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Патент USA US3074944

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3,®74,935
p
1C6
United States Patent ‘ O
W
diol andt give the G-formyl-derivative (VII). Reduction
of (V11) with sodium borohydride gives the 6-hydroxy
1
methyl derivative (VIII), which on dehydration with an
acidic reagent such as p-toluenesulf-onic acid forms the
3,074,935
6-E‘XOMETHYLENE STEROEDS AND
6-exomethylene derivative (IX). Any protecting groups
PROCESS THEREFQR
John Fried, Plainiieid, Anthony N. Nutile, Elizabeth, and
which are present at C-17, or in a side chain attached
at the C-17 position of the steroid molecule, may then
5
Glen E. Arth, Cranford, NJ, assignors to Merck & (10.,
Inc, Rahway, ‘Ni, a corporation of New Jersey
No Drawing.
‘
Patented Jan. 22, 11953
be removed.
‘
vIn the ?rst step of our' process,
the S-keto-group of
1960, Ser. No. 35,416
30 Claims. (Ci. 260-23955)
This invention is concerned generally with novel steroid
10
live. In a preferred embodiment of our invention the 3
compounds and with processes of making the same.
More particularly, it relates to novel 6-exomethylene
keto group is protected by forming the 3-ethylenedioxy
derivative. This derivative 18 prepared by reaction of a
A4-steroid-3-ones and ?-exo-methylene-AlAsteroid-3-ones
and to the process of making such compounds, starting
with
5a-steroid-3,6-dione.
In aaccordance
wit the present invention, we have
found that the novel 6-exornethylene-A4steroid-3-on'es
can be prepared from the 5ec-steroid-3,6-diones
bythea
follows, showing
process which can be represented as
rings of the steroid molecule:
the 5wsteroid-3,6-dione (protected at C-17 as indicated
above) is blocked by forming the 3-alkylenedioxy deriva
solution of the steroid in a solvent such as benzene with
15
an equivalent amount of ethylene glycol and p-toluene
sulionic acid for 8-12 hours. The product may be re
covered by washing with aqueous sodium bicarbonate,
drying and concentrating in vacuo.
The 3-keto group may also be converted into the 3
AandB
0
[/
\
CH0
O
H=CH2
v
t
O
\
tO
at
\
u
H2011
VIII
OH’
ethylenedioxy derivative by exchange dioxolanation,
1X
In this process the 5otesteroid-3,6-dione (I) in which
any oxygen function present at 0-17, or in a side chain 55
attached at the C-l7 position of the steroid molecule,
is ?rst protected with suitable protecting groups, is con
verted into the 3-alkylenedioxy-Set-steroid
alklenedioro- SOL-SlIEl‘Old (H) is
The 3
with acetyl
ene magnesium bromide to obtain the corresponding 6:»
‘which involves acid-catalyzed transfer of the ethylene
glycol portion of simple 2,2-dialltyl-1,3-dioxolanes, such
as 2,2'-dimethyl-l,S-dioxolane (acetone ethyleneketal) or,
better, 2-methyl-2-ethyl-1,3-dioxolane (but-anone ethyl
ene-ketal), with the 3-ketone steroid either in an inert
60.
have found that the‘ lower alkylenedioxy derivatives,
ethynyl-3-alkylenedioxy-5a-steroid-6p-ol
7 with
ter compound is then dehydrated, for(Ill).
example,
wherein the hydrocarbon group contains not more than '
thionyl chloride in pyridine to give the 6-ethynyl-3-al
kylenedioxy-A-‘Jsteroid (IV-A), which may be admixed
with small quantities‘ of 6-ethynyl-3-alkylenedioxy-A6
steroid (IV-B) and 6-([2]-chloro-[l,2]-propadiene)-3
'al-kylenedioxy-steroid (1V~C) and requires separation
solvent, as benzene, or simply in excess reagent.
Other cyclic ltetal derivatives can be used in our proc
ess p-for protecting a ketone group at 0-3. In general, we
seven carbon atoms, such as the ethylenedioxy, trimethyl
65
enedioxy, propylenedioxy and butylenedioxy derivatives
are suitable in carrying out the processes of our inven
tion. However, in place of using a lower alkylenedioxy
suhstituent to block or protect the keto substituent, we
therefrom. Upon hydrogenation of (IV-A) using a
can use other derivatives'readily hydrolyzable to keto,
catalyst, the corresponding 6-vinyl-derivative (V) 'is ob 70 such as a dialky-l ketal, an enol ether monothioket-al, or
tained, which ' then oxidized with osmium tetroxide to
'a dithioketal derivative for this purpose.
give the corresponding 6-diol (V1). The latter com
pound is then reacted with periodic acid to cleave the 6
' 3,074,935
The second step of our process, comprising the reac
‘tion of the 3-alkylenedioxy-5a-steroid (II) with acetylene
magnesium bromide, is brought about by adding a solu
sul?te, distilling oil’ the benzene, adding more methanol
‘
'
The prod
4
extracting
The solvent layer is
washed With Water‘, dried and concentrated in vacuo it?
give
the 6_-’([l,2]=ethanediél)=3-allc'leiiedioxy-ns-steroid
(VI), Which can ‘be used without further purification in
hours at room temperature or above.
the next step of the synthesis;
of the reaction, .the 6a-ethynyl-3-alkylenedioxy~5a-steroid= 10 In the sixth step of our
66-01 (III) is separated by chromatographing on acid
washed alumina and elution with mixtures of chloroform
and ether. The product may be puri?ed by recrystalliza
tion.
The third step of our process is effected by dehydrating
the 6a-ethynyl-3 ~'alkylenedioxy-5a-steroid-6?-ol (III),
using as the dehydrating agent thionyl chloride, or POCIQ,
in a tertiary base such as pyridine. Other tertiary bases
suitable for this purpose are the N-allryl piper'idin'es, col
the trialkylamines. The de ‘ydrat-ion is con 20
formyl-3-alkylenedioxy-A5-steroid (VII). Any contami
with the 6-ethynyl-3-alkylenedioxy-A6-steroid (IV-B)
nation of the 6-ethynyl-3-alkylenedioxy-A5-steroid (IV-A)
reaction mixture at 30°
formed as a by-product in the third step of our process will
tion of the reaction, the 6-ethynyl-3—alkylenedioxy-A5
result in the
of some 6-formyl-3-alkylenedioxy
As-steroid
in recovery
the 6th step
zene, ethyl acetate, methylene
30
drying the extracts and concentrating in vacuo. The
35
tures containing varying quantities of ether and petroleum
ether may also yield the 6-ethynyl-3-alkylenedioxy-A6-ste
derivative, (IV—C).
6-([2]chloro-[l,
2]-propadie11e)-3-alkylenedioxy-steroid
These
the 6-éthynyl-3-alkyl
enedioxy~A5-steroid (IV-A) process,
responding
6-vinyl-derivative is hydrogenated to the ‘cor
Crystallization from a solvent such as benzene gives the
6-hydroxymethyl-3~alkylenedioxy-Aisteroid.
In the eighth step of our synthesis, dehydration of the
6- ydroxymethyl-group, together with simultaneous re
45
moval of the 3~alkylenedioxyegroup, results in the con
version of the 6~hydroxymethyl~3-aikylenedioxy-A?-steroid
(VIII) into the 6-exomethylene-A4-steroid~3-one (IX).
Th"
is readily brought about by reaction of a
solution of the 6-hydroxymethyl-steroid (VIII) with p~
able solvent, such as ethyl acetate, is hydrogenated at
toluenesulfonic acid and letting the mixture stand at
temperature for several hours. The product is recovered
y pouring the mixture into ' Water and then extracting
the resulting aqueous solution with a solvent such as
chloroform or ethyl acetate.
55
The process of forming the 6-exomethylene derivatives
of the A4-steroid-3-ones is not limited to any particular
type of steroid compounds, but is an e?ective procedure
for steroid compounds generally, and includes especially
'
In the ?fth step of our reaction, the crude 6-vinyl-3
alkylenedioxy-As-steroid (V) is oxidized to the 6-([1,2]
ethanediol)
-3 -alkylenedioxy-A5-steroid ( VI) .
dation
pregnane series, provided
that any oxygen function present at C-17, or in a side
chain attached at the C—l7 position
of the' steroid mole
cule, is ?rst
'
If the synthesis is started with a 5a-pregnane-l7a,2l
'
_ diol-3,6,20-trione,
65
'
chain at the l7-carbon atom '
mation of the l7u,20,20,2l-bis-methylenedioxy
derivative,
'
'
the removal of the bis-methylenedioxy
The reaction is carried
out in a solvent such as benzene by allowing the mixture
plishedacids.
with aqueous organic acids such as formic or
acetic
In a preferred
amount of osmium tetroxide employed is approximately
'
of the crude material.
based on the total Weight
The osmate is decomposed by adding the reaction mix
ture to an aqueous methanol solution containing inorgan
exomethylene - l7a,20,20,21-bis-methylenedioxy~4-preg~
nene-B-one is heated on a steam
methylene-4-pregnene—1704,2l-diol-3,20-dione
75 75% yields. The organic phase is washed
alkali, dried and concentrated in vacuo to give the crys-
3,074,935
ylenedioxy-ll-pregnened,ll-dione by reaction with p-tolu
ene-sultonic acid at room temperature for several hours.
The
6-exomethylene-l7or,20,20,2l-bis-methylenedioxy-Ll
pregnene-3,ll-dione is transformed into 6-exomethylene
talline product. The non-crystalline material may be
recovered as the corresponding 2l-acetate aiter first
cleaving any formate esters present, suitably by reaction
with sodium methylate in methanol, recovering the prod
uct, and then acetylating with acetic anhydride and pyri
4-pregnene-l7a,2l-diol-3,l1,20-trione by heating wi
60% formic acid on a steam bath. The crystalline ma
terial is the 6-eXo-methylene-4-pregnene-17¢,2l-diol-3,ll,
O1
20-trione. The non-crystalline material is recovered as
dine.
Alternately, heating the 17oc,20,2i),21-biS-1'tl6thYl6Il?
dioxy-derivative with 50% acetic acid at 100° C. for 7
hours, followed by acetylation and chromatography,
gives the 6-exornethylene-4-pregnene-l7a,l113,21-triol-3;
the 6 - exomethylene-ll-pregnene-l7a,2l-diol-3,l1,20-tri
one-2l~acetate by reaction with sodium methoxide in
10 methanol to cleave any formate esters, recovering the
product, and acetylating with acetic anhydride in pyri
ZO-dione-Zl-acetate
in 50-60% yields.
Following the procedure set forth above, 6-exometh
y1ene-4-pregnene-l113,17a,21-triol-3,20-dione
and the cor
responding Zl-acetate are prepared by the following
steps:
l7(1,20,20,21-bis—rnethylenedioXy-3~ethylenedioxy
5u-pregnane-6,11-dione is reacted with acetylene mag
me.
Following
the corresponding 21-acetate are prepared by the follow
ing steps: 16a - methyl-l7ot-20,20,2l-bis-methylenedioxy
15
3-ethylenedioxy-5a-pregnane-6,ll-dione is reacted with
nesium bromide to give 6a-ethynyl-17a,20,20,21-bis-meth
acetylene magnesium bromide to give loot-methyl-ém
ylenedioxy - 3 - ethylenedioxy-Sa-pregnane-oti-ol-ll-one,
which is dehydrated with thionyl chloride in pyridine at
30° C. or below to form 6-ethynyl-17u,20,20,21-bis-meth
ethynyl-l7a,20,20,2l-bis-methylenedioxy - 3 - ethylenedi
oxy-5a-pregnane6B-ol-ll-one, which is dehydrated with
20
thionyl chloride in pyridine at 30° C. or below to form
l6a-methyl-6-ethynyl - l7a,20,20,21 - bis-methylenedioxy
ylenedioxy-3-ethylenedioxy-5pregnene-ll~one
and also
small quantities of 6-ethynyl-l7a,20,20,21-bis-methylene
3-ethylenedioxy-5-pregnene-ll-one and also small quan
dioxy - 3-ethylenedioxy-6-pregnene-1l-one and 6-([2]
tities of l6u-methyl-6-ethynyl-17a,20,20,2l-bis—methylene
dioxy-3-ethylenedioxy-6-pregnene-ll-one and l6a~methyl
6-([2]-chloro-[1,2]-propadiene) - l7a,20,20,2l-bis-meth
ylenedioXy-3-ethylenedioxy-5'-pregnane-ll-one.
The 16cc
methyl-6-ethynyl-17a,20,20,2l - bis - methylenedioxy - 3
chloro - [1,2] - propadiene)-l7a,20,20,2l-bis-methylene 25
dioxy-3-ethylenedioxy-5-pregnane-ll-one. The 6-ethynyl
l7a,20,20,21 - bis - methylenedioxy-3-ethylenedioXy-5—
pregnene-ll-one is separated from 6-ethynyl-l7a,20,20,
.21 — b-is-methylenedioxy-3-ethylenedioxy-?-pregnene-ll
one and 6-([Zlohloro-[1,21-propadiene)-17m,20,20,21
bis-methylenedioxy-3-ethylenedioxy—pregnane=-1l-one by
chromatography on acid-washed alumina, and puri?ed
30
ethylenedioxy-S-pregnene-ll-one is separated from the
by-products by chromatography on acid-washed alumina,
and puri?ed by recrystallization from ethyl acetate. The
160a - methyl - 6 - ethynyl-l7a,20,20,ZI-biS-methylenedi
by recrystallization from ethyl acetate. The 6-ethynyl
oxy-3-ethylenedioxy-5-pregnene-ll-one, dissolved in ethyl
l7a,20,20,21 - bis - methylenedioxy-3-ethylenedioxy-5
acetate, is then hydrogenated, using a Lindlar catalyst, at
atmospheric pressure at about 20-25° C., to give 160:
pregnene-ll-one dissolved in ethyl acetate, is then hydro
genated, using a Lindlar catalyst, at6-vinyl-17a,20,20,Z1
atmospheric pres
methyl-6-vinyl - 17a,20,20,21 - bis - methylenedioxy-3-eth
ylenedioxy-S-pregnene-ll-one. The latter compound is
sure at about 20—25° C., to give
bis - methylened-ioxy-3-ethylenedioxy-S-pregnene-ll-one.
The latter compound is oxidized with osmium tetroxide
to give 6-( [1,21-ethanediol) -17a,20,20,2l-bis-methylene
dioxy-3-ethylenedioxy-5-pregnene-ll-one, which is then
oxidized with osmium tetroxide to give l6ot-methyl-6
40
cleaved with periodic acid to yield 6-formyl-l7ct,20,20,21
bis-methylenedioxy-3-ethylenedioxy-5—pregnene-6-formyl
l7 a,20,20,2l-b-is-rnethylenedioxy-3ethylenedioxy-l l-one.
The 6-formyl-l7a,20,20,2l-bis-methylenedioXy-3-ethyl
droxy-rnethyl-l7a,20‘,20,2l-bis-methylenedioxy-3ethylene
enedioxy-S-p-regnene-ll-one is converted into the 6-hy
dioxy-S-pregnene-llB-ol, by contacting with sodium boro
hydride for about 60 hours at room temperature. The
6 - hydroxyrnethyl - l7a,20,20,21-bis-methylenedioXy-3
ethylenedioxy-S-pregnene-lle-ol is converted into 6-exo
methylene - 17 a,20,20,2l-bis-methylenedioxy-4~pregnene
ll?-ol-3-one by reaction with p-toluenesultonic acid at
room temperature for several hours.
the procedure set forth above, 160t-Tl'lBthY1-6
exomethylene-4-pregnene-l l6, l7 a,2l-triol-3 ,20-dione and
e G-exometh
([l,2]-ethanediol)-l7oc,20,20-,2l - bis - methylenedioXy-3
ethylenedioxy-S-pregnene-li-one, which is then cleaved
with periodic acid to yield l6a-methyl-6-formyl-17ot,20,
20,2l-bis-methylenedioxy - 3 - ethylenedioxy-S-pregnene
ll-one.
The 16cc - methyl-6-formyl - 17oc,20,20,21 - bis-meth
ylenedioxy-3-ethylenedioxy-5-pregnene-ll-one
is convert
ed into the 16tit-methyl-6-hydroxymethyl-17u,20,20,21-bis
methylenedioxy - 3 - ethylenedioxy-S-pregnene-l15-01, by
contacting with sodium borohydride at room temperature
for about 50 minutes. The l6oc-methyl-6-hydroxymethyl
17ot,20,20,21 - bis - methylenedioxy-3 - ethylenedioxy-S
pregnene-ll?-ol is transformed into l6ec-rnethyl-6-exo
methylene - l7a,20,20,2l- - bis - methylenedioxy-4-preg
nene-l1B-0l-3-0ne by reaction with p-toluenesultonic acid
at room temperature for several hours. The l6a-methyl
ylene - 17 a,20,20,2l-bis-methylenedioxyA-pregnene-l1(3
ol-3-one is transformed into 6-exome-thyiene-4-pregnene
1le,l7a,2i-triol-3,ZO-dione by heating with 60% formic
6-exornethylene - l7oz,20,20‘,21 - bis - methylenedioxy - 4
pregnene-ll?-ol-S-one is converted into l6oc-methyl-6-exo
methylene-4-pregnene-11B,17a,2l-triol-3,20-dione by heat
acid on a steam bath. The crystalline material is the 6
eXornethyl-ene - 4 - pregnene-llB,l7a,2l=triol-3,ZO-dione.
ing with 60% formic acid on a steam bath. The crystal
line material is the l60t-Il'l?il'lYl-6-BXOIIlGihYlBH6-4-Pi'6g
The non-crystalline material is recovered as the 6-exo 60 nene-11{3,17a,2_l-triol - 3,20 - dione. The non-crystalline
methylene-4-pregnene-115,17 a,2l-triol-3 ,ZO-dione 21-ace
material is recovered as the ldie-methyl-6-exomethylene
tate by reaction with sodium methoxide in methanol to
cleave any formate esters, recovering the product, and
4-pregnene-l
l[3,l7a,2l-triol-3,ZO-dione-Zl-acetate by reac
tion ?rst with sodium methoxide in methanol to cleave any
acetylating with acetic anhydride in pyridine.
The 6-exomethylene-4-pregnene-l7ot,2l-diol-3,11,20-tri
one and the corresponding Zl-acetate are prepared by
65
converting the 17a,20,20,2l-bis-methylenedioXy-3-ethyl
eneclioXy-Se-p-regnane-GJl-dione into the 6-£ormyl-l7a,
20,20,21 - bis-methylenedioxy-3-ethylenedioXy-5~pregnene
ll-one as indicated above. The latter compound is then 70
converted into the 6-hydr-oxymethyl-l7a,20,20,2l-bis
methylenedioxy - 3-ethylenedioxy-S-pregnene-ll-one by
contacting with sodium borohydride for about 50 minutes
at room temperature. The 6-hydrox methyl-l7ot,20,20,
21 - bis~methylenedioxy-3-ethylenedioxy-5-pregnene-1l-ol
is converted into ?-enomethylene-l7a,20,20,2labis-meth
formate esters, recovering the product, and acetylating
with acetic anhydride in pyridine.
The 161x - methyl - 6 - exornethylene - 4 - pregnene-l7et,
2l-diol-3,11,20 trione and the corresponding Zl-acetate
are prepared by converting the 17 a,20,20,2l-bis-methylene
dioxy - 3 - ethylenedioxy - Sm-pregnane-dU-dione into the
16m - 'nethyl-6-torrnyl - 17a,20,20,2l - bis - methylenedi
oxy-3-ethylenedioxy-5-pregnene-1l-one as indicated above.
The latter compound is then converted into the loot-meth
yl-6-hydroxymethyl - 17oc,2()',20,21 - bis - methylenedioxy
3-ethylenedioxy-5-pregnene-ll-one by contacting with so
3,074,935
8.
about 50 minutes at room tempera
into 2 liters of Water and extracted with ethyl acetate.
17cz,20,20,21
ethylenedioxy - 5 - pregnene-ll-ol
is converted into l6a-methyl-6-exomethylene-17a,20,20,
The organic phase is washed with water, dried over so
dium ‘sulfate and concentrated in vacuo. The crude prod
2l-bis-methylenedioxy 3,11-dione by reac
tion With p-toluenesulfonic acid at room temperature for
uct is chromatographed on acid-washed alumina.
tion with mixtures of chloroform and ether, and crys~
t-allization from ethyl acetate yields 17.2 g. (90%) of
several hours. The 16e-methyl-e-exomethylene-170:,20,
20,21~bis-1nethylenedioxy-ll-pregnene-3,1l-dione is trans
6a-ethynyl-l7a,20,20,2l-bis-methylenedioxy - 3 - ethylene
formed into the 1Gar-methyl;6-exomethylene-At-pregnene
17e,=21-diol-3,11,20-trione by heating with 60% formic
acid on a steam bath. The crystalline material is the 16er
dioxy-ja-pregnane-??-ol-ll-one, M.P. 221-230° C. A.
10
methyl-6-exomethylene - 4 - pregnene - 17a,21—di0l-3,11,
20-trione.
The non-crystalline material is recovered as
sample for analysis, after recrystallization from ethyl.
acetate, has ‘the following properties: M.P. 223-229” C.;
‘11,24 —62“ (c. 1.0 CHClg). Analysis (calculated for
dio1-3,11,20-trione 21-acetate by reaction With sodium
CWT-T3608):
C,
66.87; H, C,
7.29.66.37; H, 7.43. Found (approximately):
Example 2
covering
the product, and acetylating with acetic anhye
dride
in pyridine.
A solution of .thionyl chloride in pyridine is prepared
by adding 7.3 cc. of freshly distilled thionyl chloride to
the 16u-methyl~6-exomethylene - 4 - pregnene - 1704,21
methoxide in methanol to cleave any formate esters, re 15
36 cc. of ice cold anhydrous pyridine. This is added drop
The Mot-methyl - 17a,20,20,21 - bis - methylenedioxy
5a-pregnane-6,1l-dione used as the starting material for
Wise to a stirred solution of 6a-ethynyl_l7a,20,20,2l-bis
the above reaction can be prepared by reacting 16a-meth 20 methylenedioxy - 3 - ethylenedio-xy - 5a - pregnane - 6,8
yl-cortisone with formalin
ol-ll-one in 42 cc. of pyridine. The rate of addition is:
controlled so to maintain the temperature of the reaction
16a-rnethyl - 17a,20,20,21 - bis - methylenedroxy - 4-preg
nene-3,11-dione. The latter compound is converted into
16a-methyl - l7a,20,20,21 - ‘bis - methylenedioxy-3-eth—
ylenedioxy-S-pregnene-ll-one on reaction with ethylene
glyco and p-toluenesulfonic acid. The 16a-methyl-17u,
20,20,21-bis - methylenedioxy - 3 - ethylenedioXy-5lpreg
bis-methylenediOXy-Sa-pregnane-3,6,1l-trione. The latter
of 6-ethynyl-17ot,20,20,21-v
3 -- ethylenedioxy - 5 - pregnene
ture of the l6a-methyl-17u,20,20,2l-bis-methylenedioxy
3-ethylenedioxy-?'a-pregnane-1Lone-5,6 oxides, which on
solution gives 16oc-l'l’16thYl-l7a,20,20,2l
'
bis - methylenedioxy -
none-1 l-one is oxidized with perbenzoic acid to give a mix
reaction sequentially with formic acid and then with potas
mixture at 30°
additional 0.5
Water. The mixture is extracted with chloroform Washed.
with water, dried and concentrated in vacuo. Crystalliza
tion from other affords 2.64 g.
ll-one which decomposes at 200-207"
C. theThe
sample:
for
analysis, recrystallized from ether, has
following:
properties: decomposes at
CHCIg); U.V. A max. 233 208° C., (x1323 ~103° (c. 0 85
mp, e=12,500. Calculated
_ MEL-3407)! C, 68.92;
H, 7.28. Found: C, 68.71; H,
compound is reacted with butanone dioxolane to give 16oz
nane-6,1l-dione.
Elution with a 5:5 mixture of ether’
and petroleum ether affords 331 mg. of 6~([2]chloro-
The novel 6-exomethylene-ll-pregn-ene-ll?,l7ot,2l-triol~
3,20-dione,
the 16u-methyl-6-exomethylene-4-pregnene
3-ethylenedic-xy-S-pregnane-l l-one,
methyl - I7CL,ZO,20,21 - bis _ methylenedioxy - 5a - preg
11,8,l7a,21-triol-3,20-dione, and the 2l-acetates thereof,
[ 1,21-propadiene) - l7a,20,20,21 - bis - methylenedioxy
l76-210° C..
Two crystallizations from methanol affords an analytical
may be dehydrogenated at C-1 to form the corresponding 40 sample, which has the following properties: decomposes.
1,4-pregnadienes by the use of selenium dioxide, or by
microbiological methods.
The novel 6-exomethylene A-pregnene-Ii-ones and the
in?ammatory activity, and are
especially elfective for the
4
at 239-248° C.; (x1322 —157” (c. 0.5 CHCIg). Calculated
for C37H3507C1§ C, 63.96; H, 6.96; Cl, 7.02. Found: C,
63.67; H, 7.11; Cl, 7.06.
6-exomethylene-A1,4-pregn-adiene-3-ones possess high anti
treatment of arthritis and related diseases, since they can
be administered for their cortisone-like action in low dos
thereby minimizing undesirable side effects.
pharmaceutical compositions
ylene-A4-steroid~3-ones and containing these 6-exometh
bis - methylenedioxy - 3 - ethylenedioxy - 6
pregnene-l l-one.
Elution with ether affords an additional 0.87 g. of
é-ethynyl - l70z,20,20,21 - bis - methylenedioxy-3-ethylene
dioxy-S-pregnene-ll-one decomposing at 194-204” C.,
6-exomethylene-Alr‘i-steroid
>3-ones.
The following examples illustrate methods of carry
atotal of3.51 g. (56%).
ing out the present invention but it is to be understood
that these examples are given for purposes of illustration
‘
Example 3
A solution of 3.70 g. of 6-ethynyl-17u,20,20,2l-bis
methylenedioxy-3-ethylenedioxy-S-pregnene-l l-one in 125
Example I
cc. of 23° C. using 3.15 g. of Lindlar catalyst, which is
added in 3 separate 1.05 g. portions: (1) at the beginning
of the hydrogenation, (2)
'
A solution of ethyl magnesium bromide is prepared
from 20 g. of magnesium and 80 cc. of ethyl bromide in
(3) after about
'
200 cc. of anhydrous tetrahydrofuran. Acetylene is 60 spent catalyst.
bubbled through a solution of tetrahydrofuran for about
absorbed, the calculated amount being 189 cc.
1 hour to form a saturated solution. The ethyl magne
tion is ?ltered
'
then added at a temperature of 40-50°
vinyl~17a,.20,20,2l-bis-methylenedioxy - 3 -
S-Pregnene-II-one showed the following properties: U.V.
max. 238 mu, 11,600;
material may be used
be puri?ed by chromatography on alumina followed by
recrystallization.
70
Example 4
A solution consisting of 3.8
g. of the crude 6-vinyl
l7cz,Z0,20,21 - bis - methylenedioxy - 3 - ethylenedioxy-S
pregnene-ll-one and 1.23 g. of 0504 (60% of the amount
calculated for the pure steroid) in 100 cc. of benzene is
allowed to stand at room temperature overnight. The
3,074,935
black r-osmate is transferred to a tlask containing 150 oc.
methylenedioxy - 4-‘- pregnene - 115 - ol-Z-one, MP. 221
each of water and methane , and 15 g. each of potassium
228° C. The sample for analysis is crystallized twice
bicarbonate and sodium sul?te. The benzene is removed
by distillation, 200 cc. of methanol is added, and reflux
from ethyl acetate and has the following properties:
232° C’, 1x922 +139° (c. 1 ‘Cl-1G3); UN. 1\ max. .262
is continued for a total of 4.5 hours. The solution is
m,;., e 9,900. Analysis (calculated for GMT-13266): C,
?ltered and extracted with chloroform. The chloroform
layer is Washed with water dried and concentrated in
69.21; H, 7.74. Found:Example
C, 69.83;
7 H, 7.94.
methylenedioxy
- 3 6-([1,21-cthanediol)-17a,20,20,21-bis
_ ethylenedioxy - 5 - pregnene - 6{3—olvacuo
to give the
A suspension of 550 mg. of 6-formyl-17 u,20,20,21-bis
11-one. The unpuri?ed material may be used directly in 10 methylenedioxy - 3-ethylenedioxy-5-pregnene-1l-one, and
the next step, or it may be puri?ed by chromatography
550 mg. of sodium borohydride in 27 ml. of tetrahydro
onaluminm
furan and 5.4 ml. of water is stirred for 50 minutes at
Example 5
room temperature. The reaction mixture is poured into
ice water containing 6 g. of sodium dihydrogen phos
15 phate, extracted with chloroform, dried and concentrated.
Two crystallizations from ethyl acetate affords a sample
The 6-([1,2] - ethanediol)-17a,20,20,21-bis-methylene
for analysis of 6-hydroxymethyl-17a,20,20,21-bis-meth
dioxy-Z-ethylenedioxy-S-pregnene-ll-one is cleaved by re
ylenedioxy - 3 - ethylenedioxy - 5 - pregnene-l l-one, Ml’.
action of 3.9 g. of the crude diol With 3.8 g. of periodic
195-199” Cs, (x3323 --69 (c. 1 CHCla). Calculated for
acid in a solution consisting of 99 cc. methanol, 90 cc.
pyridine and 90 cc. of water. The solution is maintained 20 0261313608: C, 65.53; H, 7.61. Found: C, 66.01; H,
at room temperature for 45 minutes (there is an initial
7.64.
temperature rise) and poured into aqueous sodium bi;1
carbonate.
The mixture
extracted
withwater,
chloroform
the chloroform
layer is iswashed
with
dried an
concentrated.
_ of 125 {mg 0i 6'hYdTOXYm‘FthYI'170140310,
A 591mm“
Trituration with ether affords 2.2 g. of
cwde crystamne material, MR 200%“? C
21 ' bls ' methylenedloxy ' 3 ' ethylenedloxy - 5_ - 19mg
CryStaL
nene-ll-one, and 15 mg. of p-toluenesulfonic acid 18 al
?zation from ‘Ethyl acetate a?otds 030 g‘ (22%) of 6_ 25 lowed to stand at room temperature overnight. The re
formyl - 17oz,20,20,2l - bis - methylenedioxy-3-ethylenedi_
action mixture is poured ‘into aqueous sodium bicarbon
oxy_s_pregnene_n_one, which has me following womb
ties: MR 2354450 0; “D25 #1640 (Q 08 C1163);
ate solution, extracted with‘ chloroform, dried and con
centrated. Two crystallizations from ethyl acetate-ether
U v View 252
-
-
W-
11 600
mt“ e
r
afford a sample of 6 - exornethylene - 17a,20,20,21 -bis
30 methylenedioxy-4-pregnene-3,1l-dione,
Calculated for GET-13403: C, 65.80; H, 7.22. Found: C,
6574; £1,706.
MP.
197~201°
(3.; ‘x924 +201° (c. 0.9 CHClS). Calculated for
CMHWOS: C, 69.54; H, 7.30. Found: C, 69.83; H, ‘7.08.
xample 8
‘Crystallization of the mother liquors gives 60 mg. of
E _
6 - formyl - 17a,20,2(),21 - bis - methyleneclioxy - 3 -
ethylene-dioxy-6-pregnene-1l-one which decomposes at 35
A suspension of 0.26 g. of 6-exomethylene-17a,20,20,
280-310“ C, The formatlon of this compound. is pre.
21 -bis-methyleneclioxy-At-pregnene-1l?-ol-3-one in‘ 8 cc.
sumably due to the presence of some 6-ethynyl-1'7ot,20,
60% aqueous formic acid is heated on the steam bath
20,21 - bis - methylenedioxy - 3 - ethylenedioxy - 6 - Pfeg-
for 20 minutes.
nene-ll-one as a contaminant of the 6-ethynyl-17a,20,
into ice water and extracted with chloroform. The or
The resulting solution is cooled, poured
20,21 - bis - methylenedioxy - 3 - ethylenedioxy - 5 - preg- 40 ganic phase is washed with aqueous sodium bicarbonate,
nene-ll-one of Example 2. A sample for analysis, after
recrystallization from ethyl acetate, has the following
dried, and concentrated in vacuo. Crystallization from
ethyl acetate affords 30 mg. of 6-exomethylene-4-preg
properties:
dioxane); decomposes at 350° C. mp2?‘ +10° (c. 0.5
none-11B,17oc,21-triol-3,20-dione,
which has
following
properties: Ml’. 203° C.; ‘1,323 +256”
(0.5the
dioxane);
U.V. R3552“ 232 me, e 11,300
‘15
Analysis (calculated for (3261-13408): C, 65.80; H, 7.22.
Found (approximately): C, 66.19; H, 6.77.
p
Exam le 16
U.V. #1,‘:2“ 262 my, 6 10,300
To 175 mg. of the non-crystalline material is added 2
ml.
methanol
and 0.10
of at1 N
sodium
methoxide.
Theofmixture
is allowed
to ml.
stand
room
temperature
for
15 minutes under an atmosphere of nitrogen in order to
-
.
.
A solution of 0.94r g. of 6-formyl-l7a,20,20,2l-bis- “0 cleave any formate esters. After the addition of 2 drops
methylenedioxy-S~ethylenedioxy-5-pregnene~1l-one in 50
of acetic acid, the material is poured into water and ex
cc. of tetrahydrofuran is treated with a solution of 1.0 g.
tracted with chloroform. The chloroform phase is
sodium borohydride in 10 cc. of water. The two-phase
Washed with Water, dried and concentrated in vacuo.
system is stirred for 64 hours at room temperature and 55 The resulting gum is acetylated with 2.1 cc. of pyridine
then poured into 6.0 g. of sodium dihydrogen phosphate
and 2.0 cc. acetic anhydride at room temperature over
in 100 cc. of water. The suspension is extracted with
night The Solution is POllfed into ice Water and ex
ChIQI'GfQI-m and L15 Qrganic phase, is washed with Water,
tracted With chloroform. The ChlOI'O‘fOl‘l'fl P111856 is dl’i?d
dried and concentrated in vacuo. Crystallization from
and cmlcemfated in Vacllo- chromatography on 10 g’
benzene affgrdg 055 g_ (60% yield) of 6-hydyoxymeth- 60 of acid-washed alumina, elution with mixtures of chloro
yl - 17a,20,20,21 - bis - methylenedioxy - 3 - ethylensui.
form and ether and crystallization from ethyl acetate
oxy-S-pregnene-MB-ol, MI’. 220-230" C. The sample
gives 47- Ing- Of 6-@X0m6thY1eI1e-4-Pregn6n6-115.170621
for analysis after crystallization twice from benzene has
“i013,zo'dionerzl'acetate- The analytical Sample, after
the’ following properties; M_P_ 2214306., “D24 __107<>
crystallization from ethyl acetate, has the following prop
(c. 1 Touch). Calculated for cz?nmogz c, 65.25; H, 65 mice M-P- 199-204“; “D24 +293° (c- 0.8 CHEM;
8.00.
Found:ofC, 135
65.00;
A solution
mg.H,of8.22.
6-hydroxymethyl-17a,20,20,
21 - bis - methylenedioxy - 3 - ethylenedioxy - 5 - preg-
Maori 262 m’“ e “A00
U'V' Mm‘Calculated for czéHszosi C, 69-21; H, 7-74~
Found: C,
nene-llB-ol
andis 15allowed
mg. oftop-toluenesulfonic
acid in 15 70 69.00;
cc. of acetone
stand at room temperature
The H,67.63.
- exomethylene-4-pregnene-17a,21-diol-3,11,20
over night. The solution is poured into ice water and
trione and the corresponding 21-acetate are prepared
extracted with chloroform. The chloroform solution is
from the 6-exomethyleue-17a,20,20,2l-bis-methylene-di
washed with aqueous sodium bicarbonate, dried and con-
oxy-4-pregnene-3,1l-dione following the above proce
centrated. Crystallization from ethyl acetate gives 95
dures, but starting with the 6 - exomethylene - l7oc,20,20,
mg. 79% yield) of 6-exornethylene-17u,20,20,21-bis- 75 21Joismethylenedioxy-4-pregnene-3,1l-dione in place of
3,07%,935
ll
the 6 - exomethylene-l7a,20,20,21-bis-methylenedioxy-4
pregnene-l
1,6-ol-3-one.
"i2.
C., 110 mg. of 6-exometl1ylene-4-pregnene~11?,17u,21~
Example 9
To 100 mg. of 6-exomethylene-ll-pregnene-1 1,8,17et,21~
triol-3,20-dione-2l-acetate in 5 ml. of t-butanol and 0.1
triol-3,20-dione-2l-acetate is added to each ?ask as a di
methylforrnamide solution (100 mg./ml.). After a 24
5
ml. of acetic acid is added 50 'mg. of selenium dioxide.
‘The mixture is red .
50
.
trated in ‘vacuo.
1 15,17a,21-triol-3,2'G-dione-21-acetate
rectly from the concentrate. Paper chromatography of
The product is ?ltered,
?ltrate evaporated to dryness.
The residue is
the product in a system utilizing formamide as the sta~
extracted with ethyl acetate and the extract is washed suc
cessively with aqueous
tionary phase and chloroform-‘benzene (1:1) as the mo
'bile phase indicates that the product possesses a polarity
slightly greater than the substrate.
"
.15
concentrated to dryness.
from a mixture of acetone
ene - 1,4 - pregnadiene - 1
The é-exomethylene-1,4-pregnadiene-17a,2l-diol-3,11,
2‘O-trior1e ZI-acetate is prepared from the 6-exomethylene
4-pregnene-l70¢,21-di0l-3,11,2O-trione~21-acetate
following
the above procedure, but starting with the 6-exomethyl
ether gives 6-exomethyl
one~2l-acetate.
hour transformation period ‘at 28° C., the cells are cen
trifuged, followed by three ethyl acetate extracts of the
ene-4-pregnene-17a,21-diol-3,11,20-trione 21-acetate in
l - 3,20 ~ di
20
The 6 - exomethylene-1,4-pregnadiene-17a,2l-diol-3,11,
f20-trione-2l-acetate is prepared from the 6-exomethylene~
i4-pre-gnene ~~ 17a,21-diol-3,l1,20-trione ZI-acetate follow
place
of the 6-exomethylene-4-pregnene-11?,17a,2l-triol—
3,20-dione ZI-acetate.
Example 11
ing the above procedure but starting with the o-exometh
. of l6a-methyl-cortisone in 5
25 litres of chloroform is added 1300 ml. of concentrated
yIene-4-pregnene-l7a,2l-diol-3,l1,20-trione 2l-acetate in
place
of the2l-acetate.
o-exomethylene-4-pregnene-11,8,17a,21-triol~
3,20-di0ne
Example 10
Septomyxa a?im's (ATCC 6737) is inoculated from a
slant into one or more 250 ml. shake flasks containing 30
50 ml. of the following medium: 2% Edamin (Shei?eld
Farms), 5% glucose, and 0.5% corn steep liquor. After
a 48 hour incubation at 28° C., 10 mg. of o-exomethyl
crude crystalline 160t-m€thyl~17OC,2'0,20,21
*bis-methylenedioxy-4-pregnene-3,l l-dione.
ene-4-pregnene-115,17u,21-triol~3,2-0-dione is added to
35
The
of methylene chloride by addition of methanol. ‘The
yield of 16ix-methyl-17u,20,2‘0,21-ibis-methylenedioxy-4—
pregnene-3,11-dione, is ‘60-70%. It has the following
properties: M.P. 244—250° C.,
40
The o-exomethylene-1,4-pregnadiene-11B,17a,21~triol
3,20-dione
'
'
'
Descending paper chromatography of the product in a
system using formamide
substrate, but is largely
45
hydrogenation product.
whit?“ 2383 (15,200)
(Found: C, 69.29‘; H, 7.70. Calculated for C24H32O6: C,
‘69.21; H, 7.74%), [a]D +75° (in CHCl3).
A solution containing 16.2 g. of 16a-methyl-17a,20,20,
21-bis-methylenedioxy~4-pregnene-3,1l-dione dissolved in
800 cc. of benzene is re?uxed over night with 40 cc. of
ethylene glycol and 1.6 g. of p-toluene-sulfonic acid.
The cooled solution is Washed with aqueous sodium bi
carbonate solution, dried over sodium sulfate and con
Bacillus sphaerz'cus (ATCC 12488) is inoculated from
a slant into one or more 250 ml. shake ?asks containing
50 m1. of the following medium: 2% Edamin (Shef?eld
Farms), 5% glucose, and 0.5% corn steep liquor. After
centrated in Vacuo. The crude product is adsorbed on
400 g. of acid-Washed alumina from benzene. Elution
with 1:1 ether-petroleum ether yields 8.5 g. (47%) of
product, M.P. 2i00~210° C. The analytical sample of
a 24 hour incubation at 28° C., 10 mg. of é-exomethyl
ene-4-pregnene-11p’,l7e,2l-triol-3,20—dione is added to
a dimethylformamide solution (100
55
1604 - methyl-170:,20,20,21~bis-methylenedioxy-3-ethylene
dioxy-S-pregnene-ll-one is crystallized from methanol.
It has the following properties: M.P. 211—220° C.
(Found: C, 67.99; H, 8.07. Calculated for C26H36O7:
C, 67.80; H, 7.88%), [ab —80‘° (in CHCl3).
A solution containing 16.5 g. of 16a-methyl-l7a,20,2{),
The 6-exomethylene-1,4-pregnadiene-l l/3,17a,2l-triol
3,20-dione is readily crystallized from the concentrate.
Descending paper chromatography of the product in a 60 2‘1 - :bis-methylenedioxy - 3~ethylenedioxy-S-pregnene-11
system using formamide as
one and 45 meq. of penbenzoic acid in a total volume
of 350 ml. of
dark room at 25°
'
shows some of the ste
roid substrate, but is largely the somewhat more polar
A'-dehydrogenation product.
The o-exomethylene-1,4-pregnadiene-17a,21-diol-3,1l,
Z'OJtrione is prepared from the o-exomethylene-ll-preg
nene-17a,21-diol-3,11,20-trione following the above pro
starting with the 6-exomethylene-4-pregnene
17a,2l-diol-3,11,20-trione in place of the 6-exomethylene~
5% acetone in hexane, separates the isomers.
methyl - l7a,20,2\0,21 - bis-methylenedioxy-3-ethylenedi—
4-pregnene-l1o,l7a,21-triol-3,20-dione.
oxy-5,6<x-oxido - 50¢ - pregnane-l l-one has the following
properties: M.P. 236—243‘’ C. Found: C, 65.27; H, 7.45.
Corynebacrerizmz simplex (ATCC 6946) is inoculated
Caculated
(in CHCIS).for Cad-13608: C, 65.57; H, 7.61), [ab _95°
250 ml. shake ?asks contain
ing a medium having the composition: 1 g./liter yeast
extract (Difco). After an 18 hour growth phase at 28°
The mixed oxides, 1oor-rnethyl-l7m,20,20,2l-his-methyl
75
enedioxy - 3 - ethylenedioxy - 5,6“ - oxide - 5a - preg
nane-l l-one and 16Ci-mfifhyh-170t~20,20-2I-blS‘mEthYICHB
3,074,935
id
propadiene)-1'iu,20,20,2l - bis - methylenedioxy - 3 ~ eth
13
ylenedioxy-S~pregnane-ll-one and l6a-methyl-6-ethynyl
dioxy - 3 - ethylenedioxy - 5,6,6 - oxide - pregnane - 11
l7a,20,20,2l - bis - methylenedioxy - 3 - ethylenedioxy - 6
one, are dissolved in 300 cc. of 98-100% formic acid
at room temperature and allowed to stand 2.5 hours. The
pregnene-ll-one. Elution with ether gives an additional
quantity of léa-methyl-d-ethynyhl7n,20,20,21-bis-meth
ylenedioxy-3-ethylenedioxy¥5 -pregnene-l l-one.
solution is poured into water and extracted with chloro
form. The chloroform solution is washed with saturated
aqueous sodium bicarbonate solution, dried over sodium
Example 14
A solution of 3.70‘ g. of l6rx—methyl—6-ethynyl-17all),
sulfate and concentrated in vacuo.
The crude mixed formates dissolved in 1400 cc. of
20,21 - bis - methylenedioxy - 3 - ethylenedioxy - 5 - preg
methanol are re?uxed 0.5 hour under nitrogen with a 10 nene-ileone in 125 cc. of ethyl acetate is hydrogenated at
solution consisting of 27 g. of potassium hydroxide and
atmospheric pressure and 23°
' 3.15 g. of Lindlar
135 cc. of water. The cooled solution is neutralized with
catalyst, which is added in 3 separate 1.05 g. portions: (1)
37 cc. of acetic acid and concentrated in vacuo at 30° C.
at the beginning of the hydrogenation, (2) after about 20
The resulting solution is poured into Water and extracted
minutes, and (3) after about 40 minutes, after ?rst ?lter
with chloroform. The chloroform layer 15 washed with 15 ing off the spent catalyst. A total of about 177 cc. of
aqueous sodium bicarbonate solution, dried over sodium
sulfate and concentrated in vacuo. Crystallization from
hydrogen is absorbed, the calculated amount being 184
cc. The solution is filtered and concentrated to give the
methanol yields the l6a-methyl-l7a,20,20,2l-bis-methyl
1604 - methyl - 6 - vinyl - 17 a,20,20,2l - bis - methylene
dioxy-3~ethylenedioxy-5-pregnene-1l-one. The crude ma—
- enedioxy-Sa-pregnane-3,6,ll-trione.
A solution consisting of 125 mg. of p-toluene-sulfonic
20 terial may be used directly in the next step, or it may be
acid in 2 cc. of butanone dioxolane is added to a boiling
puri?ed by chromatography on alumina followed by re
suspension containing 4.94 g. of the léer-methyl-l7w20g20,
,2l-bis-methylenedioxy-S a-pregnane-3 ,6, l l-trione in
crystallization.
cc. of butanone dioxolane. After a ‘reflux period of 20
minutes, the reaction mixture is cooled quickly in an ice
A solution consisting of 3.8 g. of the crude l6m-methyl
6 - vinyl - l7<x,2(),20,21 - bis - methylenedioxy - 3 - ethyl
bath, washed with aqueous sodium bicarbonate solution,
dried over sodium sulfate and concentrated in vacuo. The
material is puri?ed by chromatography to yield the 16er
methyl - 17a,20,20,21 - bis - methylenedioxy - 3 - ethylene
dioxy-S a-pregnane-6,11-dione.
Example 1 2
Example 15
enedioxy-S-pregnenedl-one and 1.23 g. of 0504 (60%
of the amount calculated for the pure steroid) in 100' cc.
of benzene is allowed to stand at room temperature over
30 night. The black \osmate is transferred to a ?ash con
taining 159‘ cc. each of water and methanol, and 15 g.
A solution of ethyl magnesium bromide is prepared
from 20 g. of magnesium and 80 cc. of ethyl bromide in
200 cc. of anhydrous tetrahydrofuran. Acetylene is
each'of potassium bicarbonate and sodium sul?te. The
benzene is removed by distillation, 200 cc. of methanol
is added, and re?ux is continued for a total of 4.5 hours.
bubbled through a solution of tetrahydrofuran for about
1 hour to form a saturated solution. The ethyl mag
nesium bromide is then added at a temperature of 40-5 °
C. over a period of 0.5 hour to the saturated solution
The solution is ?ltered and extracted with chloroform.
The chloroform layer is washed \with water, dried and
concentrated in vacuo to give the 16e-methyl-6-([1,2]
ethanediol) - 17a,20,20,2l - bis - methylene-dioxy - 3 -~eth
ylenedioxy-S-pregncne-6?-ol-ll-one. The unpuri?ed ma
of acetylene in tetrahydrofuran, and the mixture is kept 40 terial may be used directly in the next step, or it may be
puri?ed by chromatography on alumina followed by re
at room temperature for an additional 2 hours, during
which time the acetylene how is continued. A solution
crystallization.
of 18 g. of l6a-methyl-l7a,20,20,2l-bis-methylenedioxy
3-ethylenedioxy-5a-pregnane-6,1l-dione in 200cc. of tetra
The l6a-methyl-6-( [1,2]-ethanediol)-l7oc,20,20,2l-bis
hydrofuran is then added to the acetylene magnesium
bromide for a period of 0.3 hour and the resulting solu
tion is kept
flow being
the steroid
mixture is
Example 16
methylenedioxy-3~ethylenedioxy-5 - pregnene - 11 - one is
cleaved by reaction of 3.9 of the crude diol with 3.8 g. of
at room temperature over night, the acetylene
periodic acid in a solution consisting of 90 cc. methanol,
discontinued 3.2 hours after the addition of
90 cc. pyridine and 9 cc. of Water. The solution is main
(7 hours total from the start). The reaction
tained at room temperature for 45 minutes (there is an
50
then poured into 2 liters of water and ex
initial temperature rise) and poured into aqueous sodium
tracted \with ethyl acetate. The organic phase is washed
bicarbonate. The mixture is extracted with chloroform
with water, dried over sodium sulfate and concentrated
in vacuo. The crude product is chromatcgraphed on
acid-washed alumina. Elu'tion with mixtures of chloro
and the chloroform layer is washed with water, dried and
concentrated. Trituration with ether affords a crude crys
talline material which on recrystallization from ethyl ace
form and ether, and crystallization from ethyl acetate,
tate affords the 16a-methyl-6-formyl-17m,20,20,2l-bis
yields
l6or-methyl-6wethynyl-l’loa,20,20,2l-bis-methylene
dioxy-3-ethylenedioxy-5 a-pregnane-dii-ol-l l-one.
Example 13
A solution of thionyl chloride in pyridine is prepared
by adding 7.3 cc. of freshly distilled thionyl chloride to
36 cc. of ice cold anhydrous pyridine. This is added
methylenediOxy-S-ethylenedioxy-5 -pregnene-l l-one.
Example 17
A solution of 0.94 g. of l6a-methyl-6-formyl-l7u,20,
60 20,2l-bis-methylenedioxy-3 - ethylenedioxy - 5 - pregnene
a stirred solution of l6e-methyl-l7ar-20,20,
dropwise
21 - bis - to
methylenedioxy - 3 - ethylenedioxy - 5a - preg
nane-oii-ol-llione in 42 cc. of pyridine. The rate of ad
dition is controlled so to maintain the temperature of the
l'l-one in 50 cc. of tetrahydrofuran is treated with a solu
tion of 1.0 g. sodium borohydride in 10 cc. of water.
e
two-phase system is stirred into 6.0 g. of sodium dihydro
gen phosphate in 100 cc. of water. The suspension is ex
tracted with chloroform and the organic phase is Washed
with water, dried and concentrated in vacuo. Crystalliza
reaction mixture at 30° C. or below. The solution is
tion from benzene a?ords l6m-methyl-o-hydroxymethyl
stirred an additional 0.5 hour, cooled in ice and poured
17a,20,20,2l - bis - methylenedioxy - 3 - ethylenedioxy-S
into ice water. The mixture is extracted with chloro
form, washed with water, dried and concentrated in vacuo. 70
pregnene-l
15-01.
A solution
of
Crystallization from ether allords 6e-methyl-6-ethynyl
135 mg. of 16a-methyl-l7or,20,20,21-bis
l7d_.20,20,21 - bis - methylenedioxy - 3 - cthylenedioxy
methylenedioxy-B-ethylenedioxy-5-pregnene-ll?-ol
and 15
mg. of p-toluenesulfonic acid in 15 cc. of acetone is al
S-pregnenc-ll-one.
The mother liquors
lowed to stand at room temperature overnight. The
solution is pour-ed into ice water and extracted with
are chromatographed on acid
washed alumina and eluted with mixtures of etherzpw
troleum ether to give 'l6m-methyl-6-([2]-chloro-[l,2]
3,074,935
hydrochloric acid and water, and then dried over mag
nesium sulfate. The extract is treated with activated char
coal and then concentrated to dryness. Crystallization of
the residue from a mixture of acetone and ether gives
Crystallization from ethyl acetate gives 16a-methyle6-exo
methylene-l7a,20,20,21-bis~methylenedioxy - 4 - pregnene
Example 18
l6a-methyl-6-exomethylene-1,4-pregnadicne - 1i/3,17a,21
A suspension of 550 mg. of l6a-methyl-6-formyl-l7u,
20,20,2l=bis-methylenédioxy—3 - ethylenedioxy ~ 5 - preg
'
sodium borohydride in 27
triol-3,20-dione~2l-acetate.
The 16a-methyl-?-exomethylene-1,4-pregnadiene - 17 or,
21-di0l-3,l1,20-trionc-2l-acetate is prepared from the 16cc
ml. of water is stirred for 10 methyl=6eexomethylene-4-pregnene-17a,21 - diol - 3,11,20
trione ZI-acetate following the above procedure, but start
The reaction mixture is
ing with the l6cz-rnethyl-6-exornethylene-4-pregnene-17a,
6 g. of sodium dihydro
2l-diol-3,ll,20-trione 2l~acetate in place of the l6a~meth
a sample for analysis of 16wmethyl~6-hydroxymethyl-17a,
yl-6-exornethylene-4-pregnene~1 1/3, 170:,21-t1‘l0l-3,20-di0l'1€
ZI-acetate.
‘
20,20,21-bis-methylenedioxy-3 -ethylenedioxy - 5 - preg
nene-l l-one.
I A solution of 125 mg. of 1dot-methyl-d-hydroxymethyl~
l7cr,20,20,21~bi8 - methylenedioxy - 3 - ethylenedioxy - 5
pregnene-ll-one, and 15 mg. p-toluenesulfonic acid is
Example 21
Seplomyxa a/?m's (ATCC 6737) is inoculated fromvv a
slant into one or more 250 ml. shake ?asks containing
50 ml. or" the following medium: 2% Edamin (She?ield
Farms), 5% glucose, and 0.5% corn steep liquor. After
a 48 hour incubation at 28° C., 10 mg. of 16a-methyl-6
exornetliylene - 4-pregnene-11B,17a,2l-triol-3,20-dione‘ is
afford a sample of 16a~methyl-6-exomethylene-17a,20,20,
21-bis-rnethylenedioxy-4-pregnene-3,1l-dione.
added to each shake ?ask as a dimethylformamide solu
tion (100 mg./1r1l.). After a conversion period of 24
hours, the cells are removed by ?ltration, followed by three ‘
successive extractions with equal volumes of ethyl acetate.
Example 19
A suspension of 0.26 g. of 16a—methyl-6-exomethylene—
17a,20,20,2l-bis-methylenedioxy~4~pregnene - 11/3 - ol _ 3
one in 8 cc. of 60% aqueous formic acid is heated on
the steam bath for 20 minutes. The resulting solution is
The extracts are combined and concentrated in vacuo.
The 160: - methyl~6-exomethylenc-l,4-pregnadiene-l1,6’,
17a,21-tl‘i01-3,20-di01'16 is readily crystallized from the
concentrate. Descending paper chromatography of the
product in a system using formamide as
polar A’-dehydr0genation product.
concentrated in vacuo. Crystal~
Bacillus sp/zaericus (ATCC 12488) is inoculated from
lization from ethyl acetate affords 16a-methyl~6-exometh‘
ylene-zt-pregnene-fl 1,6,l7a,21etriol-3,20~di0ne.
a slant into one or more 250 ml. shake ?asks containing
50 ml. of the following medium: 2% vEdamin (Sheffield
Farms), 5% glucose, and 0.5% corn steep liquor.
To 175 mg. of the non~crystalline material is added 2
ml. of methanol and 0.10
a 24 hour incubation at 28° C., 10 mg. of 6-exornethylene
16a - methy - 4-pregnene~l1,8,17u,21-triol-3,20-dione is
added to each shake ?ask as a dimethylforrnamide solu
tion (100 mg./rnl.).
'
'
hours, the cells are removed by ?ltration,
three successive extractions with equal volume of ethyl
45 acetate.
vacuo.
The extracts are combined and concentrated in
The 6 - exomethylene-lore-methyl-1,4-pregnadiene-1In,
l-7iz,2l,triol-3,20-dione is readily crystallized from the
concentrate. Descending paper chromatography of the
50 product in a system using formamide as the stationary
phase and chloroform as the mobile phase, shows some of
the steroid substrate, but is largely the somewhat more
and crystallization from ethyl acetate
exomethylene-4-pregnene-1 1/8,17a,21~triol-3,20-dione ace
tate.
polar A'-dehydrogenation product.
The l6a~methyl~6~eX0methylene ~ 4 - pregnene - 17a,21
and the corresponding ZI-acetate are
prepared from the 16ix-methyl-6-exomethylene-17a,20,20,
21-bis~methylenedioxy-ll-pregncne - 3,11 ~ dione following
the above procedures, but starting with the 16a-methyl-6
exomethylene-l7a,20,20,2l-bis-methylenedioxy - 4 - preg_ 60
nene-3,11~dione in place of the l6a-methyl-6-exomethyl
1
The 16st - methyl-o-exornethylene-l,4—pregnadiene-l7a,
2l-diol-3,l1,20-trione is prepared from the l6e-methyl~
6 - exomethylene - 4- regnene-l7a,2l-diol-3,l1,20-trione
following the above procedures, but starting with the 160:
methyl - 6-exomethylene-4-pregnene-17a,2l-diol-3,1 1,20
trione in place of the I6mmethyl-6~exornethylene~4-preg
none-1 1/3,17a,2l-triol—3,20-dione.
Corynebacterium simplex (ATCC 6946) is inoculated
ene-17a,20,20,21-bis-methylenedioxy-4-pregnene-1 15-01-3
one.
Example 20
ing a medium having the composition: 1 g./liter yeast
extract (Difco). After an 18 hour growth phase at 28°
To 100 mg. of 16a-methyl-o-exomethylene-4-pregnene
C., 10 mg. of 6-exomethylene~16a-methyl-4~pregnene—l1/3,
11,8,17a,21-tfiOl-3,20—dl0116 ZI-acetate in 5 ml. of t-butanol
17a,2I-triol-3,20-dione-2l-acetate is added to each shake
?ask as a dimethylformamide solution (100 mg./ml.).
After a 24 hour transformation period at 28° C., the cells
are centrifuged, followed by three ethyl acetate extracts
w
is added and the
.mixture
is re?uxed
an additional 24 hours. The product 70 of the cell-free broth. The extracts are combined and
.is ?ltered, and the ?ltrate evaporated to dryness.
concentrated in vacuo. The 6-exomethylene-l6a-methyl
residue is extracted With ethyl acetate
crystallized directly from the concentrate. Paper chro
matography of the product in a system utilizing formam
:and 0.1 ml. of acetic acid is added 50 mg. of selenium
dioxide.
'
'
1,4-pregnadiene-l1,8,171,21-triol~3,20-dione-2l-acetatc is
75
ide as the stationary phase and chloroform-benzene (1:1)
3,074,985
as the mobile phase indicates that the product possesses
10. 6 - ethynyl - 17a,20,20,2l-bis-methylenedioxy-S
a polarity slightly greater than the substrate.
The 160:, -
ethylenedioxy-6-pregnene-ll-one.
ethyl-6-exomethylene-1,4-pregnadiene-l7a,
21-diol-3,11,20-trione-2l-acetate
is prepared from the 16ozmethyl - 6-exomethylene-4-pregnene-l7u,21-diol-3,11,20trione-21-acetate following the above procedure, but
11. 6 - forrnyl - 17a,20,20,2l - biS-methylenedioXy-3
5
starting with the 16e-methyl-6-exomethylene-4-pregnene-
ethylenedioxy-S-pregnene-1l-one
12. 6 - hydroxyrnethyl - 17a,.20,20,21 - bis-methylene
di0xy-3-ethylenedioxy-S-pregnene-l15-01.
13.,6 - exornethylene - 17a,20,20,21-bis—methylenedi~
17a,21-diol-3,11,20-trione-2l-acetate,
in place of the 1611methyl
- 6 - exomethylene-4-pregnene-11?,17a,21-triol-3,
20-dione21-acetate.
oxy-4-pregnene-1l?-ol-3-one.
14. A compound selected from the
group consisting of
10 6 - exornethylene - 4 - pregnene-l113,17u,21-triol-3,20
Various changes and modi?cations may be made in
carrying out the present invention without departing from
dione and the 21-acetate thereof.
15' A compound Selected from the gYOuP Consisting 0f
the spirit and scope thereof. Insofar as these changes
6-exomethylene -1,4-pregnadiene-llB,17¢,21-triol-3,20=
and modifications are within the purview of the annexed
dione and the 21-acetate thereof.
claims, they are to be considered as part of our invention. 15
We claim:
16- 6 — hydroxymethyl — 17(130,20,ZI-biS-rh?thYlE?Bdi
oxy - 3 - ethylenedioxy - 5 - pregnene-ll-one.
1. Process for the preparation ‘of 6a-ethynyl-17a,20,20,
17- 6 - exomethylene - 1706,20,201l-bis-methylenedi
21
- bis-methylenedioxy-3-ethylenedioxy-5
a-pregnane-6pol-ll-one
which comprises reacting 17a,20,20,2l-bis-
0xy-4-preguene-3,1l-dione.
18. 16a - methyl - 6m-ethynyl-l7a,20,20,21-bis-methyl
methylene - dioxy - 3-ethylenedioxy-5a-pregnane-6,1l-di- 2O enedi0Xy-3-ethylenedioxy-5a-pregnane-6p-ol-1Lone.
one with acetylene magnesium halide.
2. Process for the preparation of 6-ethynyl-17a,2tl,20,
19. 160: - methyl - 6-ethynyl-17a,20,20,21-bis-methyl
6n6di0XY-3-6thY16I16di0XY-5-Pr6gI16I16-11-0n€.
21 - bis-methylenedioxy - 3-ethylenedi0xy-S-pregnene-ll-
one which comprises dehydrating 6u-ethynyl-17u,20,20,
2“ 16d - methyl - 6 - ethYnY1-17¢,20,2O,2l-bis-methyl
enediOXY-3-ethylenedi0XY-6-Pi‘6gnene-1LOBS
21 - bis-methylenedioxy-3-ethylenedioxy-5a-pregnane-6fg- 25 21. l6m-methyl- 6-vinyl-17a,20,20,21-bis-methylene
ol-ll-one with thionyl chloride.
dioXy-3-ethy1enediOXY-5 -r>regnene-11-<>ne
3. Process for the preparation of 6-vinyl-l7u,20,20,
22- 16¢ - m?thyl - 6 - fofmyl-?ulololl-bis—methyl
21
- bis-methylenedioxy-3-ethylenedioXy-5-pregnene-l1one which comprises hydrogenating 6-ethynyl-l7a,20,20,
enedi0XY-3-ethY1enedi0XY'5'pregnene'll'one
23- 16m - methyl - 6 - fofmyl-171119103l-bis-methyl
21
- bis-methylene-di0xy-3-ethy1enedioxy-5-pregnene-11- 30 e?edi0XY-3-ethY1enedi°XY-6-pfegnene-1l-one
one usingaLindlar catalyst,
24. 160:. - methyl - 6 - hydroxymethyl-17a,20,20,21~
4. Process for the preparation of 6—([1,2] -ethanediol)-
biS-methY1enedi0XY-3'ethylenedioxy'5'PIBgn5ne-116-01
17a,20,20,21 - bis - methylenedioxy-3-ethylenedioxy-5-
25- 16a - methyl - 6 - exom?thylene-lhlololl-bis
pregnene-ll-one which comprises oxidizing 6-vinyl-17a,
m6thY1el1edi0XY-4"Pfegnene‘115-01-34)“
nene-ll-one with osmium tetroxide_
5. Process for the preparation of 6-formyl-l7u,20,20‘,
of 160:. - methyl-6-exomethylene-4-pregnene-1l5,17u,21
trim-330410115 and the Zl?cetate thereof
one which comprises reacting 6-( [1,2]-ethanediol)-17a,
16°‘ ' methyl - 6 - exomethylwe - 1,4'PIegI1HGieI1E-11I3J7ri,
20,20,21 - bis - methylenedioxy-S-ethylenedioxy-5-preg- 35
21 - bis-methylenedioxy - 3-ethylenedioxy-S-pregnene-l1-
26- A compound Elected from the group Consisting
27- A compound Selected from the Emu? Consisting of
20,20,21 - his - methylenedioxy-3-ethylenedioxy-5-preg- 40 21"?i01'330'dmm and the 21-666mm them 5
nene-ll-one with periodic acid.
6. Process for the preparation of 6-hydroxymethyl-17a,
28' 16“ - methyl - 6 - hydl’oxymethyl - 17°‘,20,20>21'
bis'methylenedioxy - 3 - ethylenedioxy - 5 - Pfegntme-n
20,20,21
- bis-methylenedioxy-3-ethylenedioxy~S-pregnene1l/3-0l which
comprises reacting 6-formyl-l7a,2(),20‘,2l-
One29- 16°‘ " methyl ‘ 6 ' exomethylene'?u’zogo’zl'bis'
bis
- methylenedioXy-3-ethylenedioxy-5-pregnene-1l-one 45 methylenedi0Xy-4-pregnene-3,1l-dione.
with sodium borohydride for about 60 hours at room
30- A Compound selected from the group consisting of
temperature_
1611 - methyl-6-exomethylene-1,4-pregnadiene-17 ail-diel
7. Process for the preparation of 6-eXomethylene-4-
331104110116 and the zl'acetate thereof’
pregnene-l1{3,l7oc,21-triol-3,20-dione which comprises re
acting 6 - ericimetlhgllene-17220,2021-bis-methylenedioxy— 50
References Cited in the ?le of this patent
-pregnene,B-o - -one
wit a ower carboxylic acid.
8. 60¢ - ethynyl
- 17a,20,20,21-bis-methylenedioxy-3-
ethylenedioxy-Sa-pregnane-6p-ol-1l-one.
9. 6 - ethynyl - 170:,20,2O,21 - bis-methylenedioxy-3-
ethylenedioxy-S -pregnene-l l-one.
55
2,337,454
UNITED STATES PATENTS
Nobile —————————————— —— June 3’ 1958
2,980,711
001w“ --------------- -- Apr- 18, 1961
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