NO136151B - ANALOGICAL PROCEDURES FOR THE PREPARATION OF HEART-EFFECTIVE NEW CARDENOLIDES AND BUFADIENOLIDES. - Google Patents

Abstract

Analogy method for the production of cardiac active new cardenolides and bufadienolides.

Description

The present invention relates to a process for the production of new cardenolides and bufadienolides and derivatives thereof, especially with the formula,

where A means a grouping according to the formulas , A^, A- or A.; 3 4'

R<1> hydrogen, hydroxy or acetoxy,

R hydroxy or acetoxy, R amino, C1-6-alkyl-amino, £-hydroxyethylamino, C1-6-alkanoylamino, di^Cl g"31^3-) amino' di (Cj^galkanoyl) amino, N-C-^g-alkyl-N-C ^ ^-alkanoylamino, N-C^ ^-alkanoyloxyalkyl-N-C^_g-alkanoylamino, dihydroxyethylamino, di(C^ &-alkanoyloxyethyl)amino, N-P-hydroxyethyl-N-C1_6~alka-noylamino, 2-oxo-3-oxazolidinyl or ureido ;

4 1 R methyl or if A is a grouping A^ with R = hydrogen, also hydroxymethyl, formyl or carboxy, 5 4 R , if R is hydroxymethyl, formyl or carboxy, 4 (3-hydroxy or, if R is methyl, and A is a grouping A^ with R"*" = hydrogen, a- or p-hydrogen, and otherwise (3-hydrogen, or salts thereof, Compounds of formula I are obtained according to the invention in that one in a compound of the general formula

~ where Z means hydroxyimino or azido, and A, R<4> and R^ have the above meanings.

reduces the group Z to the amino group, after which, if desired, alkanoylates, alkylates and/or hydroxyalkylates or carbamoylates the amino group thus formed, and if desired cyclizes a hydroxyethylamino group to oxazolidinone, and optionally transfers the reaction product to a salt.

Particularly suitable reducing agents for the reduction of a compound of formula II are aluminum amalgam and, on noble metal catalysts, activated hydrogen.

When using aluminum amalgam as a reducing agent, it is appropriate to work at room temperature. As a reaction medium, e.g. aqueous ethanol and mixtures of dioxane, ethanol and water.

A preferred noble metal catalyst is palladium, especially palladium on a support such as calcium carbonate.

The acylation of the amino group can e.g. occur by reaction with a reactive acyl derivative, e.g. an acyl anhydride or acyl halide, in the presence of a base, such as pyridine or alkali hydroxides or carbonates.

The carbamoylation can e.g. carried out by reaction with isocyanate or by treatment with phosgene and then with ammonia.

The methylation of the amino group conveniently takes place by treatment with formaldehyde and subsequent catalytic hydrogenation. The hydroxyethylation can be carried out by reaction with ethylene oxide. A hydroxyethylamino group thus obtained can be cyclized to a 2-oxo-3-oxazolidinyl group by treatment with phosgene (method variant d). The reaction of a compound with formula IV according to process variant c) takes place under reversibility of the configuration, i.e. of a 3oc-bromide or 3oc-tosylate gives the substituted 3(3-amino compound). This reaction can be carried out in a known manner in the presence of an organic or inorganic base, such as pyridine or alkali hydroxides or carbonates.

Preferred compounds of formula I are those in which the substituent in the 3-position exhibits (3-configuration.

The compounds produced according to the invention have a positive inotropic effect on the heart muscles. The latency time as well as the time of action is shorter than for clinically used cardiac glycosides. Dosage doses for adults are 2 - 20 mg per day in consideration. In the case of animal experiments, e.g. the contraction force with respect to . the hamster's anterior chamber is increased by 50% using the compound 3(3-[ (2-hydroxyethyl)-amino]-14-hydroxy-5β,14(3-card-20 (22)-enolide with a concentration of 3.10~ <6> g/l. The hatcher dose for this compound is 1.5 mg/kg. With intravenous doses from 0.1 mg/kg, the rate of contraction of the cat's left ventricle increases by 50 - 200% without significantly changing the heart rate, and at the same time the pressure decreases in the right atrium with 1 - 1.5 cm H20.

The process products can be used as pharmaceuticals, e.g. in the form of pharmaceutical preparations containing these or their salts in admixture with a pharmaceutical, organic or inorganic inert carrier material suitable for enteral or parenteral administration, such as e.g. water, gelatin,

gum arabic, milk sugar, starch, magnesium stearate,

talc, vegetable oils, polyalkylene glycols, etc. The pharmaceutical preparations can be in solid form, e.g.

as tablets, dragées, suppositories and capsules5 or in liquid form, such as e.g. solutions, suspensions or emulsions. They are optionally sterilized and contain auxiliary substances, such as preservatives, stabilisers, wetting or emulsifying agents, and salts for changing the osmotic pressure or as buffers. They may also contain other therapeutically valuable substances.

EXAMPLE 1

Production of the output material:

a) 450 mg digitoxygenone is shaken together with 450 mg hydroxylamine hydrochloride, 900 mg sodium acetate and 45 ml 96% ethanol

30 minutes at 22°. The reaction mixture is then added with

250 ml of chloroform and washed three times each time with 50 ml of 0.5-n hydrochloric acid, 50 ml of 10% potassium bicarbonate solution and two . times with 50 ml of water. The aqueous phases are still extracted

twice with each time 100 ml of chloroform-ethanol (9:1). The combined chloroform-ethanol solutions are dried over sodium sulfate and evaporated in vacuo. The remaining crude product (474 mg) is chromatographed on 250 g of silica gel with chloroform containing 7% isopropanol. 12 ml fractions are collected. The factions

55-69 give 240 mg of crude digitoxigenone oxime A. Fractions 70-80 give 57 mg of a mixture of digitoxigenone oxime A and B. Fractions 81-98 give 150 mg of digitoxigenone oxime B. From that

crude digitoxygenone oxime A is obtained after recrystallization in methanol-ether-n-pentane 115 mg hexagonal leaves with m.p. 178 - 182°, which after longer storage shows a melting point of

234 - 238°; [ct]^<5> = +39.8° (chloroform). In a thin-layer chromatogram, this product still shows a content of approx. 2% of oxime B. Digitoxigenone oxime B is amorphous. b) 1.5 g of sodium acetate is mixed with 1.2 g of hydroxylamine hydrochloride, 20 ml of methanol are added and filtered. The filtrate obtained

is added to a solution of "2 g of digitoxygenone in 20 ml of methanol and heated for one hour at reflux. The methanol is evaporated in vacuo, the residue obtained is taken up in water and chloroform-ethanol (4:1) and the aqueous phase is extracted several times with chloroform-ethanol The combined organic solutions are washed with water, dried over sodium sulfate, filtered and evaporated.

2.04 g of crude oxime mixture is obtained, which is dissolved in acetone, added with ether and inoculated with oxime A, whereby 445 mg of digitoxygenone oxime A with a melting point of 234 - 238° crystallizes. This product is thin-layer chromatically pure.

Method according to the invention:

c) Dissolve 200 mg of the mixture of digitoxigenone-oxime A and digitoxigenone-oxime B obtained in the above manner in 2.5 ml

dioxane and 6 ml of ethanol and then added with 3.5 ml of water and 0.5 ml of 10% ammonia as well as 100 mg of crystalline

ammonium chloride. Aluminum amalgam is added to the clear solution, which is prepared fresh in the following way: 2 g of aluminum shavings are covered with 10% caustic soda and heated for 3 minutes in a steam bath, whereby strong hydrogen evolution occurs. The caustic soda is poured off, the etched aluminum is washed four times with water and then once with ethanol, then left to rest for 15 seconds with a 2% HgCl-j solution, and then immediately washed three times with water and once with ethanol. The reaction mixture is now allowed to stand

at 20° and follows the course of reduction in a thin-layer chromatogram [eluent chloroform-ethanol (10:1)]. After 2-3 days no oxime is detectable, the amalgam is washed several times with ethanol and the combined solutions are evaporated in a vacuum. The residue is evaporated twice with chloroform and once with methanol.

The residue thus obtained is treated with acetone. After evaporation, the acetone solution gives 50 mg of 3oc-amino-14-hydroxy-5|3-card-20(22)-enolide hydrochloride. The acetone-insoluble residue (150 mg)

taken up in dry hot ethanol to give 113 mg of 3|3-amino-14-hydroxy-5|3-card-20-(22)~enolide hydrochloride; m.p. 277 - 287° (decomposition): [«Id5 = +10° (in water)•

EXAMPLE 2

50 mg of 3(3-amino-14-hydroxy-5(3-card-20 (22)-enolide hydrochloride) is dissolved in 1 ml of dry pyridine, the solution is added with 0.5 ml acetic anhydride and left to stand for 16 hours at 20°. The reaction mixture with water and chloroform is then added, the chloroform phase is washed successively with dilute hydrochloric acid, dilute soda solution and water, dried over sodium sulfate, filtered and evaporated. The residue gives, after recrystallization in acetone, 37 mg of 3|3-acetamido-14-hydroxy-5(3,14|3-card-20 (22)-enolide, prisms with m.p. 281 - 287° (decomposition) $ [a] ^<5> =■ + 22° (in chloroform) .

EXAMPLE 3

150 mg of 3ct-amino-14|3-hydroxy-5|3-card-20 (22)-enolide hydrochloride in the form of the acetone-insoluble residue obtained according to example 1, section c) is dissolved in 1 ml of pyridine and 0, 5 ml of acetic anhydride and leave to stand for 16 hours at 20°. The crude acetylation product is chromatographed on 100 g silica gel with chloroform-ethanol (20:1)

as eluent. 15 ml fractions are collected. Fractions 48-52 yield 35 mg of crude 3|3-amino-14-hydroxy-5[3-card-20 (22)-enolide. from

sions 50-60 contain 30 mg of crude 3α-acetamido-14-hydroxy-5|3-card-20(22)-enolide, which is thin-layer chromatographically uniform. Melting point 156 - 177 and 253 - 257° (in acetone-ethanol after vacuum drying.

EXAMPLE 4

529 mg of the raw digitoxygenone-oxime mixture obtained according to example 1, section b) is dissolved in 50 ml of 95% ethanol and added with aluminum amalgam, which, as described above, is prepared from 1.1 g of fresh aluminum shavings. The reaction mixture is shaken for 3 days at 20° (after 2 days a further 2.5 ml of water is added). The reaction mixture is extracted in several portions with a total of 200 ml of chloroform-ethanol (5:1). The organic phases are filtered clear over a filter aid. 476 mg of colorless foam is obtained as a residue, which is dissolved in 5 ml of pyridine, 4

ml of acetic anhydride and leave to stand for 30 minutes at 20°. The work-up of the reaction mixture gives 540 mg of crude acetylation product which is crystallized in acetone-ether, gives 41.5 mg of crystalline 3P-acetamido-14-hydroxy-5(3-card-20 (22)-enolide).

Processing of the mother liquor on aluminum oxide (activity II)

with benzene/chloroform (4:1 and 3:2) gives a mixture of 3a- and 3p-acetamido-14-h<y>drox<y>-5|3-card-,20 (22)-enolide, which crystallized in acetone-ether, gives crystals with double melting point 174 -

179 and 245 - 260-.

EXAMPLE 5

Production of the starting material:

a) A solution of 1.6 g of epi-digitoxigenin in 40 ml of dry pyridine is added with a solution of 1.5 g of freshly distilled

toluenesulfonyl chloride in 10 ml of pyridine and allowed to stand for 15 hours at 20°. The reaction mixture is then acidified to pH 4 and extracted several times with chloroform. The chloroform extracts are washed with dilute sodium hydroxide, dried over sodium sulphate, filtered and evaporated. The crude product thus obtained is chromatographed on 30 g of silica gel. One elutes with chloroform/ethanol (99:1) and 1.63 g of 3-toluenesulfonyl-epi-digitoxigenin with a melting point of 155 - 156° (in acetone-ether) is obtained5

[cc]^<5> = + 35° (chloroform).

b) A solution of 500 mg of 3-toluenesulfonyl-epi-digitoxigenin in 43 ml of dimethylformamide is added with 0.7 g of sodium azide and

heated under moisture exclusion for 3 hours at 75°C. The reaction mixture is stirred for a further 16 hours at 20° and then filtered. The filtrate is added with 27 ml of water and 17 ml of chloroform-ethanol (4:1). The organic phase is separated, the aqueous phase is shaken three times with chloroform-ethanol (4:1). The. combined organic extracts are washed with water, dried over sodium sulfate and evaporated in vacuo. The residue, recrystallized in acetone, gives 235 mg of 3|3-azido-3-deoxy-digitoxigenin with m.p. 213 - 217° [ct]^<5> = +22° (chloroform).

Procedure according to the invention:

c) Dissolve 350 mg of 3f3-azido-3-deoxy-digitoxigenin in 15 ml of dioxane, 10 ml of methanol, 5 ml of water and 2 ml of 10% ammonium hydroxide, add 80 ml of ammonium chloride and shake for 17 hours in a hydrogen atmosphere with 2 g aluminum amalgam, which is prepared as described in example 1, c). Work-up of the reaction mixture according to example 3 gives 235 mg of 3j3-amino-14-hydroxy-5p-card-20 (22)-enolide hydrochloride with m.p. 277 - 287°

(fission).

EXAMPLE 6

Production of the starting material:

Analogously to example 5, one obtains from 5 g of digitoxigenin and 3 g of p-toluenesulfonic acid chloride 1.58 g of 3(3-p-toluene-sulfonyl-digitoxigenin with m.p. 145 - 147°; [<x] D = +16° (chloroform) The tosylate is converted by reaction with sodium azide into 3a-azido-3-deoxy-digitoxigenin, m.p. 165 - 167°, [oc]^~<*> = +43°

(chloroform).

Method according to the invention:

The 3a-azido compound is reduced with aluminum amalgam and the uncrystallized crude product is acetylated as in example 4. 3a-acetamido-14-hydroxy-5p-card-20(22)-enolide is obtained, which is identical to the one obtained according to example 3 the connection.

EXAMPLE 7

Preparation of the starting material:

a) Dissolve 7 g of oleandrigenin in 28 ml of pyridine, cool to 0°

and added with the cooled solution of 4.7 g of tosyl chloride

in 17.5 ml of pyridine and allowed to stand for 4 days at 0°. By successive addition of water and ice and tearing (or inoculation) of the separated oil, a crystalline, powdery precipitate is obtained which is sucked off and dissolved after washing with water in chloroform. The chloroform solution is shaken twice with water, which contains a few drops of diluted HCl, once with water,

twice with cold saturated KHCO^ solution and then with water out. After drying over Na2S0^, filtration and evaporation of the solvent in vacuo, the residue crystallizes in acetone-ether.

You receive approx. 8.8 g of crystalline 16P-acetoxy-14-hydroxy-3(3-tosyloxy-5|3,14(3-card-20 (22)-enolide. Melting point 137 -

139°. b) Dissolve 7 g of 16P-acetoxy-14-hydroxy-3P-tosyloxy-5p,14P-card-20(22)-enolide in 100 ml of dimethylformamide, add 6 g of finely powdered NaN^ and stir for 17 hours at 20° . The reaction mixture is added with 200 ml of water, made slightly acidic with acetic acid and then shaken three times with 200 ml of chloroform each time. The chloroform phase is washed twice with each time 50 ml of water,

dried over Na2SO^, filtered and evaporated in vacuo. The highly volatile dimethylformamide can, for the most part, be removed in a vacuum by repeatedly extracting the residue in benzene and expelling it. 2.5 g of 3oc-azido-16(3-acetoxy-14-hydroxy-5|3,14[3-card-20 (.22)-enolide is obtained. 1.6 g of product can be further obtained from the mother liquor. The pure product melts at 189 - 190°; [cc]^<4> = +3° - 2° (in chloroform).

Method according to the invention:

c) Dissolve 500 mg of 3cx-azido-16(3-acetoxy-14-hydroxy-5|3,14|3-card-20 (22)-enolide in 25 ml of ethanol and 10 ml of water and hydrate after

addition of 250 mg of 5% Pd/CaCO 3 as well as 25 drops of triethylamine for 1 1/2 hours at 20°. After filtering off the catalyst, it is evaporated in vacuo, the residue is taken up with dilute HCl and a little water (pH approx. 3), the acidic solution is extracted three times with

each time 30 ml of chloroform. The acidic, aqueous solution is made

alkaline with dilute NH 3 solution and extracted three times with each time 30 ml of chloroform. The extracts are washed with a little water, dried over Na 2 SO 4 , filtered and evaporated in vacuo. The remainder, 460 mg of crude amine, is dissolved in 2 ml of dry chloroform and added with as much, at 20°, in chloroform, saturated with dry HCl gas, until the solution reacts slightly acidic (pH approx. 5). The precipitated hydrochloride is filtered off, washed with chloroform and dissolved in methanol-ether. 385 mg of 3oc-amino-16|3-acetoxy-14-hydroxy-5(3,14|3-card-20 (22 )-enolide hydrochloride is obtained. [<x]^4 = +4° - 2°

(in methanol). X 214 nm.

max

EXAMPLE 8

Dissolve 1050 mg of 3α-amino-16(3-acetoxy-14-hydroxy-5β, 14|3-card-20 (22)-enolide hydrochloride in 10 ml of pyridine, add 7 ml of acetic anhydride and allow to stand for 30 minutes at 2 2°. Water is then added, left to stand for a further 30 minutes and evaporated in vacuo with several additions of benzene to dryness. The crude product is purified by chromatography on 30 g of SiO2- With chloroform-methanol (99:1) 1.1 g is obtained 3oc-acetylamino-16(3-acetoxy-14-hydroxy-5(3 ,14(3-card-20 (22)-enolide elute. M.p. 240 - 243°. X = 214 nm, Tal^4 = + 27 ° - 2° (in chloroform). max ' L JD

EXAMPLE 9

Production of the starting material:

a) Dissolve 9 g of crude 3-epi-oleandrigenin in 200 ml of pyridine, add 8 g of tosyl chloride in 30 ml of pyridine and leave to stand

at +3°. After 20 hours, the reaction mixture is taken up in 400 ml of chloroform-ether (1:3) and shaken in a separatory funnel with ice and diluted HCl until all the pyridine is removed. The organic solution is washed neutrally with water, dried over Na 2 SO 3 , filtered and evaporated in vacuo. After recrystallization in acetone-ether, 16(3-acetoxy-14-hydroxy-3a-tosyloxy-5|3,14(3-card-20 (22)-enolide) is obtained. Melting point 154 - 156°.

b) 9.45 g of 16(3-acetoxy-14-hydroxy-3oc-tosyloxy-5(3,14(3-card-20 (22)-enolide) is dissolved in 300 ml of dimethylformamide. One adds

20 g of finely powdered NaN^ and heat with stirring at 50°. After 5 hours it is filtered, the filtrate is added with 200 ml

water, neutralized with acetic acid and extracted with chloroform. Work-up of the extract yields 9.24 g of crude azide, which is chromatographed on 700 g of SiO 2 . 1.5 ml fractions are collected.

Fractions 70-90 give 2.10 g of 16(3-acetoxy-3P-azido-14-hydroxy-

- 5(3,14(3-card-20 (22)-enolide, m.p. 182 - 183°; [a]^<3> = -6° - 2°

(in chloroform).

Progress was slow according to the invention:

c) lg 16P-acetoxy-3p-azido-14-hydroxy-5(3,14(3-card-20(22)-enolide is dissolved in 50 ml ethanol and 12 ml dioxane, add

a solution of 125 mg of NH^Cl in 5 ml of water and then adds 6 g of fresh amalgamated aluminum shavings. (6 g of aluminum shavings are covered with 40 ml of 10% NaOH. As soon as, after brief heating, a strong H2 evolution occurs, it is poured from the NaOH. The shavings are washed several times carefully with distilled water and then with pure ethanol. Then add 40 ml of HgCl2 solution (2%) and swirl the vessel for 30 seconds. Then decant. Wash immediately several times with water). It is allowed to stand for 4 days at 20°. Then the shavings are poured from the solution, the shavings are thoroughly washed with ethanol. The combined solutions are centrifuged, the remaining solution is decanted, the precipitate is extracted three times with ethanol. The combined solutions are dried in vacuo at 20°. The remainder is dissolved in 25 ml of water and such highly diluted HCl that the solution reacts just like congo acid. It is then extracted several times with chloroform. The acidic aqueous solution is made alkaline with dilute NH 3 solution and extracted to exhaustion

with chloroform. The water-washed extracts are dried over Na2S0^, filtered and evaporated. Thereby 1~6(3-acetoxy-3(3-amino-14-hydroxy~5|3 ,14(3-card-20 (22)-enolide crystallizes. Melting point 167 - 177°. The hydrochloride prepared from it shows a UV -absorption X 214 nm, fal^<4> = 0° - 2° (in methanol).

* J max L J D

EXAMPLE 10

By treating 16(3-acetoxy-3(3-amino-14-hydroxy-5|3,14P-card-20 (22)-enolide hydrochloride with acetic anhydride in pyridine and chromatography of the reaction product on SiO^ with chloroform and chloroform -methanol (99:1) gives 16j3-acetoxy-3(3-acetylamino-14-hydroxy-5p,14p-card-20 (22)-enolide. Mp. 254 - 256°,

max ' L JD

X 216 nm: Fa].^<3> -10 - 2° (in chloroform),

EXAMPLE 11

Preparation of the starting material:

a) Dissolve 1.05 g of 3(3-azido-16(3-acetoxy-14-hydroxy-5(3,14P-card-20 (22)-enolide in 70 ml of methanol, add 2 ml of 25%

NH3~16sning and stands at rest at 20°. After 45 hours, neutralize with dilute acetic acid and evaporate the methanol in a vacuum. The thus precipitated precipitate is skimmed off and washed. with water. From chloroform/acetone 3(3-azido-14 ,16(3-dihydroxy-5f3 ,14(3-card-20 (22)-enolide crystallizes in prismatic small plates

(730 mg) with m.p. 227 - 229°, X = 216 nm, fal^3 + 49° <-> 2°

' max J D (in chloroform.)_

Method according to the invention:

b) 400 mg of 3(3-azido-14,16(3-dihydroxy-5(3,14(3-card-20 (22)-enolide) is dissolved in 75 ml of methanol and 12 ml of water, added with

200 mg of 5% Pd/CaCO^ as well as 25 drops of triethylamine and hydrated under normal conditions for 2 hours at 20°. The preparation of this hydration addition takes place according to example 7, section c). 3|3-amino-14,16(3-dihydroxy-5|3,14(3-card-20(22)-enolide hydrochloride is obtained. X = 216 nm. [a] =

+ 29° - 2° (in methanol.

EXAMPLE 12

500 mg of 3(3-amino-14,16|3-dihydroxy-5(3,14|3-card-20 (22))-enolide hydrochloride is introduced into 7 ml of pyridine and 0.3 ml of acetic anhydride, whereby in the course of 30 minutes a completely clear solution results. The solution stands 1 1/2 hours at 20°. It by frequent addition of benzene and evaporation of

the same residue obtained in vacuum is taken up in chloroform and

dilute HCl. The chloroform phase gives, after neutral washing with water, drying over Na2S04, filtration and evaporation, 450 mg of crude

product, which is chromatographed over 12 g of SiO2. Elution with chloroform-methanol and crystallization in chloroform-acetone gives

185 mg 3£-acetylamino-no~14,16(3-dihydroxy-5|3,14j3-card-20 (22)-enolide with m.p. 239 - 240°. X max = 216 nm, [oc]£2 = + 40° t 2° (in chloroform).

EXAMPLE 13

Production of the starting material;

al 400 mg of 16(3-acetoxy-3a-azido-14-hydroxy-5(3,14(3-card-20 (22.)-enolide is dissolved in 20 ml of methanol and added with a solution of 80 mg of K^O ^ in 4 ml of water. After 90 minutes, the solution is neutralized with acetic acid, drained and the filter residue is washed

with water. After crystallization in acetone, 3a-azidO-14,16p-dihydrOxy-5p,14P-card-20 (22)-enolide is obtained. ^ max =

215 nm; m.p. 215 - 224°; [a]£<4> + 59° - 2°.

Method according to the invention:

b) 300 mg 3a-azido-14,16(3-dihydroxy-5j3,14(3-card-20 (22) -

enolide is dissolved in 54 ml of methanol, added with 4.5 ml of water (as

contains 15 drops of triethylamine) and 150 ml of 5% Pd/CaC03.

One hydrates for 1.5 hours. Usual work-up gives 3a-amino-1-4,16f3-dihydroxy-5|3,14j3-card-20(22)-enolide. Temp. 187 - 189°. The hydrochloride crystallizes in finely tangled needles.

Cleavage mp. at 270°. [cc]<24> + 52° - 2° (in methanol).

EXAMPLE 14

Dissolve 320 mg of 3α-amino-14,16(3-dihydroxy-5(3,14(3-card-20 (22)-enolide hydrochloride in 4 ml of pyridine, add 0.115 ml of acetic anhydride) and leave to stand at 20° . After 40 minutes, 0.5 ml of water is added and evaporated with several additions of benzene in vacuum until dryness. The crude product thus obtained is chromatographed on 60 g of SiO 2 . With chloroform-ethanol (46:1) and (19:1) is obtained mixtures of non-polar substances Further elution with chloroform-ethanol (19:1) and (23:2)

gives 3α-acetylamino-14,16(3-dihydroxy-5p,14p-card-20 (22)-

enolid. Temp. 159 - 163° (in methanol-acetone). <>>s = 213 nm.

r ]24 ^ c„o * n , , ^ . max I aJD = + 60 - 2 (in chloroform-) .

EXAMPLE 15

Production of the starting material:

a) Dissolve 2.1 g of 12|3-acetoxy-14-hydroxy-3-oxo-5|3,14|3-card-20 (22)-enolide in 100 ml of 80% dioxane, cool to 0° and

is added with 15 - 20 ml of 80% dioxane, which contains 730 mg of NaBH^ and stirred at 0° for 20 minutes. 10 ml of methanol is then added, the reaction mixture is neutralized with acetic acid

and evaporate to a small volume which is extracted three times with each time 25 ml of chloroform. The extracts are washed twice with 10 ml of water each time, dried over Na 2 SO 4 , filtered and evaporated in vacuo to give 2.1 g of crude reduction product. This is chromatographed on 200 g of SiO2 with chloroform-ethanol (19:1).

One obtains 12|3-acetoxy-3a,14-dihydroxy-5|3,14|3-card-20 (22)-enolide with m.p. 200 - 203° (in acetone-ether).

b) 1.2 g of 12p-acetoxy-3a,14-dihydroxy-5p,14|3-card-20 (22)-enolide is dissolved in 10 ml of pyridine and allowed to stand with 1.2 g

tosyl chloride 4 days at 20°. You then add 10 ml of water and let it stand for 2 hours. It is then extracted three times with 20 ml of chloroform each time. The extracts are washed twice with 2-n HCl and then with water, dried over Na 2 SO 4 , filtered and evaporated in vacuo to give 1.5 crude tosylate. This is dissolved in 20 ml of dimethylformamide, added with 750 mg of NaN^ and heated for 2 hours with constant stirring at 70 - 80°. Then 15 ml of water is added, it is immediately extracted three times with each time 30 ml of chloroform, the extracts are washed three times with each time 15 ml of water, dried over Na 2 SO 4 and evaporated in vacuo. 1.6 g of crude product is obtained, which is chromatographed on 60 g of SiO2 with ethyl acetate-cyclohexane (1:1), giving 1.0 g of pure 12(3-acetoxy-3P-azido-14-hydroxy-5(3,14| 3-card-20 (22)-enolide.

Method according to the invention:

c) Dissolve 300 mg of 12|3-acetoxy-3(3-azido-14-hydroxy-5|3,14P-card-20 (22)-enolide in 50 ml of ethanol and shake with 150 mg of Pd/CaCO^

(5%) 1 1/2 hours in H2 atmosphere at 20°. The catalyst is then filtered off, the solution is evaporated in vacuo, the residue is added with dilute HCl and until a pH of approx. 3-4 and the acidic solution is extracted three times with 20 ml of chloroform each time. The acidic, aqueous solutions are made alkaline

with dilute NH3-16 solution and extracted three times with 20 ml of chloroform each time. The extract is washed with water, dried over Na2S04, filtered and evaporated in vacuo to give 293mg of crude

product. This is dissolved in 2 ml of dry chloroform and, with agitation, add enough at 20° with dry HCl gas saturated chloroform that the solution reacts slightly acidic (pH approx. 5).

The precipitated hydrochloride is filtered off, washed three times with 10 ml of chloroform each time and crystallized in methanol-ether.

270 mg of 12£-acetoxy-3(3-amino-14-hydroxy-5(3,14(3-card-20 (22)-enolide hydrochloride) is obtained, which decomposes at about 300°.

[α]20 = + 46.1° - 4° (in methanol).

EXAMPLE 16

Dissolve 100 mg of 12|3-acetoxy-3P-amino-14-hydroxy-5p,14(3-card-20 (22)-enolide hydrochloride in 1 ml of pyridine, add 0.5

ml of acetic anhydride and leave to stand for 30 minutes at 20°.

Pyridine and acetic anhydride are removed by adding benzene and evaporating the same in vacuo (repeated several times).

The residue gives from acetone-ether 3(3-acetamino-12(3-acetoxy-14-hydroxy-5(3,14|3-card-20 (22))-enolide with m.p. 241 - 257°;

[ct]£° = + 53.1° t 3° (in chloroform).

EXAMPLE 17

Production of the starting material:

a) Dissolve 2 g of digoxin in 10 ml of pyridine, cool to -10°, add at -10° with 3 g of chloroacetic anhydride, stir for 10 minutes at -10° and then add immediately with 20 ml of water. The reaction mixture is then extracted three times with 30 ml of chloroform each time. The extracts are washed in sequence with 2-n hydrochloric acid, until all pyridine has been removed, then shaken briefly with 7 ml of 10% KHCO 2 -16 solution and then washed two more times with neutral water. After drying the extracts over Na2SO4, filtration and evaporation in vacuum, the crude acylation product is dissolved in 60 ml of methanol, 15 ml of acetone and 15 ml of dioxane, added with 30 ml of 2-n H,,SO4 and boiled for 35 minutes at reflux. Then add 100 ml of water and evaporate in a vacuum at approx. 100 ml. The precipitated crystals are dissolved in chloroform, the solution is washed twice with water, dried over Na 2 SO 4 , filtered and evaporated in vacuo. 1.1 g of 12(3-chloroacetoxy-3(3,14-dihydroxy-5(3,14(3-card-20 (22)-enolide) is obtained. Mp. 269 - 2 75° (in chloroform-methanol- acetone).b) 2 g of 12P-chloroacetox<y>-3P,14-di<y>drox<y>-5<p>,14|3-card-20 (22)-enolide are dehydrated using chromic acid in acetone/dioxane to 3-ketone. This is dissolved in 63 ml of methanol, added with 2 g of KHCO 3 in 40 ml of water and allowed to stand for 16 hours at 20°. The methanol is then removed in vacuo and the aqueous solution is extracted three times each time 40 ml chloroform. The extracts are washed with 20 ml water, dried over Na2SO4, filtered and evaporated in vacuo. 1.56 g of 12(3,14-dihydroxy-3-oxo-4(3,14|3-card-20 (22)-enolide, which after recrystallization in chloroform-methanol-acetone melts at 251 - 256°. c) 1.4 g 12(3,14-dihydroxy-3-oxo-5(3,14|3-card- 20 (22)-enolide is dissolved in 25 ml of pyridine, cooled to -10°, added over the course of 30 minutes with 10 ml of a 2:1 mixture of 98% formic acid and acetic anhydride cooled to -10° and allowed to stand until delay 16 hours at -:1 0°. Extract with chloroform and work up the extracts by washing with 2-n hydrochloric acid, water and KHCO3 solution. The formyl derivative thus obtained is reduced directly with NaBH^ as described in example 15, section a) and worked up. The reduction product obtained (1.3 g) is chromatographed on 200 g of SiO 2 with chloroform-ethanol (10:1). 12(3-formyloxy-3a,14-dihydroxy-5p,14(3-card-20 (22)-enolide) is obtained, m.p. 233 - 243°.

d) Dissolve 300 g of 12P-formyloxy-3a,14-dihydroxy-5(3,14(3-card-20 (22)-enolide in 50 ml of methanol, add 300 mg of KHCO^ and

so much water that just a clear solution results. It is allowed to stand for 24 hours at 20°, then the methanol is evaporated in vacuo and the aqueous part is extracted three times with chloroform-ethanol (4:1). The organic phase is washed twice with 10 ml of water, dried over Na^SO^, filtered and evaporated in vacuo. One obtains 3a, 12(3,14-trihydroxy-5|3,14P-card-20 (22)-enolide with m.p. 259 - 264°.

e) Dissolve 1.7 g of 3a,12(3,14-trihydroxy-5(3,14|3-card-20 (22))-enolide in 20 ml of pyridine, add 1.7 g of tosyl chloride and

stand for 4 days at 23°C. After adding 20 ml of water, it is kept for 2 hours at 20° and then extracted three times with 20 ml of chloroform each time. The extracts are washed twice with 2 N HCl and then with 10% KHCO 3 solution and

water, dried over Na2S04, filtered and evaporated in vacuo.

The crude tosyl ester (2.6 g) is chromatographed on (2.6 g) 230 g SiO2 with ethyl acetate-cyclohexane (1:1), whereby 1.9 g of the tosyl ester in plates (in acetone-ether) with m.p. 152 - 154° is obtained. The tosyl ester is subjected, as described in example 15, section b) to azidolysis. The crude azide is dissolved to saponify the 12-formyloxy group in 30 ml of methanol and 30 ml of dioxane and allowed to stand with the solution of 1.6 g of KHCO^ in 50 ml of water for 20 hours at 20°. The solvent is removed in vacuo, the precipitated azide is filtered off, washed neutrally with water and chromatographed on 80 g of SiO2. Elution with chloroform-ethanol (19:1) gives 36-acido-12|3,14-dihydroxy-5|3,14(3-card-20 (22)-enolide with m.p. 245 - 249°.

Method according to the invention:

f) Dissolve 250 mg of 3£-azido-12(3,14-dihydroxy-5£,14P-card-20 (22)-enolide in 90 ml of ethanol, add 125 mg of Pd/CaCO^'

(5%) and shaken for 2 hours in an H2 atmosphere at 20°. Preparation as in example 15, section c), gives 40 mg neutral part and 200

mg crude amine, from which, as described in example 15, section c), the hydrochloride is obtained. The 3(3-amino-12|3,14-dihydroxy-5f3,14f3-card-20 (22)-enolide hydrochloride melts at 290 - 300° (under decomposition). [oc]£° = + 23° ± 3 ° (in methanol).

EXAMPLE 18

Dissolve 300 mg of 3(3-amino-12|3,14-dihydroxy-5|3,14|3-card-20 (22)-enolide hydrochloride in 3 ml of pyridine and cool to -10°, add with 1 ml acetic anhydride and allowed to stand for 20 minutes at -10 o. Then 7 ml of 2-n hydrochloric acid is added. It is extracted five times with each time 15 ml of chloroform-ethanol (4:1). The extracts are washed with 10% KHCO3 solution and then with water, dried over Na2SO4, filtered and evaporated. 286 mg of crude product, which from methanol with the addition of a little ether gives pure 3P-acetylamino-12(3,14-dihydroxy-5|3,14(3-card-20 (22)-enolide with mp 296-300°-, [a]<20> = + 21.6° - 4° (in methanol).

EXAMPLE 19

Production of the starting material:

a) Dissolve 0.8 g of 12-0-acetyldigoxigenin in 10 ml of pyridine, add 1.3 g of tosyl chloride and allow to stand for 4 days

at 20°. Processing takes place as described in example 15, section b). The crude tosyl compound is stirred in 7 ml of dimethylformamide with 500 mg of NaN^ for 3 hours at 70° and then worked up: 1.48 g of crude azide, which is chromatographed on 80 g of SiO 2 with ethyl acetate-cyclohexane (3:5). One obtains 12(3-acetoxy-3a-azido-14-hydroxy-5(3,14|3-card-20 (22)-enolide with m.p. 222 - 224°,

(in acetone-ether).

b) Dissolve 700 mg of 12P-acetoxy-3a-azido-14-hydroxy-5|3,14(3-card-20 (22)-enolide in 33 ml of methanol, add 70 mg

K^ CO^ in 3.5 ml of water and allowed to stand for 75 minutes at 23°, then neutralized with acetic acid, evaporated in vacuo, added with water and extracted three times with chloroform-ethanol (2:1): 660 mg of crude product. Chromatography on 80 g SiO 2

jned chloroform-ethanol (99:1) gives next to the starting material, 3a-azido-12(3,14-dihydroxy-5(3,1.4|3-card-20 (22)-enolide). From acetone plates with m.p. 185 - 193°.

Procedure according to the invention:

c) 160 mg 3α-azido-12(3,14-dihydroxy-5|3,14(3~card-20 (22) -

enolide is dissolved in 27 ml of ethanol and shaken with 80 mg of Pd/caCO^

(5%) 2 hours in H2 atmosphere. Workup as in example 15, section c) yields 30 mg of neutral product and 130 mg of basic product, which are transferred into the hydrochloride. 3α-amino-12(3,14-dihydroxy-5(3,14(3-card-20 (22)-enolide hydrochloride melts at 290 - 300° (decomposition);

[a]<20> = + 22.4° - 2° (in methanol).

EXAMPLE 20

Dissolve 50 mg of 3α-amino-12(3,14-dihydroxy-5|3,14(3-card-20 (22)-enolide hydrochloride in 1.0 ml of pyridine, add 0.5 ml of acetic anhydride and leave to stand for 20 minutes at -10° Then after usual work-up the obtained 3a-acetylamino-12(3,14-dihydroxy-5|3,14|3-card-20 (22)-enolide is amorphous, [cx]<21> = + 42.9° in 4° (in methanol).

EXAMPLE 21

Production of the starting material:

a) Dissolve 160 mg of 3a-amino-12(3,14-dihydroxy-5|3,14|3-card-20 (22)-enolide hydrochloride in 2 ml of pyridine, add with 1 ml

acetic anhydride and leave to stand for 16 hours at 20°. The crude product obtained after usual work-up gives 123 mg of pure 12(3-acetoxy-3a-acetylarnino-14-hydroxy-5f3,14(3-card-20 (22))enolide with m.p. 253 - 260°.

Method according to the invention:

b) Dissolve 350 mg of 12|3-acetoxy-3cx-azido-14-hydroxy-5f3,14(3-card-20 (22)-enolide in 45 ml of ethanol and shake with 175 mg

Pd/caC03 (5%) 2 hours at 20° in I^ atmosphere. Processing yields 65 mg of neutral part and 2 73 mg of crude base. The hydrochloride is amorphous. 120 mg of the crude base are dissolved in 2 ml of pyridine and 1 ml of acetic anhydride and allowed to stand for 20 hours at 20°. According to the usual work-up, 12(3-acetoxy-3a-acetylamino-14-hydroxy-5|3 ,14|3-card-20 (22)-enolide is obtained from chloroform-acetone-ether with m.p. 253 - 260°-, 0]<2>° = + 96.2° - 4° (in chloroform).

EXAMPLE 22

Dissolve 900 mg of freshly prepared 3|3-amino-14-hydroxy-5(3,14(3-card-20(22)-enolide in 100 ml of dry ethanol, add 3 g of paraformaldehyde, 600 mg of 5% Pd /CaCO^ and 1.25 ml of acetic acid and shaken for 2 2 hours in a H,, atmosphere at 20 . The catalyst and undissolved paraformaldehyde are then filtered over a filter sealed with kieselguhr and the filtrate is evaporated in vacuo. The residue is dissolved in 50 ml chloroform and shake this

with dilute hydrochloric acid out. The acidic aqueous phases are washed in chloroform, made alkaline with ammonia, extracted several times with chloroform, the extract dried over ^£50^, filtered and evaporated in vacuo: 870 mg of crude 3(3-(dimethylamino)-14-hydroxy-5( 3,14|3-card-20 (22)-enolide Crystallization in acetone gives prisms with mp 218 - 223° X = 214 nm

max

Dissolve 870 rag crude base in slightly dry chloroform and add at 20° dried HCl gas saturated chloroform dropwise, until the solution is just acidic: 835 mg colorless needles. Redissolution in methanol-ether gives 720 mg of pure 3(3-(dimethylamino)-14-hydroxy-5(3,14(3-card-20 (22)-enolide hydrochloride). M.p. 258 - 279° (decomposition) .X max = 213 nm.

Dissolve 200 mg of 3(3-(dimethylamino)-14-hydroxy-5(3,14P-card-20 (22))-enolide in 2 ml of methanol, add 0.4 ml of CH3J and leave to stand for 14 hours at 20° Then evaporated in vacuum, whereby crystallization occurs: 195 mg of crude product, which after redissolving in methanol-acetone gives 3|3-(trimethylammonium)-14-hydroxy-5(3,14P-card-20 (22) -enolide iodide in white needles with mp 251 - 254°.

EXAMPLE 23

Production of the starting material:

a) Dissolve 5 g of bufalin in 150 ml of acetone, cool to 0°, add with constant stirring dropwise with 6.5 ml of Kiliani mixture [j. Polonia et al., Heiv. 42, 1437 (1959)] and allowed to stand for 15 minutes at 0°. After adding 50 ml of water and 500 mg of Na-acetate, free from acetone^ The remaining aqueous solution is added with 5 ml of 2-n ^SO^ and shaken out four times with 50 ml of chloroform each time. The extracts are washed -with water, dried over Na^SO^, filtered and evaporated. 4.15 g of 14-hydroxy-3-oxo-5(3,14(3-bufa-20 (22)-dienolide) is obtained in prisms with a double melting point of 2o5°/232-238° (in acetone). b) lg bufalin is dissolved in 4 ml of pyridine, cooled to 0°, added with the ice-cold solution of 0.7 g of tosyl chloride in 2.5 ml of pyridine and allowed to stand for 50 hours at 0°. Then added with ice, kept for 2 hours at 20° and taken up with 50 ml of chloroform-ether (1:3). The organic phase is washed successively with 2-n HCl, water, 1-n soda solution and water, dried over Na2SO^, filtered and evaporated. 3-0-tosylbuphalin is obtained with mp 143 - 145° c) Dissolve 500 mg of 3-0-tosylbuphalin in 20 ml of dimethylformamide, add 200 mg of finely powdered NaN^° and stir at 20°.

After 18 hours it is processed. 3oc-azido-14-hydroxy-5(3,14|3-bufa-20 (22)-dienolide is obtained, m.p. 175 - 185°, "A 296 nm. [a]" = + 15 - 2 (i chloroform).

Method according to the invention:

d) Dissolve 720 mg of crude 3a-azido-14-hydroxy-5(3,14(3-buf a-20 (22))-dienolide in 250 ml of methanol-water (9:1) and hydrate after adding 300 mg of 5 % Pd+CaC03 and 20 drops of triethylamine under usual conditions in the course of 2 hours. The work-up gives 3x-amino-14-hydroxy-5(3,14(3-buf a-20 (22)-dienolide hydrochloride.

[ct]<23> = _6° <±> 2° (in methanol).

EXAMPLE 24

325 mg of 3cx-amino-14-hydroxy-5(3,14|3-buf α-20 (22)-dienolide hydrochloride is allowed to stand in 3 ml of pyridine and 2 ml of acetic anhydride for 30 minutes at 20° and then liberated in vacuum during several additions with benzene and pyridine and acetic anhydride. The residue gives from acetone 250 mg of 3a-acetylamino-14-hydroxy-5|3,14(3-bufa-20 (22)-dienolide; flat prisms with m.p. 164 - 166 o;

[a]<23> = + 24° - 2° (in chloroform).

EXAMPLE 25

Production of the output material:

a) 4 g bufalon [Example 23, section a)] dissolve in 100 ml dioxane-water (4:1), add with the solution of 1.5 g NaBH^

in 100 ml dioxane-water (4:1) and stirred for 30 minutes at 20°. It is then neutralized with acetic acid, freed in vacuum from dioxane, the remaining aqueous solution is extracted five times with 20 ml of chloroform each time and this is worked up in the usual way. 3.35 g of 3-epi-buphalin are obtained; prismatic needles with double-m.p. 257°/260-272°, (in methanol-ether).

b) 3.25 g of 3-epi-buphalin is dissolved in 70 ml of pyridine, cooled to 0°, added with the ice-cold solution of 2.8 g of tosyl chloride in 10 ml of pyridine and allowed to stand at -3°. After

65 hours add 100 g of ice, leave for 2 hours at 20°, record

then with 200 ml of chloroform-ether (1:4) and washed with 2-n HCl, water, saturated KHCO 3 solution and water, dried over Na 2 SO 4 and evaporated. 4.68 g of 14-hydroxy-3a-tosyloxy-5(3,1413-buf a-20 (22)-dienolide is obtained as a jelly-like residue.

c) Dissolve 4 g of crude 14-hydroxy-3α-tosyloxy-5|3,14(3-bufα-20 (22)-dienolide in 120 ml of dimethylformamide, add 3 g of fine

powdered NaN^ and stirred for 26 hours at 60°. It is then neutralized with acetic acid and 200 ml of water is added. The separated crystals are sieved off and washed with water. The filtrate is extracted exhaustively with chloroform. The dry residue is combined with the crystallisate and chromatographed on 50 g of SiO2. 3(3-azido-14-hydroxy-5|3,14|3-bufa-20 (22)-dienolide is obtained with m.p. 186 - 206°5 [a]<24> = -3° - 2° ( in chloroform).

Method according to the invention:

d) 800 mg 3f3-azido-14-hydroxy-5|3,14(3-buf a-20 (22)-dienolide

dissolve in 100 ml methanol and 10 ml water, add 500 mg

.5% Pd/CaCO^ and 10 drops of triethylamine and hydrated for 5 hours at 20°. The work-up gives 550 mg of crude amine, which is transferred with HCl-saturated chloroform into 3(3-amino-14-hydroxy-5|3,14(3-buf a-20 (22)-dienolide hydrochloride). V = 296 nm. fal<24> = -12°

+ o max. LJD

- 2 (in methanol).

EXAMPLE 26

From 3(3-amino-14-hydroxy-5(3,14(3-bufa-20 (22)-dienolide, one obtains by acetylation using acetic anhydride/pyridine 3(3-acetylamino-14-hydroxy-5(3 ,14|3-buf a-20 (22)-dienolide with mp 273 - 286° (in chloroform-acetone).[a]D = + 1.5° - 2° (in chloroform).

EXAMPLE 2 7

Dissolve 750 mg of 3P-amino-14-hydroxy-5(3,14(3-card-20 (22)-enolide in 7 ml of ethanol, cool to -10°, add 3 ml of a cooled to -10° solution of ethylene oxide in ethanol (130 mg/ml) and a trace of pyridine and allowed to stand for 18 hours at 20 .

It is then freed from ethanol in vacuo, the residue is taken up in 30 ml of chloroform, the extract is washed with 2-n HCl, saturated KHCO 3 solution and water, dried over Na 2 SO 4 and filtered. All aqueous phases are subsequently extracted with chloroform. The acidic solutions are cooled with ice, made alkaline with aqueous NH 2 solution and extracted three times with 20 ml of chloroform-ethanol (4:1) each time. The extracts are washed with water and diluted

NH 3 solution, dried over Na 2 SO 4 , filtered and evaporated. The residue, light yellow foam, is dissolved in a little chloroform, the solution is added with an HCl-chloroform solution (at 20° saturated with HCl gas) until just a slightly acidic reaction (litmus). The precipitated hydrochloride is filtered off and washed with dry chloroform: 510 mg of 3(3-(2-hydroxyethyl)amino]-14-hydroxy-5(3,14(3-card-20 (22)-enolide hydrochloride, which decomposes after re-dissolving in methanol-acetone at above 2 60°.

EXAMPLE 28

When treating the compounds obtained according to example 27

with acetic anhydride/pyridine and chromatography of the crude product on SiO2 with chloroform-ethanol (26:1) yields 3(3-[(acetoxyethyl)acetylamino]-14-hydroxy-5(3,14p-card-20 (22 )-enolide with mp 179 - 180° (in acetone-ether).[a]<20> + 14° 2° (in chloroform).

EXAMPLE 2 9

Dissolve 500 mg of crude 3(3-[ (2-hydroxyethyl) amino]-14-hydroxy-5|3,14(3-card-20 (22)-enolide hydrochloride in 40 ml of dry dioxane and 2 ml of dry pyridine, cooled to -10°, added with 1.015 ml of a -10° cooled solution of C0C12 in toluene (1 ml =

575 mg C0C12) and allowed to stand for 30 minutes at 20°. After this time, no more starting material can be detected.

Add a few drops of ethanol, leave for a further 30 minutes at 20° and then evaporate while adding benzene several times in a vacuum. The remainder is taken up in chloroform-

ethanol (4:1) and the aqueous phase is shaken with chloroform

ethanol (4:1). The organic phases are washed with water, dried over Na 2 SO 4 , filtered and evaporated in vacuo. The residue is chromatographed on 90 g of SiO2 with chloroform-ethanol (26:1). One obtains 14-hydroxy-3(3-(2-oxo-3-oxazolidinyl)-5(3 ,14(3-card-20 (22)-

enolide with m.p. 238 - 247°; [oc]£° = +25° - 2° [in chloroform-methanol (1:1)].

EXAMPLE 30

Production of the starting material:

a) Dissolve 2.03 g of uzarigenin in 500 ml of acetone and dehydrate analogously to example 23, section a). 1.5 g of uzarigenone is obtained,

prisms with m.p. 267 - 2 77°. 1.48 g of uzarigenone is dissolved in 50

ml of methanol while heating, is added with the solution of 1.2 g of hydroxy]amine hydrochloride and 1.5 g of crystalline Na acetate in 7.5

ml of water and boil under reflux. After 30 minutes, filter off and add water to the filtrate. The precipitate is isolated and gives a total of 1.5 g of crude 14-hydroxy-3-(hy dr ok sy lami no)-5a, 14(3-card-20 (22)-enolide. From methanol crystal leaves with m.p. 296 - 303°

(fission).

Method according to the invention:

b) 1.5 g of 14-hydroxy-3-(hydroxyimino)-5a,14|3-card-20 (22)-enolide is dissolved in 100 ml of dioxane, 120 ml of ethanol, 40 ml of methanol and 45 ml of hot water. After cooling at 20°, 30 g of fresh: amalgamated Al shavings, 1 g of NH4Cl as well as 1.5 ml of 25% NH3 solution are added and allowed to stand for 5 days at 20°. The work-up and separation into neutral and basic proportions takes place as described in example 9. The crude amine is dissolved in 15 ml of dry chloroform and added dropwise with dry HCl gas saturated chloroform at 0° until just acidic reaction. Thereby, 3(3-amino-14-hydroxy-5a, 14(3-card-20 (22)-enolide hydrochloride) is secreted.

22 + o

[a]D = + 13 - 2 (in methanol).

EXAMPLE 31

500 mg of 3P-amino-14-hydroxy-5a, 14(3-card-20 (22)-enolide hydrochloride is acetylated in 12 ml of pyridine and 9 ml of acetic anhydride 20

hours at 37 , added with benzene and evaporated in vacuo.

One obtains 150 mg of 3|3-acetylamino-14-hydroxy-5a, 14(3-card-20 (22) - enolide with m.p. 285 - 292°. [a]<22> = - 29° - 2° ( in pyridine).

EXAMPLE 32

Dissolve 2 g of freshly prepared 3(3-amino-14-hydroxy-5p ,14(3-card-20 (22) - - j i enolide in 440 ml of dry chloroform, cool at 0°, add 20 ml of ethereal HNCO solution ( according to G. Brauer, Hdb. PrMp. Anorg,Chemie, F. Enke, Stuttgart 1954, p. 508), [90 mg HNCO,ml]

at 0° and 7 drops of pyridine and allowed to stand at 0° for 32 hours. The reaction solution is then evaporated in a vacuum at approx. 30 ml

and shakes out the residue with slightly diluted HCl, whereby the HCl extract is washed with chloroform-ethanol (4:1). The organic phases are washed with 5 ml of cold saturated aqueous KHCO 3 solution and water, dried over Na 2 SO 4 , filtered and evaporated in vacuo. 1.7 g of crude product is obtained, which is chromatographed on 130 g of SiO 2 with chloroform, which contains 7.5% ethanol. The pure 3|3-ureido-14-hydroxy-5p,14(3-card-20 (22)-enolide melts at 255 - 262°.

[cx]<20> .= + 53° - 2° (in methanol).

EXAMPLE 33

Preparation of the starting material:

a) Dissolve 2.9 g of resibufogenone in 75 ml of dioxane-water (4:1), add with the solution of 1.5 g of NaBH. in 75 ml dioxane-water (4:1) and stirred for 30 minutes at 20 o. Then neutralized with glacial acetic acid and freed from dioxane in vacuum. The precipitate is filtered off, washed with water and dried at -60 o. 1.9 g of 3-epi-resibufogenin with m.p. 206 - 222° (in acetone). b) 2.5 g of 3-epi-resibufogenin is dissolved in 75 ml of pyridine and cooled at 0° and a similarly cooled solution of 2.1 g of tosyl chloride in 10 ml of pyridine is added at 0° and allowed to stand at 3°. After 4 days, ice is added, poured into 200 ml of chloroform-ether (3:1), the organic phase is washed five times with each time 50 ml of dilute HCl, once with water, three times with cold saturated KHCO^ solution and finally finally with water, dried over Na2S0^ and evaporated. The crude product thus obtained gives, after recrystallization in acetic ester ether, 3-0-tosyl-3-epi-resibufogenin with m.p. 131 - 136°.

j

c) 3.1 g of 3-0-tosyl-3-epi-resibufogenin is added to 90 ml of dimethylformamide with 2.3 g of finely powdered NaN^ and stirred for 24

hours at 60°. It is then neutralized with glacial acetic acid and added with water until blackening begins. The crystallisate is sieved off and washed with water. Chromatography on silica gel gives 3(3-azido-14,15-epoxido-5|3,14(3-bufa-20,22-dienolide) with m.p. 226 - 22 9° (in acetone).

Method according to the invention:

d) Dissolve 2.6 g of 3(3-azido-14,15-epoxido-5(3,14(3-bufa-20,22-dienolide) in 250 ml of ethanol, add 50 drops of triethylamine and 2 g of 5% ig Pd/CaC03 and shaken in an H2 atmosphere under normal pressure. After 7 hours filter from the catalyst, evaporate in vacuo at 20° (bath temperature) and the residue is taken up in chloroform. The chloroform solution is shaken out five times, each time with 20 ml of HCl acidified water and then with 20 ml of water, dried over Na2S04, filtered and evaporated. 1.8 g of crude product A are obtained. The hydrochloric acid aqueous phases are made alkaline under ice-cooling with a 10% NH3 solution and shaken four times with each time 40 - "ml chloroform. The chloroform extracts are washed with water, dried over Na2SO4 and evaporated at 20° (bath temperature): 880 mg crude 36-amino-13,15-epoxido-5(3,14|3-bufa-20,22-dienolide ( crude product B) e) The crude products A and B obtained according to the above are dissolved in 5 ml of pyridine and 3 ml of acetic anhydride and allowed to stand for 15 minutes at 20°. After addition of water, brought to dryness in vacuo during several additions of benzene: 2.7 g of crude product. This is chromatographed on 400 g of SiO2 with chloroform. 3 (3-acetyl amino-14,15-epoxido-5(3,148-bufa-20,22-dienolide) is obtained with m.p. 148 - 155° [a]<24> = + 6° - 2°

(in chloroform).

EXAMPLE 34

Preparation of the starting material:

a) Dissolve 5.8 g of cinobufagin in 125 ml of dioxane-water (4:1), add the solution of 1.75 g of NaBH4 in dioxane-water (4:1) and stir for 20 minutes at 20°. It is then neutralized with glacial acetic acid and freed from dioxane in a vacuum. The precipitate is extracted three times with chloroform, the extract is washed with water, dried over Na2SO4, filtered and evaporated in vacuo. The thus obtained

crude product is chromatographed on 500 g of SiO2 with acetic acid ethyl ester-cyclohexane-(1:1). One obtains 3-epi-cinobufagin with double-

m.p. 116 - 118°/138 - 143° (in methanol-ether) Tal<23> = -2 - 2°

L J D

(in chloroform) .

b) Dissolve 3.7 g of 3-epi-cinobufagin in 15 ml of pyridine, cool to 0°, add with a 0° cooled solution of 2.5 ml

tosyl chloride in 9 ml of pyridine and allow to stand at 3°. After 39 hours, ice is added, the precipitate is filtered off, washed with dilute HCl and water and taken up in chloroform. The chloroform solution

shaken with dilute HCl and water, dried over Na,,SO4 and

steamed. One obtains 3-O-tosyl-3-epi-cinobufagin with m.p.

161 - 166° (in chloroform-acetone 1:1).

c) Dissolve 3.96 g of 3-0-tosyl-3-epi-cinobufagin in 120 ml of dimethylformamide, add 1.6 g of NaN^ and stir for 40 hours at 60°.

It is then neutralized with glacial acetic acid and added with water until blackening begins. The precipitate is skimmed off, washed with water,

dried and chromatographed on 280 g SiO2 with acetic acid ethyl ester-cyclohexane-(1:4), 3B-azido-14,15-epoxido-16S-acetoxy-5B,14B-bufa-20,22-dienolide is obtained with m.p. 190 - 196°.

Method according to the invention:

d) Dissolve 2.05 g of 3B-azido-14,15-epoxido-16B-acetoxy-5p,14B-bufa-20, 22-dienolide in 250 ml of ethanol, add 50 drops

triethylamine and 1.0 g of 5% Pd/CaCO^ and shaken in an H2 atmosphere under normal pressure. After 3 hours, filter from the catalyst, evaporate in vacuo at 20° (bath temperature), the residue is taken up in chloroform, this is shaken three times with dilute HCl and once with water, dried over Na 2 SO 4 , filtered and evaporated in vacuo.

The residue is dissolved in 5 ml of pyridine and 3 ml of acetic anhydride and allowed to stand

to rest at 20°. After 30 minutes, pyridine is removed and

acetic anhydride by repeated addition of benzene and evaporation in vacuo. The residue is chromatographed on 200 g of SiO2 with acetic acid-ethyl ester-cyclohexane-(4:1). One obtains 3|3-acetylamino-14,15-epoxido-16f3-acetoxy-5p, 14|3-bufa-20, 22-dienolide with m.p.

185 - 197°, [a] = 0° + 2° (in chloroform).

EXAMPLE 35

3 p-[bis-(2-hydroxyethyl)amino"|- 14-hydroxy-5B,14p~card-2Q(22)-enolide. 4 g 3j3-[(2-hydroxyethyl)amino]-14-hydroxy- 5p, 14p-card-20(22) - enolide is dissolved in 50 ml of pure ethanol, cooled to -10°C, mixed with a solution prepared at -10°G of 2.4 g of ethylene oxide in 10 ml of ethanol and left to stand tightly closed at 20°C. After 65 and 90 hours respectively, a further 2.4 g of ethylene oxide in 10 ml of ethanol are added as above to the reaction solution. After a total of 5 days, evaporate in vacuum to foam. By soil chromatographic purification of 100 g SiO2 "Merck" (0.05 - 0.20 mm) with mixed chloroform-ethanol- 25%. Aqueous NH 3 solution-(99:1:0.4) (volume ratio i) as eluent yields 3.45 g according to DC. [Chloroform-ethanol-25-% aqueous NH3~16sning-(70:30:0.4)] pure 3(3-[ bis-(2-hydroxyethyl) amino ]-14-hydroxy-5(3,140-card-20 (22) - enolide. From chloroform-ether, 3.1 g of prisms are obtained with m.p. 203-205°C. A sample gave after acetylation in pyridine/acetic acid anhydride and is usually worked up in DC. [System chloroform-ethanol- (9:1 ) ] (2 times front) only one spot. [a]D = + 15.3°c + 2°c (L chloroform). ^max = 216 nm ( 1= 172O0) (in ethanol).

Hydrochloride

8.5 g of the above tertiary amine with m.p. 203-205°C is dissolved in lOO to 200 ml of dry chloroform and mixed just to a weak acid reaction (litmus) with a chloroform solution saturated at 2cfb with HCl gas. The precipitated thihydrochloride f raf Utreres

washed with dried chloroform and crystallized from anhydrous ethanol: 5.56 g of prismatic plates with m.p. 248-249°C (during solutions after centration from 238°C). After DC. [Chloroform-ethanol-25-%, aqueous NH3-16sning-(70:30:0.4)] uniform. [°0D<22> = + 12.9° + 2°

(in methanol) . } i max = 215 nm (£• = 16000) (in ethanol).

EXAMPLE 36

3p-[bis-(2-O-acetoxbutyl)-amino]-14-hydroxy-5p,14p-card-20(22)-enolide.

3.3 g of 3β-[bis-(2-hydroxyethyl)-amino]-14-hydroxy-5β, 14|3-card-20(22)-enolide with m.p. 203-205°C dissolve in 15 ml of anhydrous pyridine and 8 ml of acetic anhydride and let stand for 15 hours at 20°C. Usual work-ups yield 4 g of crude diacyl compound. Chromatograph i on 50 g SiO2 "Merck" (0.05 - 0.20 mm) with the eluent chloroform-ethanol-(99:1) gives 2.5 g. Prisms from acetoether or methanol-ether 2.4 g with m.p. 116-117°C, as according to DC.

[Chloroform-ethanol-(19:1)] (2 times front) is pure. A-max =

214 nm (£ = 17300) (in ethanol). [a]^<0> = + 11.3° + 3°C (in chloroform) .

Hydrochloride

1 g of the diacetyl compound obtained above is dissolved in 30 ml of dry chloroform and fed to a just slightly acidic reaction (litmus) to a chloroform solution saturated with HCl gas at 20°C, after which the HCl salt crystallizes out spontaneously. The crystals are filtered off and washed with dry chloroform and ether: 750 mg prisms with m.p. 192-225°c as according to DC. [Chloroform-ethanol-25%. aqueous NH3 solution-(95:5:0.5) ( 2 times front)] gives only one stain. <A>max <=> 213 ™ (6 = 17600) (in ethanol); [a]<22>=+18/6°c +3 (in methanol.

Claims (5)

1. Analogous process for the preparation of new cardiac active 3-amino derivatives of cardenolides or bufadienolides of the general formula where A means a grouping according to the formulas A-p A2, A3 and A4j R1 hydrogen, hydroxy or acetoxy, 2 3 R hydroxy or acetoxy, R amino, C-^ g-alkyl-amino, (3-hydroxyethyiamino, ^-alkanoyl amino, di (C-^_g-alkyl) amino, di(C^ ^-alkanoyl) amino, N-C^ g-alkyl-N-C^_g-alkanoylamino, N-C-^ ^-alkanoyloxyalkyl-N-C-^_g-alkanoylamino , dihydroxyethylamino, di(C^ ^-alkanoyloxyethyl) amino, N-B-hydroxyethyl-N-C-L_^-alka-noylamino, 2 -oxo-3-oxazolidinyl or ureido; 4 1 R methyl or if A is a grouping A-^ with R = hydrogen, also hydroxymethyl, formyl or carboxy, 5 4 R , if R is hydroxymethyl, formyl or carboxy, 4 B-hydroxy or, if R is methyl, and A is a grouping A1 with R<1> = hydrogen, a- or p-hydrogen, and otherwise B-hydrogen, or salts thereof, characterized in that one in a compound of the general formula where Z means hydroxyimino or azido, and A, R<4 >and R^ have the above meanings, reduces the group Z to the amino group, "whereupon, if desired, alkanoylates, alkylates and/or hydroxyalkylates or carbamoylates the amino group thus formed, and if desired cyclizes a hydroxyethylamino group to oxazolidinone, and optionally transfers the reaction product to a salt. 2. Method according to claim 1, for the production of 3p_[(2-hydroxyethyl)amino]-14-hydroxy-5p,14p-card-20(22)-enolide., characterized by reducing 3p-azido-3-deoxy -digitoxigenin to produce 3β-amino-14-hydroxy-5β/14β-card-20(22)-enolide, and then reacts this compound with ethylene oxide. 3. Method according to claim 1 for the production of 3{3-[(N-2-acetoxyethyl)acetamido]-14-hydroxy-5p-, 14(3-card-20 (22)-enolide, characterized in that it is acetylated according to claim 2 prepared 3(3-[ (2-hydroxyethyl)amino]-14-hydroxy-5p,14p-card-20(22)-enolide. 4. Process according to claim 1, for the production of 3a-acetylamino-16p-acetoxy-14-hydroxy-5p,14p-card-20(22)-enolide, characterized by reducing 3a-azido-16(3-acetoxy- 14-hydroxy-5(3,14p-card-20 (22)-enolide for the preparation lation of 3α-amino-16β-acetoxy-14-hydroxy-5β,14β-card-20(22)-enolide, and then acetylates this compound. 5. Method according to claim 1 for the production of 30-acetylamino-14,15-epoxido-5p,14p-bufa-20(22)-dienolide, characterized by reducing 3p-azido-14,15-epoxido-5p,14p -bufa-20(22)-dienolide to produce 3β-amino-14,15-epoxido-5β,14β-bufa-20(22)-dienolide, and then acetylates this compound.