CH651056A5 - SUBSTITUTED 3-AMINO STEROID DERIVATIVES AND THEIR SALTS, PREPARATION METHODS AND MEDICINAL PRODUCTS CONTAINING THEM. - Google Patents

Description

The present invention relates to new substituted 3-amino steroid derivatives as well as their salts, the methods of preparation, and the medicaments and compositions containing them.

The subject of the invention is new substituted 3-amino steroid derivatives, as well as their addition salts with mineral or organic acids, characterized in that they correspond to the general formula:

(I)

in which W represents a hydrogen atom or a hydroxyl radical or, together with X, represents an ethylidene radical, X represents an ethyl radical or a group

CH,

AT,

or fa

CH

or, together with W, a radical

OH

CH.

in this case, R3 cannot represent a radi-

and the amino radical is in position 3a, in this case, R3 cannot represent a methyl, acetyl or glycyl radical, ini) when W represents a hydrogen atom, X

CH.

represents a group

4

H

X,

and the amino radical

OH

is in position 3a, in this case, the substituents Rls R2 and R3 do not at the same time represent a methyl radical,

iiiii) when both R2 and W represent a hydrogen atom, Ri represents a methyl radical, X represents fH3

i5 a group çg whose hydroxyl is of configu-

QH

ethylidene, the wavy lines mean that the corresponding radical is in the same or other of the configurations a and ß, R] represents a hydrogen atom or a methyl radical, R2 represents a hydrogen atom or an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms and R3 represents a hydrogen atom, an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, or a radical derived from an α-amino acid or from a peptide comprising 2 or 3 α-amino acids, it being understood that:

on the one hand, at least one of R2 and R3 does not represent a hydrogen atom,

- on the other hand:

i) when both Ri represents a methyl radical, W and R2 represent a hydrogen atom and X represents the cal methyl, acyl radical, or a residue derived from a carboxylic amino acid,

ii) when at the same time R (represents a methyl radical, W represents a hydrogen atom, R2 represents a radical

CH,

| 3

methyl and X represents the radical r in this case,

/ 'V

' at

R3 cannot represent a methyl or acetyl radical,

iii) when both Ri represents a methyl radical, R2 and W represent a hydrogen atom, X represents a group

CH.

he-

pement £ H ^ whose hydroxyl is of configuration (S)

ration (R) and the amino radical is in position 3a, in this case, R3 does not represent an ethoxycarbonyl, methyl or acetyl radical,

iiiiii) when both Rb R2 and R3 represent a methyl radical and the amino radical is in position 3a, in this case, W and X cannot together represent an ethylidene radical.

In general formula I and in the following, the term alkyl radical containing from 1 to 5 carbon atoms denotes, for example, a methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl radical; the term hydroxyalkyl radical containing 2 to 5 carbon atoms denotes, for example, a hydroxyethyl or hydroxypropyl radical; the term acyl radical containing 2 to 8 carbon atoms denotes, for example, an acetyl, propionyl, n-butyryl, iso-butyryl, benzoyl or nicotinoyl radical; the term alkoxycarbonyl radical containing 2 to 8 carbon atoms denotes, for example, a methoxy, ethoxy or propoxycarbonyl radical; the residue derived from an amino carboxylic acid can be chosen from the group consisting of Ala, Val, Ival, Leu, Ile, Asp, Asn, Glu, Gin, Ser, Thr, Cys, Met, Lys, Arg, Phe, Tyr , Trp, His, and Pro, Nva, Nie, Hyp, Orn, these acids being in the D or L form, as well as by Sar and Gly, all the acids previously named being able to be N-alkylated; when R3 represents a radical derived from a peptide comprising 2 or 3 α-amino acids, these are chosen from the group consisting of the above α-amino acids.

45 It will be accepted by convention that the symbols for α-amino carboxylic acids represent these acids in their D or L configuration (for example, the term Ala means Alanine in D form or in L form).

Unless otherwise agreed, the nomenclature used in this application is the IUPAC nomenclature, the rules of which are published in particular in Biochem. J. (1972) 126, 773-780.

The addition salts with mineral or organic acids can be, for example, the salts formed with hydrochloric, hydrobromic, nitric, sulfuric, phosphoric, acetic, formic, propionic, benzoic, maleic, fumaric, succinic acids. tartaric, citric, oxalic, glyoxylic, aspartic, alkane sulfonic such as methane or ethane sulfonic, arylsulfonic, such as aci-60 of benzene or paratoluene sulfonic and arylcarboxylic.

Among the products which are subjects of the invention, there may be mentioned in particular the derivatives corresponding to formula I above, as well as their addition salts with mineral or organic acids, characterized in that, in said formula I, X represented-

CH-,

65

OH

feels a group t

: H

/ S

OH

Among these, there may be mentioned more particularly the derivatives characterized in that, in said formula I, X represents

CH,

I 3

a CH / group

Hal-R'3

651056

IV

and R2 represents an atom

10

OH

of hydrogen or a methyl radical, as well as their addition salts with mineral or organic acids and very particularly:

- (2S) 2-amino N - / (20) -20-hydroxy (5a) -pregnan-3a-yl / propanamide,

- (2S) 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / 1H-indol 3-propanamide,

- 2-amino N - / (20S) -20-hydroxy-19-nor (5a) -pregnan is -3a- yl / acetamide,

- 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / N-methyl acetamide, as well as their salts; the derivatives other than those mentioned above, described in the examples, are also very particularly retained. 20

A subject of the invention is also a process for the preparation of derivatives, as defined by formula I above,

as well as their salts, characterized in that an amine of formula II is reacted:

CH- 25

II

30

in which the wavy line, W, X and R! have the meaning already indicated, 35

- either with a halide of formula III:

Hal-R ':

III

40

in which Hai represents a chlorine, bromine or iodine atom, and R'3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, to obtain a product of formula I in which R2 represents an atom of hydrogen, R3 represents an acyl or alkoxycarbonyl radical containing 2 to 8 carbon atoms, and W, X, Rj and the wavy line have the meaning already indicated, product of formula I which is isolated and salified if desired ,

- either with an α-amino acid or a peptide comprising 2 or 3 α-amino acids whose amine function is protected by an easily cleavable group, in particular by acid hydrolysis, then treated so as to remove the protective group, in order to obtain a product of formula I in which R2 represents a hydrogen atom, R3 represents a radical derived from an α-amino acid read from a peptide comprising 2 or 355 α-amino acids, and W, X, Ri and the wavy line have the meaning already indicated, which is isolated and salified if desired,

- either with a C1-C5 alkyl or hydroxyalkyl halide containing 2 to 5 carbon atoms and whose halogen is a chlorine, bromine or iodine atom, to obtain 60 a product of formula I in which R2 and R3 represent a hydrogen atom or an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms and W, X, Rj and the wavy line have the meaning already indicated , product of formula I which is either isolated and salified if desired, or else, in the case where R3 represents a hydrogen atom, it is reacted with a halide of formula IV:

in which R "3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hai has the meaning already indicated, in order to obtain a product of formula I in which R2 represents an alkyl radical Ci-C5 or hydroxyalkyl containing 2 to 5 carbon atoms, R3 represents an alkyl radical containing 1 to 5 carbon atoms or hydroxy alkyl containing 2 to 5 atoms this carbon or acyl or acyl-oxy containing 2 to 8 carbon atoms and W, X, R] and the wavy line have the meaning already indicated, which is isolated and salified if desired, or even, in the case where R3 represents a hydrogen atom, said product of formula I is reacted with an α-amino acid or a peptide comprising 2 or 3 α-amino acids whose amine function is protected by an easily cleavable group, in particular by acid hydrolysis, then treats so as to el imine the protective group, to obtain a product of formula I in which R2 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxy alkyl containing from 2 to 5 carbon atoms, R3 represents a radical derived from an acid a- amino or a peptide comprising 2 or 3 α-amino acids and W, X, R] and the wavy line have the meaning already indicated, which is isolated and salified if desired.

Under preferred conditions for implementing the invention, the preparation process described above is carried out as indicated below.

1) The reaction of the amine of formula (II) with the halide of formula (III) is carried out in the presence of an acid-fixing agent, in particular an alkali hydroxide, an alkali carbonate (for example potassium ), an alkaline bicarbonate, an alkaline acetate, an alkaline earth carbonate, a tertiary amine (for example a trialkoylamine or pyridine) or an alkaline alcoholate (for example sodium ethylate).

The reaction can be carried out, for example, in inert solvents or suspension media such as dioxane, di-methylformamide, benzene, toluene or methylene chloride. In the case of acylation, it is also possible to use the free acid, or else a functional derivative other than a halide, such as an acid anhydride.

In the case where the free acid is used, it is necessary to carry out the amidification with an activating agent such as a carbodiimide. Other techniques can be used, such as those described for example in "The Peptides" Vol. 1, Academic Press 1979.

2) The reaction of the amine of formula II or of the product of formula I in which R3 represents a hydrogen atom and R2 represents an alkyl or hydroxy alkyl radical with the α-amino acid or the peptide whose amino function is protected by an easily cleavable protective group takes place in the presence of a condensing agent. The purpose of the condensing agent in this case is to activate the acid function of the amino acid.

As a condensing agent, a carbodiimide of formula:

A-N = C = N-B

in which A and B represent an alkyl radical containing from 1 to 8 carbon atoms optionally carrying a dialkoylamino radical or represent a cycloalkyl radical.

Mention may be made, for example, of dicyclohexylcarbodiimide or 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide, preferably the latter.

651,056

It is also possible to use a 2-chloro N-methyl pyridinium halide such as iodide.

It is also possible to use an alkyl chloroformate such as, for example, methyl, ethyl or iso-butyl chloroformate; one can also use an alkyl pyrophosphite, such as for example ethyl pyrophosphite.

As an easily cleavable protective group, the benzyloxycarbonyl group or the (Z) terbutyloxy-carbonyl group (BOC) can be used, as appropriate, as the case may be.

The invention particularly relates to a process as defined above, characterized in that the protective group is the terbutyloxycarbonyl radical.

To remove the easily cleavable protective group above, an acid such as hydrochloric acid is preferably used as the cleavage agent; one operates for example using an alkanolic solution of hydrochloric acid, or using anhydrous hydrochloric acid by bubbling in nitromethane. Acids such as paratoluene sulfonic acid, formic acid or trifluoroacetic acid can also be used. Hydrogen can also be used in the presence of palladium, for the protective group (Z) for example.

3) The reaction of the amine of formula II with the alkyl or hydroxyalkyl halide is carried out in the presence of the same acid-fixing agents and solvents as in the case of the reaction with the halide of formula III.

The hydroxy function of the hydroxyalkyl radical is advantageously blocked by an easily cleavable protective group, in particular by acid hydrolysis. This protective group is advantageously a tetrahydro-pyrannyl radical. It can be cleaved under the same conditions as those indicated above for the protective group of the amino acid.

4) The reaction of the product of formula I in which R3 represents a hydrogen atom, R2 represents an alkyl radical C, -C5 or hydroxyalkyl containing from 2 to 5 carbon atoms and W, X, Ri and the wavy line have the meaning already indicated, with the halide of formula IV is carried out according to the nature of R "3, under the conditions previously described for the reaction of the amine of formula II with the halide of formula III or with the halide of alkyl.

5) For the preparation of the products of formula I N-alkyls, it may be advantageous in certain cases to operate in a particular way:

- To prepare a product of formula (I) N-mono-methylated, one can prepare a carbamate of the amine of the product of formula (II) for example by action of an alkyl haloformate and in particular of ethyl, then reduction of the carbamate, for example using aluminum-lithium hydride;

- To prepare a product of formula (I) N-mono-ethylated, the corresponding N-acetylated derivative can be prepared and then reduced;

- to prepare the N-diethyl derivative, simply repeat the operation.

In these three cases, when X represents a group

O

II

I I

-C -CH3, it is necessary to prepare the ketal in 20, for example-

protection of amino functions does not concern cases where these amino functions are alkylated.

The derivatives of formula I with the exception of those for which R3 represents an acyl or acyloxy radical have a basic character. The addition salts of the derivatives of formula I can advantageously be prepared by reacting, in substantially stoichiometric proportions, a mineral or organic acid with said derivative of formula I. The salts can be prepared without isolating the corresponding bases.

The derivatives which are the subject of the present invention have very advantageous pharmacological properties; they are endowed in particular with remarkable immunotherapeutic properties. They are in particular capable of stimulating the immune reactions.

These properties are illustrated later in the experimental part.

These properties justify the use of the substituted 3-amino steroid derivatives of formula (I), as well as their pharmaceutically acceptable salts, as medicaments. The present application thus also relates to medicaments constituted by the substituted 3-amino steroid derivatives, as defined by formula I, as well as by their addition salts with pharmaceutically acceptable acids.

Among the drugs which are the subject of the invention, those which consist of the new substituted 3-amino steroid derivatives corresponding to formula I are preferably used in the

CH.

30

which X represents a JüH group

/%

as well as

OH

by their addition salts with pharmaceutically acceptable acids.

Among these, we especially retain those corresponding to formula I, in which X represents a group

P

: h.

- To prepare a product of formula I N-mono-or di-isopropylated it is possible to react a product of formula II with acetone in the presence of a reducing agent such as sodium cyanoboro-hydride.

It is obvious that, in the process of the invention, the

J-H, and r2 represents a hydrogen atom or a 40 bH

methyl radical, as well as their addition salts with pharmaceutically acceptable acids.

45 Among these, there are particularly the derivatives whose names follow:

- 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / propanamide,

- (2S) 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-50 yl / 1H-indole 3-propanamide,

- 2-amino N - / (20S) -20-hydroxy-19-nor (5a) -pregnan-3a-yl / acetamide,

- 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / N-methyl acetamide, as well as their addition salts with aci-

55 of pharmaceutically acceptable.

The medicaments according to the invention find their use, for example, in the treatment of autoimmune diseases resulting from a deficiency in certain lymphocytes, whether they are diseases of the connective tissue that are not specific for a gold-60 gane such as example rheumatoid arthritis, systemic lupus erythematosus or whether it is specific organ diseases such as thyroiditis, pemphigus or hemolytic anemia.

The medicaments according to the invention can thus be used as an adjuvant treatment for antibiotic therapy and anti-cancer chemotherapy.

The usual dose, which varies according to the derivative used, the subject treated and the condition in question, can be, for example, 10 mg

at 1 g per day, orally in humans, for the derivative of Example 14 used as an adjuvant for antibiotic therapy.

The invention finally relates to pharmaceutical compositions which contain at least one aforementioned derivative or one of its addition salts with pharmaceutically acceptable acids, as active principle.

As medicaments, the derivatives corresponding to formula I and their addition salts with pharmaceutically acceptable acids can be incorporated into pharmaceutical compositions intended for the digestive or parenteral route.

These pharmaceutical compositions may be, for example, solid or liquid and be in the pharmaceutical forms commonly used in human medicine, such as, for example, tablets, simple or coated, capsules, granules, suppositories, injectable preparations; they are prepared according to the usual methods. The active ingredient (s) can be incorporated therein into excipients usually used in these pharmaceutical compositions, such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous vehicles or not , fatty substances of animal or vegetable origin, paraffinic derivatives, glycols, various wetting agents, dispersants or emulsifiers, preservatives.

The starting materials of formula II, when they are not known, can be prepared as follows:

A product of formula V is reacted:

651,056

II,

10

in which Ri and the wavy line have the meaning already indicated, that

- either Yonisole,

- Either it is reduced, for example by hydrogenation in the presence of a catalyst based on rhodium, platinum or palladium, to obtain a product of formula IIB:

15

20

II

B

in which R] and the wavy line have the meaning already indicated,

25 - either hydrate for example using diborane to obtain a product of formula IIC:

in which R] er the wavy line have the meaning already indicated, with an excess of ethyl triphenylphosphonium bromide and potassium tertbutylate to obtain a product of formula VI:

VI

in which Ri and the wavy line have the meaning already indicated, which is transformed, for example, by passage to tosylate then action of sodium azide, or directly by reaction with diphenyl azidophosphate in the presence of azodicarboxylate d ethyl and triphenylphosphine in the a-zide of formula VII:

30

35

fs

I] c in which Ri and the wavy lines have the meaning already indicated, that either one isolates, or one is subjected to an oxidation for example using chromic acid, in order to obtain a product of formula IID:

II,

so in which Ri and the wavy line have the meaning already indicated,

- Either subject to cis-dihydroxylation, for example using osmic anhydride in the presence of trimethyloxami-ne, after protection or not of the amine by a fa-55 cilily hydrolyzable group such as a group trifluoroacetyl, to obtain a product of formula IIE:

VII

in which R] and the wavy line have the meaning already indicated and the configuration in 3 is opposite to that of the compounds V and VI, which are reduced, for example using lithium aluminum hydride, to obtain an amine of formula IIA:

60

65

II,

in which R represents the protective group defined above or a hydrogen atom, and Ri and the wavy line have the

651056

meaning already indicated, that either when R represents a hydrogen atom, it is isolated, or else, when R represents a protective group, a cleavage agent of said protective group is subjected, for example an alkaline hydrolysis in the case of a trifluoroacethyl group, then isolates the free amine obtained, or else one undergoes - to oxidation, for example using chromic acid, to obtain a product of formula IIF:

in which R, Rt and the wavy line have the meaning already indicated that, either when R represents a hydrogen atom, one isolates, or else, when R represents a protective group, one subjects to an agent of cleavage of said protective group, then isolates the free amine obtained, or else it is reduced, for example using sodium borohydride, to obtain a product of formula IIG:

in which R, Ri and the wavy lines have the meaning already indicated, that either when R represents a hydrogen atom is isolated or when R represents a protective group, it is subjected to a cleavage agent of said protective group, then isolates the free amine obtained.

Examples of preparations of products of formula (II) appear below in the experimental part.

The following examples illustrate the invention without, however, limiting it.

Example 1

(2S) 2-amino N-j (20S) -20-hydroxy 5a-pregnan-3a-yllpropan-amide (hydrochloride and base)

Stage A: 2- / 1,1-dimethyl ethoxy carbonylamino / N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / propanamide

1.92 g of (20S) 3a-amino 5a-pregnan-20-ol are dissolved and

8

2.27 g of tertbutyloxycarbonyl-L-Alanine (BOC-L-Alanine) in 60 cm3 of chloroform and 12 cm3 of pyridine. The solution is stirred at 0 ° C / + 5 ° C under an inert atmosphere and 1.14 g of 1-ethyl 3- (3-dimethylaminopropyl) carbo-5 diimide hydrochloride is added. After 1 hour 15 minutes, 1.15 g of 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide hydrochloride are again added, stirring is continued for 50 minutes at 0 ° C / -5 ° C, brought to dry, taken up in an aqueous solution of sodium hydrogen carbonate, extracted with ethyl acetate, washed with a saturated aqueous solution of sodium chloride and then with a normal aqueous solution of hydrochloric acid and again with an aqueous solution saturated with sodium chloride, dried, brought to dryness, crystallized from isopropyl ether, drained, washed with isopropyl ether, dried at 60 ° C. under reduced pressure and obtained the expected product. M = 171 ° C.

Stage B: hydrochloride and base of (2S) 2-amino Nj (20S) -20-hydroxy 5a-pregnan-3a-yl / propanamide variant 1: 1.55 g of the product obtained are suspended under an inert atmosphere in stage A, in 100 cm3 of nitro-methane, bubbled hydrochloric acid for 10 minutes, stirred at 20 ° -25 ° C for 35 minutes, removed excess hydrochloric acid, drained, washed with ethyl ether, dried at 60 ° C under reduced pressure, recrystallized from methanol and obtained 990 mg of expected hydrochloride. F ~ 220 ° C.

variant 2: 2.7 g of the product obtained in Stage A are dissolved under an inert atmosphere in 10 cm3 of ethanol, 40 cm3 of hydrochloric ethanol (3.5N) are added, the mixture is stirred for 8 hours, brought to dryness, taken up in 1 ethyl acetate, ice, drain, wash with ethyl acetate, dry at 60 ° C under reduced pressure, recrystallize from methanol and obtain 2.2 g of the expected hydrochloride. F at 220 ° C.

Preparation of the base:

35 1.74 g of hydrochloride are dissolved in 100 cm3 of t-trahydrofuran at 30% water, 4 cm3 of 2N sodium hydroxide are added, brought to dryness, taken up in 100 cm3 of chloroform, washed with water, dried, brought to dryness, recrystallizes from ethyl acetate and obtains 1.27 g of the expected base. Mp 224 ° C. 40 By operating according to a process analogous to that described above, the derivatives described in the table below were prepared.

Example number

Variant

Acid starting Amine

R3

F

2

3

4

5

I + remark

1 1 1

BOC-L-Glutamic acid

BOC-L-Serine

E JC-L-Phenyl-alanine

BOC-L-Tryptophan ■

O ^ CH-CH2-CH2-COOH (S)

nh2

cAcïï-ch2oh (s)

ne2

o ^ ch-ch - / 'y (s) nh2

<AçH-CH2-rç-d0 (S)

2 1 H

220 ° C (hydrochloride)

210 ° C (hydrochloride)

240 ° C (hydrochloride)

135 ° C (base)

9

651,056

Example number

Variant

Starting Amino Acid

R3

F

6

7

2

cf3cooh

BOC-L-Proline

BOC-Glycyl Glycine

O ^ ClT ^ (S)

1 H 0 ^ CH2-NH-CO-CH2-NÏ ^

275 ° C (hydrochloride) (240 ° C (hydrochloride (200 then 252 ° C (base

Note for example # 2:

The second acid function of L-Glutamic acid is blocked in the form of a benzyl ester; it is released by catalytic hydrogenation in acetic acid in the presence of palladium, before cleavage of the BOC.

Example 8

N-j (20S) -20-hydroxy 5a-pregnan-3a-yll 3-pyridine carbox-

amide

960 mg of (20S) 3a-amino 5a-pregnan-20-ol and 813 mg of nicotinic acid are dissolved under an inert atmosphere in 30 cm 3 of chloroform and 6 cm 3 of pyridine, 582 mg of 1-ethyl 3- hydrochloride are added. (3-dimethylaminopropyl) carbodiimide. After 3 hours 30 minutes, with stirring, 291 mg of 1-ethyl 3- (3-dimethylaminopropyl) carbodiimide hydrochloride is added, stirred for 16 hours, brought to dryness, taken up in 30 cm3 of water, ice, drained, washed with water. water, dried at 80 ° C under reduced pressure, recrystallized from methanol and obtained 920 mg of the expected product. Mp 258 ° C.

ja / D = + 14.5 ° ± 1 ° (c = 1% Pyridine).

Example 9

2-amino hydrochloride N - / (20S) 17a, 20-dihydroxy 5 a-pregnan 3a-ylj acetamide

By operating according to a process analogous to that described in Example 1 (variant 2) from BOC-Glycine and (20S) 3a-amino 5a-pregnan 17a, 20-diol, the expected product is obtained. F ~ 200 ° C.

/ a / D = + 4 ° ± l ° (c = 1.5% Ethanol95 °). The starting (20S) 3a-amino 5a-pregnan-17a, 20-diol can be prepared as follows:

Stage 1: (Z) (5a) pregn-17 (20) -èn3ß-ol

59.4 g of triphenylethylphosphonium bromide are added to 16.1 g of potassium tert-butoxide in solution in 160 cm3 of tetrahydrofuran, stirred for 30 minutes, 23.2 g of epiandrosterone, stirred for 15 hours, poured into ice water, extracted with ethyl acetate, washed with water, dried, distilled to dryness, purified by chromatography on silica (eluent: cyclohexane-ethyl acetate 7-3), crystallized from methanol, ice, drain, dry and obtain 23.1 g of the expected product. Mp 160 ° C.

Stage 2: (Z) 3a-azido (Sa) -pregn-17 (20) -ene

1.92 g of ethyl azodicarboxylate and 3.02 g of diphenyl azidophosphate are added to a solution of 1.66 g of the above product in 30 cm 3 of benzene and 5 cm 3 of tetrahydrofuran, stirred in the bath ice, add in 20 minutes a solution of 2.88 g of triphenylphosphine in 30 cm3 of benzene, stir again for 40 minutes at 10 ° C, wash with a 2N hydrochloric acid solution, then with water, dry and distill when dry, purify by chromatography on silica (eluent: hep-tane then heptane-ethyl ether 1-1), and obtain 1.67 g of crystals of the expected product. M = 114 ° C after recrystallization from methanol.

Stage 3: (Z) 5a-pregn 17 (20) -ene-3a-amine (and hydrochloride) 14.5 g of (Z) 3a-azido (5a) -pregn - 17 (20) are dissolved in 290 cm3 of tetrahydrofuran ) -ene as obtained above, stirred while heating at 25-27 ° C, added in 1 hour 800 mg of lithium aluminum hydride, stirred for another 1 hour, removed excess hydride by methanol, filter, wash the filtrate with an aqueous solution of Seignette salt then with a saturated aqueous solution of sodium chloride, dry, dry distillate and obtain 13.1 g of crystals of the expected amine. F ~ 90 ° C.

The base is dissolved in 150 cm3 of ethyl acetate and 30 cm3 of methylene chloride, 27 cm3 of 1.7N hydrochloric ethyl acetate are added, drained, washed with ethyl acetate and dried under pressure reduced the crystals obtained and obtained 13.2 g of crystals of the hydrochloride of the expected product. F>

30 300 ° c-

/ a / D at 1% in pyridine, at 10% water = + 38.5 ° ± 1.5 °.

Stage 4: Nj (Z, 5a) -pregn 17 (20) -en 3a-yl / trifluoroacetamide 16.5 g of the hydrochloride obtained in Stage 3 above are suspended under an inert atmosphere in 165 cm3 of chloride methylene and 16.5 cm3 of pyridine, cooled to approximately 5 ° C., introduced 16.5 cm3 of trifluoroacetic anhydride in 5 minutes, stirred for 15 minutes at room temperature, distilled to dryness under reduced pressure, added 200 cm3 of water, wring, 40 washes with water, dries under reduced pressure and obtains 18.1 g of crystals. M = 204 ° C.

Stage 5: N - / (20S) 17a, 20-dihydroxy (5a) -pregnan-3a-yl / trifluoroacetamide 45 18.1 g of the product obtained above are dissolved in 100 cm3 of methyl ethyl ketone under an inert atmosphere, 9 g are added of trimethylamine N-oxide dihydrate as well as a solution of 360 mg of osmium tetroxide in 71 cm3 of methyl ethyl ketone, stirred for 2 hours at reflux, allowed to cool, 50 added 200 cm3 of 10% sodium thiosulfate solution in 1 , stirred for 30 minutes at room temperature, decanted, washed with water, dried over magnesium sulphate, filtered, distilled to dryness under reduced pressure, purified the oil obtained by chromatography on silica (eluent: benzene-acetate ethyl 7-3), and 55 obtains 14 g of the expected product; F = 172 "C then 192" C.

Stage 6: (20S) 3a-amino 5a-pregnan-17a, 20 diol

4 g of the product obtained above are dissolved under an inert atmosphere in 20 cm 3 of methanol, 8 cm 3 of sodium hydroxide solution are added, stirred for 1 hour 30 minutes, added 50 cm 3 of water, stirred for 10 minutes, drained, washed with water, dry under reduced pressure at 40 ° C and obtain 3 g of the desired product. Mp 210 ° C.

Example 10

65 2-amino hydrochloride N- / (20S) -20-hydroxy 19-nor (5a) -pregnan-3a-yll acetamide

By operating according to a process analogous to that described in Example 1 (variant 1) from BOC-Glycine and (20S)

651,056

3-a-amino-19-nor (5a) -pregnan-20-ol, the expected product is obtained. F ~ 270 ° C with sublimation.

/ a / D + 39.5 ° ± 1.5 ° (c = 1% methanol). The starting (20S) 3a-amino-19-nor (5a) -pregnan-20-ol can be prepared as follows:

By operating according to a process identical to that described in Example 9 (stages 1,2,3) for the preparation of (Z) 5a-preg-17 (20) -èn-3a-amine, it was prepared from 19 Nor-epi-androsterone la (Z) 5a-19 nor-pregn 17 (20) -èn-3a-amine.

156 mg of sodium borohydride in 5 cm 3 of tetrahydrofuran are suspended under nitrogen, a solution of 0.5 cm 3 of boron trifluoride etherate in 2.5 cm 3 of tetrahydrofuran is added dropwise at + 5 ° C. stir for 1 hour in an ice bath, add 296 mg of (Z) 5a-19 nor-pregn 17 (20) -en-3a-amine dissolved in 3 cm3 of tetrahydrofuran, stir for 1 hour 30 minutes at room temperature, cool to ice bath, slowly add 2 cm3 of 6N sodium hydroxide, stir for 5 minutes at room temperature, decant, extract the aqueous phase with tetrahydrofuran, wash the organic phase with water, add 4 cm3 of 5N sodium hydroxide, 2 cm3 of water oxygenated to 110 volumes, stirred for 45 minutes, extracted with ethyl acetate, washed with water, dried and distilled to dryness under reduced pressure. The dry extract is taken up in 10 cm3 of methanol with 5 cm3 of N hydrochloric acid, heated for 30 minutes in a water bath at 50 ° C., poured into a saturated solution of sodium bicarbonate, extracted with methylene chloride, washed with water, dry, evaporate under reduced pressure and obtain 257 mg of crystals of the expected product. F ~ 190 ° C.

Example 11

2-amino hydrochloride N- / 17a-hydroxy 20-oxo 5a-pregnan-3a-ylj acetamide

By operating according to a process analogous to that described in Example 1 (variant 2) from BOC-Glycine and 3a-amino 17a-hydroxy 5a-pregnan-20-one, the expected product is obtained. F> 300 ° C.

/ a / D = + 50 ° ± 1 ° (c = 1% Ethanol 95 °).

The starting 3a-amino 17a-hydroxy 5a-pregnan-20-one can be prepared as follows:

By perchromic oxidation of the product obtained in stage 5 of Example 9, N- (17a-hydroxy-20-oxo-5a-pregnan-3a-yl) trifluoroacetamide is prepared (F = 178 ° C and then 186 ° C), treated as indicated in step 6 of Example 9 to obtain 3a-amino 17a-hydroxy 5a-pregnan-20-one. M = 216 ° C (after crystallization from water).

Example 12

2-amino hydrochloride N - / (20R) -20-hydroxy 5a-pregnan-3a-yl / acetamide

By operating according to a process analogous to that described in Example 1 (variant 1) from BOC-Glycine and (20R) 3a-amino 5a-pregnan-20-ol, the expected product is obtained. M = 210 ° C then 260 ° C.

/ a / D = + 22 ° ± 1 ° (c = 0.8% pyridine at 10% water).

Example 13

Nl (20S) -20-hydroxy (5a) -pregnan-3a-ylj ethyl carbamate 5 g of (20S) 3a-amino 5a-pregnan-20-ol are added dropwise over 10 minutes under an inert atmosphere in 500 cm3 of methylene chloride to 15 cm3 of ethyl chloroformate and 45 cm3 of methylene chloride, stirred for 30 minutes, added 20 cm3 of sodium hydroxide, stirred again for 30 minutes, decanted, extracted with methylene chloride, washed with water, dry, filtrate, evaporate to dryness under reduced pressure, take up in ethyl ether, spin dry and obtain 5.7 g of the expected product. F ~ 198 ° C.

/ a / D = +25 ° C ± 1.5 ° (c = 1% methylene chloride).

10

Example 14

2-amino hydrochloride N - / (20S) -20-hydroxy 5a-pregnan-3a-ylj N-methyl acetamide

By reduction of the product of Example 13, 5 (20S) 3a-methylamino 5a-pregnan-20-ol (mp = 170 ° C.) described by Vetter and Coli in Bull was prepared. Soc. (1963) 1324, which was reacted with BOC-Glycine according to a process analogous to that described in Example 1 (variant 2) to obtain the expected product, from which the hydrochloride is prepared. F ~ 250 ° C.

10

Example 15

2 (S) 2-amino N-methylN - / (20S) 20-hydroxy-5a-pregnan-3a-yl / propanamide hydrochloride.

The (20S) 3a-methylamino 5a-pregnan-20-ol 15 hydroxy-5a-pregnan-3a-yl / propanamide is reacted.

The (20S) 3a-methylamino5a-pregnan-20-ol is reacted with BOC-L-Alanine according to a process analogous to that described in Example 1 (variant 1) to obtain the expected product, the hydrochloride of which is prepared . M> 270 ° C.

20

Example 16

2 (R) 2-amino hydrochloride N-methyl N - / (20S) 20-hy-droxy-5a-pregnan-3a-yl / propanamide.

The (20S) 3a-methylamino-5a-pregnan-20-ol 25 is reacted with BOC-L-Alanine according to a process analogous to that described in Example 1 (variant 1) to obtain the expected product, of which prepare the hydrochloride. (Sublimation at around 260 ° C).

30 Example 17

(20S) 3a-ethylamino hydrochloride 5a-pregnan-20-ol.

The (20S) 3a-amino 5a-pregnan-20-ol is reacted with ethyl iodide in the presence of sodium carbonate, to obtain the base of the expected product. Mp 129 ° C after recrystallization in ethyl acetate.

Formation of the hydrochloride: the hydrochloride is prepared by adding hydrochloric ethyl acetate and the expected product is obtained. M> 270 ° C.

- »o Example 18

2-amino N-ethyl hydrochloride N-j (20S) 20-hydroxy 5a-pregnan 3a-yl / acetamide.

The base of the product of Example 17 is reacted with BOC-L-Glycine according to a process analogous to that described in Example 1 (variant 1), to obtain the expected product, the hydrochloride of which is prepared. F ~ 230 ° C.

Example 19

(20S) 3a - / (2-hydroxyethyl) amino / 5a-50 pregnan-20-ol hydrochloride.

7 g of (20S) 3a-amino 5a-pregnan-20-ol and 7 g of sodium carbonate are suspended in 100 cm3 of anhydrous dioxane, 6.6 cm3 of 2/2-bromoethoxy / tetrahydropy-ran are added and refluxed for 24 hours under an inert atmosphere 55. The mixture is cooled to room temperature, diluted with water, extracted with ether, dried, distilled to dryness under reduced pressure, purified by chromatography on silica (eluent: chloroform-metha-nol-acetic acid 85-5-10) and collects 7.7 g of yellow oil.

60 Hydrolysis of the protective group.

3 g of the above product are dissolved in 30 cm3 of ethanol and 15 cm3 of 2N hydrochloric acid, stirred at reflux for 1 hour and 30 minutes, cooled, poured onto 100 cm3 of aqueous sodium bicarbonate solution, ice for 15 minutes, wring, wash with water, dry at 40 ° C under reduced pressure, dissolved in 350 cm3 of methylene chloride to 10% methanol, filter, concentrate to about 50 cm3, add 100 cm3 of acet

11

651,056

ethyl tate, concentrate in half and wring out the precipitate. It is dissolved in 100 cm3 of methanol, filtered, concentrated to approximately 30 cm3, added 70 cm3 of ethyl acetate, concentrated by half, wrung, dried at 40 ° C under reduced pressure and obtained 1.65 g of base of the expected product. Mp 215 ° C.

Hydrochloride formation.

1.5 g of the above base are dissolved in 40 cm3 of methanol, a 1.6 N hydrochloric ethyl acetate solution is added until the pH is acid, 60 cm3 of ethyl acetate is added, concentrated under reduced pressure, wrung, dried at 40 ° C under reduced pressure and obtained 1.5 g of the expected product. M> 260 ° C.

Example 20

a-dimethylamino N - / (20S) 20-hydroxy 5a-pregnan-3a-yl / acetamide.

The procedure is carried out as indicated in Stage A of Example 1, using 2.552 g of (20S) 3a-amino 5a-pregnan-20-ol and 3.350 g of N, N-dimethyl glycine hydrochloride. 4 g of crude product are obtained, containing the expected derivative and the derivative

0, N-disubstituted. By saponification with sodium hydroxide in methanol, the expected product is obtained. Mp 206 ° C.

/ a / D = + 29 ° ± 1 (C = 1% CHC13).

Example 21

Tablets corresponding to the formula have been prepared:

- (2S) 2-amino N - / (20S) -20-hydroxy hydrochloride 5a-pregnan-3a-3ayl / propanamide 20 mg;

- excipient q.s. for a tablet finished at 100 mg. (Detail of excipient; lactose, starch; talc, magnesium stearate).

Example 22

Tablets corresponding to the formula have been prepared:

- 2-amino hydrochloride N - / (20S) -20-hydroxy 5a-pregnan-3a-yl / N-methyl acetamide 15 mg;

- excipient q.s. for a tablet finished at 100 mg. (Details of excipient: lactose, starch, talc, magnesium stearate).

Pharmacological study

1. Anaphylactic shock adjuvant Principle:

The administration to animals of a product capable of stimulating the activity of the immune systems results in an increase in shock in response to the administration of an antigen to which the animal was sensitized.

Male mice weighing 30 to 35 g are sensitized intra-plantar with serum albumin from beef. 8 days later, they receive this antigen intravenously. Under conditions of minimum sensitization, the control animals do not make a fatal shock during this last administration.

The product to be tested is injected intra-plantar, mixed with the antigen: if this product is an adjuvant, it will increase awareness and this will result in fatal shock when administered intravenously.

The dose which causes mortality equal to or greater than 50% of the animals is retained as the active dose.

Results:

Example product

1

2 4

Dose per animal in mg

5

5

6

2

7

1

8

5

5 9

1

10

0.5

13

5

14

0.5

2. Test of rosettes with red blood cells of sheep Principle:

The administration to animals of a product capable of stimulating the activity of the immune systems results in an increase in their capacity to react to the injection of an immunogenic product.

Male rats aged 3 months are sensitized intraperitoneally with sheep erythrocytes (day 0) 7 days later (day 7) their spleen is removed and the splenocytes are brought into contact with sheep erythrocytes : we then count the percentage of leukocytes around which the erythrocytes have formed rosettes.

The product to be studied is administered orally daily from day -1 to day 1.

The dose of product which approximately multiplies by 2 the percentage of rosettes observed in the control animals is considered as an immunostimulating dose.

30

Results:

)

Example product

35 1

5

6 8

10

40 11

14

Dose per animal in mg / kg per os

1

2> 5

5 5 5 10

3. Acute toxicity study 45 The lethal doses LD0 of the various compounds tested were evaluated after oral administration in mice.

The maximum dose causing no mortality after 8 days is called DL0.

The results are as follows:

50

0.5 <1 5

Product of

DL0in the example

55 1

200

2

^ 400

3

^ 400

4

400

5

^ 1000

60 6

^ 800

7

^ 400

9

> 600

10

> 400

11

5 * 1000

65 12

Ss 400

13

> 400

14

^ 200

VS

Claims (13)

651,056 2 CLAIMS 1. Substituted 3-amino steroid derivatives, as well as their addition salts with mineral or organic acids, characterized in that they correspond to general formula I: CH. represents a cH group and the amino radical / \ OH is in position 3a, in this case, the substituents RIs R2 and R3 do not at the same time represent a methyl radical, iiiii) when, at the same time R2 and W represent a hydrogen atom, Rj represents a methyl radical, X represents in which W represents a hydrogen atom or a hydroxyl radical or, together with X, represents an ethylidene radical, X represents an ethyl radical or a group 15 CH Ì- /% 3 a group H of which the hydroxyl is of configu - "'OH CH. AT OR CH, I 3 CH '\ OH or, together with W, an ethylidene radical, the wavy lines signify that the corresponding radical is in one or other of the configurations a and ß, R] represents a hydrogen atom or a methyl radical, R2 represents a hydrogen atom or an alkyl radical containing 1 to 5 carbon atoms or hydroxy alkyl containing 2 to 5 carbon atoms and R3 represents a hydrogen atom, an alkyl radical containing 1 to 5 carbon atoms or hydroxyalco- y containing 2 to 5 carbon atoms, an acyl or alkoxycarbonyl radical containing 2 to 8 carbon atoms, or a radical derived from an α-amino acid or from a peptide comprising 2 or 3 α-amino acids, being understood that: on the one hand at least one of R2 and R3 does not represent a hydrogen atom, - on the other hand: i) when both R] represents a methyl radical, W and R2 represent a hydrogen atom and X represents the radical ration (R) and the amino radical is in position 3a, in this case, r3 does not represent an ethoxycarbonyl, methyl or acetyl radical, iiiiii) when both Rb R2 and R3 represent a methyl radical and the amino radical is in position 3a, in this case, W and X cannot together represent an ethylidene radical. 2. Substituted 3-amino steroid derivatives according to claim 1, as well as their addition salts with mineral or organic acids, characterized in that X represents a group. CH, 1 3 30 pement CH / \ h 3. 3-amino substituted steroid derivatives according to claim 2, as well as their addition salts with mineral or organic acids, characterized in that X represents a group 40 CH, I 3 CH and R2 represents a hydrogen atom or *OH ÇH3 cal r> in this case, R3 cannot represent a / ^ O methyl, acyl radical, or a residue derived from an amino carboxylic acid, ii) when at the same time R] represents a methyl radical, W represents a hydrogen atom, R2 represents a methyl radical and CH, | 3 X represents the radical r, in this case, R3 cannot '\> not represent a methyl or acetyl radical, iii) when, at the same time, Ri represents a methyl radical, R2 and W represent a hydrogen atom, X represents a ch3 çjj group including configuration hydroxyl / * OH (S), and the amino radical is in position 3a, in this case, R3 cannot represent a methyl, acetyl or glycyl radical, iiii) when W represents a hydrogen atom, X a methyl radical. 4. One of the derivatives of formula I according to claim 1, the names of which follow: 45 - (2S) 2-amino N - / (20S) -20-hydroxy (5a-pregnan-3a-yl / propanamide, - (2S) 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / 1H-indol 3-propanamide, - 2-amino N - / (20S) -20-hydroxy-19 nor (5a) -pregnan-50 3a-yl / acetamide, - 2-amino N - / (20S) -20-hydroxy (5a) -pregnan-3a-yl / N-methyl acetamide, as well as their addition salts with mineral or organic acids. 5. Method for preparing the derivatives of formula I according to claim 1, in which R2 represents a hydrogen atom, R3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms and W, X, Rj and the line corrugated have the meaning already indicated, as well as 6o their addition salts with mineral or organic acids, characterized in that an amine of formula is reacted: H- .X R, ' 65 II in which the wavy line, W, X and R] have the meaning already indicated, with a halide of the formula: Hal-R'3 III in which Hai represents a chlorine, bromine or iodine atom and R'3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, and if desired, the product of formula I obtained is salified. 6. Process for the preparation of the derivatives of formula I according to claim 1, in which R2 represents a hydrogen atom, R3 represents a radical derived from an examined acid or from a peptide comprising 2 or 3 α-amino acids and W , X, R, and the wavy line have the meaning already indicated, as well as their addition salts with mineral or organic acids, characterized in that an amine of formula is reacted: 651,056 in which the wavy line, W, X and R! have the meaning already indicated, with an alkyl halide Q-C5 or hydroxyalkyl containing 2 to 5 carbon atoms and the halogen of which is a chlorine, bromine or iodine atom, j to obtain a product of formula I in which R2 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms and R3 represents a hydrogen atom, said product is reacted with a halide of formula 10 Hal-R, (IV) II in which the wavy line, W, X, and R] have the meaning already indicated, with an α-amino acid or a peptide comprising 2 or 3 α-amino acids whose amine function is protected by an easily cleavable group, then processes so as to eliminate the protective group and, if desired, the product of formula I obtained is salified. 7. Process for preparing the derivatives of formula I according to claim 1, in which R2 and R3 represent a hydrogen atom or an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms and W , X, Rj and the wavy line have the meaning already indicated, as well as their addition salts with mineral acids in organic, characterized in that an amine of formula is reacted: ? H in which R3 "represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 to 15 carbon atoms, or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hai represents a chlorine atom, from bromine or iodine, and if desired, the product of formula I obtained is salified. 9. Process for the preparation of derivatives of formula I according to claim 1, in which R2 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, R3 represents a radical derived from a α-amino acid or a peptide comprising 2 or 3 α-amino acids and W, X, R! and the wavy line have the meaning already indicated, as well as their addition salts with mineral or organic acids, characterized in that an amine of formula is reacted: H- X 30 35 II II in which the wavy line, W, X and R! have the meaning already indicated, with an alkyl halide Q-C5 or hydroxyalkyl containing 2 to 5 carbon atoms and the halogen of which is a chlorine, bromine or iodine atom, and, if desired, the salt is salified product of formula I obtained. 8. A process for the preparation of the derivatives of formula I according to claim 1, in which R2 represents an alkyl radical C1-C5 or hydroxyalkyl containing 2 to 5 carbon atoms, R3 represents an alkyl radical containing 1 to 5 carbon atoms or hydroxyalkyl containing 2 to 5 carbon atoms, or acyl or acyloxy containing 2 to 8 carbon atoms and W, X, R] and the wavy line have the meaning already indicated, as well as their addition with mineral acids or organic, characterized in that an amine of formula is reacted: : a3 ^ II in which the wavy line, W, X and Rj have the meaning already indicated, with an alkyl halide Q-C5 or hydroxyalkyl containing 2 to 5 carbon atoms and the halogen of which is a chlorine, bromine or of iodine, 40 to obtain a product of formula I in which R2 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 atoms this carbon and R3 represents a hydrogen atom, reacting said product of formula I with an α-amino acid or a pep-45 tide comprising 2 or 3 α-amino acids whose amine function is protected by an easily cleavable group, then treated so as to remove the protective group, and, if desired, the product of formula I obtained is salified. 10. The method of claim 6 or 9, characterized in 50 that the amine function of 1 a-amino acid or peptide comprising 2 or 3 a-amino acids, involved, is protected by an easily cleavable group by acid hydrolysis. 11. Medicines, characterized in that they consist of 3-amino steroid derivatives of formula I according to 55 claim 1, as well as by their addition salts with pharmaceutically acceptable acids. 12. Medicines according to claim 11, constituted by the 3-amino steroid derivatives substituted according to one of claims 2 or 3, as well as by their addition salts with 60 pharmaceutically acceptable acids. 13. Medicines according to claim 11, constituted by the 3-amino steroid derivatives substituted according to claim 4, as well as by their addition salts with pharmaceutically acceptable acids. 65 14. Medicines according to one of claims 11,12 and 13, in the form of a pharmaceutical composition. 651056 3-amino substituted steroid derivatives and their salts, processes for their preparation and medicaments containing them.