JPH0247479B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel 3-amino substituted steroid derivatives and salts thereof, methods for their production, uses of these novel derivatives as drugs, and compositions containing them. The subject of the present invention is novel 3-amino substituted steroid derivatives and their addition salts with mineral or organic acids, which have the general formula It is characterized by that it corresponds to, and the symbol in the formula

[Formula] indicates that the 5th position of the steroid skeleton is in the configuration α, W indicates a hydrogen atom or a hydroxyl group, or together with X indicates an ethylidene group, X is an ethyl group or

[Formula] or

[Formula] or together with W represents an ethylidene group, the wavy line means that the corresponding group is in one or the other of the configurations α and β, R ₁ represents a hydrogen atom or a methyl group, R ₂ is a hydrogen atom, 1 to 5 carbon atoms
_R3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, a carbon represents an alkoxycarbonyl group having 2 to 8 atoms, a nicotinoyl group or a group derived from an α-amino acid or from a peptide having 2 or 3 α-amino acids, while R ₂ and R ₃
does not represent a hydrogen atom, and on the other hand: (i) R ₁ represents a methyl group, W and R ₂ represent a hydrogen atom, and X represents a group;

When representing [Formula], R ₃ cannot represent a methyl or nicotinoyl group or a group derived from an aminocarboxylic acid, (ii) R ₁ represents a methyl group, W represents a hydrogen atom,
R ₂ represents a methyl group and X represents a group

In the case of [Formula], R ₃ cannot represent a methyl group, (iii) R ₁ represents a methyl group, R ₂ and W represent a hydrogen atom, and X represents a group.

[Formula], where the hydroxyl is in the (S) configuration and the amino group is in the 3α position, R ₃ cannot represent a methyl or glycyl group, and (iv) W represents a hydrogen atom. and X is the group

[Formula] and when the amino group is at the 3α position, the substituents R ₁ , R ₂ and R ₃ cannot simultaneously represent a methyl group, (v) R ₂ and W represent a hydrogen atom, R ₁ represents a methyl group, and X represents a group

[Formula], where the hydroxyl is in the (R) configuration and the amino group is in the 3α position, R ₃ cannot represent an ethoxycarbonyl or methyl group, and (vi) R ₁ , R When ₂ and R ₃ represent a methyl group and the amino group is in the 3α position, W and X together cannot represent an ethylidene group. In the general formula and the following explanation, carbon atom 1
The term alkyl having up to 5 carbon atoms means, for example, the methyl, ethyl, propyl, isopropyl, butyl, isobutyl or bentyl group; the term hydroxyalkyl having 2 to 5 carbon atoms means, for example, the hydroxyethyl or hydroxypropyl group. and the term alkoxycarbonyl having 2 to 8 carbon atoms means, for example, methoxycarbonyl, ethoxycarbonyl or propoxycarbonyl, groups derived from aminocarboxylic acids include Ala, Val, IVal, Leu, Ile. ,
Aap, Aan, Glu, Gln, Ser, Thr, Cys, Met,
Lys, Arg, Phe, Tyr, Trp, His & Pro,
It can be selected from the group consisting of NVa, Mle, Hyp, Orn, these acids are present in the D or L form, and can also be selected from the group consisting of Sar and Gly, all of the above The acid may be N-alkylated and when R ₃ represents a group derived from a peptide with 2 or 3 α-amino acids, it is selected from the group consisting of the above α-amino acids. The symbol for α-aminocarboxylic acids refers to these acids as D
It will be generally recognized that the alanine is designated in the configuration or in the L configuration (for example, the symbol A1a refers to the D or type alanine). Other than the general nomenclature used in this specification, the nomenclature used in this specification is IUPAC nomenclature, the rules of which are inter alia published in Biochemical Journal, Vol.
It is described on page 780 (1972). Addition salts with mineral or organic acids are, for example, hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, acetic acid, formic acid, propionic acid, benzoic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, oxalic acid, glyoxylic acid. acids and aspartic acid, alkanesulfonic acids (e.g. methanesulfonic acid or ethanesulfonic acid),
Salts formed with arylsulfonic acids (eg benzenesulfonic acid or p-toluenesulfonic acid) and arylcarboxylic acids can be used. Among the compounds constituting the subject matter of the present invention, in particular derivatives corresponding to the above formulas and addition salts with mineral or organic acids, in which X is a radical.

Examples include those characterized by showing the following formula: Among these, more specifically, in the above formula, X is a group.

[In the formula, Hal represents a chlorine, bromine or iodine atom, and _R'3 represents an alkoxycarbonyl group having 2 to 8 carbon atoms]

A compound of the formula in which R ₂ represents a hydrogen atom, R ₃ represents an alkoxycarbonyl group having 2 to 8 carbon atoms, and W, X, R ₁ and the wavy line have the above meanings The compound of this formula can be isolated and salted if necessary, or can be combined with nicotinic acid, or with an αr amino acid, or with two or three groups in which the amine function is protected by a group that can be easily removed, especially by acid hydrolysis. of α
- reacting with a peptide having an amino acid and then treating the product to remove the protecting group,
R ₂ represents a hydrogen atom, R ₃ represents a group derived from nicotinic acid, an α-amino acid, or a peptide having two or three α-amino acids, and W, X, R ₁ and the wavy line are as described above. A compound of the formula having the meaning of , by reacting with a halogenated hydroxyalkyl which is a bromine or iodine atom,
Formulas in which R ₂ and R ₃ represent a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, or a hydroxyalkyl group having 2 to 5 carbon atoms, and W, X, R ₁ and the wavy line have the above meanings. A compound of this formula is obtained, and a compound of this formula is isolated and a salt is formed if necessary, or when R ₃ represents a hydrogen atom, a compound of the formula Hal-R ₃ [wherein R ₃ has 1 to 5 carbon atoms] represents an alkyl group, a hydroxyalkyl group having 2 to 5 carbon atoms, or an alkoxycarbonyl group having 2 _to 8 carbon atoms, and Hal has the above meaning.
an alkyl group having ~5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, where R ₃ is an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms; A compound of _the formula representing an acyloxy group having 2 to 8 acyloxy groups and in which W _, , the above compound of the formula is combined with nicotinic acid or with an α-amino acid or with two or three groups in which the amine function is protected by a group that is easily releasable, especially by acid hydrolysis.
react with a peptide having α-amino acids,
The product is then treated to remove the protecting groups so that R ₂ represents an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, and R ₃ is from nicotinic acid or A compound of the formula representing a group derived from an α-amino acid or from a peptide having two or three α-amino acids and in which W, X, R ₁ and the wavy line have the above meanings is obtained and isolated. It is characterized by generating salt as necessary. Under suitable conditions for carrying out the present invention,
The above manufacturing method is carried out as follows. (1) The reaction of the amine of the formula with the halide of the formula is carried out using an acid fixative, especially an alkali metal hydroxide, an alkali metal carbonate (e.g. potassium carbonate),
Alkali metal bicarbonates, alkali metal acetates,
It is carried out in the presence of alkaline earth carbonates, tertiary amines (eg trialkylamines or pyridines) or alkali metal alcoholates (eg sodium ethylate). The reaction can be carried out, for example, in an inert solvent or suspending medium, such as dioxane, dimethylformamide, benzene, toluene or methylene chloride. In the case of acylation, free acids can also be used or functional derivatives other than halides, such as acid anhydrides. If the free acid is also used, it may be necessary to carry out the conversion to the amide by means of an activating agent such as a carbodiimide. For example, "The Peptide", Volume 1, Academic Press (1979)
Other techniques can also be used, such as those described in . (2) The amine of the formula or the compound of the formula in which R ₃ represents a hydrogen atom and R ₂ represents an alkyl or hydroxyalkyl group and nicotinic acid or an α-amino acid, or the amine function can be easily separated. The reaction with a peptide protected by a protecting group is carried out in the presence of a condensing agent. in this case,
The purpose of the condensing agent is to activate the acid function of the amino acids. As a condensing agent, the formula A-N=C=N-B [wherein A and B represent an alkyl group having 1 to 8 carbon atoms, preferably a dialkylamino group, or The carbodiimides shown below can be used. Mention may be made, for example, of dicyclohexycarbodiimide or 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, preferably the latter. Halides of 2-chloro-N-methylpyridinium, such as iodides, can also be used. Alkyl chloroformates can also be used, for example methyl, ethyl or isobutyl chloroformate, and also alkyl pyrophosphites, for example ethyl pyrophosphite. As easily removable protecting groups, for example benzyloxycarbonyl groups or (Z) tert-butyloxycarbonyl groups (BOC) can be used, as desired depending on the case. The subject of the invention is primarily the above-mentioned process, characterized in that the protecting group is a tert-butyloxycarbonyl group. To remove easily removable protecting groups, preferably an acid such as hydrochloric acid is used as the removing agent, the procedure being carried out e.g. using an alkanolic solution of hydrochloric acid or by bubbling anhydrous hydrochloric acid into nitromethane. It is done by twisting. It is also possible to use acids such as p-toluenesulfonic acid, formic acid or trifluoroacetic acid. As for the protecting group (Z) it is also possible to use, for example, hydrogen in the presence of palladium. The reaction of the amine of formula (3) with the alkyl or hydroxyalkyl halide is carried out in the presence of the same acid fixative and solvent as in the reaction with the halide of formula. The hydroxy function of the hydroxyalkyl group is
It is advantageously protected by a protecting group which is easily removable, especially by acid hydrolysis. Advantageously, this protecting group is a tetrahydropyranyl group. This can be removed under the same conditions as described above for protecting groups on amino acids. (4) R ₃ represents a hydrogen atom, and R ₂ represents a carbon atom of 1 to 5
represents an alkyl group having 2 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, and W,
The reaction of a compound of formula in which X, R ₁ and the wavy line have the above meanings with a halide of formula R″ ₃
Depending on the nature of the compound, the reaction of the amine of the formula with the halide of the formula or with the alkyl halide is carried out under the same conditions as described above. (5) For the preparation of N-alkylated compounds of the formula, it may be advantageous in some cases to operate in the following specific manner: - for the preparation of the N-monomethylated compounds of the formula. In order to prepare a carbamate of an amine of the formula compound, for example by the action of an alkyl haloformate, in particular an ethyl haloformate, this carbamate is then reduced, for example with lithium aluminum hydride, to produce an N-monoethylated compound of the formula To prepare the corresponding N-acetylated derivative and then reduce it, it is sufficient to repeat the operation to produce the -N-diethylated derivative. In these three cases, X is a group

When representing the formula, it is necessary to prepare a ketal in position 20, for example by the action of ethylene glycol, and then, after reduction, to hydrolyze this ketal, for example with a mineral acid. To prepare N-monopropylated or N-diisopropylated compounds of formula -, a compound of formula can be reacted with acetone in the presence of a reducing agent such as, for example, sodium cyanoborohydride. In the method of the invention, the protection of the amine function is
It is very clear that these amine functions are not relevant if they are alkylated. The derivatives of the formula are basic, except when R ₃ represents an acyloxy group. Addition salts of derivatives of the formula may advantageously be prepared by reacting mineral or organic acids in approximately stoichiometric proportions with the derivatives of the formula. These salts can be prepared without isolating the corresponding base. The derivatives which form the subject of the present invention have very interesting pharmacological properties, in particular remarkable immunotherapeutic properties. In particular, they can stimulate the immune response. Examples of these properties will be given in the experimental section below. These properties justify the use of the 3-amino substituted steroid derivatives of formula, as well as their pharmaceutically acceptable salts, as drugs. The subject of the invention is therefore also the use as medicaments of the 3-amino substituted steroid derivatives defined by the formula as well as their addition salts with pharmaceutically acceptable acids. Among the drugs constituting the subject matter of the present invention, preferably in novel 3-amino substituted steroid derivatives corresponding to the formula

Conceivable are drugs that are characterized by the formula: and their addition salts with pharmaceutically acceptable acids. Among these, especially

Those corresponding to the formula represented by the formula and in which R ₂ represents a hydrogen atom or a methyl group, as well as addition salts thereof with pharmaceutically acceptable acids are considered. Furthermore, among these, in particular derivatives with the following names are considered: 2-amino-N-[(20S)-20-hydroxy-
(5α)-pregnane-3α-yl]-propanamide, (2S)-2-amino-N-[(20S)-20-hydroxy-(5α)-pregnane-3α-yl]-1H-indole-3- Propanamide, 2-amino-N
−[(20S)-20-hydroxy-19-nor-(5α)−
pregnane-3α-yl]-acetamide, and 2-amino-N[(20S)-20-hydroxy-
(5α), pregnane-3α-yl]-N-methylacetamide, and addition salts thereof with pharmaceutically acceptable acids. The drug of the invention is used, for example, in the treatment of autoimmune diseases resulting from a deficiency of certain lymphocytes,
Whether the disease is a connective tissue disease, such as rheumatoid arthritis, or a non-specific disease, such as systemic lupus erythematosus, or a specific disease, such as thyroiditis, acne scars or hemolytic anemia. Used in any organ disease. Therefore, the drug according to the present invention can be used as an adjunct to antibiotic therapy or anticancer drug therapy. The usual dosage may vary depending on the derivative used, the patient treated and the condition in question, but for example in the case of the derivative of Example 14 used as an adjunct to antibiotic therapy, 100 mg daily by oral route in humans.
It can be from mg to 1 g. Finally, a subject of the invention is a pharmaceutical composition which contains as active ingredient at least one of the above-mentioned derivatives or one pharmaceutically acceptable addition salt with an acid. As drugs, derivatives corresponding to the formula and their addition salts with pharmaceutically acceptable acids can be incorporated into pharmaceutical compositions for the gastrointestinal or parenteral route. These pharmaceutical compositions may, for example, be solid or liquid and may be presented in the pharmaceutical forms currently used in human medicine, such as plain tablets, dragee-coated tablets, gelatin capsules, granules, suppositories and injectable preparations. These can be manufactured by conventional methods. The active ingredients include excipients commonly used in these pharmaceutical compositions, such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous vehicles,
It can be blended with animal or vegetable fatty substances, paraffin derivatives, glycols, various wetting agents, dispersing agents or emulsifying agents and preservatives. If the starting material of the formula is unknown, it can be prepared as follows: [wherein R ₁ and the wavy line have the above meanings] is reacted with excess ethyltriphenylphosphonium bromide and potassium tert-butyrate to form the formula
In the formula R ₁ and the dashed line have the meanings given above, the product is obtained, for example by passing it through a tosylate and then by the action of sodium azide or directly in the presence of ethyl azodicarboxylate and triphenylphosphine. By reacting with diphenyl azidophosphate, the formula [wherein R ₁ and the dashed line have the meanings given above and the configuration at position 3 is reversed with respect to the configuration of the compound and] and this is reduced with, for example, lithium aluminum hydride to form the azide of the formula _A [wherein R ₁ and the dashed line have the meanings given above] are obtained and isolated, or reduced by hydrogenation in the presence of catalysts based on rhodium, platinum or palladium, to give the formula _B [wherein R ₁ and the dashed line have the meanings given above] or by hydration, for example with diborane, of the formula
_C [wherein R ₁ and the wavy line have the meanings given above] are obtained and isolated or subjected to oxidation, for example with chromic acid, to give the product of formula _D or, after protection of _the amine with an easily hydrolysable group such as, for example, a trifluoroacetyl group, or without protection, Upon cis-dihydroxylation with osmic anhydride in the presence of methyloxylamine, formula _E [In the formula, R represents the above-mentioned protecting group or hydrogen atom,
R ₁ and the wavy line have the meaning given above], which is isolated when R represents a hydrogen atom or by alkaline hydrolysis when R represents a protecting group, e.g. in the case of a trifluoroacetyl group. or subjecting the protective group to the action of a reagent which removes said protecting group and then isolating the resulting free amine, or
or upon oxidation with e.g. chromic acid, the formula _F [wherein R, R ₁ and the wavy line have the meanings given above] is obtained, which is isolated when R represents a hydrogen atom, or when R' represents a protecting group, the said protecting group is removed. Subjected to the action of a reagent for elimination, the resulting free amine is then isolated or reduced, for example with sodium borohydride, to form the formula _G [In the formula, R, R ₁ and the wavy line have the above-mentioned meanings] A reagent which isolates the product when R represents a hydrogen atom or removes the protecting group when R represents a protecting group. The free amine obtained is then isolated. An example of the preparation of a compound of formula is given in the experimental section below. The invention is illustrated by, but not limited to, the following examples. Example 1 (2S)-2-amino-N-[(20S)-20-hydroxy-(5α)-pregnane-3α-yl]-propanamide (hydrochloride and base). Step A: 2-[1,1-dimethyl-ethoxycarbonylamino]-N-[(20S)-20-cidroxy-(5α)-pregna-3α-yl]-propanamide. 1.92 g (20S)-3α-amino-(5α)-pregnane-20-ol and 2.27 g tert-butyloxycarbonyl-L-alanine (BOC-L-alanine) in 60 mg chloroform and 12 ml pyridine. dissolved in. This solution was heated at 0°C under an inert atmosphere.
Stir at ~+5°C and add 1.14 g of 1-ethyl-3-
(Dimethylaminopropyl)-carbodiimide hydrochloride was added. After 1 hour and 15 minutes, 1.15 g of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride was added again and stirring was continued for 50 minutes at 0°C to +5°C until the whole was dried and acidified. Taken in an aqueous solution of sodium carbonate, extracted with ethyl acetate, washed with a saturated aqueous solution of sodium chloride, then diluted with 1 ml of hydrochloric acid.
Washing with aqueous solution and again with saturated aqueous sodium chloride solution, drying after drying, crystallization from isopropyl ether, separation, washing with isopropyl ethyl and drying under reduced pressure at 60 °C gives the expected product. I got it. Melting point = 171℃. Step B: (2S)-2-amino-N-[(20S)-20-
Hydroxy-(5α)-pregnane-3α-yl]
Propanamide. Variant 1: Under an active atmosphere, 1.55 g of the product obtained in step A are suspended in 100 ml of nitromethane, hydrogen chloride is bubbled through for 10 minutes, and the whole is
Stir for 35 minutes at 20-25°C, remove excess hydrochloric acid, and separate the residue, wash with ethyl ether and dry at 60°C under reduced pressure, then recrystallize from methanol to give the expected hydrochloride salt. Obtained 990mg.
Melting point = 220° C. Variant 2: Under an inert atmosphere, 2.7 g of the product obtained in step A are dissolved in 10 ml of ethanol and 40
ml of hydrochloric acid ethanol (3.5N) was added, the whole was stirred for 8 hours, dried, taken up in ethyl acetate,
After cooling, separation and washing with ethyl acetate, drying under reduced pressure at 60° C. and recrystallization from methanol yielded 2.2 g of the expected hydrochloride salt. Melting point=
220℃. Preparation of base Dissolve 1.74 g of hydrochloride in 100 ml of tetrahydrofuran containing 30% water, add 4 ml of 2N sodium hydroxide, dry the whole, take up in 100 ml of chloroform, wash with water, dehydrate and dry, Recrystallization from ethyl acetate gave 1.27 g of the expected base. Melting point = 224℃. The derivatives listed in the table below were produced by the same method as above.

[Table] Note for Example No. 2: After protecting the second acid function of L-glutamic acid in the form of benzyl ester and liberating it by catalytic hydrogenation in acetic acid in the presence of palladium, BOC is released. I let it happen. Example 8 N-[(20S)-20-hydroxy-(5α)-pregnane-3α-yl]-3-pyridine carboxide. Under an inert atmosphere, 960 mg of (20S)-3α-amino-(5α)-pregnan-20-ol and 813 mg of nicotinic acid were dissolved in 30 ml of chloroform and 6 ml of pyridine, and 582 mg of 1-ethyl -3-(3
-dithylaminopropyl)-carbodiimide hydrochloric acid was added. After 3.5 hours, 291 mg of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride was added under stirring and the whole was stirred for 16 hours.
After drying, taking up in 30 ml of water, cooling, decomposition and washing with water, drying under reduced pressure at 80° C. and recrystallization from methanol gave 920 mg of the expected product.
Melting point = 258℃. [α] _D = +14.5° ± 1° (c = 1% pyridine). Example 9 2-Amino-N-[(20S)-17α,20-dihydroxy-5α-pregnane-3α-yl]acetamide hydrochloride. BOC-glycine and (20S)-3α-amino-5α-
Starting from pregnane-17α,20-diol and proceeding in a manner similar to that described in Example 1 (variant 2), the expected product was obtained. Melting point = 200
℃. [α] = +4° ± 1° (c = 1.5% ethanol 95°)
. Starting (20S)-3α-amino-5α-pregnane-
17α,20-diol can be produced as follows. Step 1: (Z)-(5α)-Pregnane-17(20)-en-3β-ol 59.4 g of triphenylethylphosphonium bromide are added to a solution of 16.1 g of potassium tert-butyrate in 160 ml of tetrahydrofuran and the whole is dissolved. Stir for 30 minutes,
Add 23.2g of epiandrosterone for a total of 15
Stirred for minutes, poured into ice water, extracted with ethyl acetate, washed with water, dried, distilled to dryness and purified by chromatography on silica (eluent: cyclohexane/ethyl acetate 7:3). Recrystallization from methanol, cooling, separation and drying yielded 23.1 g of the expected product. Melting point = 160℃. Step 2: (Z)-3α-azido-(5α)-pregna-
17(20)-Ene 1.92 g of ethyl azicarboxylate and 3.02 g of diphenyl azidophosphate are added to a solution of 1.66 g of the above product in 30 ml of benzene and 5 ml of tetrahydrofuran and the whole is stirred in an ice bath and 30 ml of benzene is added. A solution of 2.88 g of triphenylphosphine in the solution was added over 20 minutes, and the whole was stirred again for 40 minutes at 10°C.
After washing with a hydrochloric acid solution and then with water, it is dried and distilled to dryness and purified by chromatography on silica (eluent: heptane followed by heptane/ethyl ether 1:1) to give the expected product. 1.67 g of crystals were obtained. Melting point after recrystallization from methanol = 114°C. Stage 3: (Z)-5α-pregna-17(20)-en-
3α-amine (and hydrochloride). 14.5 g of (Z)-3α-azido-(5α)-pregna-17(20)-ene obtained above was dissolved in 290 ml of tetrahydrofuran, and the whole was stirred while heating.
At 25-27°C, 800 mg of lithium aluminum hydride was added over 1 hour, the whole was stirred again for 1 hour, and after removing excess hydride with methanol,
The residue was filtered and the liquor was washed with an aqueous solution of Rothsiel's salt and then with a saturated aqueous solution of sodium chloride,
Dehydrated and distilled to dryness to obtain 13.1 g of crystals of the expected amine.
I got it. Melting point = 90℃. Add this base to 150 ml of ethyl acetate and methylene chloride.
After adding 27 ml of 1.7N hydrochloric acid in ethyl acetate, the whole was separated and washed with ethyl acetate, and the resulting crystals were dried under reduced pressure to give 13.2 g of hydrochloride crystals of the expected product. I got it. Melting point >300℃.
[α] _D = +38.5゜±1.5゜ (1 in pyridine containing 10% water
%). Step 4: N-[(Z,5a)-pregna-17(20)-en-3α-yl]-trifluoroacetamide. In an inert atmosphere, 16.5 g of the hydrochloride obtained in step 3 above was dissolved in 165 ml of methylene chloride and 16.5 g of pyridine.
ml, and after cooling the whole to about 5°C,
16.5 ml of trifluoroacetic anhydride was added over 5 minutes, the whole was stirred at room temperature for 15 minutes and distilled to dryness under reduced pressure, then 200 ml of water was added and the whole was separated, washed with water and dried under reduced pressure. 18.1 g of crystals were obtained. Melting point = 204℃. Step 5: N-[(20S)-17α,20-dihydroxy-
(5α)-pregnane-3αyl]trifluoroacetamide. The product obtained above under an inert atmosphere
Dissolve 18.1g in 100ml of methyl ethyl ketone,
Osmium tetroxide in 9 g of dihydrated trimethylamine N-oxide and 71 ml of methyl ethyl ketone
Add 360 mg of solution and stir the whole under reflux for 2 hours, allow to cool and add 10% of sodium thiosulfate in water.
After adding 200 ml of the solution and stirring the whole at room temperature for 30 minutes, it was decanted, washed with water and dried over magnesium sulfate, filtered and evaporated to dryness under reduced pressure, and the resulting oil was chromatographed on silica. Purified by graphie (eluent: benzene/ethyl acetate 7:
3) to obtain 14 g of the expected product. Melting point = 172℃
Then 192℃. Step 6: (20S)-3α-amino-5α-pregnane-
17α,20-diol. Under an inert atmosphere, the product 4 obtained above
Dissolve g in 20 ml of methanol, add 8 ml of sodium hydroxide solution and stir the whole for 1.5 hours.
ml of water was added, the whole was stirred for 10 minutes, separated, washed with water, and dried under reduced pressure at 40°C to give 3 g of the desired product.
I got it. Melting point = 210℃. Example 10 2-amino-N-[(20S)-20-hydroxy-
19-Nor-(5α)-pregnane-3α-yl]acetamide hydrochloride. BOC-glycine and (20S)-3α-amino-19-
Starting from nor-(5α)-pregnan-20-ol and proceeding in a manner similar to that described in Example 1 (variant 1), the expected product is obtained. Melting point = 270℃. Involves sublimation. [α] _D = +39.5゜±1.5゜(
c.
= 1% methanol). Starting (20S)-3α-amino-19-nor-(5α)-
Pregnane-20-ol can be produced as follows. Proceeding in the same manner as described in Example 9 (steps 1, 2 and 3) for the preparation of (Z)-5α-pregna-17(20)-ene-3α-amine, (Z)
-5α-19-nor-pregna-17(20)-en-3α
-amines were prepared starting from 19-nor-epiandrosterone. Under nitrogen, 156 ml of sodium borohydride were suspended in 5 ml of tetrahydrofuran, a solution of 0.5 ml of boron triborate etherate in 2.5 ml of tetrahydrofuran was added dropwise at +5°C, and the whole was placed in an ice bath for 1 hour. (Z)-5- in 3 ml of tetrahydrofuran with stirring.
α-19-nor-pregna-17(20)-ene-3α-
A solution of 296 mg of amine was added and the whole was stirred at room temperature for 1.5 hours, cooled in an ice bath, 2 ml of 6N sodium hydroxide solution was added slowly and the whole was stirred at room temperature for 5 minutes, decanted and the aqueous phase was extracted with tetrahydrofuran, the organic phase was washed with water, 4 ml of 5N sodium hydroxide solution and 2 ml of 110 times the volume of hydrogen peroxide were added, the whole was stirred for 45 minutes, extracted with ethyl acetate, washed with water, and dehydrated. Distilled to dryness under reduced pressure.
Add the dry extract to 10 ml of methanol containing 5 ml of 1N hydrochloric acid.
Dissolved in water, heated in a 50 °C water bath for 30 minutes, poured into a saturated solution of sodium bicarbonate, extracted with methylene chloride, washed with water, dried and evaporated under reduced pressure to give 257 mg of expected product as crystals. I got it. Melting point = 190℃. Example 11 2-Amino-N-[17α-hydroxy-20-oxo-5α-pregnane-3α-yl]acetamide hydrochloride. Starting from BOC-glycine and 3α-amino-17α-hydroxy-5α-pregnan-20-one,
Working up in the same manner as described in Example 1 (variant 2), the expected product was obtained. Melting point >300℃.
[α] _D = +50° ± 1° (c = 1% ethanol 95°). The starting 3α-amino-17α-hydroxy-5α-pregnan-20-one can be prepared as follows. Oxidation of the product obtained in step 5 of Example 9 with perchromic acid produced N-(17α-hydroxy-20-oxo-5α-pregnan-3α-yl)-trifluoroacetamide (melting point = 178℃ then 186
C), which was treated as shown in Step 6 of Example 9 to give 3α-amino-17α-hydroxy-5α-pregnan-20-one. Melting point = 216°C (after crystallization from water). Example 21 2-amino-N-[(20R)-20-hydroxy-
5α-pregnane-3α-yl]acetamide hydrochloride. BOC-glycine and (20R)-3α-amino-5α-
Starting from pregnane-20-ol and proceeding in a manner analogous to that described in Example 1 (variant 1), the expected product was obtained. Melting point = 210℃ then
260℃. [α] _D = +22° ± 1° (c = 0.8% pyridine containing 10% water). Example 13 Ethyl N-[(20S)-20-hydroxy-(5α)-pregnane-3α-yl]-carbamate (20S)-3α-amino-5α-pregnane in 500 ml of methylene chloride under an inert atmosphere. A solution of 5 g of -20-ol was added dropwise to 15 ml of ethyl chloride and 45 ml of methylene chloride over 10 minutes, the whole was stirred for 30 minutes, 20 ml of 1N sodium hydroxide was added and the whole was stirred again for 30 minutes, After decanting and extraction with methylene chloride, washing with water, drying and filtering, evaporation to dryness under reduced pressure, dissolution with ethyl ether and separation afforded 5.7 g of the expected product. Melting point = 198℃. [α] _D = +25° ± 1.5° (c = 1% methylene chloride). Example 14 2-amino-N-[(20S)-20-hydroxy-
5α-pregnane-3α-yl]-N-methylacetamide hydrochloride. Due to the ring element of the product of Example 13, Wötzter et al.
(20S)-3α-methylamino-5α-pregnane-20 as described in Bull.Soc. (1963), p. 1324.
-ol (melting point = 170°C) was prepared and used in Example 1
BOC-
Reaction with glycine gave the expected product and its hydrochloride was prepared. Melting point = 250℃. Example 15 2(S)-2-amino-N-methyl-N-
[(20S)-20-hydroxy-5α-pregnane-
3α-yl]propanamide hydrochloride. (20S)-3α-methylamino-5α-pregnan-20-ol was reacted with BOC-alanine in a manner similar to that described in Example 1 (variant 1) to give the expected product, the hydrochloride of which Manufactured. Melting point>
270℃. Example 16 2(R)-2-amino-N-methyl-N-
[(20S)-20-hydroxy-5α-pregnane-
3α-yl]propanamide hydrochloride. (20S)-3α-Methylamino-5α-pregnan-20-ol was reacted with BOC-L-alanine in a manner similar to that described in Example 1 (variant 1) to give the expected product, whose hydrochloric acid produced salt. (about
Sublimation at 260℃). Example 17 (20S)-3α-ethylamino-5α-pregnan-20-ol hydrochloride. (20S)-3α-amino-5α-pregnane-20-
The base of the expected product was obtained by reacting the ol with ethyl iodide in the presence of sodium carbonate. Melting point after recrystallization from ethyl acetate = 129°C. Formation of the hydrochloride salt: The hydrochloride salt was prepared by addition of ethyl acetate to give the expected product. Melting point>
270℃. Example 18 2-amino-N-ethyl-N-[(20S)-20-
Hydroxy-5α-pregnane-3α-yl]acetamide hydrochloride. The base of the product of Example 17 was reacted with BOC-L-glycine in a manner similar to that described in Example 1 (variant 1) to give the expected product and its hydrochloride salt prepared. Melting point = 230℃. Example 19 (20S)-3α-[(2-hydroxyethyl)amino]-5α-pregnan-20-ol hydrochloride. (20S)-3α-amino-5α-pregnane-20-
7 g of ol and 7 g of sodium carbonate were suspended in 100 ml of anhydrous dioxa, 6.6 ml of 2-[2-promethoxy]-tetrahydropyran was added and the whole was refluxed for 24 hours under an inert atmosphere. After cooling it to room temperature, diluting it with water, and extracting it with ether,
Dried and distilled to dryness under reduced pressure and purified by chromatography on silica (eluent: chloroform/methanol/acetic acid 85:5:10) to collect 7.7 g of a yellow oil. Hydrolysis of protecting group 3 g of the above product was added to 30 ml of ethanol and 2N hydrochloric acid.
Dissolve in 15 ml and stir the whole thing under reflux for 1.5 hours.
Cool, pour into 100 ml of an aqueous solution of sodium bicarbonate, cool for 15 minutes, separate and wash with water at 40 °C.
The mixture was dried under reduced pressure, dissolved in 350 ml of methylene chloride containing 10% methanol, concentrated to about 50 ml by filtration, 100 ml of ethyl acetate was added, the whole was concentrated to half volume, and the precipitate was separated. Add this to methanol
The solution was filtered and concentrated to about 30 ml, the whole was concentrated to half by adding 70 ml of ethyl acetate, separated and dried under reduced pressure at 40° C. to give 1.65 g of the expected product base. Ta. Melting point = 215℃. Formation of hydrochloride Dissolve 1.5 g of the above base in 40 ml of methanol,
A 1.6N solution of ethyl acetate with hydrochloric acid was added until acidic pH, 60 ml of ethyl acetate was added, the whole was concentrated under reduced pressure, separated and dried under reduced pressure at 40 °C to obtain 1.5 g of the expected product. Ta. Melting point >260℃. Example 20 α-dimethylamino-N-[(20S)20-hydroxy-5α-pregnane-3α-yl]acetamide. The procedure was carried out as described in Step A of Example 1 using 2.552 g of (20S)-3α-amino-5α-pregnan-20-ol and 3.350 g of N,N-dimethylglycine hydrochloride. The expected derivative is O,N-
4 g of a crude product containing a disubstituted derivative was obtained.
Saponification with sodium hydroxide in methanol gave the expected product. Melting point = 206℃.
[α] _D = +29° (c = 1% CHCl ₃ ). Example 21 Compressed tablets were manufactured according to the following formulation. (2S)-2-Amino-N-[(20S)-20-hydroxy-5α-pregnan-3α-yl)propanamide hydrochloride...20mg Enough amount of excipient to obtain one compressed tablet of the right amount ……
100mg (excipient details: lactose, starch, talc, magnesium atearate). Example 22 Compressed tablets were manufactured according to the following formulation. 2-amino-N-[(20S)-hydroxy-5α-
Pregnane-3α-yl]-N-methylacetamide hydrochloride...15mg Enough excipient to obtain one compressed tablet of the left amount...100
mg (excipient details: lactose, starch, talc, magnesium stearate). Pharmacological considerations (1) Anaphylactic shock adjuvant principle: Administration of a compound capable of stimulating the activity of the immune system to an animal results in an increase in the number of shots in response to administration of the antigen to which the animal has been sensitized. It will be done. Male mice weighing 30-35 g were sensitized by intrafoot administration of bovine serum albumin. Eight days later the mice were challenged with the antigen by intravenous route. Under minimal sensitization conditions, control animals did not receive a lethal shot at the time of the last dose. The test compound was mixed with the antigen and injected via the intraplantar route. If the compound is an adjuvant, it increases sensitization and results in a lethal shock upon intravenous administration. The active drug dose resulting in 50% animal mortality (AD ₅₀ ) was determined. The results are shown in the table below. Examples 1, 2, 4-10, 13 as test compounds
and 14 compounds and the following compounds A, B, C and D for comparison were used. Compound A: 3β-(acetylamino)pregnan-20-one Compound B: 3β-(methylamino)pregnane
20−on Compound C: Funtumin Compound D: N-glycidylfuntumin

Table 2. Test principle for rosettes with sheep red blood cells. The administration to animals of compounds capable of stimulating the activity of the immune system is manifested by an increase in its ability to respond to injections of the immune compound. Three month old male rats are sensitized with sheep red blood cells by the intraperitoneal route (day 0). 7 days later (7
day), the spleen is removed and the splenocytes are contacted with sheep red blood cells. The ratio of white blood cells around which red blood cells have formed a rosette is then counted. The compounds to be studied are administered orally daily from day -1 to day +1. The proportion of rosettes observed in control animals was approximately 2
The dose of the compound doubled is considered the immunostimulatory dose.

[Table] 3 Examination of acute toxicity Lethal dose of each different compound tested
LD ₀ was determined after administration by oral route in mice. The maximum dose that does not cause any lethality at the end of the 8th day is called LD ₀ . The results were as follows.

【table】

Claims (1)

[Claims] 1. General formula Corresponding to this formula, the symbol [formula] indicates that the 5th position of the steroid skeleton is in the configuration α, W indicates a hydrogen atom or a hydroxyl group, or together with X indicates an ethylidene group, and X An ethyl group or a group of the formula [Formula] or [Formula] or together with W indicates an ethylidene group, the wavy line means that the corresponding group is in one or the other of the configurations α and β, and R ₁ Represents a hydrogen atom or a methyl group, R ₂ is a hydrogen atom, 1 to 5 carbon atoms
_R3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, a carbon represents an alkoxycarbonyl group having 2 to 8 atoms, a nicotinoyl group or a group derived from an α-amino acid or from a peptide having 2 or 3 α-amino acids, while R ₂ and R ₃
does not represent a hydrogen atom, and on the other hand: (i) When R ₁ represents a methyl group, W and R ₂ represent a hydrogen atom, and X represents a group [formula], R ₃ is methyl or nicotinoyl group or a group derived from an aminocarboxylic acid, (ii) R ₁ represents a methyl group, W represents a hydrogen atom,
When R ₂ represents a methyl group and X represents a group [formula], R ₃ cannot represent a methyl group, (iii) R ₁ represents a methyl group, and R ₂ and W represent a hydrogen atom. , X represents a group [formula], where the hydroxyl is in the (S) configuration and the amino group is in the 3α position, R ₃ cannot represent a methyl or glycyl group, and (iv) W represents a hydrogen atom, X represents a group [formula], and the amino group is at the 3α position, the substituents R ₁ , R ₂ and R ₃ cannot simultaneously represent a methyl group, and (v) R When ₂ and W represent a hydrogen atom, R ₁ represents a methyl group, and X represents a group [formula], where hydroxyl is in the (R) configuration and the amino group is in the 3α position, R ₃ cannot represent an ethoxycarbonyl or methyl group; (vi) if R ₁ , R ₂ and R ₃ represent a methyl group and the amino group is in the 3α position, then W and X together represent an ethylidene group; A novel 3-amino-substituted steroid derivative and its addition salt with a mineral acid or organic acid, which are characterized in that they are incapable of exhibiting 2. A novel 3-amino-substituted steroid derivative according to claim 1, characterized in that X represents a group [formula], and its addition salt with a mineral acid or organic acid. 3. A novel 3-amino-substituted steroid derivative according to claim 2, characterized in that X represents a group [formula] and R ₂ represents a hydrogen atom or a methyl group, and its mineral acid or organic acid; addition of salt. 4 The derivative of formula 20S)-20-hydroxy-(5α)-pregnane-3α-yl]-1H-indole-3-propanamide, 2-amino-N
-[(20S)-20-hydroxy-19-nor-(5α)-
pregnane-3α-yl]acetamide, and 2-amino-N-[(20S)-20-hydroxy-
The derivative according to claim 1, which is (5α)-pregnane-3α-yl]-N-methylacetamide and its addition salt with a mineral acid or an organic acid. 5 General formula Corresponding to this formula, the symbol [formula] indicates that the 5th position of the steroid skeleton is in the configuration α, W indicates a hydrogen atom or a hydroxyl group, or together with X indicates an ethylidene group, and X An ethyl group or a group of the formula [Formula] or [Formula] or together with W indicates an ethylidene group, the wavy line means that the corresponding group is in one or the other of the configurations α and β, and R ₁ Represents a hydrogen atom or a methyl group, R ₂ is a hydrogen atom, 1 to 5 carbon atoms
_R3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, a carbon represents an alkoxycarbonyl group having 2 to 8 atoms, a nicotinoyl group or a group derived from an α-amino acid or from a peptide having 2 or 3 α-amino acids, while R ₂ and R ₃
does not represent a hydrogen atom, and on the other hand: (i) When R ₁ represents a methyl group, W and R ₂ represent a hydrogen atom, and X represents a group [formula], R ₃ is methyl or nicotinoyl group or a group derived from an aminocarboxylic acid, (ii) R ₁ represents a methyl group, W represents a hydrogen atom,
When R ₂ represents a methyl group and X represents a group [formula], R ₃ cannot represent a methyl group, (iii) R ₁ represents a methyl group, and R ₂ and W represent a hydrogen atom. , X represents a group [formula], where the hydroxyl is in the (S) configuration and the amino group is in the 3α position, R ₃ cannot represent a methyl or glycyl group, and (iv) W represents a hydrogen atom, X represents a group [formula], and the amino group is at the 3α position, the substituents R ₁ , R ₂ and R ₃ cannot simultaneously represent a methyl group, and (v) R When ₂ and W represent a hydrogen atom, R ₁ represents a methyl group, and X represents a group [formula], where hydroxyl is in the (R) configuration and the amino group is in the 3α position, R ₃ cannot represent an ethoxycarbonyl or methyl group; (vi) if R ₁ , R ₂ and R ₃ represent a methyl group and the amino group is in the 3α position, then W and X together represent an ethylidene group; A method for producing a 3-amino-substituted steroid derivative and its addition salt with a mineral acid or organic acid, characterized in that it cannot exhibit the following formula: [In the formula, _the wavy line, W _,
R′ ₃ represents an alkoxycarbonyl group having 2 to 8 carbon atoms] R ₂ represents a hydrogen atom, R ₃ represents an alkoxycarbonyl group having 2 to 8 carbon atoms, and Obtaining a compound of the formula in which W _,
Alternatively, reacting with nicotinic acid or with an α-amino acid or with a peptide having two or three α-amino acids protected in the amine function, particularly by a group that can be easily removed by acid hydrolysis, and then protecting the product. R ₂ represents a hydrogen atom and R ₃ is removed from nicotinic acid or α-
A compound of the formula representing a group derived from an amino acid or from a peptide having _two or three α-amino acids and in which W, R ₂ as well as
A compound of the formula in which R ₃ represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, or a hydroxyalkyl group having 2 to 5 carbon atoms, and W, X, R ₁ and the wavy line have the above meanings is obtained. , isolate a compound of this formula and form a salt if necessary, or
Or, when R ₃ represents a hydrogen atom, the formula Hal-R″ ₃ [wherein R″ ₃ is an alkyl group having 1 to 5 carbon atoms, a hydroxyalkyl group having 2 to 5 carbon atoms, or a hydroxyalkyl group having 2 to 5 carbon atoms] represents an alkoxycarbonyl group having ~8 atoms, Hal has the above meaning], and _R2 is 1 carbon atom.
an alkyl group having ~5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, where R ₃ is an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms; A compound of _the formula representing an acyloxy group having 2 to 8 acyloxy groups and in which W _, , the compound of the formula is combined with nicotinic acid, or with an α-amino acid, or with two or three amino acids protected by a group in which the amino function is easily releasable, especially by acid hydrolysis.
react with a peptide having α-amino acids,
The product is then treated to remove the protecting groups so that R ₂ represents an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, and R ₃ is from nicotinic acid or represents a group derived from α-amino acids or from a peptide having 2 or 3 α-amino acids, and W;
The above-mentioned manufacturing method is characterized in that a compound of the formula in which X, R ₁ and the wavy line have the above-mentioned meanings is obtained, and this is isolated and, if necessary, a salt is produced. 6 General formula Corresponding to this formula, the symbol [formula] indicates that the 5th position of the steroid skeleton is in the configuration α, W indicates a hydrogen atom or a hydroxyl group, or together with X indicates an ethylidene group, and X An ethyl group or a group of the formula [Formula] or [Formula] or together with W indicates an ethylidene group, the wavy line means that the corresponding group is in one or the other of the configurations α and β, and R ₁ Represents a hydrogen atom or a methyl group, R ₂ is a hydrogen atom, 1 to 5 carbon atoms
_R3 represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 2 to 5 carbon atoms, a carbon represents an alkoxycarbonyl group having 2 to 8 atoms, a nicotinoyl group or a group derived from an α-amino acid or from a peptide having 2 or 3 α-amino acids, while R ₂ and R ₃
does not represent a hydrogen atom, and on the other hand: (i) When R ₁ represents a methyl group, W and R ₂ represent a hydrogen atom, and X represents a group [formula], R ₃ is methyl or nicotinoyl group or a group derived from an aminocarboxylic acid, (ii) R ₁ represents a methyl group, W represents a hydrogen atom,
When R ₂ represents a methyl group and X represents a group [formula], R ₃ cannot represent a methyl group, (iii) R ₁ represents a methyl group, and R ₂ and W represent a hydrogen atom. , X represents a group [formula], where the hydroxyl is in the (S) configuration and the amino group is in the 3α position, R ₃ cannot represent a methyl or glycyl group, and (iv) W represents a hydrogen atom, X represents a group [formula], and the amino group is at the 3α position, the substituents R ₁ , R ₂ and R ₃ cannot simultaneously represent a methyl group, and (v) R When ₂ and W represent a hydrogen atom, R ₁ represents a methyl group, and X represents a group [formula], where hydroxyl is in the (R) configuration and the amino group is in the 3α position, R ₃ cannot represent an ethoxycarbonyl or methyl group; (vi) if R ₁ , R ₂ and R ₃ represent a methyl group and the amino group is in the 3α position, then W and X together represent an ethylidene group; immunotherapeutic properties characterized by containing as an active ingredient at least one 3-amino substituted steroid derivative and its addition salt with a pharmaceutically acceptable acid. A pharmaceutical composition comprising: