Steroid compounds and processes thereof

Novel steroids of the formula ##STR1## Also there are provided processes for preparing such compounds. The new compounds are useful as intermediates in the preparation of known cardenolides; various of the compounds have other uses as blocking agents.

This invention relates in one embodiment to new steroid compounds, and in a 
further embodiment, to processes for preparing steroid compounds. 
More particularly, according to one embodiment of the present invention, 
there are provided new compounds of the formula 
##STR2## 
wherein R.sub.1 to R.sub.6 and X - Y have the meanings defined below, and 
wherein the dotted lines represent optional double bonds. In the 
embodiment where the 3 and 5 positions of these compounds are saturated, 
the substituents R.sub.1 and R.sub.6 may either be in the alpha or beta 
positions. 
For a greater understanding of the present invention, the novel compounds 
may be classified as being of one of two groups of compounds, in which 
according to one embodiment, the componds of Formula 1 have saturated A, 
B, C and D -- rings and in which R.sub.1 represents either hydroxy or OAc, 
R.sub.2 represents CH.sub.2 OH or CHO, R.sub.3 is H, R.sub.4 and R.sub.5 
are Ch.sub.2 --O, R.sub.6 is either R.alpha. or 5.beta.-H, and X and Y 
together represent 
##STR3## 
alternatively, the compounds according to a further embodiment are those 
compounds in which R.sub.1 represents H, O.dbd., HCO.sub.2, 
(CH.sub.3).sub.3 CCO.sub.2, Cl.sub.3 CCO.sub.2, (C.sub.2 H.sub.5 O).sub.2 
P(O)CH.sub.2 CO.sub.2, 
##STR4## 
HC.tbd.C-CO.sub.2, H.sub.2 C.dbd.CH-CO.sub.2, (HO)(CH.sub.3)CH CO.sub.2, 
(HO)(CH.sub.3).sub.2 CCO.sub.2, HO.sub.2 C(CHOH).sub.2 CO.sub.2, HO.sub.2 
C(CH.sub.2).sub.2 CO.sub.2, HO.sub.2 CCH.sub.2 CO.sub.2, CH.sub.3 O, 
(CH.sub.3).sub.3 CO, C.sub.6 H.sub.5 CH.sub.2 O, HC.tbd.CCH.sub.2 O, 
tetrahydropyran-2-yloxy, 3.alpha., 9.alpha.-oxido and 
3-hydroxy-3.alpha.,9.alpha.-oxido, R.sub.2 represents HCO.sub.2 CH.sub.2, 
(CH.sub.3).sub.3 CCO.sub.2 CH.sub.2 ; Cl.sub.3 CCO.sub.2 CH.sub.2 ; 
(C.sub.2 H.sub.5 O).sub.2 P(O)CH.sub.2 CO.sub.2 CH.sub.2 ; 
##STR5## 
HC.tbd.C--CO.sub.2 Ch.sub.2, H.sub.2 C.dbd.C--CO.sub.2 CH.sub.2, 
(HO)(CH.sub.3)CHCO.sub.2 CH.sub.2, (HO)(CH.sub.3).sub.2 CCOOCH.sub.2, 
HO.sub.2 C(CHOH).sub.2 CO.sub.2 CH.sub.2, HO.sub.2 C(CH.sub.2).sub.2 
CO.sub.2, HO.sub.2 CCH.sub.2 CO.sub.2 CH.sub.2, CH.sub.3 OCH.sub.2, 
(CH.sub.3).sub.3 COCH.sub.2, C.sub.6 H.sub.5 CH.sub.2 OCH.sub.2, 
HC.tbd.CCH.sub.2 OCH.sub.2, tetrahydropyran-2'-yloxymethyl, CH.sub.2 Cl, 
Ch.sub.3, 8,19-oxido, 19.fwdarw.8-lactone, 19.fwdarw.8-lactol, and 
6,19-oxido; R.sub.3 represents H, CO.sub.2 H, CO.sub.2 CH.sub.2 C.sub.6 
H.sub.5 ; CO.sub.2 CH.sub.3, CO.sub.2 CH.sub.2 CH.sub.2 OH, CO.sub.2 
CH.sub.2 -C.tbd.CH and CN; R.sub.4, R.sub.5, represent CH.sub.2 -O, 
R.sub.4 represents CH.sub.3 ; R.sub.5 represents OH, OCH.sub.2 C.sub.6 
H.sub.5, OCH.sub.3, OCH.sub.2 CH.sub.2 OH and OCH.sub.2 -C.tbd.CH, R.sub.6 
represents H, Cl, Br or 5.alpha.-OH, X - Y represents the groups defined 
above or the group 
##STR6## 
or CH--CH.sub.2 ; and wherein double bonds may be present in the 
2,4,6,8(14)-; 4,6,8(14)-; 4,6,8(9)-; 4,6-; 4-; 5; 5,7-; and 
3,5,7-positions. 
Particularly preferred embodiments of the present invention are compounds 
which can be expressed as follows: 
A steroid of the formula 
##STR7## 
wherein the dotted lines represent covalent bonds which may or may not be 
present; wherein the group at the 17 .beta.-position R.sub.3 is H, 
CO.sub.2 H, CO.sub.2 CH.sub.2 C.sub.6 H.sub.5, CO.sub.2 CH.sub.3, CO.sub.2 
CH.sub.2 CH.sub.2 OH or CN, wherein, in the case of there being a bond 
between R.sub.4 and R.sub.5, the latter represent 
##STR8## 
respectively, wherein, in the case of the absence of a bond between 
R.sub.4 and R.sub.5, R.sub.4 is CH.sub.3 and R.sub.5 is selected from the 
R.sub.3 group; wherein in the steroid nucleus R.sub.1 is H, OH, 
##STR9## 
with the proviso that R.sub.1 is not O, OH or an acyloxy group, in the 
case the 17.beta.-group 
##STR10## 
represents an unsubstituted butenolide ring 
##STR11## 
wherein, in the case of there being a bond between R.sub.2 and R.sub.7 or 
R.sub.2 and R.sub.8 the latter represents a 6,19-oxide, an 8,19-oxide or a 
19 .fwdarw. 8 lactone group, respectively; wherein, in the case of there 
being no bond between R.sub.2 and R.sub.7, or R.sub.2 and R.sub.8, R.sub.2 
stands for CH.sub.2 OH, CHO, 
##STR12## 
for H and R.sub.8 stands for OH or H wherein, in the case of the 
5-position being saturated, there may either be present a 5.beta.-or 
5.alpha.-hydrogen atom; wherein, as regards the substitutent in the 
steroid nucleus at the 14- or 15-position, X-Y stands for 
C(.beta.-OH)-CH.sub.2, C(.beta.-OH)-CHBr, C(.beta.-OH)-CHCl, 
C(.alpha.-OH)-CH.sub.2, a .beta.-oxide, an .alpha.-oxide, a double bond, 
C(.alpha.-H)CH.sub.2 or C(.beta.-H)-CH.sub.2. 
Other preferred embodiments of the compounds of the above formula are those 
compounds wherein the steroid is selected from the group consisting of 
carda-14,20(22)-dienolide; carda-4,20(22)-dienolide group; 
19-oxygenated-14-hydroxy-5.alpha., 14.beta.-card-20(22)-enolide; or 
19-oxygenated-14-hydroxy-5.beta., 14.beta.-card-20(22)-enolide. 
Other embodiments relate to compounds of the above formula in which the 
steroid contains a member selected from the group consisting of 6,19-oxido 
substituent; 8,19-oxido substituent; 3.beta.-acetoxy substituent; 22-cyano 
substituent; 22 -benzyloxycarbonyl substituent; 3.beta.-acetoxy 
substituent; 19-formyloxy substituent; 19-hydroxy substituent; 
22benzyloxycarbonyl substituent; card-20(22)-enolide; 14,15 .beta.-oxido 
group, 19- formyloxy substituent; 14-hydroxy substituent; 14.beta., 
19-dihydroxy substituents; 3.beta.-acetoxy substituent; or 3.beta., 
19-diacetoxy substituents. 
A still further embodiment of the compounds of the above formula is where 
there is provided a 3-deoxy steroid having an R.sub.1 substituent in the 
3-position which is a hydrogen atom and in which the 19-position carries 
an oxygen function; and in other embodiments, and in such a case, the 
steroid preferably contains the group in which the substituent R.sub.1 at 
the 3-position is a hydrogen atom and in which the 19-position carries an 
oxygen function. Still further, preferably the crotonic acid or crotonic 
acid derivative carries as group R.sub.3 a carboxylic acid group or a 
derivative thereof. In this group of compounds, preferably the 
8.beta.-position is linked to the 19-position by an oxygen atom and in 
which A-B stands for a double bond or an .alpha.-oxide; there may be 
further included an oxide in the 6,19-position or in the 14,15-position - 
and there may be a hydroxy group in the 14.beta.- and a halogen atom in 
the 15.alpha.-position. 
In accordance with a further aspect of the present invention, there are 
provided processes for preparing the above compounds and compounds 
generally of the formula 
##STR13## 
wherein R.sub.1 through R.sub.6, X and Y have the above-defined meanings 
and additionally may represent, when the A, B, C, and D rings of the 
compounds are saturated, compounds in which X and Y are 
##STR14## 
or C.dbd.CH.sub.2, R.sub.1 is OH or Ac, R.sub.2 is CH.sub.2 OH or CHO, 
R.sub.3 is H and R.sub.4,5 is CH.sub.2 O; in accordance with this 
embodiment of the invention, the process is selected from the group 
consisting of 
(a) treating a compound of the formula 2 
##STR15## 
wherein R.sub.1, R.sub.2, R.sub.6, X and Y and the dotted lines are as 
defined above, and R.sub.7 is OH; with an .alpha.-substituted acetic acid 
to form a compound of the formula(3) 
##STR16## 
wherein R.sub.1, R.sub.2, R.sub.6, X and Y and the dotted lines are as 
defined above and wherein Z is a substituent chosen from those enhancing 
the acidity of the adjacent methylene group; and finally treating the 
compound of the formula (3) with a base to form a compound of the formula 
(1), in which R.sub.1, R.sub.2, R.sub.3, R.sub.6, X and Y and the double 
bonds are as defined above and in which R.sub.4 and R.sub.5 are CH.sub.2 
--O; 
(b) treating a compound of the formula (2) in which R.sub.7 is OH, 
OCOCH.sub.3 or H, with an alkali alkoxyacetylide and then with an alcohol 
or water to form an intermediate compound of the formula (4) 
##STR17## 
wherein R.sub.7 is OH or H and in which alkyl is CH.sub.2 H.sub.5, 
CH.sub.3 or CH.sub.2 C.sub.6 H.sub.5, subjecting the latter compound to 
acid treatment to form a compound of the formula (1) in which R.sub.3 is H 
and R.sub.4, and R.sub.5 is CH.sub.2 --O-- when R.sub.7 in the compound of 
formula (4) is OH, or in which R.sub.3 is H, R.sub.4 is CH.sub.3 and 
R.sub.5 is OCH.sub.2 H.sub.5, OCH.sub.3 or OCH.sub.2 C.sub.6 H.sub.5 when 
R.sub.7 in the compound of formula (4) is H; 
(c) treating a compound of the formula (1) in which R.sub.1, R.sub.2, 
R.sub.6, X and Y and the dotted lines are as defined above, while R.sub.4 
and R.sub.5 represent CH.sub.2 --O and R.sub.3 represents CO.sub.2 H, 
CO.sub.2 CH.sub.2 C.sub.6 H.sub.5 ; CO.sub.2 CH.sub.3 ; CO.sub.2 CH.sub.2 
CH.sub.2 OH or CO.sub.2 CH.sub.2 -C.tbd.CH, with zinc and a carboxylic 
acid to form a compound of the formula (1), in which R.sub.3 represents a 
member selected from the group consisting of CO.sub.2 H, CO.sub.2 CH.sub.2 
C.sub.6 H.sub.5, CO.sub.2 CH.sub.3, CO.sub.2 CH.sub.2 CH.sub.2 OH and 
CO.sub.2 CH.sub.2 --.tbd.CH; R.sub.4 is CH.sub.3, and R.sub.5 is OH, 
OCH.sub.2 CH.sub.2 C.sub.6 H.sub.5, OCH.sub.3, OCH.sub.2 CH.sub.2 OH or 
OCH.sub.2 -C.tbd.CH. 
In greater detail of the above processes, the conversion of the 21-alcohols 
of the formula (2), where R.sub.7 is OH, to the corresponding compounds of 
formula 1 by process (a) is most preferably carried out by treatment of 
the 21-alcohols with an .alpha.-substituted acetic acid, such as 
(diethylphosphono) acetic acid, (benzyloxycarbonyl) acetic acid or 
cyanoacetic acid in the presence of a carbodiimide (for example, 
dicyclohexylcarbodiimide and a water immiscible solvent such as methylene 
chloride or benzene, and subsequent treatment of the resulting reaction 
mixture containing 21-acylate (3) as an intermediate with a water stable 
base- for example aqueous potassium hydroxide or aqueous or anhydrous 
t-butylamine. The reaction is preferably carried out at room temperature, 
though higher or lower temperatures may be employed if desired. 
In an alternate embodinent of the above process (a), the 21-alcohol may be 
heated between 50 to about 150.degree. C. with the corresponding 
.beta.-substituted acetic acid in the presence of a ketonic solvent, sich 
as acetone or methyl isobutyl ketone. Where the radical Z is CO.sub.2 H, 
the resulting 21-hemimalonate may be then esterified by a suitable 
conventional technique - such as for example, to treatment with an 
appropriate diazoalkane, such as diazomethane or diazotoluene to yield 
compounds of the formula (3) in which Z is CO.sub.1 alkyl. Similarly, in a 
related embodiment of process (a), the 21-alcohol may be treated with a 
derivative of an .alpha.-substituted acetic acid, such as an acid 
chloride, ZCH.sub.2 COCl anhydride, (ZCH.sub.2 CO).sub.2 O, mixed 
anhydrides, ZCH.sub.2 CO--O--OCR, wherein Z is as defined above and R is 
(CH.sub.3).sub.3 C, F.sub.3 C or Cl.sub.3 C, or ester, ZCH.sub.2 CO.sub.2 
alkyl, wherein alkyl is CH.sub.3, C.sub.2 H.sub.5, CH.sub.2 C.sub.6 
H.sub.5 or CH.sub.2 -C.tbd.CH. In the case where the carboxylic acid 
derivative is an acid chloride, or an anhydride or mixed anhydride, the 
treatment may be carried out at room temperature in the presence of a mold 
base such as pyridine; in the case where the carboxylic acid derivative is 
an ester the conversion to the 21-acylate may be brought about by heating 
the 21-alcohol with the ester at a temperature of between about 50.degree. 
to 220.degree. C. The base treatment of process (a) may be preferably 
carried out with a strong aqueous base such as 1 - 80% aqueous potassium 
or sodium hydroxide. 
The treatment of the 21-alcohol or 21-acetate of process (b) with an alkali 
alkoxy acetylide may be carried out as described by F. Sondheimer, 
Chemistry in Britain, 1, 454 (1965); preferably it may be carried out at 
room temperature in an anhydrous ether such as diethyl ether or 
tetrahydrofuran and as the alkali alkoxy acetylide lithium methoxy. 
Alternatively, ethoxyacetylide may be employed. The subsequent acid 
treatment may also be carried out at room temperature. Dilute aqueous 
strong inorganic acids, such as hydrochloric, sulfuric or perchloric acid 
are preferably employed as the acid; while as the solvent for the 
intermediate steroidal 20-alkoxyethylnyl-20-alcohol of formula 4 a 
water-immiscible solvent, such as benzene, either or methylene chloride, 
is preferably used. 
The treatment of the 22-substituted cardenolide of process (c) with zinc 
and a carboxylic acid may be preferably carried out at room temperature in 
the presence of an inert solvent such as toluene, methylene chloride or 
ethyl acetate and the carboxylic acid formic acid, acetic acid and 
substituted acetic acids such as trimethylacetic or trifluoroacetic acid 
may be employed. 
The various processes of the present invention possess several unexpected 
and advantageous features. Thus, with regard to process (a) it is 
unexpected that the esterification of the 21-alcohol to compounds of 
formula (2), in which Z is CO.sub.2 H, gives improved yields when ketonic 
solvents, such as methyl isobutyl ketone, are used and that the 
esterification proceeds in a highly selective manner, yielding practically 
only the 21-hemimalonate, in which one of the 2-carboxylic acid groups of 
the malonic acid employed as the reagent remains still unreacted. 
With regard to process (a) it is also unexpected that the conversion of the 
21-(dialkylphosphono)acetates of formula (3), Z being (alkyl)P(O) to the 
corresponding cardenolides of formula (1), R.sub.3 being H, R.sub.4, 
R.sub.5 being CH.sub.2 O can be effected by a base of only moderate basic 
strength, such as an aqueous alkali hydroxide or alkylamine since this 
conversion is a phosphonate modification of the Wittig reaction (see for 
example L. F. Fieser and M. Fieser, John Wiley and Sons, Inc., 1967, p. 
1319 and 251) and hence, according to the prior art of carrying out Wittig 
reactions, requires a very strong base such as n-butyl lithium, sodium 
hydride and potassium t-butoxide (ibid. p 1319), as one of the reagents. 
In contrast to the moderate bases of this invention, the latter, very 
strong bases have in common that they are rapidly destroyed by water. 
It is further unexpected that the above conversion of formula (3) to 
formula (1) can be accomplished by employing a water-immiscible, non-polar 
solvent, such as benzene as the solvent for the steroid. As is well known 
to those skilled in the art, reactions being brought about by the action 
of a chemical reagent are generally greatly enhanced when the reagent is 
soluble in the solvent employed for the dissolution of the reactant. 
It is also unexpected and a novel feature of process (a) that the 
conditions developed for the conversion of compounds of formula (3), Z 
being (alkyl)P(O), to compounds of formula (1), R.sub.3 being H, are also 
very suitable for the conversion of compounds of formula (3), where Z is 
CN and CO.sub.2 alkyl to compounds of formula (1), where R is CN and 
CO.sub.2 alkyl. The latter type of conversion is no longer a Wittig 
reaction but resembles the Knoevenagel condensation (see for example R. C. 
Denney "Denney Organic Reactions", Butterworth, London 1969, p 50). 
The wide scope of process (a) owing to which it can be applied to 21-esters 
of formula (3) in which Z is not only (alkyl)P(O) but also CN and CO.sub.2 
alkyl, is a special advantage of this process. 
It is another advantage of the process (a) of this invention that the 
conversion of (3) to (1) can readily be brought about by bases of only 
moderate strength as this allows the transformation of reactants 
possessing groups which would be chemically changed if the usual stronger 
bases of a Wittig reaction were used. 
It is a further advantage of process (a) that the formation of the 
phosphonoacetate (3) from 21-alcohol (2) and the conversion (3) to (1) can 
be carried out in one vessel and that no intermittent evaporations of 
solvents are used in the two conversions are required. It is yet another 
advantage that, since the moderately strong bases employed in this 
invention are not affected by water, no special precautions have to be 
observed to ensure that the conversion of (3) to (1) is carried out under 
anhydrous conditions. The conversion of (3) to (1) via (2) can thus be 
carried out in a manner which is considerably more simple and economical 
than the previous procedure of choice of converting 21-hydroxy-20-ones of 
the pregnane series to cardenolides (see W. Eberlein, J. Nickl, J. Heider, 
G. Johns and H. Machleidt, Chem. Ber., 105, 3686 (1972). who prepared 
22-halo, alkyl- and alkoxycardenolides). 
It is an advantage of process (b) that the conversion of the 
alkyloxyacetylene adduct (4) to the corresponding 20(22)-en-23-oic acid 
esters or cardenolides of formula (1), respectively, can be brought about 
treatment of a solution of (4) in a water immiscible inert solvent, (e.g. 
benzene, hexane, or ether) with a dilute aqueous solution of a mineral 
acid, such as perchloric, hydrochloric or sulfuric acid. By contrast in 
the previous method of converting the adduct (4) to the ester (1) the 
latter are treated with a mixture of an alcohol and aqueous sulfuric acid. 
Since alcohols in the presence of acid are not inert solvents, e.g. they 
may cause an exchange of the alkoxy group in esters 1 (F. S. Khristulas, 
M. B. Gorovich and N. K. Abubakirov, Khimya Prirodnikh Soedinenii, 5, 545 
(1970) p 551), the method of this invention employing inert solvents for 
the above conversion has a greater synthetic utility. Further enhanced 
utility derives from the fact that in the method of this invention the 
reaction products can be more readily isolated from the reaction mixture, 
i.e. by the simple procedure of extracting the organic, water immiscible 
phase with water till the extracts are neutral and subsequent evaporation 
of the inert solvent. Yet further utility derives from the fact that the 
method is more applicable to such compounds of formula (4), which are not 
soluble or only partially soluble in the acidic aqueous alcohol employed 
in the prior art. As is well known, steroids are generally more soluble in 
water immiscible organic solvents than in aqueous alcohols. 
With regard to process (c), it is submitted that there is no prior art 
which teaches the conversion of cardenolides into the corresponding 
22-alkoxycarboxyl-21-methyl-20(22)-en-23-oic acids or the corresponding 
23,24-dioc acids, such as represented by formula (1), where R.sub.3 is an 
alkoxycarbonyl, R.sub.4 is methyl group and R.sub.5 is the hydroxy group 
of a carboxylic acid. It is a special advantage of process (c) that it 
provides stereo-specific routes towards (22-E) and (22-Z) 22-substituted 
20(22)-enes. By contrast, no such stereo-specific routes are provided for 
by the previous methods of preparing 21-methyl-22-substituted 20(22)-enes 
the stereochemistry of which is generally left unspecified (see, for 
example, F. Sondheimer and F. S. Khristulas, cited above, and also M. 
Okada and Y. Saito, Chem. Pharm. Bull., 16, 2223 (1968)). 
Many of the products of the present invention of formula (1) have been 
found to have smaller properties to the compounds of forumla 
##STR18## 
wherein R.sub.8 is chosen from the group of H, CH.sub.2 OH, CH.sub.2 
--O--CO-- NH--C(CH.sub.3).sub.3, CH.sub.2 Oalkyl and CH.sub.2 Oacyl, 
wherein alkyl is tetrahydropyranyl, lower alkyl, preferably methyl, or a 
substituted methyl wherein the substituent is selected from the group 
consisting of phenyl, halogen, preferably chlorine and bromine, methoxy, 
CH.sub.2 .dbd.CH and HC.tbd.C, and wherein acyl represents a group 
selected from those consisting of acetate, lower trialkyl acetates wherein 
the lower alkyl group is preferably methyl or ethyl, monohalo acetates and 
trihalo acetates, preferably wherein the halogen is chlorine, fluorine and 
bromine. The compounds of the above formula and their glycosides are known 
for treatment of cardiac insufficiency, as for example disclosed in 
Angewandte Chemie vol. 9, No. 5, pp 321-332, and Fieser and Frieser, Chap. 
20, "Steroids", Reinhold, NY 67. In addition, the compounds of formula (1) 
may in some cases be also used as intermediates for the preparation of 
compounds of the above given formula, by converting the compounds of 
formula (1) by conventional techniques, or alternatively, by methods 
similar to those described in the following Examples. Still further, 
certain of the compounds of formula (1) -- i.e. those without a 14.beta. 
oxygen, may be useful as modifiers of cardiatonic compounds - e.g. by 
suppresing selectively their toxic activity. Those compounds not having a 
3 oxygenated substituent have similar properties to the known compounds or 
3-deoxy analogs, 17 analogs in which the 17 substituent is a 
20(22)-en-23-oic acid ester derivative or the 22 substituted buteneolide 
as described in, e.g., Annular Reports in Medicinal Chemistry, 1970, page 
174 and the references therein; W. Eberlein et al, Chem. Ber Vol 105, 3686 
(1972); J.S. Boutaery, R. Thomas, Australian Journal of Chemistry, Vol. 
24, 2723 (1971).

EXAMPLE 1 
A mixture, prepared by successive addition of 3.942 ml of a 10% solution of 
diethylphosphono!acetic acid (1.3 moles per mole of 21-hydroxy-20-ketone) 
in benzene and 10.05 ml of a 0.2 molar solution of 
dicyclohexylcarbodiimide (1.3 moles per mole of 21-hydroxy-20-ketone) in 
benzene to a solution of 600 mg of 
3.beta.-acetoxy-21-hydroxy-8,19-oxido-5.alpha.-pregn-14-en-20-one in 30 ml 
of benzene, was stirred under nitrogen at room temperature for 18 minutes, 
whereupon a small fraction was withdrawn and added to 1 volume of water. 
Evaporation of the organic phase gave 
3.beta.-acetoxy-21-diethylphosphono!acetoxy-8,19-oxido-5.alpha.-pregn-14- 
en-20-one as evidenced by tlc analysis and subsequent transformations. 
The remaining major reaction mixture was stirred under nitrogen with 15 ml 
of 50% aqueous potassium hydroxide for 15 minutes at room temperature, 
whereupon 75 ml of a mixture of water and ice was added. Acidification of 
the mixture with acetic acid-water 1:10, followed by dilution with 
ether-methylene chloride 4:1, five extraction of the organic phase with 
1/4 volume of water, evaporation at reduced pressure and drying at high 
vacuum gave a residue which was stirred with 30 ml of methylene chloride 
under nitrogen for 15 minutes. The undissolved solid was then removed by 
filtration. Concentration of the filtrate, followed by addition of hexane 
till a faint turbidity appeared, filtration through diatomaceous earth, 
concentration of the filtrate at reduced pressure with intermittent 
addition of hexane and ether, standing at -5.degree. C for 2 hours and 
filtration gave 524 mg of 
3.beta.-acetoxy-8,19-oxido-5.alpha.-carda-14,20(22)-dienolide, mp 
214,220.degree.-221.degree. C. Recrystallisation from methanol water gave 
a purified sample, mp 219,222.degree.- 224.degree. C. uv (MeOH) 219 m.mu. 
EXAMPLE 2 
A mixture, prepared by successive addition of 17.7 mg of 
diethylphosphono!acetic acid (2.6 moles per mole of 3,21-diol), 0.175 ml 
of benzene and 0.455 ml of a 0.2 molar solution of 
dicyclohexylcarbodiimide (2.6 moles per mole of 3,21-diol) in benzene to a 
suspension of 12mg of 3.beta. 
,21-dihydroxy8,19-oxido-5.alpha.-pregn-14-en-20 -one in 0.175 ml of 
benzene, was stirred under nitrogen at room temperature. Tlc analysis on a 
sample withdrawn after 20 minutes indicated that all starting material had 
been converted to 3.beta. 
,21-didiethylphosphono!acetoxy-8,19-oxido-5.alpha.-pregn-14-en-20-one. 
After 35 minutes of stirring 0.115 ml of 50% aqueous potassium hydroxide 
was added and stirring was continued for 30 minutes, whereupon 0.455 ml of 
water was added. The mixture was then filtered, the phases were separated, 
the aqueous phase of the filtrate was acidified. Filtration of the 
resulting precipitate yielded 
3.beta.-diethylphosphono!acetoxy8,19-oxido-5.alpha.-carda-14,20(22)-dieno 
lide, uv(MeOH) 219 m.mu., ir (KBr) 3070, 1777, 1748, 1628, 1110, 1050, 
1003, 926, 890, 860 and 818 cm.sup.-1. 
EXAMPLE 3 
A mixture, prepared by successive addition of 3.35 ml of a 0.2 molar 
solution of dicyclohexylcarbodiimide (1.3 moles per mole of 
21-hydroxy-20-one) in benzene to a solution of 130 mg of benzyl 
hemimalonate (1.3 moles per mole of 21-hydroxy20-one) and 200 mg of 
3.beta.-acetoxy-21-hydroxy-8,19-oxido-5.alpha.-pregn-14-en-20-one in 10 ml 
of benzene, was stirred under nitrogen at room temperature. Tlc analysis 
on a sample withdrawn after 15 minutes indicated that all starting 
material had been converted to 
3.beta.-acetoxy-21-benzoyloxycarbonyl!acetoxy8,19-oxido-5.alpha.-pregn-14 
-en-20-one. 
The mixture was then stirred under nitrogen for 2-1/4 hours with 2.5 ml of 
10% aqueous potassium hydroxide, whereupon 4.0 ml of water and 2.0 ml of 
acetic acid-water 1:10 and 2 volumes of ether was added. The mixture was 
agitated and the fine, undissolved precipitate was removed by filtration. 
The organic phase of the biphasial filtrate was extracted 5 times with 
water and then evaporated. Recrystallisation of the residue from 
ether-hexane gave 216 mg of a precipitate which after a further 
recrystallisation from the same solvent system gave 3.beta. 
-acetoxy-22-benzyloxycarbonyl-8,19-oxido-5.alpha.-carda-14,20(22)-dienolid 
e, mp 174, 176.degree.-178.degree.C, uv (MeOH) 212 and 229 m.mu.. 
EXAMPLE 4 
When 40 mg of 
3.beta.-acetoxy-21-hydroxy-8,19-oxido-5.alpha.-pregn14-en-20-one as 
subjected to reaction conditions, which were essentially the same as those 
described in Example 3 except that cyanoacetic acid instead of benzyl 
hemimalonate was used, the 21-cyanoacetate of the starting material was 
obtained as the intermediate product, which after the base-treatment and 
chromatographic separation on silica gel G coated glass plates with ethyl 
acetate-benzene 1:1 as the eluant gave 
3.beta.-acetoxy-22-cyano-8,19-oxido-5.alpha.-carda-14,20(22)-dienolide, uv 
(MeOH) 210 and 234 m.mu., and 
22-cyano-3.beta.-hydroxy-8,19-oxido-5.alpha.-carda-14,20(22)-dienolide, uv 
(MeOH) 209 and 234 m.mu.. 
EXAMPLE 5 
When 23 mg of 3.beta.,19-diacetoxy-21-hydroxy-5.alpha.-pregn-14-en20-one 
was subjected to reaction conditions, which were essentially the same as 
those described in Example 1, except that, instead of 1.3 moles, 1.5 moles 
per mole starting material of diethylphosphono!acetic acid and 
dicyclohexylcarbodiimide were used, the 21-diethylphosphono!acetate of 
the starting material was obtained as an intermediate. The subsequent 
base-treatment, followed by chromatographic purification of the cruude 
product on silica gel coated glass plates, with ethyl acetate-benzene 1:7 
as the eluant, and digestion of the purified material with pentane 
afforded 3.beta.,19-diacetoxy-5.alpha.-carda-14,20(22)-dienolide, mp 
155.degree.-162.degree. C. 
EXAMPLE 6 
When 97.38 mg of 19-acetoxy-21-hydroxy-pregna-4,6-diene-3,20dione was 
subjected to reaction conditions, which were essentially the same as those 
described in Example 1, except that 2.6 moles per mole of starting 
material of diethylphosphono!acetic acid and dicyclohexylcarbodiimide 
were used instead of 1.3 moles each, the 21-diethylphosphono!acetate of 
the starting material was obtained as an intermediate. The subsequent 
base-treatment, and chromatographic purification of the crude product on 
silica gel G coated glass plates, ethyl acetate-benzene 2:1 being the 
eluant, afforded 19-acetoxy-3-oxo-carda-4,6,20(22)-trienolide, mp 
180,197.degree.-199.degree., uv (MeOH) 218 and 285 m.mu.. 
EXAMPLE 7 
When 172.2 mg of 8,19-oxido-21-hydroxypregn-4-en-3,20-dione was subjected 
to reaction conditions which were essentially the same as those described 
in Example 6, except that the intermediate 21-diethylphosphono! acetate 
of the starting material was isolated and separated from residual starting 
material by chromatography on silica coated glass plates and then 
subjected to the base-treatment, recrystallisation of the final crude 
product from ethyl acetate-pentane gave 36.3 mg of a crystalline material 
consisting essentially of 
3-oxo-8,19-oxido-5.alpha.-carda-14,20(22)-dienolide containing a more 
polar alcohol as an impurity. Treatment of the latter product with 
pyridine-acetic anhydride at +5.degree. C. for 21/2 days, followed by 
addition of water, filtration and recrystallisation of the precipitate 
collected with methylene chloride-ether gave 18 mg of the purified 
product, mp 227, 228.degree.-231.degree. C., ir(NUJOL) 3105 (weak), 1778, 
1745, 1670, 1622, 1375, 1305, 1258, 1045, 1020, 966, 890, 882 and 815 
cm.sup.-1. 
EXAMPLE 8 
When 400 mg of 3.beta.-acetoxy-21-hydroxypregn-5-en-20-one was subjected to 
reaction conditions, which were essentially identical to those described 
in Example 1, 336 mg of 3.beta.-acetoxycarda-5,20(22)-dienolide, mp 168, 
169.degree.-173.degree. C., was obtained via the 
21-diethylphosphono!acetate of the starting material. 
EXAMPLE 9 
When 166.25 mg (0.5 millimoles) of 3.beta.-21-dihydroxy-pregn-5-en-20-one 
was subjected to reaction conditions, which were essentially the same as 
those described in Example 2, the base-treatment of the intermediate 
3.beta.,21-didiethylphosphono!-acetoxy-pregn-5-en-20-one gave 
3.beta.-diethylphosphono! acetoxycarda-5,20(22)-dienolide, ir(KBr) 2970, 
1785, 1755, 1730, 1635, 1260, 1250, 1145, 1120, 1070, 1025, 900, 875 and 
790 cm.sup.-1. 
EXAMPLE 10 
When 400 mg of 3.beta.-acetoxy-21-hydroxypregn-5-en-20-one was treated 
under reaction conditions which were essentially the same as those 
described in Example 3 except that the intermediate 
3.beta.-acetoxy-21-benzyloxycarbonyl!acetoxy-pregn-5-en-20-one was 
isolated and purified by precipitation with hexane from a methylene 
chloride solution, dissolution of the crude final product in methylene 
chloride, following by precipitation with hexane, gave 551 mg of 
3.beta.-acetoxy-22-benzyloxycarbonyl!-carda-5,20(22)-dienolide as 
evidenced by the comparison with a sample of another batch, which had 
uv(MeOH) 216 and 230 m.mu.. 
EXAMPLE 11 
A mixture, prepared by successive addition of 118.8 mg of cyanoacetic acid 
(1.3 moles per mole of 21-hydroxy-20-ketone) and 6.96 ml of 0.2 molar 
solution of dicyclohexylcarbodiimide (1.3 moles per mole of 
21-hydroxy-20-ketone) in benzene to a solution of 400 mg of 
3.beta.-acetoxy-21-hydroxypregn-5-en-20-one in 20 ml of benzene, was 
stirred under nitrogen. Tlc analysis on a sample within drawn after 80 
minutes showed that all of the starting material had been converted to the 
corresponding 21-cyanoacetate. The mixture was stirred with 6.0 ml of 10% 
aqueous potassium hydroxide under nitrogen for 10 minutes, whereupon 160 
ml of water and 200 ml of ether was added. The lower, aqueous phase was 
separated, acidified with 9.0 ml of 2N aqueous sulfuric acid and extracted 
with ether. The ether solution was extracted with water till the aqueous 
extracts were neutral, dried with sodium sulfate and evaporated at reduced 
pressure. Dissolution of the residue obtained in methylene chloride, 
addition of hexane till the solution became slightly turbid, filtration 
through diatomaceous earth, concentration of the filtrate with 
intermittent addition of hexane, standing at -5.degree. C and filtration 
gave 355 mg of 3.beta.-acetoxy-22-cyanocarda-5,20(22) dienolide, mp 
200.degree.-215.degree., uv(MeOH)241 m.mu., as verified by ir- and 
nmr-spectroscopy. 
Agitation of 34.4 mg of the above compound, dissolved in 3.45 ml of 
benzene, with 3.45 ml of 50% aqueous potassium hydroxide for 3 hours, 
followed by acidification with 2N aqueous sulfuric acid and isolation of 
the steroidal product by a procedure which was essentially the same as the 
one outlined above, gave 3.beta.-hydroxy-22-cyanocarda-5,20(22) dienolide 
mp 211.degree.-225.degree., uv(MeOH) 239 m.mu., ir (KBr) 3550, 2225, 1760, 
1640, 1065, 1045, 1025 and 760 cm.sup.-1. 
EXAMPLE 12 
When 50 mg of 21-hydroxy-8,19-oxido-5.alpha.-pregn-14-en-20-one was 
subjected to reaction conditions which were essentially the same as those 
described in Example 1, 44 mg of 8,19-oxido-5.alpha.-carda-14,20(22) 
-dienolide, mp 177, 179-184.degree. C, uv (CH.sub.3 OH) 218 m.mu. was 
obtained via the 21-diethylphosphono!acetate of the starting material. 
EXAMPLE 13 
When 20 mg of 21-hydroxy-8,19-oxido-5.alpha.-pregn-14-en-20-one was 
subjected to reaction conditions which were essentially the same as those 
described in Example 4, 21-cyanoacetoxy8,19-oxido-5.alpha.-pregn-14-ene 
was obtained as the intermediate and 
22-cyano-8,19-oxido-5.alpha.-carda-14,20(22) dienolide, ir (Kbr) 2220, 
1774, 1625 and 1572 cm.sup.-1 as the final product. A methanolic solution 
of the latter compound had uv 237 m.mu.; after addition of a small amount 
of aqueous potassium hydroxide it had uv 256 mu. 
EXAMPLE 14 
A mixture of 250 mg of 3.beta.-acetoxy-8,19-oxido-5.alpha.-carda-14,20(22) 
-dienolide, 7.5 g of zinc, 18.75 ml of toluene and 6.25 ml of formic acid 
was shaken for 16 hours whereupon the supernatant phase was decanted. The 
residue was shaken briefly with 25 ml of benzene, which was then decanted. 
The benzene extraction was repeated another 4 times. The decanted 
supernatant liquids were filtered, combined and evaporated at reduced 
pressure. The residue was dissolved in methylene chloride and hexane was 
added to the solution till it became faintly turbid. The mixture was 
filtered through diatomaceous earth and the filtrate was concentrated at 
reduced pressure with intermittent addition of hexane and ether. Standing 
of the mixture at -5.degree. C., followed by filtration gave 237 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14,20(22) -dienolide, uv(MeOH) 
218 m.mu., ir(KBr) 3065 (weak), 1795, 1767, 1740, 1730, 1640, 1255, 1207, 
1170, 1141, 1081, 1059, 1030, 911, 895, 867, 815 and 745 cm.sup.-1. TLC 
analysis showed that the reaction proceeds via 
3.beta.-acetoxy-19-hydroxy-5.alpha.-carda-14,20(22) -dienolide. 
EXAMPLE 15 
Reduction of 70 mg of 8,19-oxido-5.alpha.-carda-14,20(22)-dienolide with 
zinc in presence of formic acid under conditions which were essentially 
the same as those described in Example 14, afforded 39.8 mg of 
19-hydroxy-5.alpha.-carda-14,20(22)-dienolide 19-formate, mp 
131,135-136.5.degree. C., via 19-hydroxy-5.alpha.-carda-14,20(22) 
-dienolide. 
EXAMPLE 16 
A mixture of approximatly 38 mg of 3.beta.-acetoxy-8, 
19-oxido-5.alpha.-carda-14,20(22)-dienolide, 3.8 ml of glacial acetic 
acid, 0.97 ml of water and 77 mg of zinc dust was stirred briefly at room 
temperature and then with external heating by an oil-bath having a 
temperature between 65.degree. to 69.degree. C. After 5 minutes of 
stirring in the oil-bath, 388 mg of zinc was added slowly during 3 minutes 
followed by 3 further slow additions during 3-4 minutes of the same 
quantity of zinc after 36, 227 and 254 minutes respectively. The mixture 
was then filtered and the precipitate was washed with acetic acid-water 
4:1 and ethyl acetate. The filtrate was concentrated 3 times at reduced 
pressure to 1/2 volume with intermittent addition of 3 .times. 30 ml of 
water. The resulting mixture was neutralized with a 1/2 saturated aqueous 
sodium bicarbonate and left to stand at +5.degree. C for 21/2 days. 
Filtration gave 28 mg of a product containing 
3.beta.-acetoxy-19-hydroxy-5.alpha.-carda-14,20(22)-dienolide as the major 
compound as evidenced by tlc. The latter material, 0.11 ml of pyridine and 
0.056 ml of acetic anhydride was then left to stand under nitrogen for 16 
hours at room temperature, whereupon it was diluted with hexane-ether 2:1 
and filtered. The filtrate was concentrated at reduced pressure with 
intermittent addition of toluene. Chromatography of the resulting resin on 
silica gel G coated glass plates with ethyl acetate-benzene 1:4 as the 
eluant, followed by digestion with pentane of the fraction isolated, gave 
10 mg of 3.beta.,19-diacetoxy-5.alpha.-carda-14,20(22)-dienolide, mp 
162,163.degree.-164.degree., which as evidenced by its ir-spectrum and 
melting point, was found to be identical with 
.beta.-anhydrocoroglaucigenin 3,19-acetate, mp 161.degree.-163.degree., 
(A. Hunger and T. Reichstein, Helv., 35, 1073 (1952). 
EXAMPLE 17 
A mixture of 30 mg of 3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14,20(22) 
-dienolide, 1.2 ml of acetone and 0.6 ml of a solution of 60 mg of 
N-bromoacetamide in 1 ml of water was stirred under nitrogen in the dark 
in an ice-bath for 97 minutes whereupon 18 ml of ice-water and 1.2 ml of a 
1/2 saturated aqueous sodium bisulfite solution was added. After 30 
minutes of further stirring, filtration, followed by drying at high vacuum 
afforded 
3.beta.-acetoxy-15.alpha.-bromo-19-formyloxy-14.beta.-hydroxy-5.alpha.-car 
d-20(22)-enolide; ir(KBr) 3425 (broad), 3295 (sharp), 3070 (weak), 1735, 
1727, 1719, 1710, 1705, 1619, 1565, 1370, 1355, 1239, 1115, 1021, 901, 
890, and 882 cm.sup.-1 ; the ir-spectrum of a chloroform solution had a 
strong peak at 3570 instead of the peaks at 3425 and 3295 cm.sup.-1. 
EXAMPLE 18 
When 11 mg of 3.beta.,19-diacetoxy-5.alpha.-carda-14,20(22)-dienolide was 
subjected to reaction conditions which were essentially the same as those 
in the preceding example, filtration of the aqueous suspension gave 
3.beta.,19-diacetoxy-15.alpha.-bromo-14.beta.-hydroxy-5.alpha.-carda-(20(2 
2)-enolide; uv (MeOH)217 m.mu.; ir (NUJOL), 3470 (broad), 3390 (weak), 3360 
(weak), 3090 (weak), 1769, 1735, 1720, 1620, 1450, 1368, 1355, 1237, 1025, 
951, 880 and 865 cm.sup.-1. 
EXAMPLE 19 
A mixture, prepared by addition of a freshly made solution of 180 mg of 
N-bromoacetamide in 3.0 ml of water to 6.0 ml of acetone and 150 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14,20(22) -dienolide, was 
stirred in the dark under nitrogen with external cooling by an ice-bath 
for 2 hours whereupon 90 ml of ice-water and 6.0 ml of a half-saturated 
aqueous sodium bisulfite was added and stirring was continued for another 
hour. Filtration gave 
3.beta.-acetoxy-15.alpha.-bromo-19-formyloxy-14.beta.-hydroxy-5.alpha.-car 
d-20(22)-enolide, which was dissolved in 30 ml of methylene chloride and 
shaken for 16 hours with a mixture of 3.0 ml of pivalic acid-methylene 
chloride 1:10, 60 ml of water and Raney nickel; the latter had been 
freshly prepared from 9.0 g of a 50% nickel-aluminum alloy. Addition of 
ether-methylene chloride 4:1, followed by filtration through filter-pulp, 
extraction of the organic phase with half-saturated aqueous sodium 
bicarbonate and water, drying with sodium sulfate and evaporation at 
reduced pressure, combination of the crude product obtained with that of 
another reaction in which 50 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14,20(22) -dienolide were used 
as the starting material, and several recrystallizations of the combined 
products from ether-hexane gave 50.65 mg of product, mp 
232,234.degree.-239.degree.. Further purification of 24 mg of this 
material by chromatography on Silica gel G coated glass plates with ethyl 
acetate-benzene 1:2 as the eluant gave 19 mg of a fraction, which after 
recrystallization from ether-hexane yielded 16.76 mg of purified 
coroglaucigenin 3-acetate 19-formate, mp 248.5.degree.-250.2.degree. C. 
The mother liquors, when subjected to chromatography as described above, 
afforded further amounts of the latter compound as well as several 
fractions amounting to approximately 20 mg of 
3.beta.-acetoxy-19-formyloxy-14.beta.-,15.beta.-oxido-5.alpha.-card-20(22) 
-enolide. 
EXAMPLE 20 
When 
15.alpha.-bromo-3.beta.,19-diacetoxy-14.beta.-hydroxy-5.alpha.-card-20(22) 
-enolide, which had been freshly prepared from 11 mg of 
3.beta.,19-diacetoxy-5.alpha.-carda-14,20(22)-dienolide as described in 
Example 18, was treated under reaction conditions, which were essentially 
similar to that described in the preceding example except that the 
reduction with Raney Nickel was carried out in an atmosphere of hydrogen 
instead of nitrogen, chromatography of the crude product on silica gel G 
coated glass plates with ethyl acetate-benzene 1:2 as the eluant, followed 
by recrystallization from ether-pentane, gave 
3.beta.,19-diacetoxy-14.beta.-hydroxy-5.alpha.-card-20(22)-enolide 
(coroglaucigenin diacetate), mp 207.degree.-212.degree. C.; 
coroglaucigenin diacetate prepared by A. Hunger and t. Reichstein, Helv., 
35, 1073 (1952) p.1097 from coroglaucigenin had mp 210.degree.-214.degree. 
and 216.degree.-219.degree. C. 
EXAMPLE 21 
3.beta.-Acetoxy-19-formyloxy-15.alpha.-bromo-14.beta.-hydroxy-5.alpha.-card 
-20(22)-enolide was prepared from 25 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha. -carda-14,20(22)-dienolide as 
described in Example 17. A methanolic solution of the freshly filtered 
bromohydrin was then stirred with a molar excess of concentrated aqueous 
ammonia at room temperature under nitrogen for several hours and then left 
at -5.degree. C for 16 hours, whereupon the base was neutralized with 
acetic acid in ethyl acetate and the mixture was evaporated at reduced 
pressure. The residue obtained was extracted 3 times with ether-methylene 
chloride 4:1 and the combined extracts were chromatographed graphed on 
silica gel G coated glass plates using ethyl acetate-benzene 1:2 as the 
eluant. Recrystallization of the major fraction from ether-hexane gave 
11.36 mg of 
3.beta.-acetoxy-19-formyloxy-14.beta.,15.beta.-oxido-5.alpha.-card-20(22)- 
enolide mp 213.degree.-215.degree. C. 
EXAMPLE 22 
A mixture, consisting of 
15.alpha.-bromo-3.beta.,19-diacetoxy-14.beta.-hydroxy-5.alpha.-cardenolide 
, which had been freshly prepared from 2.6 mg of 
3.beta.,19-diacetoxy-5.alpha.-carda-14,20(22)-dienolide, by the method 
described in Example 17, and 0.13 ml of t-butylamine was left to stand 
under nitrogen for 30 minutes whereupon it was evaporated at reduced 
pressure. Treatment of the residue with ether and water, repeated 
extraction of the organic phase with water, evaporation and 
recrystallization from ether-pentane gave 
3.beta.,19-diacetoxy-14.beta.,15.beta.-oxido-5.alpha.-card-20(22)-enolide, 
mp 194, 200.degree.-204.degree. C. 
EXAMPLE 23 
A mixture, consisting of 
3.beta.-acetoxy-19-formyloxy-15.alpha.-bromo-14.beta.-hydroxy-5.alpha.-car 
d-20(22)-enolide, which had been freshly prepared from 20 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14, 20(22)-dienolide as 
described in Example 17 and was still wet, 1 ml of t-butylamine and 0.5 ml 
of water was stirred for 1 hour in an atmosphre of nitrogen, whereupon it 
was concentrated at reduced pressure with intermittent addition of hexane 
and water. Filtration yielded 8.0 mg of crude 
3.beta.-acetoxy-19-hydroxy-14.beta., 
15.beta.-oxido-5.alpha.-card-20(22)-enolide. Recrystallization from 
hexane-methylene chloride gave the purified sample, mp 
227.degree.-229.degree. C. 
EXAMPLE 24 
A mixture, consisting of 7 ml of methanol and crude 
3.beta.-acetoxy-19-formyloxy-14.beta.-hydroxy-5.alpha.-card-20(22)-enolide 
, prepared from 150 mg of 
3.beta.-acetoxy-19-formyloxy-5.alpha.-carda-14,20(22)-dienolide as 
described in Example 19, was stirred under nitrogen at room temperature; 
4.9 ml of methanol-2% aqueous potassium hydroxide 20:1 was then added 
during 1/2 hour. After 2 hours 0.245 ml of glacial acetic acid-ethyl 
acetate 1:50 was added and the mixture was evaporated at reduced pressure. 
Extraction of the residue with ethermethylene chloride 4:1, followed by 
extraction of the organic phase with water, drying with sodium sulfate 
evaporation and chromatography on silica gel G coated glass plates with 
ethyl acetate-benzene 1:1 as the eluant gave 37 mg of 
3.beta.-acetoxy-14.beta.,19-dihydroxy-5.alpha.-card-20(22)-enolide 
(coroglaucigenin 3-acetate), mp 233.degree.-234.degree.. 
EXAMPLE 25 
Oxidation of 4 mg of coroglaucigenin 3-acetate with t-butyl chromate in 
t-butanol-carbon tetrachloride 1:6 by a method which was essentially the 
same as the one described by A. Katz, Helv. 35, 487 (1957), p.490, gave 
corotoxigenin 3-acetate, mp 200, 221.degree.-229.degree., after 
recrystallization of the crude product with ether-hexane. The irspectrum 
of the latter compound was identical to that obtained by A. Hunger and T. 
Reichstein, Helv. 35, 1073 (1952), who obtained corotoxigenin 3-acetate, 
mp 227.degree.-234.degree. and 200.degree.-230.degree. from corotoxigenin. 
The t-butyl chromate was prepared as described by K. Hensler and A. 
Wettstein, Helv., 35, 284 (1952). 
EXAMPLE 26 
A mixture of 6 mg of coroglaucigenin 3-acetate 19-formate, 0.3 ml of 
t-butylamine and 0.3 ml of water was shaken under nitrogen at room 
temperature for 24 hours whereupon it was evaporated at reduced pressure 
with intermittent addition of glacial acetic acid-water 5:1. The residue 
was disintegrated in 0.2 ml of water and the resulting suspension was 
filtered. The precipitate was recrystallized from methanol-ether yielding 
3.7 mg of crude coroglaucigenin, mp 227.degree.-230.degree. . 
Chromatography on silica gel G coated glass plates with ethyl acetate as 
the eluant, followed by recrystallization from methanol-ether, gave the 
purified sample; mp 241.degree.-242.degree., 244.degree.; ir(KBr) max. 
3610, 3350 3098, 1781, 1750, 1738, 1620, 1445, 1370, 1339, 1309, 1301, 
1175, 1148, 1135, 1075, 1035, 1025, 1019, 959, 891, 887, 880, 869, 790, 
780, 741 and 698 cm.sup.-1. 
EXAMPLE 27 
A mixture, consisting of 45 mg of 3.beta.-acetoxy-carda-5,20(22)-dienolide, 
9.0 ml of methanol and 0.9 ml of 2% aqueous potassium hydroxide was 
stirred for 4 hours whereupon another lot of 0.9 ml of 2% aqueous 
potassium hydroxide was added. The mixture, in which a precipitate had 
formed was left to stand at -5.degree. C for 16 hours and was then 
filtered. The precipitate was washed with water-methanol 1:3 and then with 
water. The filtrate was concentrated and filtered yielding a second 
precipitate. Both precipitates were combined and chromatograhed on silica 
gel G coated glass plates using ethyl acetate-benzene 1:4 as the eluant. 
The fraction having rf 0.2-0.3 was recrystallized from ether-hexane and 
yielded 5.7 mg of 3.beta.-hydroxy-carda-5,20(22)-dienolide, ir(KBr) 3460 
(broad),1805, 1730, 1620, 1190, 1170, 1135, 1108, 1070, 1052, 1030, 985, 
965, 905, 870, 812, 745 and 712 cm.sup.-1. 
The latter 3.beta.-hydroxycardenolide was also obtained, as indicated by 
tlc analysis, when the 3.beta.-diethylphosphono!-acetoxy analog of the 
starting material was similarly hydrolysed with methanol and 2% aqueous 
potassium hydroxide. 
EXAMPLE 28 
When 25 mg of 19-formyloxy-5.alpha.-carda-14,20(22)-dienolide was oxidized 
with N-bromo acetamide as outlined in Example 17, the corresponding 
14.beta.-hydroxy-15.alpha.-bromo analog was obtained. The freshly filtered 
and still wet bromohydrin, when reduced with Raney nickel as described in 
Example 17, gave a crude product which was purified by chromatography on 
silica gel G coated glass plates with ethyl acetate-benzene 1:1 as the 
eluant. Recrystallization of the fraction having rf 0.3-0.4 from 
ether-hexane gave 8.4 mg of 
14.beta.,19-dihydroxy-5.alpha.-card-20(22)-enolide 19-formate, mp 
195.degree.-198.degree. C, ir(KBr) 3565, 3415 (broad), 3330 (shoulder), 
3090 (small) 1785, 1755, 1742, 1737, 1730, 1721, 1715, 1628 and 1175 
cm.sup.-1. A less polar fraction isolated was recrystallized from 
hexane-methylene chloride are yielded 
19-formyloxy-14,15.beta.-oxido-5.alpha.,14.beta.-cardenolide, mp 
190.degree.-192.degree. C. 
EXAMPLE 29 
A freshly prepared mixture, consisting of 11.2 ml of anhydrous 
tetrahydrofuran, 0.187 ml of redistilled ethoxyacetylene and 1.12 ml of 
1.95M methyl lithium in ether was added to a solution of 50 mg of 
3.beta.-acetoxy-21-hydroxy-8,19-oxido-5.alpha.-pregn-14-en-20-one in 8.7 
ml of tetrahydrofuran which was protected by an atmosphere of nitrogen. 
After stirring under nitrogen for 4 hours the solvents were evaporated at 
reduced pressure. The residue was treated with wet ether and the ethereal 
phase was extracted several times with water till the aqueous extracts 
were no longer basic. Evaporation yielded a resin consisting mainly of 
20-ethoxyethynyl-3.beta.,20, 
21-trihydroxy-8,19-oxido-5.alpha.-pregn-14-ene as evidenced by tlc 
analysis and the subsequent transformations. The latter product was then 
vigorously stirred with 3.5 ml of benzene and 1.75 ml of 2N aqueous 
sulfuric acid under nitrogen for 4 hours, whereupon the reaction mixture 
was diluted with benzene, and extracted successively with water and half 
saturated aqueous sodium bicarbonate. The benzene solution was dried over 
sodium sulfate and filtered through a column of 70 mg of aluminium oxide. 
Washing of the column with ethyl acetate afforded 21 mg of a product, 
which solidified with hexane and consisted essentially of 
3.beta.-hydroxy-8,19-oxido-5.alpha.-carda-14,20(22)-dienolide. Subsequent 
standing with 0.084 ml of pyridine and 0.042 ml of acetic anhydride for 20 
hours in a nitrogen atmosphere, followed by addition of 20 volumes of 
water, extraction with ether, extraction of the ethereal phase with water 
and evaporation afforded the corresponding 3-acetate, which was then 
reduced with zinc and formic acid in presence of toluene under conditions 
which were similar to those described in Example 14. Chromatography of the 
total product obtained on silica gel G coated glass plates with ethyl 
acetate-benzene 1:4 as the eluant gave 9.7 mg of 
3.beta.,19-dihydroxy-5.alpha.-carda-14,20(22)-dienolide 3-acetate 
19-formate as a white solid, which had an ir spectrum identical to the 
product of Example 13. 
When 3.beta.,21-dihydroxy- or 
3.beta.21-diacetoxy-8,19-oxido-5.alpha.-pregn-14-ene instead of 
3.beta.-acetoxy-21-hydroxy-8,19-oxido-5.alpha.-pregn-14-ene were reacted 
with ethoxyacetylene as described above, 20-ethoxyethynyl-3.beta.,20, 
21-trihydroxy-8,19-oxido-5.alpha.pregn-14-ene was obtained. 
EXAMPLE 30 
When 3.beta.-acetoxy-8,19-oxido-5.alpha.-pregn-14-en-20-one was reacted 
with ethoxyacetylene as described in the preceding example, a compound 
considered to be 
20-ethoxyethynyl-3.beta.,20-dihydroxy-8,19-oxido-5.alpha.-pregn-14-ene was 
obtained; similarly 
8,19-oxido-3.beta.-tetrahydropyran-2-yloxy!-5.alpha.-pregn-14-en-20-one 
gave a compound considered to be 
20-ethoxyethynyl-20-hydroxy-8,19-oxido-3.beta.-tetrahydropyran-2-yloxy-5.a 
lpha.-pregn-14-ene. The latter two ethoxyacetylene adducts, when treated 
with aqueous sulfuric as described in the preceding example, gave ethyl 
3.beta.-hydroxy-8,19-oxido-24-nor-5.alpha.-chol-20(22)-en-23-oate, uv max. 
222 m.mu., as evidenced by tlc analysis. 
EXAMPLE 31 
When 32 mg of 
14.beta.-hydroxy-3.beta.-pyran-2-yloxy!-8,19-oxido-5.alpha.-pregnan-20-on 
e was reacted with ethoxyacetylene similarly as described in Example 29, a 
product containing 
20-ethoxyethynyl-3.beta.,14.beta.-dihydroxy-8,19-oxido-5.alpha.-pregnan-20 
-one was obtained, which after treatment with aqueous sulfuric acid, 
similarly as described in Example 29 except that ethanol was used as the 
solvent, gave a product containing ethyl 
3.beta.,14.beta.-dihydroxy-8,19-oxido-24-nor-5.alpha.-chol-20(22)-en-23-oi 
c acid. Acetylation with acetic anhydride-pyridine 1:2 followed by 
chromatography of the crude product on silica gel G coated glass plates 
with ethyl acetate-benzene 1:4 as the eluant gave ethyl 
3.beta.-hydroxy-8,19-oxido-24-nor-5.alpha.-chol-20(22)-en-oate, uv max. 
231 m.mu., treatment of this product with selenium dioxide in boiling 
benzene as described by N. Danieli, Y. Mazur and F. Sondheimer, J. Am. 
Chem. Soc., 84, 875 (1962) followed by chromatography on silica gel G 
coated glass plates with ethyl acetate-benzene 1:4 as the eluant gave a 
fraction considered to be 8,19-oxidouzarigenin acetate, uv max. 218 m.mu.. 
A further, more polar fraction had also uv max. 218 m.mu. and was 
considered to be 14.alpha.-hydroxy-8,19-oxidouzarigenin 3-acetate (cf F. 
Sondheimer, Chemistry in Britain, 1, 454 (1965), p. 459). 
EXAMPLE 32 
A mixture of 200 mg of 3.beta.-acetoxy-21-hydroxy-pregn-5-en-20-one, 10 ml 
of methyl isobutyl ketone, 2.0 g of malonic acid and 2.0 g of calcium 
chloride was heated under nitrogen at 84.degree. C. for 2 days, whereupon 
it was evaporated at reduced pressure with intermittent addition of 
benzene. The residue obtained was treated with water and p the resulting 
precipitate was filtered and washed well with water. The precipitate was 
dissolved in methylene chloride and the solution was concentrated with 
intermittent addition of pentane. The supernatant petroleum ether phase 
was decanted and the precipitation from pentane was repeated twice. The 
remaining residue was dissolved in ethermethylene chloride, treated with 
charcoal and filtered through diatomaceous earth. Concentration of the 
filtrate at reduced pressure with intermittent addition of hexane and 
ether, followed by filtration, gave 192 mg of 
3.beta.,21-dihydroxypregn-5-en-20-one 3-acetate 21-hemimalonate. 
A solution of the latter compound in ether-diazotoluene, which was prepared 
by mixing briefly 22.3 mg of N-benzyl-N'-nitro-N-nitrosoguanidine, 0.040 
ml of potassium hydroxide-water 1:1 and 1 ml of ether with external 
cooling by an ice-acetone bath, was then left to stand at -50.degree. C. 
for 15 minutes. Evaporation at reduced pressure gave an off-white solid 
consisting mainly of 
3.beta.-acetoxy-21-bensyloxycarbonyl!acetoxy-pregn-5-en-20-one. Treatment 
of part of the latter product with 20 parts t-butylamine for 25 minutes at 
room temperature, followed by evaporation, gave a product consisting 
mainly of 3.beta.-acetoxy-22-benzyloxycarbonyl!-carda-5,20(22)-dienolide 
as evidenced by tlc analysis. 
EXAMPLE 33 
A mixture of 200 mg of 3.beta.-acetoxy-21-hydroxypregn-5-en-20-one, 10 ml 
of dimethyl malonate and 2.0 g of calcium chloride was heated in an 
oil-bath having a temperature of 84.degree. C. for 4 days under nitrogen 
whereupon 20 ml of toluene was added. The reaction was then arranged for 
distillation at atmospheric pressure and the bath temperature was raised 
to 160.degree. C. during 1.5 hours. Subsequently the temperature of the 
bath temperature was lowered to 84.degree. and the mixture was subjected 
to distillation at high vacuum for 1.5 hours. The resulting solid residue 
was dissolved in 15 ml of ether, 2.5 ml of hexane and some charcoal was 
added. Filtration through diatomaceous earth, followed by concentration of 
the filtrate with intermittent addition of hexane and ether gave a 
suspension which after filtration gave 
3.beta.-acetoxy-21-methoxycarbonyl!acetoxy-pregn-5-en-20-one. 
EXAMPLE 34 
A mixture of 220 mg of the product of the preceding Example and 2.2 ml of 
t-butylamine-benzene 1:10 was left to stand at room temperature under 
nitrogen for 35 minutes, whereupon it was concentrated at reduced pressure 
with intermittent addition of benzene. Hexane was then added till the 
solution became slightly turbid. Subsequent filtration through 
diatomaceous earth and concentration of the filtrate at reduced pressure 
with intermittent addition of hexane gave resinous precipitate, which was 
dissolved in ether and precipitated from hexane. The precipitation from 
hexane was repeated once more yielding finally 127 mg of a beige solid 
consisting mainly of 
3.beta.-acetoxy-22-methoxycarbonylcarda-5,20(22)-dienolide. Chromatography 
on silica gel G coated glass plates with ethyl acetate-benzene 1:6 as the 
eluant, followed by recrystallization from hexane gave a purified sample, 
mp 100, 112.degree.-120.degree., uv (MeOH) 229 m.mu.; ir(KBr) 1770, 1725, 
1655, 1240 and 1030 cm.sup.-1. A methanolic solution of the sample, after 
addition of a small amount of aqueous potassium hydroxide, rapidly changed 
from uv 229 to 235, 286 (major) m.mu.. During the chromatographic 
development a pink colour, which appears typical for 
22-alkyloxycarbonyl!-card-20(22)-enolides, developed. 
EXAMPLE 35 
A mixture, prepared by addition of 0.1875 ml of anhydrous tetrahydrofuran 
to 30 mg of methyldiethylphosphono!-acetate, cooling in an ice-bath, 
addition of 2.2 mg of sodium hydride-mineral oil 1:1, warming to room 
temperature and addition of 27.75 mg of 
21-hydroxy-6,19-oxidopregn-4-en-3-one, was stirred at room temperature for 
20 hours, whereupon a mixture of 30 mg of methyl 
diethylphosphono!acetate. 0.1875 ml of anhydrous tetrahydrofuran and 2.2 
mg of sodium hydride-mineral oil 1:1 was added and stirring was continued 
for another 2 hours. Evaporation at reduced pressure followed by addition 
of 5 ml of ethyl acetate, 5 extractions with water and evaporation of the 
extracted organic phase at reduced pressure gave a product which, after 
chromatography on silica gel G coated glass plates with ethyl 
acetatebenzene 1:1 as the eluant and recrystallisation of the fraction 
having rf 0.35 from ether-pentane gave 
3-oxo-6,19-oxidocarda-4,20(22)-dienolide, uv(MeOH) 225 (broad) m.mu. which 
had an ir spectrum identical to that of the product of Example 7. 
EXAMPLE 36 
A mixture was prepared by successive addition, against a stream of 
nitrogen, of 76 mg of sodium hydride-mineral oil 1:1, a solution of 0.340 
ml of diethyophosphono!-acetonitrile in 1.2 ml of tetrahydrofuran and 73 
mg of 5.alpha.-chloro-3.beta. ,21-diacetoxy-6,19-oxidopregnan-20-one in 6 
ml of tetrahydrofuran to 5.2 ml of tetrahydrofuran; during the additions 
the reaction vessel was cooled by an ice-bath. The ice-bath was then 
removed and the mixture was stirred under nitrogen at room temperature for 
20 hours. One half of the reaction mixture was then evaporated at reduced 
pressure and the residue obtained was left to stand with 5.7 ml of 2N 
aqueous hydrochloric acid in the refrigerator overnight. The precipitate 
which had formed was filtered off and washed with water and ether yielding 
3.beta.-acetoxy-5.alpha.-chloro-6,19-oxido-23-iminocard-20(22)-enolide 
hydrochloride, uv(MeOH) 237 m.mu., ir(KBr) 3375, 1735, 1675, 1605, 1450, 
1370, 1245, 1230, 1095, 1035, 1025 and 920 cm.sup.-1. 
A fraction amounting to 1/4 of the original reaction mixture was then 
evaporated and the residue obtained was treated with 0.57 ml of 
concentrated hydrochloric acid and 0.57 ml of ether with external cooling. 
Heating of the separated acidic aqueous phase at 70.degree. C. under 
nitrogen and subsequent filtration gave 
5.alpha.-chloro-3.beta.-hydroxy-6,19-oxidocard-20(22)-enolide, uv(MeOH) 
218 m.mu., ir(KBr) 3430 (broad), 1775, 1750, 1725, 1620, 1170, 1050, 1030, 
1025, 1000, 920, 860 and 795 cm.sup.-1. 
EXAMPLE 37 
When 33.4 mg of 5.alpha.-chloro-3.beta.,21-diacetoxy-6,19-oxidopregn-20-one 
was subjected for 21 hours to the reaction condition of the preceding 
Example, evaporation of the reaction mixture at reduced pressure followed 
by treatment of the residue obtained with ether and 2N aqueous 
hydrochloric acid and evaporation of the ethereal phase gave a material, 
uv (MeOH) 222 m.mu., which contained 
3.beta.,21-diacetoxy-5.alpha.-chloro-20-cyanomethylene-6,19-oxidopregnane 
as the major steroid, as evidenced by tlc analsyis. 
EXAMPLE 38 
A mixture of 130 mg of 
3.beta.-acetoxy-8,19-oxido-5.alpha.-22-benzyloxycarbonyl!carda-14,20(22)- 
dienolide, 3.9 g of zinc dust, 9.75 ml of toluene and 3.25 ml of 90% formic 
acid was shaken for 16 hours under nitrogen, whereupon an additional lot 
of 3.9 g of zinc and of 1.3 ml of 90% formic acid was added. After 2 days 
of further shaking the supernatant liquid phase was decanted, 7.5 ml of 
benzene-methylene chloride 1:1 was added to the remaining residue, the 
mixture was shaken briefly and the supernatant liquid was decanted. After 
4 further extractions with benzene-methylene chloride 1:1 and 5 subsequent 
extractions with methylene chloride the combined supernatant liquids were 
evaporated at reduced pressure. Dissolution of the resulting residue, 
followed by addition of pentane till a faint turbidity appeared, 
filtration through diatomaceous earth, concentration of the filtrate at 
reduced pressure with intermittent addition of pentane, standing at 
-5.degree. C for 30 minutes, decantation of the supernatant liquid and 
drying at high vacuum gave 111 mg of a residue which contained 
3.beta.-acetoxy-22-benzyloxycarbonyl-19-formyloxy-24-nor-5.alpha.-chola-14 
,20(22)-dien-23-oic acid (probably as the olefinic Z-isomer) and also 
3.beta.-acetoxy-22-benzyloxycarbonyl-19-formyloxy-5.alpha.-carda-14,20(22) 
-dienolide as evidenced by tlc. 
Heating of 100 mg of the residue between 110 to 125.degree. C in an 
evacuated tube for 225 minutes followed by chromatographic separation on 
silica gel G coated glass plates with ethyl acetate-benzene 1:20 as the 
eluant gave 35 mg of 
3.beta.-acetoxy-19-formyloxy-24-nor-5.alpha.-chol-20(22)-en-23-oic acid 
(probably the olefinic Z-isomer) as a colourless resin; uv (MeOH) 218 and 
227 (shoulder; no change after basification of the methanolic sample 
solution with potassium hydroxide) m.mu.; nmr (CDCl.sub.3) 8.16, 7.40, 
6.55, 5.84, 5.18, 5.08, 4.4, 3.13, 2.01 and 0.78 ppm; m/e 534, 443 and 
337. The chromatographic separation also yielded, as the more polar 
fraction, 15 mg of 
3.beta.-acetoxy-19-formyloxy-22-benzyloxycarbonyl-5.alpha.-carda-14,20(22) 
-dienolide; m/e 576 (weak), 485 (medium), 483 (medium) and 467 (strong); uv 
(MeOH) 216, 230 and 249 (minor peak) m.mu., 288 (major peak) m.mu. after 
basification of the methanolic sample solution with aqueous potassium 
hydroxide. 
EXAMPLE 39 
A mixture of 300 mg of 
3.beta.-acetoxy-22-benzyloxycarbonylcarda-5,20(22)-dienolide, 9.0 g of 
zinc dust, 22.5 ml of toluene and 7.5 ml of 90% formic acid was shaken 
under nitrogen for 16 hours, whereupon an additional lot of 9 g zinc dust 
and 3.0 ml of 90% formic acid was added. Shaking was continued for 24 
hours, whereupon yet another lot of 9 g of zinc dust and 3.0 ml of 90% 
formic acid was added. The mixture was then filtered, the precipitate was 
washed well with methylene chloride and the combined filtrates were 
evaporated at reduced pressure. The residue obtained was treated with 
methylene chloride and the resulting suspension was filtered. The filtrate 
was concentrated with intermittent addition of hexane and then left to 
stand at room temperature under nitrogen till the supernatant liquid had 
clarified. Decantation gave a residue containing mainly an acid considered 
to be 
(20(22)Z)-3.beta.-acetoxy-22-benzyloxycarbonyl-24-nor-chola-5,20(22)-dien- 
23-oic acid in addition to some starting material. A mixture consisting of 
4.0 ml of an ethereal solution of the latter compound and 4.0 ml of an 
ethereal solution of diazomethane was then left to stand at 0.degree. C 
for 20 minutes. Subsequent tlc analysis indicated that still substantial 
amounts of the steroidal carboxylic acid remained and an additional 4 ml 
of the ethereal diazomethane solution was added. The mixture was then left 
to stand at 0.degree. C for another 20 minutes, whereupon it was 
evaporated at reduced pressure. Chromatography of the greenish foam on 
silica gel G coated glass plates with ethyl acetate-benzene 1:20 gave 55 
mg of a more polar fraction as a resin consisting of a mixture of methyl 
(20(22)Z)- and (20(22)E) 
acetoxy-22-benzyloxycarbonyl-24-norchola-5,20(22)-dien-23-oate; uv(MeOH) 
213 and 235 ;l (unchanged after basification of the methanolic sample 
solution with aqueous potassium hydroxide) m.mu.; nmr (CDCl.sub.3) 7.19, 
5.21, 5.18, 5.1, 4.55, 3.55, 3,50, 2.5, 2.6, 1.0, 0.68, 0.59 and 0.59 ppm. 
The chramatography also afforded 6.6 mg of a less polar fraction as a 
resin, which was considered to be a diazomethane adduct of the above 
methyl enoates. 
EXAMPLE 40 
A solution of 60 mg of 
3.beta.-acetoxy-22-benzyloxycarbonylcarda-5,20(22)-dienolide in 12.0 ml of 
ethanol was stirred at room temperature in an atmosphere of hydrogen in 
presence of 6 mg of 5% palladium on charcoal for one hour, whereupon it 
was filtered through paper-pulp in presence of nitrogen. Evaporation at 
reduced pressure gave a foam. A solution of 9/10 of the latter in 9 ml of 
ethanol was then stirred at room temperature in an atmosphere of hydrogen 
in presence of 6 mg of 5% palladium on charcoal for 50 minutes 
(rehydrogenation). Isolation of the steroidal material as outlined above 
and treatment of the residue obtained with ether-hexane gave a white solid 
consisting essentially of 
3.beta.-acetoxy-22-carboxylcarda-5,20(22)-dienolide. A small fractiion of 
the latter product, when heated in an evacuated tube at 
137.degree.-142.degree. for 20 minutes gave 3.beta.-acetoxycardenolide as 
evidenced by tlc analysis. 
EXAMPLE 41 
A mixture of 40 mg of the product of the latter reaction, when treated 
repeatedly with zinc dust as described in Example 39, gave a product 
containing 2,2-3.beta.-acetoxypregn-5-en-20-ylidene!malonic acid. 
Subsequent treatment of the latter with 0.8 ml of ether and 2.4 ml of an 
ethereal diazomethane solution, which was freshly prepared as described in 
Aldrich Chemical Catalog 14, 1969-1970, at 0.degree. for 1-2 hours, 
concentration at reduced pressure to 1/2 of the original volume, addition 
of hexane, further concentration with intermittent addition of hexane, 
evaporation and chromatography of the residue obtained on silica gel G 
coated glass plates, using ethyl acetate benzene as the eluant, gave a 
fraction which after recrystallisation from ether-hexane gave 2.5 mg of 
dimethyl 2,2-3.beta.-acetoxypregn5-en-20-ylidene!malonate, uv (MeOH) 219 
and 232 (no change after addition of aqueous potassium hydroxide to the 
methanolic sample solution) m.mu.; m/e 412, 397, 380 and 365. 
EXAMPLE 42 
A mixture of 70 mg of 21-acetoxy-8,19-oxido-5.beta.-pregn-14-en-20-one, 
12.2 ml of anhydrous tetrahydrofuran and 17.6 ml of 
tetrahydrofuran-ethoxyacetylene -- 1.95N methyl lithium in ether 
11.2:0.245:1.12 was stirred under nitrogen for about 2 hours, whereupon it 
was evaporated at reduced pressure with minimal exposure to air. The 
residue was dissolved in 35 ml of ether and extracted five times with 17.5 
ml of water in an atmosphere of nitrogen. Evaporation at reduced pressure 
gave a product considered to consist essentially of 
20.xi.,21-dihydroxy-20.xi.-ethoxyethynyl-8,19-oxido-5.beta.-pregn-14-ene, 
which was stirred for 40 minutes with 6.95 ml of ethanol and 1.39 ml of 2N 
aqueous sulfuric acid under nitrogen, whereupon 35 ml of water was added. 
Extraction of the aqueous mixture with ether and methylene chloride, 
followed by repeated extraction of the combined organic phases with water, 
evaporation and preparative thin layer chromatography (silica gel G, ethyl 
acetate - benzene 1:4) of the residue obtained gave 26.32 mg of a yellow 
solid consisting essentially of 
8,19-oxido-5.beta.-carda-14,20(22)-dienolide as evidenced by tlc analysis 
and by the reactions of the following Examples. 
EXAMPLE 43 
A mixture of 660 mg of zinc dust, 22 mg of 
8,19-oxido-5.beta.-carda-14,20(22)-dienolide, 1.65 ml of toluene and 0.55 
ml of 90% formic acid was shaken at room temperature for 19 hours, 
whereupon 2.2 ml of ethyl acetate and 1.1 ml of water was added. Shaking 
was continued for 0.5 hours, whereupon the mixture was filtered. Repeated 
extraction of the filtrate with water, evaporation of the organic phase, 
preparative thin layer chromatography (silica gel G, ethyl acetate-benzene 
1:4), extraction of the major product from the scraped silica gel powder 
with methanol-ethyl formate 4:1, evaporation of the extract at reduced 
pressure, treatment of the residue with methylene chloride, filtration and 
evaporation of the filtrate gave 16.22 mg of 
19-formyloxy-5.beta.,8.beta.-carda-14,20(22)-dienolide as a white solid, 
which was used for the next reaction. 
EXAMPLE 44 
When 12 mg of 19-formyloxy-5.beta.-carda-14,20-(22)-dienolide was oxidized 
with N-bromoacetamide under conditions which were essentially the same as 
those outlined in Example 17, the corresponding 
14.beta.-hydroxy-15.alpha.-bromo analog was obtained, the freshly filtered 
and still wet bromo hydrin was reduced with Raney nickel under the 
conditions described in Example 17. The crude product was purified by 
chromatography on silica gel G coated glass plates with ethyl 
acetatebenzene 1:4 as the eluant. The fraction, rf 0.45-0.5, 0.82 mg, was 
considered to consist of 
19-formyloxy-14,15.beta.-oxido5.beta.,14.beta.-card-20(22)-enolide. The 
fraction rf 0.20-0.25, 6.65 mg consisted of 
19-formyloxy-14-hydroxy-5.beta.,14.beta.-card-22(22)-enolide and a small 
amount of an impurity of similar rf value. The latter fraction, 0.325 ml 
of methylene chloride, 195 mg of zinc dust and 0.65 ml of methylene 
chloride saturated with 90% formic acid was shaken under nitrogen for 55 
minutes. Filtration, followed by dilution of the filtrate with ether, 
extraction of the organic phase with water, half-saturated aqueous sodium 
bicarbonate and water, evaporation of the organic solvent, chromatography 
of the residue as outlined above and recrystallisation of the major 
fraction with methylene-chloride-pentane gave 4.75 mg of purified 
19-formyloxy-14-hydroxy-5.beta.,14.beta.-card-20(22)-enolide, mp 
158.degree.-158.5.degree. C, ir(KBr) 3871, 3463, 3100, 2945, 2910, 2890, 
2880, 1786, 1750, 1718, 1635, 1628, 1481, 1456, 1381, 1350, 1311, 1270, 
1185, 1089, 1072; 1049, 1032, 991, 961, 912, 860 and 700 cm.sup.-1 ; m/e 
402, 384, 356 and 325.