DE3427090A1 - 7 alpha -Thio-substituted 19-hydroxy-15 beta ,16 beta -methylene-3-oxo-17 alpha -pregn- 4-ene-21,17-carbolactones, process for their preparation and their use as pharmaceuticals - Google Patents

Abstract

17 alpha -Thio-substituted 19-hydroxy-15 beta ,16 beta -methylene-3-oxo-17 alpha -pregn- 4-ene-21,17-carbolactones of the general formula I <IMAGE> in which C1...C2 is a single or double bond and R is a lower saturated alkyl radical with 1 to 4 carbon atoms or a lower saturated aliphatic acyl radical with 2 to 4 carbon atoms, are described. The compounds have a strong antialdosterone activity with, at the same time, the absence of endocrinological side effects.

Description

The invention relates to new thio-substituted in the 7a-position 19-Hydroxy-15 ß j 16ß-methylene-3-oxo-17a-pregn-4-en-21,17-carbolactone, their use and processes for their production and pharmaceutical products containing these substances Preparations according to the claims.

If R is a lower saturated alkyl radical, it represents a methyl, ethyl, propyl or ii-butyl group, the methyl and ethyl radicals being preferred. R stands for a lower saturated aliphatic acyl radical represents the acetyl, propionyl or butyryl radical; the acetyl and propionyl radical are preferred.

The new compounds of general formula I have the properties and effects of aldosterone or deoxycorticosterone to reverse or reverse the excretion of sodium and potassium salts. The compounds according to the invention are therefore suitable for the treatment of certain forms of hypertension, edema and primary aldosteronism and other aldosterone-related endo-

crinological disorders. They can also be used as diuretics. The new active ingredients have opposite the well-known spironolactone has the advantage of not showing any of the known endocrinological side effects, because they have neither an affinity for the androgen nor for the gestagen receptor.

During treatment with spironolactone, undesirable endocrine side effects are often caused by the anti-androgenic and gestagenic activity caused by spironolactone. So you watch for longer Ongoing treatment of male patients with spironolactone the occurrence of gynecomastia (Smith, W.G.,

The Lancet 1962, p. 886; Mann, NM, JAMA 1963, p. 778; Clark, E., JAMA 1965, p. 157; Greenblatt, DJ, JAMA 1973, p. 82) and impotence (Greenblatt, DJ, JAMA 1973, p. 82), which is attributed to the antiandrogenic side effect of this active ingredient (Steelman, SL et al, Steroids 1963, p. Kk9; Schane , II.P., J. of Clinical Endocrinology and Metabolism 1978, p. 69I) ■

Those in women taking spironolactone treatment occurring side effects such as atnenorrhea and cycle irregularities on the other hand, the gestagenic side effects of spironolactone are blamed. Both side effects can be tested in animal experiments as well as in vitro using the receptor binding test with the Andregen- Detect or Gestajren receptor.

It was an object of the present invention to provide compounds which the spironolactone in the The anti-aldostson effects are comparable or superior and at the same time reduced, but if possible no endocrine disruptions Show side effects.

The antialdosterone effect was determined in the test model by Hollmann (G. Ilolltnann et al., Tubular effects and rena-Ie elimination of spironolactones, Naunyn-Schmiedebergs Arch. Exp. Path. Pharmak. 2 ^ 7 (1964 :), p. 419? P. Marx, Renal effects of d-aldosterone and its antagonist spironolactone, Diss. Med. Fac. FU Berlin, 1966) determined and measured.

The androgen receptor binding test is carried out as follows :

The androgen receptor (protein) contained in the cytosol of a homogenate of rat prostate binds dihydrotesto-

steron (DHT) with high affinity but low capacity. If this receptor is loaded with ³ H-DHT in the presence of the compound to be tested, it depends on the concentration and on the binding affinity of the compound to be tested how much ³ H-DHT is displaced by the receptor. After separating the receptor-bound DHT from the non-bound DHT, the binding can be determined in percent and this value is plotted against the logarithm of the molar concentration of the test substance. The concentration of the test substance that is required to completely displace the reference substance from the receptor is now determined. The competition factor (KF) as a measure of the binding strength is defined as the ratio of the concentration of the test substance to the concentration of the reference substance, so that a high KF value indicates low binding strength, a low but high affinity.

The anti-androgenic effect is determined for compounds that do not themselves have an androgenic effect, but because of their high affinity for dying the body's own Completely or partially displace androgen from the receptor, as is the case with spironolactone to a certain extent is observed. A high competition factor in the androgen and gestagen receptor test is therefore desirable.

The gestagen receptor binding test is carried out in the same way using cytosol from rat uterus homogenate.

The following table summarizes the relative values of the antialdosterone potency (with spironolactone = 1) and the competition factors im Androgen receptor test (K) and in the gestagen receptor test (Kpv of spironolactone (A) and those according to the invention Connections using the example of

7c £ -Acetylthio - 1 9-hydroxy- 1 5ß ι 1 6ß-methylene-3-oxo · 'l-en-21,17-carbolactone (B)

TABEL

link relative
Antialdosterone
effect Competitive
factor A. 1 ^K A ^K G B. 1 - 2 8.9 21

The table shows that the compound according to the invention, none of the known undesirable endocrine disruptors Has side effects because it has neither an affinity for the androgen nor for the gestagen receptor. the Antialdostrone activity is stronger than that of spironolactone.

The compounds of the formula I according to the invention are prepared according to claim 5 per se known methods. The reaction of compounds of the formula II is particularly suitable with the corresponding thioic acid acyl-SH in the presence of a protic solvent such as methanol or ethanol, also mixed with water at a temperature from room temperature to the boiling point of the reaction mixture. After suitable work-up, the compounds according to the invention are obtained in this way of formula I with R meaning an acyl

. S *

restes. However, it is also possible to carry out the addition of the thiocarboxylic acid without a solvent.

For the preparation of the compounds according to the invention of Formula I with R meaning a lower alkyl radical is added to the compound of the formula II alkanethiol, by standing at a temperature of -1O ° C to 0 ° C in a polar solvent such as methanol or ethanol in the presence of an organic base such as piperidine or pyridine in the suspension of the compound of the formula II introduces the alkanethiol and stirs at a temperature of 10 C to 30 C until after thin-layer chromatography Check the reaction has ended. For working up, it is precipitated in water, worked up and - if necessary - by column chromatography cleaned.

The Δ double bond can be introduced chemically or microbiologically. Suitable chemical Dehydration tools for 1,2-dehydration are for example Selenium dioxide, 2,3-dichloro-5,6-dicyanobenzoquinone, Chloranil, thallium triacetate or lead tetraacetate.

Suitable microorganisms for 1,2-dehydration are for example schizomycetes, especially those of the Arthrobacter genera, such as A. simplex (ATCC 694: 6); Bacilli, such as B. lentus (ATCC I3805) or B. sphaericus (ATCC 7055) »Pseudomonas such as P. aeruginosa (IFO 35Op); Flavobacterium such as for example F. flavescens (IFO 3058) etc.

The 1,2-dehydrogenation is preferably carried out chemically For this purpose, the 1,2-dihydrosteroid is in a suitable Solvent with the dehydrating agent heated to 6O to 12O ° C over a longer period of time. Suitable solvents are, for example, dioxane, toluene, benzene, tetrahydrofuran, tert. Butanol and mixtures of these solvents.

The compounds according to the invention can per se known methods of galenics can be formulated into drugs for oral or parenteral administration » They can be dosed and administered in the same way as the commercially available spironolactone.

For human use, the dosage is 20 up to 500 mg per day.

The for the preparation of the compounds according to the invention required starting compound can be synthesized as follows:

Preparation of the starting compound

19-Hydroxy ~ 15β, 16 ~ methylene-3-oxo-17a ~ pregna ~ 4,6-diene ~ 21,17-carbold ^. 'Tori (H):

1. To a solution of 114 ο 3.beta.-acetoxy ~ 15SS, 16ss ~ 5 ~ methylene-androst-5-or -1 · 7-one in 2 ^ ¹ - 1 ether 1 71 .1 g of N-Brotnsuccinimid and A solution of 57 ml of 70% perchloric acid in 570 ml of water is added and the mixture is stirred for 2.5 h at room temperature and 1 h at ice bath temperature. The deposited precipitate is filtered off with suction, washed neutral with ice-cold ether, with water and dried at room temperature. 149 g of 3β-acetoxy-5-bromo-6β-hydroxy-15β, 16β-methylen-5aandrostan-17-one, contaminated with the corresponding 5,6β-epoxide and also succinimide and N-bromosuccinimide, are obtained.

It is used to remove the nonsteroidal impurities the precipitate was taken up in 1.49 l DMF ^ with 447 g anhydrous sodium acetate was added and the mixture was stirred at room temperature for 20 h. It is then precipitated in ice water, the precipitate filtered off, washed with water, taken up in methylene chloride, dried and evaporated. It will be 75 g crude 3ß-acetoxy-5,6ß-epoxy-15ß, 16ß-methylene ~ 5ß ~ androstane ~ 17-on received. These 75 g of epoxy are in 750 ml of acetic acid stirred with 150 g of lithium bromide for 16 h at room temperature. After ice water precipitation, the precipitate is filtered off with Water washed and dried. 86 g of 3β-acetoxy-5-bromo-6β-hydroxy-15β, 16β-methylen-5a-androstan-17-one are obtained. obtain. One recrystallized from diisopropyl ether / acetone Sample melts at 185.1 ° C.

2. To a boiling solution of 44 g of 3ß-acetoxy ~ 5 ~ bromo-6ßhydroxy-15ß, 16β ~ methylen-5a ~ androstan «-17-one in 1.8 l benzene 22 g of iodine and 48.4 g of lead (IV) acetate are added and the mixture is refluxed for 30 minutes. For work-up, the chamfered one washes Reaction solution successively with sodium hydrogen sulfite, sodium hydrogen carbonate solution and water. After this

45.6 g of a mixture of 3ß-acetoxy-5, 6ß-epoxy-15ß, 16ß-methylene-5ß-androstan-17-one and 3ß-acetoxy-5-bromo-15ß, 16ß-methylene-6ß, 19- epoxy-5a-androstan-17 ~ one obtained. This product mixture is dissolved in 684 ml of acetic acid, 68.4 ml of water and 91.2 g of zinc dust are added and the mixture is stirred at room temperature for 30 minutes. For working up, the zinc is filtered off with suction and washed thoroughly with methylene chloride. The filtrate is i. Vac. largely concentrated, the residue taken up in ether, washed with sodium hydrogen carbonate solution, dried and evaporated. After chromatography on silica gel to 10.8 g of 3.beta.-acetoxy-19-hydroxy-15SS, 16ss-methylene-5-androsten-17-one obtained ⁰ C of melting point 208.5-10.

12.5 g of 5,6β epoxide are obtained as a by-product.

3. To a solution of 4.5 g of 3β-acetoxy-19-hydroxy-15β _S 16β-methylene-5-androsten-17-one in 37.5 ml of dimethylformamide, 1.162 g of imidazole and 2.363 g of tert-butyldimethylsyl chloride are added and the mixture is stirred at room temperature for 20 hours. The reaction solution is then stirred into ice water, and the precipitate which has separated out is filtered off with suction, washed with water and dried. 5.75 g of 3 [beta] -acetoxy-19-tert-butyldimethylsilyloxy-15 [beta], 16 [beta] -methylene-5-androsten-17-one with a melting point of 153-159 ° C. are obtained.

4. To an ice-cold solution of 4.5 g of 3β-acetoxy 19 ~ tert-butyl dimethy lsilyloxy-15β, 16β-methy1 en-5-and rust-17-one in 67.5 ml of absolute tetrahydrofuran, 18 g of potassium ethoxide are added under argon then 6.75 ml dist. Propargyl alcohol was added dropwise. The reaction solution is then stirred for 1.5 hours at room temperature, then stirred into ice water, neutralized with acetic acid, and the precipitate which has separated out is filtered off with suction, washed with water and dried. 4.75 g of 19-tert-butyldimethylsi lyloxy-17a- (3-hydroxy-1-propynyl) -15β, 16β-methylene-5-androstene-3β, 17β-diol are obtained. A sample recrystallized from acetone melts at 208.4 ⁰ C.

To a solution of 4.6 g of 19-tert-butyldimethylsilyloxy-17o> (3 ~ hydroxy-1-propynyl) -15ß, 16ß-methylene-5-androstene-3ß, 17β-diol in 104 ml of methanol and 104 ml of tetrahydrofuran are about 4.7 g of moist Raney nickel catalyst and hydrogenated with hydrogen until 2 equivalents are absorbed. For work-up, the catalyst is filtered off and evaporated the filtrate to dryness. 4.6 g of 19-tert-butyldimethylsilyloxy-17a- (3-hydroxypropyl) ~ 15 ~, 16 ~ -methylene-5-androstene ~ 3 ~, 17 ~ diol are obtained obtain.

To a boiling solution of 4.6 g of 19-tert-butyldimethylsi IyI oxy-17a- (3-hydroxypropy I) -15β, 16β-methylene-5-aηdrosten-3β, 17β-diol in 280 ml of toluene and 9.4 ml of cyclohexanone are added 2.35 g of aluminum isopropoxide, dissolved in 25 ml of toluene, and heated for 8.5 h with slow distillation. For working up, the cooled reaction solution is diluted with ether, washed with dilute sulfuric acid and water, dried over sodium sulfate and evaporated. The residue is chromatographed on silica gel and 2.59 g of 19-tert-butyldimethylsilyloxy-15β, 16β-methylene-3-oxo-17a-pregn-4-ene-21,17-carbolactone with a melting point of 147.6 ° C. are obtained after trituration with diisopropyl ether. UV (MeOH): ε ₂₄₂ = 14,350.

3 g of tetrabutylammonium fluoride are added to a solution of 3.0 g of 19-tert-butyldimethylsilyloxy-15β, 16β-methylene-3-oxo-17a-pregn-4-ene-21,17-carbolactone in 30 ml of absolute tetrahydrofuran and for 30 min Left to stand at room temperature. The reaction solution is then diluted with ether, washed with water, dried and evaporated. The residue is chromatographed on silica gel and after Diisopropyletherverreibung be 1:41 g 19-hydroxy-15SS, 16ss-methylene-3-oxo-17a-pregn-4-ene-21,17-carbolactone of melting point 207-08 ⁰ C. UV (MeOH): ε ₂₄₂ = 14,750.

8.1.4 g of 19 ~ hydroxy-15 [beta], 16 [beta] -methylene ~ 3 ~ oxo-17a-pregn-4-ene ~ 21,17-carbolactone are left to stand in 6 ml of pyridine and 3 ml of acetic anhydride for 2 h at room temperature. For work-up, in Ice water was precipitated, the precipitate was filtered off, washed with water and dried. 1.6 g of 19-acetoxy-IS3, 16ßmethylene-3-oxo-17a-pregn-4-ene-21,17-carbolactone are obtained obtained as a crude product.

9. To a solution of 1.6 g of 19-acetoxy-15β, 16β-methylene-3-oxo-17a-pregn ~ 4-en-21,17-carbolactone 1.6 ml of dioxane / conc. in 16 ml of dioxane and 1.6 ml of triethyl o-formate are added. Sulfuric acid solution (prepared from 4.5 ml of dioxane and 0.16 ml of conc. Sulfuric acid) and stirred for 30 min at room temperature after. The reaction solution is neutralized by adding 0.5 ml of pyridine, then diluted with ether, with water washed, dried and evaporated. About 2 g of 19-acetoxy-3-ethoxy-15β, 16β-methylene-17α-pregna-3,5-diene-21,17-carbolactone are the residue obtained as a crude product.

2 g of crude ^ -acetoxy-S-ethoxy-loess, 16ß-methylene-17o> pregna-3,5-diene-21,17-carbolactone are dissolved in 40 ml of acetone and, with ice cooling, successively with 0.31 ml of pyridine, 1 «425 g anhydrous sodium acetate, 4 ml water and 1.06 M N-bromosuccinimide offset. The mixture is then stirred for 30 minutes while cooling, and the reaction solution is diluted with ether, with water washed, dried and evaporated. 2.3 g of 19-acetoxy-6 ^ -bromo-15ß, 16ß-methylene-3-oxo-17a-pregn ~ 4-en-21,17-carbolactone was obtained as a crude product.

2.3 g of crude 19-acetoxy-6 ^ -bromo-15ß, 16ß-methylene-3-oxo-17apregn-4-ene-21,17-carbolactone are dissolved in 23 ml of dimethylformamide with 842 mg of lithium bromide and 1,055 g of lithium carbonate for 1 hour at 100 ⁰ C stirred. For work-up, the reaction solution is diluted with ether, washed with water, dried and evaporated. The residue is chromatographed on silica gel and 1.01 g of 19-acetoxy-15β, 16β-methylene-3-oxo-17a-pregna-4,6-diene-21,17-carbolactone are obtained as an oil. UV (MeOH): & _2Q2 = 19,700.

10. 215 mg of potassium carbonate are added to a solution of 650 mg of 19 ~ acetoxy-15 ~, 16 ~ -methylene-3 ~ oxo-17a-pregna ~ 4,6-diene-21,17-carbolactone in 65 ml of methanol and the mixture is stirred for 2 hours at room temperature after. For work-up, it is diluted with methylene chloride, washed with water, dried and evaporated. 410 mg of 19-hydroxy-15β, 16β-methylene-3-oxo-17oc-pregna-4,6-diene-21,17-carbolactone are obtained as an oil. UV (MeOH): ε ₂₈₄ = 22,900.

example 1

1 ml of thioacetic acid are added to a solution of 1.0 g of 19-hydroxy-15 [beta], 16 [beta] -methylene-3-oxo-17o-pregna-4,6-diene-21,17-carbolactone in 20 ml of methanol and 4 ml of water and added for 24 h left to stand at room temperature. The reaction solution is then diluted with ether, washed with sodium hydrogen carbonate solution and water, dried and evaporated. The residue is chromatographed on silica gel and, recrystallized from diisopropyl ether / acetone, 520 mg of 7o> acetylthio-19-hydroxy ~ 15β, 16β-methylene-3-oxo-17a-pregn-4-ene-21,17-carbolactone from Melting point 252.1 ⁰ C obtained. UV (MeOH): ε ₂₃₈ = 16 050.

Example 2

Methanethiol is introduced almost to saturation into an ice-cold solution of 1.0 g of 19-hydroxy-15β, 16β-methylene-3-oxo ~ 17a-pregna-4,6-diene-21,17-carbolactone in 20 ml of methanol and 1 ml of piperidine. After storage at room temperature overnight, it is diluted with ether, washed with water, dried and evaporated. The residue is chromatographed on silica gel. 370 mg of 19-hydroxy-15 [beta], 16 [beta] -methylene-7a-methylthio-3-oxo-17a-pregn-4-ene-21,17-carbolactone with a melting point are recrystallized from diisopropyl ether / acetone Obtained 229.7 ° C.
UV (MeOH): ε _{£ 41} = 12 150.

Example 3

250 mg of 7a-acetylthio ~ 19 ~ hydroxy ~ 15 ~, 16 ~ methylene ~ 3 ~ oxo ~ 17a-pregn-4 ~ ene-21,17 ~ carbolactone are dissolved in 10 ml of dioxane with 250 mg of 2,3 ~ dichloro-5, o -dicyan-p-benzoquinone stirred for 24 h at 100 ⁰ C. For work-up, the reaction solution is diluted with ether, washed with sodium hydrogen carbonate solution and water, dried and evaporated. The residue is chromatographed on silica gel and there are 40 mg of 7a-acetylthio-19 ~ hydroxy ~ 15 ~, 16 ~ -methylene-3-oxo ~ 17a>»pregna-1,4-diene> -21, 17-carbolactone as an oil obtain. UV (MeOH): E ₂₄₃ = 15,400.

Claims (7)

Claims 7α-thio-substituted 19-hydroxy-15β, 16β-methylene-3-oxo-1 7 «-pregn - '+ - en-21,1 7-carbolactone of the general formula I. SR .CH, (I) where C " ... C _{n is} a single or double bond and R is a lower saturated alkyl radical with 1 to k carbon atoms or a lower saturated aliphatic acyl radical with 2 to t carbon atoms» 2) 7a-acetylthio-19-hydroxy-15ß, 16ß-methylene-3-oxo-1? A- pregn-'i-en-21,17-carbolactone 3) 19-Hydroxy-15 [beta], 16 [beta] -tethylene-7a-methylthio-3-oxo-17apregn- ^z 1-en-21,17-carbolactone k) 7α-acetylthio-1 9-hydroxy-15β, 16β-methylene-3-oxo-1 7ctpregna-1, k- diene-21,17-carbolactone 5) Process for the preparation of compounds of the general formula I. characterized in that in a known manner a) for the preparation of compounds of the general formula I with C ... C "meaning a CC single bond and R a lower saturated acyl radical with 2 to k carbon atoms,
to a compound of formula II HO (II) one Thiocarboic acid of the general formula III, R ¹ -SH (III) where R 'denotes a lower, saturated acyl radical having 2 to k carbon atoms, adds or b) for the preparation of compounds of general formula I with C ^ ... C _o meaning a CC- I ti * Single bond and R of a lower, saturated alkyl radical with 1 to k carbon atoms, to a compound of the formula II an alkylthiol of the general formula IV R '' -SII (IV) when R ″ denotes a lower, saturated alkyl radical with 1 to ^l i carbon atoms, is added or c) for the preparation of compounds of the general formula I with C " ... C _n meaning a C =C double bond and R having the meaning given in claim 1 a compound of the general formula I in which C ... C stands for a CC single bond and R has the meaning given above, is dehydrated, 6) Medicines based on a compound according to claims 1 to k. 7) Use of a compound according to claims to k for the production of medicaments.