Preparation of [1,2-3H]17-alpha-methyltestosterone and of some unlabelled derivatives.

Full text of DOI:10.1677/joe.0.0350121

OF
PREPARATION
AND OF
[1,2-3H]17\g=a\-METHYLTESTOSTERONE
SOME UNLABELLED
DERIVATIVES
C. H. GRAY
R. V.
and
QUINCEY
Chemical
Medical
Pathology, King's College Hospital
Department of
School,
5
S.E.
London,
3
February
(Beceived
1966)
a
of
metabolism
the
facilitate
of
To
study
(MeT),
[1,2-3H]
17\g=a\-methyltestosterone
of MeT
This
were
MeT and four unlabelled derivatives
of these
describes
the
prepared.
report
preparation
compounds.
This
tritiation
was
the
prepared
[1,2-3H]17\g=a\-Methyltestosterone.
by partial
17\g=a\-methyl-17\g=b\-hydroxy-androst-1(,4a\tx=req-\
of
Radiochemical
dien-3-one
the steroid
Amersham,
Centre,
Bucks.)
from Ciba
A
at
hundred
mg.
of
a
Laboratories,
Horsham,
Sussex).
(obtained
for
5
12
in
ml. of
5
ml. dioxan and shaken
hr.
22\s=deg\
dissolved in
was
50
10
of
the
tritium\p=m-\hydrogen mixture
in
charcoal.
on
%
palladium
(5 c)
presence
an
mg.
specific
18
of
of
recovered with
steroid
were
activity
Eighty
average
mc/mg.
mg.
a
isolated
in
was
yield by
[1,2-3H]MeT
subsequently
paper
%
chromatography
10\p=n-\15
:
formamide
of
material
the
in
of
the tritiated
cyclohexane
portions
(Savard,
system
of Bush
and then
the A
in
thus obtained
(Bush,
1952).
1953)
system
was
two
of
further in
one
purified
added
MeT
[1,2-3H]MeT
ways.
[l,2-3H]MeT
to
of
the
385
unlabelled
was
of
1900
and the
specific
activity
were
fio
(1)
mg.
three times from methanol. The
diluted material
of
recrystallized
second and third
steroid
material isolated from the
constant
and
the
was
recrystallization
4-15
171
obtained.
material
of the
were
were
was
/¿c/mg;
mg.
[l,2-3H]MeT
left
and
purified
2-5
with unlabelled
to
a
and
diluted
mc
specific
(2)
activity
at 22°
1-0
in acetic
of
to
in
were
order
anhydride
reagents,
overnight
fic/ftg.
acetylate
pyridine
of
After
the
the diluted
steroid
was
evaporation
impurities.
any
A
the Bush
in
chromatography
purified by
system.
to
both methods
in
be
obtained
when
appeared
[l,2-3H]MeT
by
radiochemically pure
A
:
were
the Bush
formamide
or
portions
cyclohexane
MeT
rechromatographed
systems.
Unlabelled derivatives
This
of
5
from
was
prepared
mg. 17a-methyl-
l7ct-Methyl-5ß-androstane-3a,17ß-diol.
Steraloids
from
(obtained
Ltd.,
Croydon,
17/?-hydroxy-5/?-androstan-3-one
Surrey)
3-oxo
with sodium
reduction of the
was
stereoselective
borohydride
by
group
chromatography
(Barton,
in the Bush
A
The
of
After
purified
system
product
by
(ABMR
1953).
on
with
reaction
located
and
and
acid.
was
+
by
0-70)
paper
phosphomolybdic
mm.
at
vacuum
about
the
infrared
'
sublimation
elution
i.r.
108°,
mercury)
(0-01
'
a
was
Perkin-Elmer Infracord
measured in
3400
of the steroid
spectrophoto-
spectrum
the
a
meter.
at
cm.-1 and
absence of
contained at
least
An
in the
region
peak
peak
one
absorption
1750-1650
the
cm.-1 showed that
compound
group.
hydroxyl group
not
and did
contain
an oxo

was
oxidation
from the
acetic
This
prepared by
(obtained
llot-Methyl-nß-hydroxy-androst-4:-en-3,ll-dione.
2-5
of
mg.
17a-methyl-ll/?,17/?-dihydroxy-androst-4-en-3-one
in
with chromium trioxide
Co.,
50%
Upjohn
Kalamazoo,
aqueous
Michigan)
The
was
acid
Studer
&
Dobriner,
Shaw
&
Schneider,
1955).
(Lieberman, Katzenellenbogen,
was
of
and
L
in
product
purified by chromatography
Gray
(1965)
system
and
on
u.v.
of
vacuum
soda-fluorescence
located
Bush
(ABMB
steroid
by absorption
light
by
paper
of the
of
was
After elution and
sublimation the i.r.
0-47).
spectrum
an
3400
of
at
indicated
the
presence
indicated that the
cm."1
measured. An
and
cmr1
at 1600
peak
absorption
1690
at
cm.-1 and
com¬
peaks
hydroxyl
group
11-oxo
an
a
contained
and A4-3-oxo
pound
group
group.
was
This
hydro¬
prepared by catalytic
17a-Methyl-5ß-androstane-3a, 11/?, llß-triol.
4
the method
of
17a-methyl-ll/?, 17/?-dihydroxy-androst-4-en-3-one by
génation
mg.
of
the
with sodium boro-
of Gabbard
and
reduction
&
product
Segaloff
by
hydrogénation
not
(1962)
was
The
of the
of
which
u.v.
purified by
hydride.
in the
with alkaline
product
chromatography
was
and
with sodium
acid but
located
on
did
absorb
L
system (ABMR
light
paper
0-30)
m-dinitrobenzene. Reduction of this
borohydride
product
not
alkaline
a
with
of
which reacted with
phosphomolybdic
compound
yielded
in
the L
0-16
which
an
and
had
of
m-dinitrobenzene
(ABMR
BF
+0-67).
system
a
with
acetic
of the steroid
After
and
was
elution,
acetylated
major portion
anhydride
acetate
at 3400
measured after
at
at
and the
of the
i.r.
was
vacuum
22°,
pyridine
spectrum
indicated
cm.-1
the
about 105°. An
sublimation
peak
absorption
presence
1250
and at
cm.-1
1740
1730
one
and
at
an
cm.-1
from
of at
least
indicated that
cm."1,
group.
peaks
hydroxyl
group
the
contained
compound
acetoxy
This
was
compound
17ß-triol.
17ct-Methyl-5ß-androstane-3a,,l6a,,
prepared
from
3-8
(obtained
17a-methyl-16a, 17/?-dihydroxy-androst-4-en-3-one
Upjohn
mg.
of
was
for the
the
the method used
Co., Kalamazoo,
17a-methyl-
Michigan) by
preparation
of
The
purified
u.v.
hydrogénation
by
11/?,
product
5/?-androstane-3a,
17/?-triol.
not
L
of
It did
absorb
but
the
in
light
(ABMB
chromatography
—0-40).
system
sodium
Reduction of this
with
with alkaline m-dinitrobenzene.
located
was
borohydride yielded
m-dinitrobenzene and which had
product
not
a
acid but
which reacted with
phosphomolybdic
0-40
compound
an
of
in the Bush B5
system
treatment with
acetic
with alkaline
BF
was
acetate of this
at 22°
The
prepared by
compound
after
(Bush,
1952).
vacuum
at
about
the i.r.
sublimation
and
95°,
and,
anhydride
spectrum
pyridine
3400
the
cm."1 indicated that
at
was measured. An
an
com¬
absorption peak
1725
at 1740
at
cm.-1 and
and
contained
cm.-1,
group.
peaks
pound
group
hydroxyl
one
at
least
1250
the
of
cm.-1 indicated
acetoxy
presence
were confined
Losses
to
those
were
about
Yields of
incurred in
steroids
50%.
largely
purified
the
purification procedures.
REFERENCES
1027-1040.
J. chem. Soc.
D. H. R.
Barton,
pp.
(1953).
J.
A.
Biochem.
370-378.
I. E.
50,
(1962).
Bush,
(1952).
J.
Chem.
655-656.
33-55.
R. B.
H.
&
27,
Gabbard,
org.
Segaloff,
D. A.
Endocrin.
J.
C.
&
33,
Shaw,
(1965).
Gray,
K.
J. biol. Chem.
P. E.
&
E.
Dobriner,
(1955).
Schneider, R., Studer,
R.,
Lieberman, S.,
87-91.
205,
Katzenellenbogen,
biol. Chem.
J.
457-477.
K.
202,
Savard,
(1953).