JPH0439479B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel 14-aminosteroids and their therapeutic applications, and also to methods for their production. French patent application No. 2464270 describes compounds of the 14-aminosteroid type, in particular the oxy derivatives of 14-aminoandrostane and 14-amino-21-norpregnane. Steroid alkaloids of the pregnane and androstane series substituted in the 14-position by an amino group are also known, for example 14β-aminopregnane-3β,20α-diol as described by A. Astier et al. (Bull. Soc. Chim.no.9−10.P.1581～
1582, 1976). Other
14β-Aminopregnane and 14β-aminoandrostane are also described by A. Astyar et al. (Tetrahedron, Vol. 34, P. 1481-1486, 1978).
It is stated by However, neither the pharmacological properties nor the pharmaceutical applications of these derivatives have been described. French patent application no.
All are described and introduced as having immunotherapeutic properties that can be applied as drugs to treat autoimmune diseases resulting from certain lymphocyte deficiencies. Research carried out in the applicant's company surprisingly revealed that 14-aminosteroids, specifically pregnane-3,20-diol and pregnane-3,12,20-triol types substituted in the 14-position with an amino group, It was possible to confirm that the derivative has clear muscle inotropic properties. The present invention therefore relates to new medicaments based on 14-aminosteroids with active myotropic activity, which can be applied in particular as cardiotonic agents for the treatment of heart failure. The present invention also provides new products such as 14-aminopregnane-3,20-diol and 14-aminopregnane-3,20-diol.
3,12,20-triol and 14-amino-21-
Norpregnane-3,12,20-triol 14
- Concerning aminosteroids and also processes for their production. The new drug based on the present invention has the following general formula () as an active ingredient: (wherein R represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, such as a methyl group, an ethyl group or an isopropyl group, and R ₁ represents a hydrogen atom or a hydroxyl group)14
-Contains aminosteroids. The present invention also relates to pharmaceutically acceptable salts of 14-aminosteroids of general formula () obtained by reaction with mineral or organic acids according to methods customary in the art. The acid used can be selected from hydrochloric acid, oxalic acid, tartaric acid, fumaric acid, lactic acid, phosphoric acid, p-toluenesulfonic acid, formic acid, hydrobromic acid, maleic acid, sulfamic acid, and the like. The present invention also relates to the general formula () (wherein R represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, and R ₁ is a hydrogen atom or a hydroxyl group, and R and R ₁ are simultaneously The present invention relates to novel 14-aminosteroids represented by 14-aminosteroids (not hydrogen atoms) and also to their salts with acids. 14-Aminosteroids of the general formula () have several asymmetric carbon atoms in their molecules, especially 3
-, 5-, 14-, 17- and 21-positions and can therefore exist in different stereoisomeric forms, the novel compounds according to the invention are those in which R ₁ is a hydroxyl group. While they can be one of these stereoisomers if
When R ₁ is a hydrogen atom, - at the 3-position
When the OH group and the _-NH2 group at the 14-position have the β configuration and the hydrogen atom at the 17-position has the α configuration, 20 irrespective of whether the hydrogen atom at the 5-position has the α or β configuration It is assumed that the OH group at the -position consists of a stereoisomer having a β configuration. More specifically, the present invention relates to the general formula ()
(wherein R is a methyl group)
14-Relating to aminosteroids. In this formula, the _-NH2 group in the 14-position and the hydrogen atom in the 5-position can have the α or β configuration, preferably the β configuration. Similarly, the -CHROH group in the 17-position can have a 17α or 17β configuration. The -OH group in position 3 and the -OH group represented by R ₁ preferably have a β configuration. When R is not a hydrogen atom, - at the 20-position
The OH group can have the α or β configuration and when R ₁ is a hydrogen atom it preferably has the β configuration. The invention, of course, relates to these various isomers, isolated or in mixtures.
Unless otherwise indicated, 5 based on conventional nomenclature.
The hydrogen atoms in the - and 17-positions occupy the α configuration in the examples below. The compound of general formula () is the following general formula () (wherein R represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms and R ₁ represents a hydrogen atom or an alkyl group), The reduction reaction followed by acetylation gives the following general formula () 3- and 20- (wherein R is a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, R ₁ is a hydrogen atom or an acetyl group, and Ac is an acetyl group) position and when R ₁ is a hydroxyl group, a 3,14,20-trihydroxysteroid O-acetylated at the 12-position is formed and corresponding by reaction with triazohydrogen acid/boron trifluoride complex salt.
The 14-azide derivative is formed and then reduced with a metal hydride or by catalytic hydrogenation. reduction followed by acylation to produce di-O-acetylated trihydroxysteroids when R ₁ is a hydrogen atom, and when R ₁ is a hydroxyl group,
The first step, consisting of forming tri-O-acetylated tetrahydroxysteroids, is described, for example, in Organic Reactions in Steroid Chemistry, published by Van Nostrand Reinhold (1972).
It can be carried out with hydrides by the technique described by J. Fried and JA Edwards.
For example, it is possible to use aluminum or boron dihydrides, such as sodium borohydride or lithium aluminum hydride, or alternatively other reducing agents such as hydrogen in the presence of a catalyst. The acetylation reaction making it possible to obtain acetylated steroids of the general formula () can be carried out based on customary techniques, for example reaction with acetic anhydride in an organic solvent. In a second step, the derivative of general formula () is converted into the corresponding 14-azido derivative by reaction with a solution of triazohydrogen acid/boron trifluoride complex in benzene. 14- by tertiary azide group
Substitution of the hydroxyl group in the 3-position, 12-position (as appropriate) and 20-position previously protected by acetylation results in good yields of other hydroxyl groups. The third step, consisting of converting the 14-azido derivative into the corresponding 14-amino derivative, is carried out by catalytic hydrogenation followed by removal of the acetyl protecting group, or more specifically, as a metal hydride. This is carried out by reaction with a reducing agent such as lithium aluminum hydride to form a 14-aminosteroid of formula (). The starting ketone of general formula () can be prepared by applying the method described in N. Danieli, Tetrahedron, 22, p. 3189 (1966).
For example, when R ₁ is a hydroxyl group, 3,12-diacetoxy-14-hydroxyethianic acid is a methyl-
It can be stored in organometallic compounds such as lithium. The reaction is advantageously carried out by gradually preparing a solution of methyl-lithium in ether at a temperature below 10° C. and 3,12-diacetoxy-14-hydroxy-3,12-diacetoxy-14-hydroxy-lithium in tetrahydrofuran in the presence of sodium hydride.
This can be carried out by adding it to a solution of 5β-ethianic acid. In the case of the preparation of derivatives in which R is hydrogen, the reaction can also be carried out directly with a reducing agent such as lithium aluminum hydride. The etianic acid derivatives used as starting materials are described in the literature (D.Taylor, J.
Chem.Soc., 1953.P.3325). When R ₁ is a hydrogen atom, 3-acetoxy-14-hydroxyethianic acid is treated in the same manner as organometallic compounds. 14-Aminosteroids of the general formula () in which R and R ₁ represent hydrogen atoms can also be prepared by the method described in French Patent No. 2,464,270. Similarly, Astyar et al. [Bull.Soc.Chim., no. 9-10, P. 1581-1582
(1976)] is also represented by the general formula (14), where R is a lower alkyl group and more precisely a methyl group.
- Can be applied to obtain aminosteroids. The method according to the invention allows, under sufficient conditions, to obtain various stereoisomers of the 14-aminosteroid derivative of the general formula (), in particular the 3-position and the 12-position.
derivatives in which the hydroxyl group in position (suitably) and the hydrogen atom in the 5-position have a β configuration, preferably an amino group in the 14-position with a β configuration;
Derivatives can be prepared in which the hydroxyl group in the 20-position can have an α or β configuration when R is an alkyl group. The following examples illustrate the invention, but the invention is not limited thereto.
The structure of the obtained product can be seen in the infrared spectrum, NMR
Confirmed by spectrum and mass spectrum. Example 1 a Preparation of 3,20-di-O-acetyl-5β-pregnane-3β,4β,20-triol 20-oxo-5β-pregnane-3β obtained by the method of N. Danieli et al. (described above) , 6 g of 14β-diol are dissolved in 30 ml of methanol, then 1.5 g of potassium borohydride is added in portions and left to stand for about 30 minutes to accelerate the reaction. The methanol is removed by evaporation, the residue is extracted with ethyl acetate and the extract is washed with water. The residue was treated with 16.5 ml of acetic anhydride in 35 ml of pyridine overnight. After hydrolysis and washing with citric acid and water, the two isomers 20α are separated by chromatography on a silica column.
and 20β (7 g, yield 95
%). b 14β-amino-5β-pregnane-3β,20α-
Preparation of the diol 210 mg of the 20α isomer of diacetyltriol obtained as indicated above are dissolved in a solution of 16.5 ml of triazohydrogen acid in benzene, the mixture is then stirred for 5 minutes and freshly distilled boron 0.4 ml of trifluoride etherate reagent was added.
After a reaction time of 10 minutes, the reaction mixture is poured into a mixture of crushed ice and aqueous ammonia solution. Extraction is performed with benzene, and the extract is washed with water and dried. The residue was then dissolved in 20 ml of tetrahydrofuran and 54 mg of lithium aluminum hydride
Add. After heating at reflux for 1 hour, the mixture was hydrolyzed and the neutral fraction of aminopregnanediol 80 mg and the basic fraction 77 mg
(yield 49%). Melting point mp=258℃. Example 2 14β-amino-5β-pregnane-3β,20β-diol 9 g of the 20β isomer of diacetyltriol prepared as shown in Example 1a are dissolved in 600 ml of a benzene solution of triazohydrogen acid and The boron trifluoride etherate is added after a reaction time of 5 minutes and the same procedure as in Example 1b is carried out. As a result, 14-amino-5β-pregnane-3β,
2.3 g of 20β-diol are obtained (yield 32%). Crystallization is carried out from ethyl acetate. Melting point mp=196℃.
IR spectrum (Nujol) ν=3600~2100, 1585,
1030cm ^-1 . Example 3 14α-amino-5β-pregnane-3β,20α-diol The procedure given in Example 1 is followed. The desired product is isolated from the 14β-amino-5β-pregnane-3β,20α-diol isomer by chromatography on a silica column of the mother liquor from crystallization of the main product. Crystallization is carried out from a mixture of ethyl acetate and methanol. Melting point mp=188℃. Example 4 14α-amino-5β-pregnane-3β,20β-diol The procedure of Example 2 is followed. The desired product is isolated by chromatography on a silica column of the mother liquor from the crystallization of the 14β-amino-5β-pregnane-3β,20β-diol isomer. The product is crystallized from isopropanol. Melting point mp=216℃. Example 5 14β-amino-5β, 17βH-pregnane-3β,
20α-diol 20-oxo-5β, 17βH-pregnane-3β,
0.5 g of 14β-diol is reduced with 120 mg of potassium borohydride in 3 ml of methanol. After the reaction, alrylation was carried out with acetic anhydride in pyridine using conventional techniques to give di-O-acetyl-5β, with a yield of 95%.
17βH-pregnane-3β,14β,20α-triol is obtained. 0.4 g of diacetyltriol obtained as shown above was added to 30 ml of a benzene solution of triazohydrogen acid.
and then add 0.6 ml of boron trifluoride etherate. After a similar treatment as in Example 1, 0.1 g of lithium aluminum hydride is reacted with the residue in 10 ml of tetrahydrofuran. After hydrolysis, extraction from ethyl acetate and crystallization, 0.25 g of 14β-amino-5β,17βH-pregnane-3β,20α-diol are obtained (80% yield).
Melting point mp = 200-201℃. Example 6 14β-amino-5α-pregnane-3β,20α-diol The procedure of Example 1 is carried out using the 5α isomer,
Leave for about 5 hours to facilitate reaction with boron trifluoride etherate. As in Example 1, after treatment with lithium aluminum hydride and reduction, the residue is taken up in ethyl acetate and the mixture is then washed with 10% hydrochloric acid solution. The acetic acid is then removed by evaporation and the acid phase is neutralized with sodium carbonate and extracted with chloroform. After filtration, the liquid was dried and after crystallization from ethyl acetate, 14β-amino-5α-
Pregnane-3β,20α-diol is obtained with a yield of over 50%. Melting point mp=246℃. Example 7 14β-amino-5α-pregnane-3β,20β-diol Starting from the procedure of Example 1a, pregnane diol
The 5α isomer is used and the 20β isomer of the diacetyltriol obtained is isolated. The same treatment as in Example 1b is then carried out. 14β-amino-5α-pregnane-3β, which is thereby recrystallized from diisopropyl ether,
20β-diol is obtained. Melting point mp=213℃. Example 8 14β-amino-5β-pregnane-3β,20-diol This compound is prepared as shown in Example 6 of French Patent No. 2464270 in the name of the applicant's company. Example 9 14β-amino-5β-pregnane-3β, 12β, 20β
-Triol 1.1 g of lithium aluminum hydride was added to 20-oxo-5β-pregnane-3β, 12β, 14β-triol and the corresponding O
- Add to a solution in 130 ml of tetrahydrofuran containing 8 g of the mixture of acylated derivatives. This mixture was obtained by reacting a solution of methyl-lithium in ether with 3β,12β-diacetoxy-14β-hydroxy-5β-ethianic acid in tetrahydrofuran under a nitrogen atmosphere at a temperature below about 10°C. be. Ethyl acetate is added cold to the tetrahydrofuran and water mixture. After filtering and evaporating the solvent, 5β-pregnane-3β, 12β,
7.6 g of 14β,20β-tetrol are collected in the form of a colorless powder. The product is dissolved in 120 ml of methylene chloride and 9.2 ml of acetic anhydride and 1.2 g of 4-dimethylaminopyridine are added to the solution. The reaction mixture is stirred for several hours and then washed with a solution of sodium carbonate and the aqueous phase is extracted with methylene chloride. After drying, 3,12,20-tri-O-acetyl-
9.3 g of 5β-pregnane-3β, 12β, 14β, 20ξ-tetrol is obtained. The two isomers 20α and 20β are separated by chromatography on Mel 7H60 silica using a mixture of methylene chloride and acetone (96.5/3.5) as eluent. 3,12,20-tri-O-acetyl-5β-pregnane-3β,12β,14β,20β-tetrol is recrystallized from ether (melting point, 216° C.). Dissolve 5.7 g of this product in 330 ml of an approximately 10M solution of triazohydrogen acid in benzene. Then 7.3 ml of freshly distilled boron trifluoride are added and the mixture is stirred for about 25 minutes. Alkalinization with a mixture of aqueous ammonia and crushed ice, extraction with benzene, washing with water,
After drying over sodium sulfate and evaporation, the residue
5.8 g was obtained, which was dissolved in 300 ml of ethanol. This solution was mixed with palladium-calcium carbonate (5
% concentration) in the presence of 2.9 g at room temperature under a hydrogen atmosphere for 48 hours. The catalyst is removed and the liquid is evaporated to dryness to give 5.5 g of a colorless foam. Dissolve this in toluene. This solution is washed several times with a 5% strength solution of sulfamic acid. The precipitate formed is removed and purified to obtain 0.5 g of 3,12,20-tri-O-acetyl-14β-amino-5β-pregnane-3β,12β,20β-triol. The acid aqueous phase is made alkaline with sodium carbonate and extracted with methylene chloride to give 0.75 g of the same product. Melting point, mp=211℃
(isopropyl ether). 3,12,20-tri-O obtained as above
-acetyl-14β-amino-5β-pregnane-
0.2 g of 3β,12β,20β-triol is saponified in 150 ml of a 1N methanolic solution of sodium hydroxide by stirring the suspension for 2 hours at ambient temperature, and then heating it under reflux for 30 minutes. After evaporating the solvent, the residue is extracted with chloroform. This gives 0.15 g of 14β-amino-5β-pregnane-3β,12β,20β-triol. It can be recrystallized from a mixture of ethyl acetate and methanol. Melting point, mp=
238℃. IR spectrum (Nujol) ν=3400, 3330,
3270, 3180, 3100, 2660, 1615, 1600cm ^-1 Based on the present invention represented by the above general formula ()
14-Aminosteroids have valuable pharmaceutical properties;
Specifically, it has active muscle inotropic activity. These properties are shown below, but it is important to note that 14-aminosteroids of general formula () and also their pharmaceutically acceptable salts can be used in human and animal therapy as drugs to treat heart failure. It shows. The toxicity tests carried out showed that the acute toxicity of the 14-aminosteroids according to the invention is relatively low and in particular considerably lower than that of conventional digitalis-based compounds such as digitoxin and digoxin. In vitro muscle inotropic effects were demonstrated in isolated guinea pig hearts and isolated guinea pig auricles based on Langendorff's method.
The obtained results show that the 14-aminosteroid according to the invention can be applied in the treatment of heart failure. According to an example, in the case of 14β-amino-5β-pregnane-3β,12β,20β-triol, the Langendorff test in isolated hearts showed that for doses between 1 and 30 μg/ml, The number shows an increase of approximately 30-40%. In tests on isolated guinea pig auricles, a 150% increase in contractile force was found for a concentration of 10 ^-5 g/ compared to control values. These results are consistent with the administration of propanolol,
After the demonstration of the direct myocardial action of the compound and its effect on the adrenergic effect, it has been confirmed by changing the experimental conditions. 14β-amino-21-nor-5β-pregnane
Comparable results have been obtained using 3β,20β-diols. The results of experimental measurements of the muscle inotropic effects of the compounds according to Examples 1, 2, 5 and 8 of the present invention are shown in the following table. In the experiment, stimulation was applied at 3.3 Hz to the auricle of guinea pigs to which each compound was administered, and changes in muscle contraction were expressed as a percentage of muscle contraction when no compound was administered.

[Table] From the results in the table, it can be seen that the compounds according to the present invention have excellent muscle inotropic activity. Derivatives of the general formula () and their pharmaceutically acceptable salts can be prepared in a conventional manner, i.e. by diluting the active ingredient in a suitably selected pharmaceutically acceptable carrier, e.g. tablets, gelatin capsules, coatings. It can be administered in the form of tablets, suppositories, injections or syrups. According to an embodiment, the tablets can be prepared by mixing a derivative of the general formula () or one of its salts with one or more solid diluents such as lactose mannitrate, starch, polyvinylpyrrolidone, magnesium stearate, talc, etc. It can be prepared by If appropriate, the tablets may contain several layers placed around the core according to conventional techniques to ensure sustained release or delayed effect of the active ingredient. The coating may consist of one or more layers of polyvinyl acetate, carboxymethylcellulose or cellulose acetophthalate, for example. The derivatives according to the invention can also be administered in the form of syrups or solutions for oral ingestion. This can be done, for example, by dissolving the derivative of formula () or one of its pharmaceutically acceptable salts in water or glycerol and, where appropriate, incorporating customary additives such as sweeteners or antioxidants. It is obtained by Injectable solutions can be prepared according to known techniques and include, for example, a derivative of formula () or its pharmaceutical It consists of a type of salt that can be tolerated. Suitable additives such as preservatives may be added if appropriate. The dosage will be determined by the physician based on the chosen method of administration, severity of treatment dissatisfaction, duration of treatment, etc. For example, in the case of oral administration to humans, the dosage can be between 0.005 and 5 mg/Kg.

Claims (1)

[Claims] 1 General formula () (wherein R represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, and R ₁
represents a hydrogen atom or a hydroxyl group)
A therapeutic agent for heart failure, characterized by containing a 14-aminosteroid or a pharmaceutically acceptable salt thereof. 2. The heart attack therapeutic agent according to claim 1, wherein R is a hydrogen atom or a methyl group. 3. The therapeutic agent for heart palsy according to claim ₂ , wherein the -NH2 group at the 14-position has an α configuration. 4. The therapeutic agent for heart palsy according to claim ₂ , wherein the -NH2 group at the 14-position has a β configuration. 5 General formula () (In the formula, R is a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, and R ₁ is a hydrogen atom or a hydroxyl group, and R and R ₁ are not hydrogen atoms at the same time, When R ₁ is a hydrogen atom, when the -OH group at the 3-position and the -NH ₂ group at the 14-position have the β configuration, and when the hydrogen atom at the 17-position has the α configuration, the -OH group at the 20-position A 14-amino steroid having a β configuration. 6. The 14-aminosteroid according to claim 5, wherein R is a methyl group. 7 formula () (wherein R is a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms, and R ₁ is a hydrogen atom or a hydroxyl group)
The 14-dihydroxysteroid is reduced and then acetylated to give the formula () (In the formula, R has the same meaning as above,
R ₁ is a hydrogen atom or an acetyl group, and
Ac is an acetyl group) to form an O-acetylated 3,14,20-trihydroxysteroid, and react the compound of formula () with a triazohydrogen acid/boron trifluoride complex to form the corresponding 14 - general formula (), characterized in that it forms an azide derivative and then reduces it with a metal hydride or by catalytic hydrogenation; A method for producing a 14-amino steroid represented by the formula (wherein R and R ₁ have the same meanings as above).