution
1. Molecular formula; C19H28O2. 2. Presence of tetracyclic system: The molecular formula of parent HC is C19H32 & this corresponds to the general formula CnH2n-6. 3. Presence of α, - unsaturated keto group: As testosterone is very sensitive to alkali with the λmax = 240nm confirms the presence of α, unsaturated keto group. Further because of this facts it is suggested that testosterone may be structurally related to progesterone.
D. Ilangeswaran 34

4. Oxidation: In 1935 K. David oxidised testosterone to a diketone namely androst-4ene-3,17-dione and its structure is well known. This diketone also obtained during the Oppenauer oxidation of dehydroepiandrosterone. The formation of diketone can be explained if the structure I is assigned to testosterone.

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OH H H O
Testosterone (I)

H

Oxidation

O H H H

O
Oppenauer oxn.

O
Androst-4-ene-3,17-dione

H H HO
Dehydroepiandrosterone

H

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5. Synthesis: Ruzicka & Butenandt (1935)

H H HO
Cholesterol

H
(i) Ac2O

H

(ii) Br2

H AcO Br Br

H

Cholesteryl acetate dibromide

O O H
CrO3 - AcOH (i) Zn - AcOH

H H HO
Dehydroepiandrosterone

H Br Br

H

(ii) HOH

H

AcO

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OH
(i) Ac2O (ii) Na - C3H7OH

OCOPh H
(i) PhCOCl

H H H

AcO

(ii) mild hydrolysis (CH3OH/NaOH)

H HO

H

OCOPh H
Oppenauer Oxidation Hydrolysis

OH H

H O

H

H O

H

KOH

Testosterone

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