IE52188B1 - New 3-amino steroid compounds,their production and pharmaceutical compositions containing them - Google Patents

Abstract

The title compounds of formula I and their salts are, with certain exceptions, new: W is H or OH; X is ethyl, 1-hydroxyethyl, or acetyl; or W + X = ethylidene; R, is H or Me; R2 and R3 are each H, alkyl or hydroxyalkyl, R3 alternatively being acyl (including alkoxycarbonyl and radicals derived from amino acids and peptides), and at least one of R2 and R3 being other than H. Preparation: from the corresponding primary amines (R2 = R3 = H) Use: treatment of autoimmune diseases.

Description

PATENT APPLICATION BY (71) ROUSSEL-UCLAF, A FRENCH BODY CORPORATE, OF 35 BOULEVARD DES INVALIDES, PARIS 7EME, FRANCE.

Price 90p The present invention relates to new 3-amino steroid compounds, their production» their use as medicaments and pharmaceutical compositions containing them.

Accordingly, the invention provides a compound which is a steroid of formula I: or a hydroxyl radical or, together withX, represents an 10 ethylidene radical, X represnts an ethyl radical or a OH or, together with W, an ethylidene radical, the wavy lines signify that the corresponding radical is in configuration σ or p, represents a hydrogen atom or a methyl radical, Eg represents a hydrogen atom or an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms and Rj represents a hydrogen atom, an alkyl radical containing 1 to 5 carbon atoms, a hydroxyalkyl radical containing 2 to 5 carbon atoms, or an acyl radical corresponding to an e-amino 10 carboxylic acid, an N-alkylated a-amino carboxylic acid or to a peptide containing 2 or 3 e-amino carboxylic acid or N-alkylated α-amino carboxylic acid units, with the proviso that at least one of Rg and R^ does 15 not represent a hydrogen atom, and with ihe further provisos:. i) when at the same time R^ represents a methyl radical, W and Rg each represent a hydrogen atom and X represents the radical CH^ , then R^ then R3 cannot represent methyl or an acyl radical corresponding to an o-amino carboxylic acid or an N-alkylated α-amino carboxylic acid, ii) when at the same time R-^ represents a methyl radical, W represents a hydrogen atom, Rg represents a methyl radical and X represents the radical , then Rj cannot represent a methyl radical, iii) when at the same time R^ represents a methyl radical, Rg and W each represent a hydrogen atom, X CH, represents a group /K whose hydroxyl is of (S) configuration and the -NRgR^ radical is at position 5?» then R^ cannot represent a methyl or glycyl radical, iv) when W represents a hydrogen atom, X represents CH, a group CH and the -NR„R, radical is at 2 ? OH position 3ct, then the substituents Rp Rg, and R^ do not each represent at the same time a methyl radical, v) when at the same time Rg and W each represent a hydrogen atom, R^ represents a methyl radical, X represents a group CH^ whose hydroxyl is of CH / \ OH (R) configuration and the -NRgRj radical is at position 5σ, then Rj does not represent a methyl radical, and vi) when at the same time Rp Rg and R^ each b represent a methyl radical and the -NRgRj radical is at position Jo, then W and X cannot together represent an ethylidene radical.

The α-amino carboxylic acid residue which can represent can be N-alkylated. The N can be mono- or di-alkylated e.g. by alkyl containing 1 to 5 carbon atoms. Similarly some or all of the o-amino carboxylic acid units in the peptide residue which R^ can represent can be N-alkylated, the N being mono- or di-alkylated e.g. by alkyl containing 1 to 5 carbon atoms.

In formula I and in the following the term alkyl radical containing 1 to 5 carbon atoms includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl; the term hydroxyalkyl radical containing 2 to 5 carbon atoms includes hydroxyethyl or hydroxypropyl; the acyl radical corresponding to an α-amino carboxylic acid can be a radical corresponding for example to Ala, Val, Ival, Leu, lie, Asp, Asn, Glu, Gin, Ser, Thr, Cys, Met, Lys, Arg, Phe, Tyr, Trp, His, Pro, Nva, Nle, Hyp, Orn, these acids being in the D or L form or Sar or Gly; the acyl radical corresponding to an N-alkylated o-amino carboxylic acid can be a radical corresponding to an N-alkylated derivative of any of these specified o-amino carboxylic acids; when represents an acyl radical corresponding to a peptide containing 2 or 3 o-amino carboxylic or N-alkylated o-amino carboxylic acid units, the units can he for ejtample of the o-amino carboxylic acids and N-alkylated O-amino carboxylic acids referred to above.

By convention, the symbols of the a-amino carboxylic -acids represent these acids in their D or L configuration (for example, the term Ala signifies alanine in D form or in L form).

Unless indicated otherwise, the nomenclature used in the present Specification is the IUPAC nomenclature, whose rules are published especially in Biochem. J. (1972) 126, 773-780.

The salts of the steroid can be addition salts with mineral or organic acids for example hydrochloric, hydrobromic, nitric, sulphuric, phosphoric, acetic, formic, propionic, benzoic, maleic, fumaric, succinic, tartaric, citric, oxalic, glyoxylic or aspartic acids, alkane sulphonic acids such as methane or ethane sulphonic acids, arylsulphonic acids such as benzene or paratoluene sulphonic acids or arylcarboxylic acids.

Among the present compounds, there may be mentioned especially those wherein X represents the group CH, I CH / \ Among these, there may he mentioned particularly the compounds wherein X represents the group CH and R. represents a hydrogen atom / '-OH or a methyl radical. Particular compounds are specified in the Examples and include: (2S) 2-amino N-[(20S)-20~hydroxy (5or)-pregnan-3a-yl] propanamide; (2S) 2-amino N-[(20S)-20-hydroxy (5u)-pregnan-3a-ylJ ΙΗ-indol 3-propanamide; 2-amino N- [ (20S)-20-hydroxy-19-nor(5«)-pregnan-Jcr-yl] acetamide; 2-amino H-[(20S)-20-hydroxy (5a)-pregnan-3cx-yl] N-methyl acetamide; and their salts.

The invention also provides a process for preparing the present compounds» which process comprises reacting an amine of formula II: either with an α-amino carboxylic acid, an N-alkylated a-amino carboxylic acid or a peptide containing 2 or 3 α-amino carboxylic acid or N-alkylated α-amino carboxylic acid units, the amine function of these acids or units being protected if necessary by an easily10 cleavable protecting group substituted thereon, in which case the protecting group is subsequently removed, to obtain a steroid of formula I in which Rg represents a hydrogen atom, Rg represents an acyl radical corresponding to an α-amino carboxylic acid, an N-alkylated α-amino carboxylic acid or to a peptide containing 2 or 3 α-amino carboxylic acid or N-alkylated α-amino carboxylic acid units and W, X, R-j and the wavy line are as defined above, which steroid is salified if desired, or with an alkyl halide containing 1 to 5 carbon atoms or a hydroxyalkyl halide containing 2 to 5 carbon atoms, in both cases the halogen being a chlorine, bromine or iodine atom, to obtain a steroid of formula I, in which Rg and Rj are the same or different and each represents a hydrogen atom or an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms and W, X, R^ and the wavy line are as defined above, which steroid of formula I is salified if desired, and optionally where Rj represents a hydrogen atom the steroid is reacted (a) with a halide of formula IV: Hal-R5 IV in which R''^ represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing -2 to 5 carbon atoms and Hal is as defined above, to obtain a steroid of formula I in which R2 represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms, Rg represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms and W, X, R-] and the wavy line are as defined above, which steroid is salified if desired, or (b) with an σ-amino carboxylic acid, an N-alkylated α-amino carboxylic acid or a peptide containing 2 or 3 α-amino carboxylic acid or N-alkylated α-amino carboxylic acid, units, the amine function of these acids or units being protected if necessary by an easily cleavable protecting group substituted thereon, in which case the protecting group is subsequently removed, to obtain a steroid of formula I in which R^ represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms, R^ represents an acyl radical corresponding to an a-amino carboxylic acid, an N-alkylated α-amino carboxylic acid or to a peptide containing 2 or 3 a-amino carboxylic acid or N-alkylated σ-amino carboxylic acid units and W, X, R^ and the wavy line are as defined above, which steroid is salified if desired, or with an alkyl haloformate to obtain a carbamate of the amine of formula II,which is then reduced to obtain a steroid of formula I in which R^ represents a methyl group, which steroid is salified if desired, or with an acetylating agent to obtain the N-monoacetylated steroid, which is then reduced to obtain a steroid of formula I in which R^ represents an ethyl group, which steroid is salified if desired, and the steroid is optionally reacted again with an acetylating agent and then reduced, to obtain a steroid of formula I in which Rn and R, each represents an ethyl group, which steroid is salified if desired, or with acetone in the presence of a reducing agent, to obtain a steroid of formula I in which Rg represents isopropyl and R^ represents a hydrogen atom or isopropyl, which steroid is salified if desired.

In the reaction of the amine of formula II with the α.-amino carboxylic acid, N-alkylated α-amino carboxylic acid or peptide, it is usually necessary to protect the amino function in the acid or the peptide unit, so that the amine of formula II reacts with the acid function to produce the desired compound. When the amino function is already protected by being 218 8 Ν,Ν-dialkylated, no further protection is necessary. The easily cleavable protecting group is especially such a group which can be removed by acid hydrolysis.

Under preferred conditions for carrying out the present process, b it has one or more of the following features: 1) In the reaction of the amine of formula II with the acetylating agent, the acetylating agent is preferably an acetyl halide of formula HalR'g in which Hal is as defined above and R'g represents an acetyl radical; in that event,the reaction is preferably carried out in the presence of an acid fixing agent such as an alkali metal hydroxide, an alkali metal (for example potassium) carbonate, an alkali metal bicarbonate, an alkali metal acetate, an alkaline-earth carbonate, a tertiary amine (for example a trialkylamine or pyridine) or an alkali metal alcoholate (for example sodium ethylate). The acetylating agent can be, however, the free acid R'jOH or an acid functional derivative other than the halide, especially the acid anhydride (R'gJgO· When the free acid is used, it is preferred to carry out the conversion into an amide with an activation agent such as a carbodiimide. Other techniques can be used such as those described for example in The Peptides Vol.l, Academic Press 1979.

The reaction with the acetylating agent which is the acid R^OH or an acid functional derivative thereof can be carried out for example in inert solvents or suspension media such as dioxan, dimethylformamide, benzene, toluene or methylene chloride. 2) The reaction of the amine of formula 11 or of the steroid of formula I in which R^ represents a hydrogen atom and Rg represents an. alkyl or hydroxyalkyl radical, with the a-amino carboxylic acid N-alkylated a-amino carboxylic acid or peptide, protected as necessary, preferably takes place in the presence of -a condensation agent, which activates the acid function of the acid or peptide.

As condensation agent there may be used a carbodiimide of formula : A-N=C«N-B in which A and B represent -an alkyl radical containing 1 to 8, e.g. 1 to 5 carbon atoms, possible bearing a dialkylamino radical or they represent a cycloaikyl radical.

There may he used, for example, dicyclohexylcarbodiimide or 1-ethyl 3-(3-dimethylaminopropyl) carbodiimide, preferably the latter.

A 2-chloro N-methyl pyridinium halide such as the 5 iodide can also be used.

An alkyl chloroformate such as, for example, methyl, ethyl or isobutyl chloroformate can also be used; an alkyl pyrophosphite such as, for example, ethyl pyrophosphite can also be used.

As easily-cleavable protecting group there may be used, as desired according to the compound, for example the benzyloxycarbonyl group or the (Z) tertbutyloxycarbonyl group (BOC).

It is especially preferred that the protecting group is the tertbutyloxycarbonyl radical.

To eliminate the easily-cleavable protecting group, there is used preferably as cleaving agent an acid such as hydrochloric acid; an alkanolie solution of hydrochloric acid can be employed or anhydrous hydrochloric acid can be employed by bubbling it into nitromethane containing the protected substance. Acids such as paratoluenesulphonic acid, formic acid or trifluoroacetic acid can also be used. Hydrogen in the presence of palladium can also be used for the protecting group (Z) for example. 3) The reaction of the amine of formula II with the alkyl or hydroxyalkyl halide preferably takes place in the presence of the same acid fixing agents and solvents as in the case of the reaction with the acetylating agent.

The hydroxy function of the hydroxyalkyl radical is advantageously blocked by a protecting group which is easily cleavable especially by acid hydrolysis.

This protecting group is advantageously a tetrahydropyranyl radical. It can be cleaved under the same conditions as those discussed above for the group protecting the amino acid. 4) The reaction of the steroid of formula I in which R, represents a hydrogen atom, R3 represents 2 an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms and W, X, R^ and the wavy line are as defined above with the halide of formula IV is preferably carried out under the conditions discussed above for the reaction of the amine of formula II with the acetyl ati ng agent or with the alkyl halide.

) For the preparation of the N-alkylated steroids of formula I, one can preferably proceed as follows: - to prepare a N-monomethylated steroid of formula I, a carbamate of the amine of formula II can be prepared for example by the action of an alkyl haloformate and especially an ethyl haloformate, then reduction of the carbamate for example with the aid of lithium aluminium hydride; - to prepare a N-monoethylated steroid of formula I, the corresponding N-acetylated steroid can be prepared then reduced; - to prepare the N-diethylated steroid, the procedure can he repeated.

In these three cases under item (5), when X II represents a group -C-CH^, it may he necessary to 10 prepare the ketal at position 20, for example by the action of ethylene glycol, then, after reduction, to hydrolyse the said ketal, for example with the aid of a mineral acid. - to prepare a N-mono- or di-isopropylated steroid of 15 formula I, a steroid of formula II can be reacted with acetone in the presence of a reducing agent such as sodium .cyanoborohydride.

In the present process, protection of the amine functions is clearly not necessary when these amine functions are alkylated.

The steroids of formula I are basic in nature and hence form salts with acids. The addition salts of the steroids can advantageously be prepared by reacting, in substantially 53188 stoichiometric proportions, a mineral or organic acid with the steroid. The salts can be prepared without isolating the corresponding bases.

The present compounds possessvery interesting 5 pharmacological properties; they are endowed especially with remarkable immunotherapeutic properties. They are, in particular, capable of stimulating immune reactions.

These properties indicate the use of the steroids of formula I and their pharmaceutically acceptable salts as medicaments for use in humans or animals.

Use in humans is of especial interest.

Accordingly the invention provides the present compounds or their pharmaceutical compositions for use in a method of treatment of the human or animal body by therapy’.

Preferably, the present compounds are the steroids of formula I in which X represents a group and their addition salts with pharmaceutically- acceptable acids. Among these 20 compounds, especially preferred i.re the steroids of formula I in which X represents CH, a group CH Z OH and Eg represents a hydrogen atom or a methyl radical, and their addition salts with pharmaceutically acceptable acids.

Particularly preferred is: 2-amino N-[(20S) - 20-hydroxy (5o)-pregnan -3»-yl3 propanamide; (2S) 2-amino N-i(20S)-20-hydroxy (5p)-pregnan-3cc-yl] ΙΗ-indole 3-propanamide; 2-amino N-[(20S)-20-hydroxy-19-npr (5o')-pregnan-3a-ylJ acetamide; or 2-amino N[(20S)-20-hydroxy (5a)-pregnan-3a-yl] N-methyl acetamide; or an addition salt of any of these with a pharmaceutically acceptable acid.

The present medicaments are useful, for example, in the treatment of autoimmune diseases resulting from a deficiency in certain lymphocytes, whether they be non-specific diseases of the connective tissue of an organ such as, for example, rhumatoid arthritis or systemic lupus erythematous or whether they be specific diseases of an organ such as thyroiditis, pemphigus or haemolytic anaemia.

The present medicaments can, therefore, be used as adjuvant treatment of antibiotherapy and of anticancer chemotherapy.

The usual dose, which depends on the compound used, the subject treated and the complaint concerned, can be, for example, from 10 mg to 1 g per day, by oral route in man, e.g. for the product of Example 14 used as adjuvant to antibiotherapy.

The invention provides pharmaceutical compositions which contain at least one of the present compounds as active principle.

The present compounds can be incorporated in pharmaceutical compositions intended for the oral or parental route.

The pharmaceutical compositions can be, for example, solid or liquid and can be presented in the pharmaceutical forms currently used in human medicine such as, for example, plain or sugar-coated compressed tablets, gelatin capsules, granules, suppositories and injectable preparations; they can be prepared according to the usual methods. The active compound or compounds can be incorporated in excipients usually employed in pharmaceutical compositions. Such excipients may he solid or liquid as appropriate to the pharmaceutical form chosen, and may include a wide range of organic and inorganic solids, and aqueous and non-aqueous liquids; examples include talc, gum arahic, starch, lactose, magnesium stearate or fatty substances of animal or vegetable origin such as cocoa butter, paraffin derivatives or glycols. These excipients may be compounded with one or more wetting, dispersing or emulsifying agents and/or one or more preservatives.

The starting amines of formula II,· when they are not known, can he prepared as follows; in which and the wavy line are as defined above, is reacted with an excess of ethyl triphenylphosphonium bromide and of potassium tert-butylate to obtain a product of formula VI: in which R^ and the wavy line are as defined above, which is converted, for example, by conversion to the tosylate then the action of sodium azide or. directly by reaction with diphenyl azidophosphate in the presence of ethyl azodicarboxylate and triphenylphosphine, into in which R^ and the wavy line are as defined above, and the configuration at position 3 is inverted with respect to that of the compounds V and VI, which is reduced, for example with the aid of lithium aluminium $3188 in which R^ and the wavy line are as defined above,which - either is used itself, - or is reduced for example by hydrogenation in the presence of a catalyst based on rhodium, platinum or in which R^ and the wavy line are as defined above, - or is hydrated for example with the aid of diborane to obtain a product of formula ΙΙθ: OH II, in which and the wavy lines are as defined above, which can be used · itself or subjected to oxidation for example with the aid of chromic acid, to obtain a in which and the wavy line are as defined above, - or is subjected to cis-dihydroxylation, for example with the aid of osmic anhydride in the presence of trimethyloxamine, after protection or non-protection of the amine by an easily-hydrolysable group such as a trifluoroacetyl group, to obtain a product of formula in which R represents a protecting group defined above or a hydrogen atom, and R^ and the wavy line are as defined above, which either, when R represents a hydrogen atom, can he used itself, or, when R represents a protecting group, is subjected to an agent for cleaving the said protecting group, for example by alkaline hydrolysis in the case of a trifluoroacetyl group, then the free amine obtained can be used itself, or is subjected to oxidation, for example with the aid which either, when R represents a hydrogen atom, can be used itself, or, when R represents a protecting group, is subjected to an agent for cleaving the said protecting group, then the free amine obtained can be used itself, or, is reduced, for example with the aid of sodium borohydride, to obtain a product of formula Hg.: R in which R, R-|_ and the wavy lines are as defined above, which either, when R represents a hydrogen atom, can he used itself, or, when R represents a protecting group, is subjected to an agent for cleaving the said protecting group, to produce the free amine which can be used.

Examples of the production of amines of formula II appear hereinafter.

The following Examples illustrate the invention, Example 12 illustrating the preparation of an intermediate which is reacted in Example 13 to produce a compound according to the invention.

Example 1 s (23) 2-amlno H-[(20S)-20-hydroxy 5«pregnan-5e-yl3 propanamide (hydrochloride and base).

Stage A ; 2-El.l-dimeth.vl ethoxy carbonylamino] H-[(20S)-20-h.vdroxy (5a)-pregnan-5g-yl3 propanamide. 1.92 g of (20S) Je-amino 5«-pregnan-20-ol and 2.27 β of tertbutyloxycarbonyl-L-alanin (BOO-Lalanine) were dissolved in 60 cm^ of chloroform and 12 cm^ of pyridine. The solution was agitated at 0° C to +5° 0 under inert atmosphere and 1.14 g of 1-ethyl 3-(3-dimethylaminopropyl) carbodiimide hydrochloride were added. At the end of 1 hour 15 minutes, 1.15 g of 1-ethyl 3-(5-dimethylaminopropyl)carbodiimide hydrochloride were added, the agitation was maintained for 50 minutes at 0° C to +5° C, the mixture was brought to dryness, taken up with an aqueous solution of sodium acid carbonate, extracted with ethyl acetate, washed withjg saturated aqueous •solution of sodium chloride then with a normal aqueous solution of hydrochloric acid and again with a saturated aqueous solution of sodium chloride, dried, brought to dryness, crystallized from isopropyl ether, separated, washed with isopropyl ether and dried at 60° C under reduced pressure. The title product of this Stage was obtained.

M. Pt. - 171° C.

Stage Β : Hydrochloride and base of (2S) 2-amino N-[20S)~ -20-hydroxy 5°-pregaan-3g-yl3 propanamide, variant 1 : Under inert atmosphere, 1.55 ε of the product obtained in Stage A were suspended in 100 cm^ of nitromethane, hydrogen chloride was bubbled in for 10 minutes, the mixture was agitated at 20° - 25° C for 35 minutes, the excess hydrochloric acid was eliminated, the residue was separated, washed with ethyl ether, dried at 60° C under reduced pressure and recrystallized from methanol. 990 mg of the title hydrochloride of this Stage were obtained. M.Pt - 220° C variant 2 : Under inert atmosphere, 2.7 B oi the product obtained in Stage A was dissolved in 10 cm^ of ethanol, cm^ of hydrochloric ethanol (3.5N) were added, the mixture·..was agitated for 8 hours, brought to dryness, taken up with ethyl acetate, chilled, separated, washed with ethyl acetate, dried at 60° C under reduced pressure and recrystallized from methanol. 2.2 g of the title hydrochloride of this Stage were obtained. M.Pt. - 220° C. Preparation of the base : z 1-74-g of the hydrochloride were dissolved in 100 cm 3 of tetrahydrofuran containing 30# of water, 4 cm of 2N sodium hydroxide were added, the mixture was brought to dryness, taken up with 100 cm^ of chloroform, washed with water, dried, brought to dryness and recrystallized from ethyl acetate. 1.27ε of the base were obtained. M.Pt · 224° 0 ’By proceeding analogously to the process described above, the steroids (and their hydrochlorides) listed in the Table below were prepared.

E No. of Example Variant Starting amino acid Melting Point 2 1 and BOC-L-glutamic cAch-ch--ch,-cooh (S: 220°C see note acid 1 έ 2 nh2 (hydrochloride) below 3 1 BOC-L-aerine O^CH-CH-OH (S) 210°C 1 2 HH2 (hydrochloride) 4 1 BOC-L-phenyl- cAcH-CH,-(A\ (s) 240°C . alanine1 2 ν—y NH27 (hydrochloride) 5 1 BOC-L-trypto- phan O^CH-CH./^Zi^S) ι 2 N H 135°C (base) 6 2 BOC-L-proline o^chJ (s) 275°C 1 H ;hydrochloride) 7 CF.COOH BOC-glycyl . O^CH^NH-CO-CHp-NH, 3 glycine ( 240°C hydrochloride £00 then 2^C ( base 53188 Note regarding Example 2 The second acid function of L-glutamic acid was blocked by forming its benzyl ester; it was released by catalytic hydrogenation in acetic acid in the presence of palladium, before cleavage of the BOC.

Example 8 2-amino N-[j20S) 17a,20-dihydroxy 5a-pregnan 3a-yl] acetamide hydrochloride By proceeding analogously to Example 1, variant 2, but starting with BOC-glycine and (20S) 3a-amino 5a-pregnan 17a,20-diol, the title product was obtained.

M.Ft. - 200° C Co]D - +4° i 1° (0=1.5% ethanol 95°).

The starting (20S) 3“-amino 5“-pregnan-17“,20-diol can be prepared as follows: Stage 1 : (Z) (3 a) pregn-17 (20)-en 3B-ol 59.4 g of triphenylethylphosphonium bromide was added to 16.1 g of potassium tert-butylate in solution in 160 cm^ cf. tetrahydrofuran, the mixture was agitated for 30 minutes, 23.2 g of epiandrosterone were added, the mixture was agitated for 15 minutes, poured into iced water, extracted with ethyl acetate, washed with water, dried, distilled to dryness, purified by chromatography on silica (eluant : cyclohexane / ethyl acetate, 7 ί 3 by volume), crystallized from methanol, chilled, separated and dried, 23.1 g of the title product of this Stage were obtained.

M.Pt. = 160° C.

Stage 2: (Z) 3°-azido (5“)-pregn-17(20)-ene 1.92 g of ethyl azodicarboxylate and 3·θ2 g of diphenyl azidophosphate were added to a solution of 1.66 g of the product above in 3θ cm^ of benzene and 5 cm^ of tetrahydrofuran, the mixture was agitated in an ice bath, a solution of 2.88 g of triphenylphosphine in 30 cm^ of benzene was added over 20 minutes, the mixture was agitated further for 40 minutes at 10° C, washed with a 2N solution of hydrochloric acid, then with water, dried and distilled to dryness, and purified by chromatography on silica (eluant : heptane then heptane / ethyl ether, 1 : 1 by volume). 1.67 6 of crystals of the title product arfe obtained.

M.Pt. « 114° 0 after recrystallization from methanol.

Stage 3 : (Z) 5 14.5g of (Z) 3a-azido (5“)-pregn-17(20)-eae z as obtained above were dissolved in 290 cor of tetrahydrofuran, the mixture was agitated, with heating, at 25 - 27° C,' 800 mg of lithium aluminium hydride were added over 1 hour, the mixture was agitated further for 1 hour, the excess hydride was eliminated with methanol, the residue was filtered, the filtrate was washed with an aqueous solution of Seignette salt then with a saturated aqueous solution of sodium chloride, dried and distilled to dryness. 13.1 g of crystals of the title amine of this Stage were obtained, M.Pt Ί go0 C.

The base was dissolved in 150 cm^ of ethyl acetate and 30 cm^ of methylene chloride, 27 on? of l.?N hydrochloric ethyl acetate were added, the mixture was separated and washed with ethyl acetate and the crystals obtained were dried under reduced pressure. 13·2 g of crystals of the hydrochloride were obtained.

M.Pt > 500° c. [α]ρ at 1% in pyridine containing 10% of water = +38.5° £ 1.5°· Stage 4 j N-CfZ.5tt)-nregn 17(20)-en 3a-vl] trifluoroacetamide.

Under inert atmosphere, 16.5 g the hydrochloride obtained in Stage 3 above were suspended in 3 165' cm of methylene chloride and 16.5 of pyridine, o 3 the mixture was cooled to about 5 C, 16.5 cm of 10 triflouroacetic anhydride were introduced over 5 minutes, the mixture was agitated for 15 minutes at ambient temperature and distilled to dryness under reduced pressure, 200 cm^ of water were added, the mixture was separated, washed with water and dried under reduced pressure. 18.1 g of crystals were obtained.

M.Pt. « 204° 0..

Stage 5 : N-C(20S) 17g.20-dihyd.roxy (5g)-uregnan-3a-yl] trifluoroacetamide.

. Under inert atmosphere, 18.1 g of the product 20 obtained in Stage 4 above were dissolved in 100 cm^ of methylethylketone, 9 g of dihydrated trimethylamine N-oxide were added together with a solution of 360 mg of osmium tetroxide in 71 cm^ of methylethylketone, the mixture was agitated for 2 hours at reflux and allowed ’ to cool, 200 cm^ of 10% solution of sodium thiosulphate ί I Ε ί ί in water were added, the mixture was agitated for 30 p minutes at ambient temperature, decanted, washed with i; water, dried on magnesium sulphate, filtered and r distilled to dryness under reduced pressure. The oil obtained was purified by chromatography on silica (eluant : benzene / ethyl acetate, 7 : 3 by volume). 14 g of the title product of this Stage were obtained. p M.Pt. rl?2°C then 192°C.

Stage 6 : (20S) 3e-amino 5o-pregnan-17o,20 diol.

Under inert atmosphere, 4 g of the product obtained above were dissolved in 20 cm^ of methanol, 8 cm^ of sodium hydroxide solution were added, the mixture was agitated for 1 hour 30 minutes, 50 cnr of water were added, the mixture was agitated for 10 minutes, separated, washed with water and dried under reduced pressure at 40°C. 3 g of the title product of this Stage were obtained..

M.Pt = 210°C.

Example 9 : 2-amino N-[(20S)-20-hydroxy 19-nor I. (5o)-pregnanr3ct-yl] acetamide hydrochloride.

Proceeding analogously to Example 1, variant 1, but startihg with BOC-glycine and (20S) 3a-amino-19-nor (5o)-pregnan-20-ol, the title product was obtained.

M.Pt 270°C with sublimation. [o3d +59.5° + 1.5° (c.ljimethanol).

The starting (20S) 3a-amino-19-nor (5o)-pregnan-20-ol can be prepared, as follows: Proceeding analogously to Example 8, Stages 1,2 and 3, for the preparation of (Z) 5o-pregn-17(20)-en-3o5 amine, hut starting with 19-nor-epiandrosterone, (Z) 5c-19 nor pregn 17(20)-en-3or-amine was prepared.

Under nitrogen, 156 mg of sodium borohydride were suspended in 5 cm^ of tetrahydrofuran, a solution of 0.5 cm^ of boron trifluoride etherate in 2.5 cm^ of tetrahydrofuran was added drop by drop at +5°C, the mixture was agitated for 1 hour in an ice bath, 296 mg of (Z) 5o-19 nor-pregn 17(20)-en-3o-amine in solution in 3 cm5 of tetrahydrofuran were added, the mixture was agitated for 1 hour 30 minutes at ambient temperature 3 and then cooled in an ice bath, 2 cm of 6N sodium hydroxide were added slowly, the mixture was agitated for 5 minutes at ambient temperature and decanted, the aqueous phase was extracted with tetrahydrofuran, the 3 organic phase was washed with water, 4 cm' of 5U 3 sodium hydroxide and 2 cm' of hydrogen peroxide of 110 volumes were added, and the mixture was agitated for 45 minutes, extracted with ethyl acetate, washed with water, dried and distilled to dryness under reduced pressure. The dry extract was taken up in 10 cm^ of methanol with 5 cm^ 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, dried and evaporated under reduced pressure. 257 mg of crystals of the expected product were obtained. M.Pt - 190°C. Example 10 : 2-amino N-[17»-hydroxy 20-oxo 5»pregnan-3ft-yl] acetamide hydrochloride.

Proceeding analogously to Example 1, variant 2, but starting with BOO-glycine and 3a-amino 17<*-hydroxy 5o-pregnan-20-one, the title product was obtained.

M.Pt. > 300°C. [α]β = +50° + l°(c=l% ethanol 95°).

The starting 3a-amino 17a-hydroxy 5»-pregnan-20-one can he prepared as follows; By perchromic oxidation of the product obtained in Stage 5 of Example θ> N-(17»-hydroxy-20-oxo-5a-pregnan3a-yll trifluoroaeetamide was prepared (M.Pt = 178°C then 186°0), which was treated as described in Stage 6 of Example 8, to obtain 3c-amino l?tt-hydroxy 5a-pregnan-20-one. M.Pt. = 216°C (after crystallization from water). Example Π ; 2-amino N-[(20R)-20-hydroxy 5apregnan-3a-yl] acetamide hydrochloride.

Proceeding analogously to Example 1, variant 1, hut starting with BOC-glycine and (20R) 3a-amino 5o-pr®gnan-20-ol, the title product was obtained.

M.Pt = 210°C then 260°C. [σ]ρ = +22°_+ 1° (c « 0.0% pyridine containing 10% of water). Example!2 ; Ethyl N-C(20S)-20-hydroxy (5K)-pregnan-5o-yl] carbamate Drop by drop over 10 minutes, under inert atmosphere, g of (20S) 3e-amino 5«-pregnan-20-ol in solution in 500 cm^ of methylene chloride were added to 15 om^ of ethyl chloroformate and 45 cm^ of methylene chloride, the mixture was agitated for 50 minutes, 20 cnr of N sodium hydroxide were added, the mixture was agitated further for 30 minutes, decanted, extracted with methylene chloride, washed with water, dried, filtered, evaporated to dryness under reduced pressure, taken up with ethyl ether and separated. 5.7 S°f the title product were obtained.

M.Pt - 198°C [o)D = +25°+ 1.5° (o = 1% methylene chloride).

Example 13 i 2-amino N-[(20S)-20-hydroxy 5»-prsgnan-5ct-yl] N-methyl acetamide hydrochloride.

By reduction of the product of Example 12, (20S) 3o-methylamino 5n-pregnan-20-ol (M.Pt e 170°C), described by Vetter and colleagues in Bull. Soc.(1963) 1324, was prepared. This was reacted with BOC-glyoine by a process analogous to that of Example 1, variant 2, to obtain the title hydrochloride. M.Pt a/ 25O°C.

Example 14 : 2(S) 2-amino N-methyl N-[(20S) 20-hydroxy-5o-pggKnan-5c-yl3 propanamide hydrochloride. (20S) 3a-methylamino 5«-pregnan-20-ol was reacted with BOC-L-alanine by a process analogous to that of Example 1, variant 1, to obtain the title hydrochloride M.Pt> 270°C.

Example 15 ; 2(R) 2-aaino N-methyl N-[(20S) 20-hydroxy-5o-pregnan-5c-yl] propanamide hydrochloride. (20S) 3a-methylamino-5 (20S) Ja-amino 5a-pregnan-20-ol was reacted with ethyl iodide in the presence of sodium carbonate, to obtain the base of the title product. M.Pt = 129°C after recrystallization from ethyl acetate.

The hydrochloride was prepared by the addition of hydrochloric ethyl acetate. M.Pt.^> 27O°C.

Example 17J, 2-amino N-ethyl N-[(20S) 20-hydroxy 5o-pregnan 5ft-yl3 acetamide hydrochloride.

The base of the product of Example 16 was reacted with BOC-L-glycine by a process analogous to that of Example 1, variant 1, to obtain the title hydrochloride. M.Pt - 250°C. ί Example ιβ .· (20S) 3o-[(2-hydroxyethyl) amino] 5P-pregnan-20-ol hydrochloride. g of (20S) 3a-amino 5a-pregnan-20-ol and 7 g of sodium z carbonate were suspended in 100 cnr of anhydrous dioxan, X 6.6 cm of 2-[2-bromoethoxy] tetrahydropyran were added and the mixture was refluxed for 24 hours under inert atmosphere. The resultant mixture was cooled to ambient i temperature, diluted with water, extracted with ether, dried, distilled to dryness under reduced pressure and purified by chromatography on silica (eluant ; chloroform / methanol / acetic acid, 85 : 5 s 10 by volume). 7.7 g of yellow oil were collected.

Hydrolysis of the protecting group g of the product prepared as above were dissolved z . z in 30 cnr of ethanol and 15 cm of 2N hydrochloric acid, the mixture was agitated under reflux for 1 hour 30 minutes, z cooled, poured into 100 cm of aqueous sodium bicarbonate solution, chilled for 15 minutes, separated, washed with water, dried at 40°C under reduced pressure, dissolved in 350 cm^ of methylene chloride containing 10% of 3 methanol, filtered and concentrated to about 50 cm, 100 cm' of ethyl acetate were added, the mixture was concentrated by half and the precipitate was separated. It was z disolved in 100 cm of methanol, the solution was z z filtered and concentrated to about 30 cnr, 70 cnr of ethyl acetate was added, the mixture was concentrated by half, separated and dried at 40°C under reduced pressure. 1.65 6 oi the expected base were obtained.

M.Pt = 215°C.

Formation of the hydrochloride. 1.5 g of the base prepared as above were dissolved in 40cm^of methanol, a 1.6N solution of hydrochloric ethyl acetate was added until the pH was acid, 60 cm^ of ethyl acetate were added, the mixture was concentrated under reduced pressure, separated and dried at 40°C under reduced pressure. 1.5 g of the expected hydrochloride were obtained. M.Pt 7 260°C.

Example 19 : ot-dimethylamino N-[(20S) 20-hydroxy 5a-pregnan-jfr-ylj acetamide.

Proceeding analogously to Stage A of Example 1, but using 2.552 g of (20S) 3®-amino 5o-pregnan-20-ol and 3.350 g of Ν,Ν-dimethyl glycine hydrochloride, produced 4 g of crude product containing the title product and its Ο,Ν-disubstituted derivative. By saponification with sodium hydroxide in methanol, the title product was obtained.

M.Pt = 206°C. [ot)D = +29° + 1° (C r 1% CHClj).

Example 20 : Compressed tablets were prepared, of formula: (2S) 2-amino N-[(20S)-20-hydroxy 5&-pregnan-3o-yl] propanamide hydrochloride....................... 20 mg; excipient q.s. for one compressed tablet up to .................................................100 mg.

(Detail of the excipient : lactose, starch, talc, magnesium stearate).

Example Compressed tablets were prepared of formula; 2-amino N-C(20S)-hydroxy 5o-pregnan-3o-yl) N-methyl acetamide hydrochloride .......................... 15 mg; excipient q.s. for one compressed tablet up to ... 100 mg. (Detail of the excipient; lactose, starch, talc, magnesium stearate).

Pharmacological study 1. Anaphylactic shock adjuvant Principle ; The effect of administering to animals a product capable of stimulating the activity of the immune systems is measured by an increase in the shock in response to the administration of an antigen to which the animal has been sensitized.

Male mice weighing 30 to 35 ε ar® sensitized by intra-plantar route with beef serum albumin. days later they receive this antigen by intravenous route. Under conditions of minimum sensitization the control animals do not suffer any mortal shock at the time of this last administration.

The product to he tested is injected by intraplantar route, mixed with the antigen: if this product is an adjuvant it will increase the sensitization and the result of this will be a mortal shook at the time of administration by intravenous route.

The dose which causes mortality in 5θ% or more of the animals is considered as the active dose.

Results Product of Example Dose per animal in mg 1 0.5 2 4l 4 5 5 5 6 2 7 1 8 1 9 0.5 12 5 13 0.5 The products in this table and the following 2 tables are in the form of their hydrochlorides except for the products of examples 5 and 12 which are in the form of their bases. 2. Test of rosettes with red corpuscles from sheep.

Principle : The effect of administering to animals a product capable of stimulating the activity of the immune systems is measured by an increase in their capacity for reaction to the injection of an immunogenic product.

Male rats, 3 months old, are sensitized hy intra-peritoneal route with erythrocytes from sheep (day 0). 7 days later (day 7) their spleens are removed and the splenocytes are put into contact with erythrocytes from sheep ; the percentage of leucocytes around which the erythrocytes have formed rosettes is then counted.

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

The dose of product which multiplies by about 2 the percentage of rosettes observed in the control animals is regarded as immunostimulating dose.

Results Product of Example Dose per animal in mg/kg 1 per os 1 5 2 6 >5 9 5 10 5 13 10 3. Study of the acute toxicity The lethal doses LDq of the products tested were evaluated by administration orally to the mouse.

The maximum dose not causing any mortality at the end of 8 days is called LDq.

The results are as follows: Product of Example LDq in mg/Kg 1 200 2 >400 3 >400 4 >400 5 >1000 6 >800 ' 7 >400 8 >600 9 >400 10 >1000 11 >400 12 >400 13 >200

Claims (6)

CLAIMS R3 or a salt thereof, in which W represents a hydrogen atom or a hydroxyl radical or, together with X, represents an ethylidene radical, X represents an 5 ethyl radical or a group ch3 ch3 C or CH or, together with W, an ethylidene radical, the wavy lines signify that the corresponding radical is in configuration tcorβ , R1 represents a hydrogen atom or a methyl radical, Rg represents a hydrogen atom 10 or an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms and R3 represents a hydrogen atom, an alkyl radical containing 1 to 5 carbon atoms, a hydroxyalkyl radical containing 2 to 5 carbon atoms or an acyl radical corresponding to an a-amino carboxylic acid, an N-alkylated cC-amino carboxylic 5 acid crtoapeptide containing 2 or 3 OC-amino carboxylic acid, or N-alkylated cc-amino carboxylic acid units, with the proviso that at least one of Rg and Rg does not represent a hydrogen atom, 10 and with the further provisos: i) when at the same time R1 represents a methyl radical, W and Rg each represent a hydrogen atom and X represents the radical then R3 cannot represent methyl 15 or an acyl radical corresponding to an a-amino carboxylic acid or an N-alkylated cc-amino carboxylic acid, ii) when at the same time R1 represents a methyl radical, W represents a hydrogen atom, Rg represents on a methyl radical and X represents the radical CH. , then Rg cannot represent a methyl radical, iii) when at the same time R1 represents a methyl radical, Rg and W each represent a hydrogen atom, X CH, represents a group whose hydroxyl is OH of (S) configuration and the -NRgRg radical is at position 3oc·, then Rg cannot represent a methyl. or glycyl radical, iv) when W represents a hydrogen atom, X’ represents CH, I 3 a group ,CH 7 OH position 3ec> then the substituents R^, R?J and Rg dp not each represent at the same time a methyl radical, v) when at the same time Rg and W each represent a hydrogen atom, R^ represents a methyl radical, X and the -NRgRg radical is at represents a group CHwhose hydroxyl is of CH /\ OH (Rl configuration and the -NRgRg radical is at position 3cc, then Rg' does not represent a methyl radical, and vi) when at the same time R1, Rg and Rg each represent a methyl radical and the -NRgRg radical is at position 3cc» then W and X cannot together represent an ethylidene radical.

1. Wherein 2. -amino N-C(20S)-20-hydroxy (5cc)-pregnan-3®-yl] N-methyl · acetamide, ; or a pharmaceutically acceptable salt of any of these. 13. A compound claimed in any one of claims 1 to 9 or a composition I claimed in claim 11 or 12, which compound or composition is for use in j a method of treatment of the human O r animal body by therapy. i f 5 14. A process for preparing a compound as claimed in claim 1, substantially as described herein with reference to Examples 1 to 19. 15. A compound as claimed in claim 1 whenever prepared by a process as claimed in any of claims 10 to 14. 2-amino N-[(20S)-20-hydroxy-19-nor(5ec)-pregnan-3«-ylJ acetamide, or 2 wherein X I?2 represents a hydrogen atom or a methyl radical.

-2. A compound according to claim CH, I 3 X represents a group CH ' ^OH

3. A compound according to claim CH, I 3 represents a group CH and / ''OH

4. A steroid claimed in claim 1 which is named in any one of the Examples. 5. A salt of the steroid claimed in claim 4. 6. (2S)2-amino N-[(20S)-20-hydroxy (5cc)-pregnan-3 7. (2S) 2-amino N-[(20S)-20-hydroxy (5oc )-pregnan-3 8. 2-amino N-[(20S)-20-hydroxy-19 nor (5cc.)-pregnan-3cv-yl] acetamide or a salt thereof. 9. ' 2-amino N-[(20S)-20-hydroxy (5oc-)-pregnan-3ac-yl] N-methyl acetamide or a salt thereof. 10. Process for preparing a compound claimed in any one of claims 1 to 9, which process comprises reacting an amine of formula II: in which the wavy line, W, X and are as defined in claim 1, either with an α-amino carboxylic acid, an N-alkylated cOamino carboxylic acid or a peptide containing 2 5 or 3 10 subsequently removed, to obtain a steroid of formula I in which Rg represents a hydrogen atom, R^ represents an acyl radical corresponding to an cc-amino carboxylic acid, an N-alkylated oC-amino carboxylic acid or to a peptide containing 2 or 3 15 QC-amino carboxylic.acid or N-alkylated cc-amino carboxylic acid units and W, X, R 1 and the wavy line are as defined above, which steroid is salified if desired, or with an alkyl halide containing 1 to 5 carbon 2θ atoms or a hydroxyalkyl halide containing 2 to 5 carbon atoms, in both cases the halogen being a chlorine, bromine or iodine atom, to obtain a steroid of formula I in which Rg and R^ are the same or different and each represents a hydrogen atom or an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to '5 carbon atoms and V, X, R 1 and the wavy line are as defined above, which steroid of formula I is salified if desired, 5 and optionally where Rj represents a hydrogen atom the steroid is reacted (a) with a halide of formula IV: Hal-R 3 IV in which R 3 represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical 10 containing 2 to 5 carbon atoms and Hal is as defined above, to obtain a steroid of formula I in which Rg represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical 15 containing 2 to 5 carbon atoms, R^ represents an alkyl radical containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms or an alkoxycarbonyl radical containing 2 to 8 carbon atoms and W, X, R 1 and the wavy line are as defined 20 above, which steroid is salified if desired, or (b) with aneC-amino carboxylic acid, an N-alkylated «-amino carboxylic acid or a peptide containing 2 or 3 «-amino carboxylic acid or N-alkylated 5 of these acids or units being protected if necessary by an easily cleavable protecting group substituted thereon, in which case the protecting group is subsequently removed, to obtain a steroid of formula I in which R^ represents an alkyl radical 10. containing 1 to 5 carbon atoms or a hydroxyalkyl radical containing 2 to 5 carbon atoms, R^ represents an acyl radical corresponding to an «-amino carboxylic acid, an N-alkylated qC-amino carboxylic acid or to a peptide containing 2 or 3 15 carboxylic acid or N-alkylated tc-amino carboxylic acid units and W, X, R 1 and the wavy line are as defined above, which steroid is salified -if desired, or with an alkyl haloformate to obtain a carbamate of the amine of formula II, which is then reduced to 20 obtain a steroid of formula I in which R^ represents a methyl group, which steroid is salified if desired, or with an acetylating agent to obtain the N-monoacetylated steroid, which is then reduced to obtain a steroid of formula I in which R^ represents 52188 an ethyl group, which steroid is salified if desired, and the steroid is optionally reacted again with an acetylating agent and then reduced, to obtain a steroid of formula I in which Rg and Rg each 5 represents an ethyl group, which steroid is salified if desired, or with acetone in the presence of a reducing agent, to obtain a steroid of formula I in which Rg represents isopropyl and Rg represents a hydrogen 10 atom or isopropyl, which steroid is salified if desired. 11. A pharmaceutical composition comprising a pharmaceutically-acceptable excipient together with a compound claimed in any ere of claims 1 to 9 or 15 prepared by a process claimed in claim 10. 12. A pharmaceutical composition comprising a pharmaceutically-acceptable excipient together with (2S) 2-amino N-[(20S)-20-hydroxy (ScJ-pregnan-Sttf-yl] propanamide, 20 (2S) 2-amino N-[60S)-20-hydroxy (5 1H-indol 3-propanamide,

5. 1

6. A pharmaceutical composition substantially as described herein with reference to Examples 20 and 21.