FR2494698A1 - NOVEL SUBSTITUTED 3-AMINO STEROID DERIVATIVES, THEIR SALTS, PREPARATION METHOD, MEDICAMENT APPLICATION AND COMPOSITIONS COMPRISING THE SAME - Google Patents

Abstract

NEW SUBSTITUTED 3-AMINO STEROID DERIVATIVES AS WELL AS THEIR ADDITIONAL SALTS WITH MINERAL OR ORGANIC ACIDS, OF FORMULA: (CF DRAWING IN BOPI) WH OR OH OR WX ETHYLIDENEX ETHYL ,, OR XW ETHYLIDENER H OR CHR H, ALCOYL CC, HYDROXYALCOYL C-CR H, ALCOYL CC OR HYDROXYALCOYL CC, ALCYL CC, ALCOXYCARBONYL CC, AMINO-ACYL OR PEPTIDIC REMAINDER, THEIR PREPARATION PROCESS, THEIR APPLICATION AS MEDICINAL PRODUCTS AND THE COMPONENTS CONTAINING THEM.

Description

The present invention relates to novel derivatives

  substituted 3-amino compounds and their salts, the method of

  preparation, the application as drugs of these new

  derived calves and compositions containing them.

  The subject of the invention is novel 3-amino substituted steroid derivatives and their addition salts with them.

  inorganic or organic acids, characterized in that they meet

  to general formula I: wherein W represents a hydrogen atom or hydroxyl or together with X represents a radical X represents an ethyl radical, or a group CH3 fH3 or CH or, together with W, a rad, / OH a ethylidene radical,

Ethylidene

  ne, the wavy features mean that the corresponding radical

  is in either of the configurations i and, R1.

  represents a hydrogen atom or a methyl radical, R2 represents

  hydrogen or an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms.

  carbon atoms and R3 represents a hydrogen atom, a

  alkali metal containing from 1 to 5 carbon atoms or hydroxyalkyl

  cleaving from 2 to 5 carbon atoms, an acyl or alkoxycarbonyl radical

  closing of 2 to 8 carbon atoms, or a radical derived from an amino acid or a peptide containing 2 or 3 amino acids, it being understood that:

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

  hydrogen volume, - on the other hand: i) when both R 1 represents a methyl radical, 14 and R 2 represent a hydrogen atom and X represents the radical CH 3 1 3 / C <>, in this case R 3 does not can not represent a radical o

  methyl, acetyl, or a residue derived from an amino carboxylic acid

than,

  ii) when both R 1 represents a methyl radical, W represents

  is a hydrogen atom, R2 represents a methyl radical and W

2 2494698

  CH and X represents radical C, in this case R3 can not represent a methyl or acetyl radical, iii) when both R1 represents a methyl radical, R2 and W represent a hydrogen atom, X represents a CH3 group; H%. whose hydroxyl is of configuration (S) and the OH amino radical is in position 3 W, in this case, R3 can not represent a methyl radical,

  iiii) when both R2 and W represent a hydrogen atom

  CH 1 3 gene, X represents a CH group and the amino group -OH is in the 3 "position, in this case at least one of the substituents R 1, R 2 and R 3 does not represent a methyl radical and

  iiiii) when both R2 and W represent a hydrogen atom

  gene, R1 represents a methyl radical, X represents a CH3 group

  CH whose hydroxyl is of configuration (R) and the radical

  / OH cal amino is in position 3 ", in this case, R3 does not represent

an ethoxycarbonyl radical.

  In the general formula I, and in the following, the term "alkyl radical" containing from 1 to 5 carbon atoms means, for example, a methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl radical; the term hydroxyalkyl radical containing

  from 2 to 5 carbon atoms means, for example, a radical radical

  droxyethyl or hydroxypropyl; the term acyl radical containing

  2 to 8 carbon atoms means, for example, an acetic radical,

  tyl, propionyl, n-butyryl, isobutyryl, benzoyl or nicotinoyl,

  term alkoxycarbonyl radical containing from 2 to 8 carbon atoms

  bone is, for example, a methoxy, ethoxy or

  poxycarbonyl; the residue derived from an amino carboxylic acid may be selected from the group consisting of Ala, Val, Ival, Leu, Ile, Asp, Asn, Glu, Gln, Ser, Thr, Cys, Met, Lys, Arg, Phe, Tyr , Trp, His, and Pro, Nva, Nle, Hyp, Orn, these acids being in the D or L form, as well as by Sar and Gly; when R3 represents

  a radical derived from a peptide comprising 2 or 3 c-amino acids

3 2494698

  born, these are selected from the group consisting of

cides (-amines above.

  It will be accepted by convention that the symbols of the acids o-

  amino carboxylic acids represent these acids under their

  D or L (for example, the term Ala means Alanine under

form D or form L).

  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.

  Addition salts with mineral or organic acids

  may be, for example, the salts formed with chloroacids

  hydrochloric, hydrobromic, nitric, sulfuric, phosphoric, acetic, formic, propionic, benzoic, maleic, fumaric,

  succinic, tartaric, citric, oxalic, glyoxylic, aspar-

  tick, alkane sulfonic acids such as methane or ethane

  sulfonic, arylsulfonic, such as benzene or

  ratoluene sulfonic and arylcarboxylic.

  Among the products which are the subject of the invention, mention may be made especially of the derivatives corresponding to formula I above, and also their addition salts with mineral or organic acids, characterized in that, in said formula I, X represents a CH3 OH group

  Among these, we can cite more particularly the de-

  characterized in that in said formula I, X represents

CH

  a group CH and R2 represents a hydrogen atom

  "H gene or a methyl radical, as well as their addition salts with mineral or organic acids and more particularly:

  2-amino N - [(2S0)] - 20-hydroxy (5 ") - pregnan-3" -ylpropane-

formamide,

  (2S) 2-Amino N - [(20S) -20-hydroxy (5U) -pregnan-3a-yl] -1H-

indol 3-propanamide,

  2-Amino N - [(20S) -20-hydroxy-19-nor (5α-pregnan-3-yl] acyl]

  2-amino-N - [(20S) -20-hydroxy (50) -pregnan-3 "-yl-N-methylacetamide, as well as their salts;

4 2494698

  derivatives described in the examples.

  The subject of the invention is also a method for preparing

  derivatives, as defined by the formula I above, and their salts, characterized in that an amine of formula II CH R is reacted in which the wavy line> W, X and R have the meaning already indicated, - either with a halide of formula III Hal-R'3 III in which Hal represents a chlorine, bromine or iodine atom, and R '3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 atoms of carbon, to obtain a product of formula I in which R 2 represents a hydrogen atom, R 3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, and W, X, R 1 and the wavy line have the

  meaning already indicated, a product of formula I which is isolated

  and salt if desired, - either with an amino acid or a peptide comprising 2 or 3

  amino acids whose amine function is protected by a group

  easily cleavable especially by acid hydrolysis, then

  treated in order to eliminate the protective group, to ob-

  to contain a product of formula I in which R 2 represents a hydrogen atom, R 3 represents a radical derived from a d-amino acid or a peptide comprising 2 or 3 amino-acids, and W, X, R 1 and corrugated line have the meaning already indicated, which is isolated and salified if desired,

  or with a C1-C5 alkyl halide or hydroxyalkyl

  closes from 2 to 5 carbon atoms and whose halogen is a chlorine, bromine or iodine atom, to obtain a product of

  wherein R 2 and R 3 represent a hydrogen atom

  gene or an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms and W, X, R1 and the corrugated line have the meaning already indicated,

  product of formula I that either isolates and salifies if

  or, in the case where R3 represents a hydrogen atom,

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  said product of formula I is reacted with a halide of formula IV: Hal-R "IV in which R" 3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, carbon or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hal has the meaning already indicated, to obtain a product of formula I in which R 2 represents a C 1 -C 5 alkyl or hydroxyalkyl radical containing from 2 to 5 carbon atoms, R 3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 atoms

  carbon or acyl or acyloxy containing from 2 to 8 carbon atoms.

  and W, X, R 1 and the wavy line have the meaning already indicated, which is isolated and salified if desired, or even in the case where R 3 represents a hydrogen atom, said reaction product is reacted with formula I with a 0-amino acid or a peptide comprising 2 or 3 amino acids whose amine function is protected by an easily cleavable group including

  by acid hydrolysis and then treated in such a way as to eliminate the

  protecting agent to obtain a product of formula I in the

  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 an α-amino acid or from a peptide comprising 2 or 3 acids; <-amines and W, X, R1 and the wavy line have the meaning already indicated, which one

isolates and salifies if desired.

  Under preferred conditions of implementation of the invention, the process of preparation above described, is characterized in that: - the reaction of the amine of formula II with the halide of

  Formula III is carried out in the presence of a fixing agent of

  acids, in particular alkali hydroxides, alkali carbonates (potassium carbonate), alkaline bicarbonates, alkali acetates, alkaline earth carbonates, tertiary amines (eg trialkylamines, pyridine)

  or alkali alkoxides (sodium ethoxide).

  The reaction can be carried out for example in solvents

  or inert suspension media such as aliphatic ketones

  dioxane, dimethylformamide, benzene or toluene. In the case of acylation, it is also possible

6 2494698

  use the free acid, or else another functional derivative

such as an acid anhydride.

  Another way of grafting the radical R'3 is to first convert, in the compound of formula II, the amino group to obtain the corresponding alkaline compound, by reacting it in an inert solvent such as dioxane, dimethylformamide, benzene or toluene, with

  an alkali metal, alkali hydrides or alkali metal amides

  (in particular sodium or sodium compounds) at a temperature between 0 and 150 C, and then add the agent

  acylant / chloride or acid anhydride).

  In the case where the free acid is used, it is necessary to

  to activate it by the presence of condensation agents

  that a carbodiimide or bis-alkyl amides of sulphuric acid

generous.

  The reaction of the amine of formula II or of the product of formula

  In which R3 represents a hydrogen atom and R2 represents

  An alkyl or hydroxyalkyl radical with the amino acid or peptide whose amino function is protected by an easily cleavable protecting group takes place in the presence of a condensing agent. to activate the

acid function of the amino acid.

  As a condensing agent, a carbodi

mide 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 radical

  dialkylamino or represent a cycloalkyl radical.

  There may be mentioned, for example, dicyclohexylcarbodiimide or

  1-ethyl 3- (3-dimethylaminopropyl) carbodiimide, preferably

this, the latter.

  It is also possible to use an alkyl chloroformate such as

  for example, methyl, ethyl or isobutyl chloroformate

  tyle; it is also possible to use an alkyl pyrophosphite, as

  for example ethyl pyrophosphite.

  As an easily cleavable carrier group, one can

  use, for example, the carbobenzyloxyl group or the group

carboterbutyloxyl

  The subject of the invention is in particular a method characterized

7 2494698

  in that the protecting group is the carbotubular

  tyloxyl. In order to eliminate the easily cleavable protective group above, an acid such as hydrochloric acid is preferably used as the cleavage agent; for example, an alkanolic solution of hydrochloric acid is used, or

  anhydrous hydrochloric acid by bubbling through the

  tromethane. It is also possible to use acids such as

  paratoluene sulphonic acid, formic acid or trifluoro-

  acetic. It is also possible to use hydrogen in the presence of palladium.

  The reaction of the amine of formula II with the halogenide of

  alkyl or hydroxyalkyl is 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 reaction of the product of formula I in which R3 represents

  is a hydrogen atom, R2 is a C1-C5 alkyl or hydroxyalkyl radical having 2 to 5 carbon atoms and W, X, R1

  and the wavy line have the meaning already indicated with the

  The process 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 alkyl halide.

  For the preparation of N-alkylated products of formula I, it may be advantageous in certain cases to operate in a particular way: to prepare a product of formula I N-mono-ethyl it is possible to

  prepare the corresponding N-acetyl derivative and then reduce it.

  To prepare the N-diethyl derivative, it suffices to repeat the operation

  ration to prepare a product of formula I N-mono-or di-iso

  propylated a product of formula II can be reacted with the

  ketone in the presence of a reducing agent such as cyanoborohydride

sodium.

  The derivatives of formula I with the exception of those for

  which R 3 represents an acyl or acyloxy radical have a basic character. The salts can advantageously be prepared

  of the derivatives of formula I, by reacting, in

  substantially stoichiometric portions, a mineral or organic acid with said derivative of formula I. The salts may be

  prepared without isolating the corresponding bases.

  The derivatives, object of the present invention, have very interesting pharmacological properties; they are endowed notably with remarkable immunotherapeutic properties. They

  in particular are able to stimulate the immune reactions

  tary. These properties are illustrated further in the experimental section.

  These properties justify the use of sterile derivatives

  substituted 3-amino radicals, as well as their salts, as

drugs.

  The present application is thus also intended to

  cation, as medicaments, substituted 3-amino steroid derivatives, as defined by formula I, as well as

  their addition salts with the pharmaceutically acceptable acids

tables.

  Among the medicinal products which are the subject of the invention, the medicaments characterized in that they consist of the new 3-amino substituted steroid derivatives are preferably retained.

  corresponding to formula I, in which X represents a group

  CH3 ment H as well as by their addition salts with H

  pharmaceutically acceptable acids.

  Among these, particular mention is made of those corresponding to formula I, in which X represents a fH 3 CH group, and R 2 represents a hydrogen atom or a bH radical.

  methyl, as well as their addition salts with the pharmaceutical acids.

ceutically acceptable.

  Among these, we particularly remember, the following derivatives:

  2-amino N - [(20S) -20-hydroxy (5e) -pregnan-3 "-yl / propan]

formamide,

  (2S) 2-amino N - [(20S) -20-hydroxy (5 ") - pregnan-3U-yl / 1H-

indole 3-propanamide,

  2-amino N - [(20S) -2o-hydroxy-19-nor (5m) -pregnan-30-yl]

  tamide, 2-amino-N - [(20S) -20-hydroxy (5e) -pregnan-3-yl] -N-methylacetamide,

9 2494698

  as well as their addition salts with the pharmaceutical acids

acceptable.

  The medicaments according to the invention find their use,

  for example, in the treatment of autoimmune diseases resulting from

  so much of a deficiency in certain lymphocytes, whether they are non-specific organ connective tissue diseases such as rheumatoid arthritis, lupus erythematosus

  systemic or whether they are specific diseases of an organism

  such as thyroiditis, pemphigus or haemolytic anemia.

  The products of the invention can thus be used

  as an adjuvant treatment for antibiotic and chemotherapy

anti-cancer therapy.

  The usual dose, which varies according to the derivative used, the

  treated jet and the condition in question may be, for example, 10 mg to 1 g per day, orally in humans derived from

  Example 14 as an adjunct to antibiotic therapy.

  Finally, the subject of the invention is the pharmaceutical compositions

  ticks which contain at least one aforementioned derivative or one of its addition salts with pharmaceutically acceptable acids,

as an active ingredient.

  As medicaments, the derivatives corresponding to formula I and their addition salts with pharmaceutically acceptable acids

  acceptable can be incorporated into pharmaceutical compositions.

  pharmaceuticals for the digestive or parenteral route.

  These pharmaceutical compositions may be, for example,

  solid or liquid and be in the form of pharmaceuticals

  ticks commonly used in human medicine, such as tablets, single or coated tablets, capsules, granules, suppositories, injectables; they are prepared according to the usual methods. The active ingredient (s) can be incorporated into excipients usually employed in these pharmaceutical compositions, such as talc,

  gum arabic, lactose, starch, magnesium stearate

  sium, cocoa butter, aqueous or non-aqueous vehicles, fatty substances of animal or vegetable origin, paraffinic derivatives,

  glycols, the various wetting, dispersing or emulsifying agents

stimulants, the preservatives.

  The starting materials of formula II, when they are not known, can be prepared as follows. A product of formula V: V HO 'in which R1 and the corrugated line have the meaning already indicated, is reacted with an excess of ethyl triphenylphosphonium bromide and potassium tertbutylate to give a product of formula VI: R

1 VI

  in which R1 and the wavy line have the meaning already

  indicated, which can be transformed, for example, by

  sylate then action of sodium azotine, or directly by

  action with diphenyl azidophosphate or ethyl azodicarboxylate to the azide of formula VII: VII N3

  in which R1 and the wavy line already have the meaning

  indicated, which is reduced, for example by means of hydride of

  luminium-lithium, to obtain an amine of formula VIII: ## STR1 ## in which R 1 and the corrugated line have the meaning already indicated, that

  or hydrate for example with diborane to obtain

  a product of the formula IIA: ## STR2 ## in which R 1 and the wavy lines have the meaning already indicated, whether it is isolated, or it is subjected to

  oxidation for example using chromic acid, to ob-

hold a formula IlB product:

"B

2 3

  in which R1 and the wavy line have the already indicated meaning, which is isolated and salified if desired,

  or the product of formula VII is subjected to a cis-dihydro-

  xylation, for example with trimethyloxamine, preferably

  after protection of the amine by an easily hydrolysable group such as a trifluoroacetyl group to obtain a product of formula II CH

R1 OH II-

R

  in which R represents the protective group defined below.

  above or a hydrogen atom, and R1 and the wavy line have the meaning already indicated, that or, when R represents

  a hydrogen atom is isolated, or when R represents

  a protective group, is subjected to a cleavage agent of said protecting group, for example an alkaline hydrolysis in the case of a trifluoroacetyl group, then isolated, or is subjected to said product of formula IIc to oxidation, for example using chromic acid, to obtain a product of formula IID CH3 RoN D in which R, R1 and the corrugated line have the meaning

  already indicated that, or when R represents an atom of hy-

  or when R represents a protecting group, it is subjected to a cleavage agent of said group

= 12 2494698

  protector, then isolates the obtained free amine, or

  for example using sodium borohydride to obtain

  to hold a product of formula IIE: CH3 iH3 R1, oooH HI IE H R in which R, R1 and the wavy lines have the meaning

  already indicated, that either when R represents an atom of hy-

  it is isolated and salified if desired, or when R

  has a protecting group, is subjected to a cleavage agent of said protecting group, and then isolates the free amine

obtained.

  An example of such a preparation is given below in the

Experimental part.

  The following examples illustrate the invention without however

* limit it.

  Example 1: (2S) 2-Amino N - [(20S) -20-hydroxy-5'-previn-3% -vl / Dropanamide (hydrochloride and base)

  Step A: 2- / 1,1-dimethylethoxycarbonylamino / N - / (20S) -20-

  hydroxy (5 ") - pregnan-3-yl / propanamide 1.92 g of (205) 3-chloro-5-pregnan-20-ol and 2.27 g of tert-butyloxycarbonyl-L-alanine (BOC-LAlanine) are dissolved in

  cm3 of chloroform and 12 cm3 of pyridine. The solution is stirred

  at 0 ° C / + 5 ° C under an inert atmosphere and add 1.14 g of chlorine

  1-ethyl 3- (3-dimethylaminopropyl) carbodiimide hydrate. the

  After one hour 15 minutes, 1.15 g of hydrochloride were again added.

  1-ethyl 3- (3-dimethylaminopropyl) carbodiimide, stirred for 50 minutes at 0 ° C / + 5 ° C, brought to dryness, taken up with an aqueous solution of sodium hydrogen carbonate, extracted with sodium acetate ethyl acetate, washed with a saturated aqueous solution of

  sodium chloride and then with a normal aqueous solution of

  hydrochloric acid and again with a saturated aqueous solution

  dissolved in sodium chloride, dried, evaporated to dryness, crystallized from isopropyl ether, filtered off, washed with isopropyl ether,

  dry at 60 ° C. under reduced pressure and obtain the expected product.

of. PF = 171 C.

  Stage B: hydrochloride and (2S) 2-amino N - / (20S) -base

  hydroxyl 5-pregnan-3-yl / propanamide No. 13 Lt7 variant 1: The suspension is suspended under an inert atmosphere,

  1.55 g of the product obtained in Stage A, in 100 cm3 of nitromethane.

  thane, bubbled hydrochloric acid for 10 minutes

  stirred at 20 ° -25 ° C. for 35 minutes, eliminating the excess of

  hydrochloric acid, drained, washed with ethyl ether, dried at reduced pressure under reduced pressure, recrystallized from methanol and

  990 mg of expected hydrochloride. PFO - '220 C.

  variant 2: 2.7 g of the product are dissolved under an inert atmosphere

  obtained in stage B in 10 cm3 of ethanol, adds 40 cm3 of ethanol

  hydrochloric acid (3.5N), stirred for 8 hours, brought to dryness, taken up in ethyl acetate, ice, filtered, washed with ethyl acetate, dried at 60 ° C. under reduced pressure, recrystallized

  in methanol and obtains 2.2 g of the expected hydrochloride.

PFO - 2200C.

Preparation of the base:

  1.74 g of hydrochloride are dissolved in 100 cm3 of tetrahydryl.

  drofuran 30% water, 4 cm3 of 2N sodium hydroxide, brought to dryness, taken up in 100 cm3 of chloroform, washed with water, dried, brought to dryness, recrystallized in ethyl acetate and obtained 1, 27 g

of the expected base. PF = 224 C.

  By operating according to a process similar to that described above

  above, the derivatives described in the table below have been prepared.

  below. I (s) N Amino Acid of R PF Example Starting Variant 3 2 1 + Acid BOC-L-OHCH-C 2 -COOH (S) 220 C note Glutamic NH2 (hydrochloride) 3 1 BOC-L-Serine 0 H-CH 2 OH (S) 2100 C 2 (hydrochloride) 4 1 BOC-L-Phenyl-CH-CH (S) 240 C alanine NH 2 (hydrochloride) 1 BOC-L-Trypto-) CH-CH (s) 135C phan NH 2 nv (base)

  _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ H _ _ _ _ _ _ _ _ _ _ _ _ _ _

14 2494698

  Note for Example No. 2: The second acid function of LGlutamic acid is blocked in the form of benzyl ester; it is liberated by catalytic hydrogenation in acetic acid in the presence of palladium,

before the cleavage of the BOC.

  Example 8: N - [(2OS) -20-hydroxy-5d-preanan-30-yl] -3-pyridine carboxamide 960 mg of (20S) 3-amino-5-pregnan-20ol and 813 mg of acid are dissolved under an inert atmosphere Nicotine in 30 cm3 of

  chloroform and 6 cm3 of pyridine, added 582 mg of hydrochloride

  1-ethyl 3- (3-dimethylaminopropyl) carbodiimide. After

  3 hours 30 minutes, 291 mg of 1-ethyl hydrochloride 3- (3-

  dimethylaminopropyl) carbodiimide, stirred for 16 hours, brought to dryness, taken up in 30 cm3 of water, ice, filtered, washed with water, dried at 80 ° C. under reduced pressure, recrystallized from

  methanol and 920 mg of the expected product. PF = 258 C.

  / "/ D = + 14.5 + 1 (c = 1% pyridine).

  Example 9: 2-Amino N - [(20S) 17o, 20-dihydroxy-5-pregnan-3-yl] -acetamide hydrochloride Using a procedure analogous to that described in Example 1 (variant 2) using BOC- Glycine and (20S)

  3 U-amino 5o-pregnan 17o, 20-diol gives the expected product.

PFo 2000C.

  / M / D = +4 + 1 (c = 1.5% ethanol 95).

  The starting (20S) 3c-amino 5c-pregnan-17α, 20-diol can be prepared as follows: Step 1: (Z) (5x) pregn-17 (20) -en 35-ol 59.4 g of triphenylethylphosphonium bromide to 16.1 g of potassium tert-butoxide in solution in 160 cm3 of tetrahydrofuran, stirred for 30 minutes, added 23.2 g of epiandrosterone, stirred for 15 hours, poured into water N Amino Acid of R F PF Example Starting Variant R3 6 2 BOC-L-Proline H (S) 2750C A 0A (Hydrochloride)

& H 2400C 7 CF3COOH BOC-Glycyl O CH-NCO-CEH - N -Glycine 2_NC0-CH2N hydrochloride

Glycine '200 then 252 base

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  iced, extracted with ethyl acetate, washed, dried, distilled to dryness, purified by chromatography on a silica column (eluent: cyclohexane-ethyl acetate 7-3), distilled to dryness, taken up in methanol, ice, drain, air dry and obtain 23.1 g of the expected product. Mp = 160 ° C. Stage 2: (Z) 3 "-azido (5'-pregn .. -17 (20) -ene 1.92 g of ethyl azodicarboxylate and 3.02 g are added

  of diphenyl azidophosphate to a solution of 1.66 g of

  above in 30 cc of benzene and 5 cc of tetrahydro-

  furan, stirred in an ice bath, adds in 20 minutes a solution

  triphenylphosphine in 30 cm3 of benzene, stirred

  core for 40 minutes at 10 ° C., washing, drying and dry-distilling the organic phases, purified by chromatography on silica

  (eluent: heptane then heptane-acetone 1-1), distilled and ob-

  holds 1.67 g of crystals - mp = 106 ° C - of the expected product.

  Mp = 114 ° C. after recrystallization from methanol.

  Stage 3: (Z) 5 "-pregn 17 (20) -ene-3t-amine (and hydrochloride) 14.5 g of (Z) 3a-azido (54) -pregn 17 (14.4 g) are dissolved in 290 cm3 of tetrahydrofuran ( 20) -ene as obtained above, stirred by heating at 25-27 ° C., 800 mg of lithium aluminum hydride are added to the water bath for 1 hour, the mixture is stirred for a further hour, and the excess is removed. hydride with methanol, filter, washes, dries,

  distilled to dryness and obtained 13.1 g of crystals of the amine

due. PF p-90 C.

  The base is dissolved in 150 cm3 of ethyl acetate and

  cm3 of methylene chloride, add 27 cm3 of acetate acetate

  1.7N hydrochloric acid, drained, washed and dried under reduced pressure the crystals obtained and obtained 13.2 g of crystals of the

  hydrochloride of the expected product. PFO 300 C.

  / to / D at 1% in pyridine, at 10% water = +38.5 + 1.5.

  Stage 4: N - / (Z, 5c) -pregn 17 (20) -En 3 "-yl / trifluoroacetamide

  16.5 g of the chlorine are suspended under an inert atmosphere.

  hydrate obtained in Stage 3 above, in 165 cm3 of chlorine

  methylene chloride and 16.5 cm3 of pyridine,

  5 C, 16.5 cm3 of trifluoroacetic anhydride was introduced in 5 minutes.

  stirred for 15 minutes at room temperature, distilled to dryness under reduced pressure, 200 cm3 of water

  the water, dried under reduced pressure and obtained 18.1 g of crystals.

PFo = 204 C.

  Step 5: N - / (20S) 17α, 20-dihydroxy (5 <) -pregnan-3α-yl / trifluoromethyl

  Roacetamide 18.1 g of the product obtained are dissolved under an inert atmosphere.

  above in 100 cc of methyl ethyl ketone, add 9 g of N

  trimethylamine dihydrate and a solution of 360 mg of osmium tetroxide in 71 cm3 of methyl ethyl ketone, stirred for 2 hours under reflux, allowed to cool, 200 cm3 of 10% sodium thiosulphate solution in water was added, stirred 30 minutes at room temperature, decanted, washed with water, dried over

  magnesium sulphate, filter, distilled to dryness under pressure

  extraction, purifies the oil obtained by chromatography on silica (eluent: benzene-ethyl acetate 7-3), and obtains 14 g of

  expected product; Mp = 172 ° C. and then 192 ° C.

  Step 6: (20S) 3α-amino 5oc-pregnan-17α, diol

  4 g of the product obtained under an inert atmosphere are dissolved under an inert atmosphere.

  in 20 cm3 of methanol, add 8 cm3 of sodium hydroxide solution

  of, stirred for 1 hour and 30 minutes, added 50 cm3 of water, stirred

  10 minutes, drain, wash, dry under reduced pressure at

   C and gets 3 g of base of the sought product. PF = 210 C.

  Example 10: 2-Amino N - [(2 OS) -20-hydroxy-19-nor (5 ") pregnan-3" -1-acetamide hydrochloride Using a procedure analogous to that described in Example 1 (variant 1 ) from BOC-Glycine and (20S)

  3 "-amino-19-nor (5") - pregnan-20-ol gives the expected product.

of. PFF 270 C with sublimation.

/ </ D +39.5 + 1.5 (c = i% methanol).

  The (20S) 3-amino-19-nor (50) -pregnan-20-ol starting material can be prepared as follows: By following a procedure identical to that described in

  Example 9 (Steps 1,2,3) for the preparation of the (Z) 5 "-preg-

  3017 (20) -en-3-amine, was prepared from Nor-epian

  drosterone la (Z) 5 -19 nor-pregn 17 (20) -en-3o-amine.

  156 mg of sodium borohydride in 5 ml of tetrahydrofuran are suspended under nitrogen, a solution of 0.5 ml of boron trifluoride etherate is added dropwise at +5 ° C.

  in 2.5 ml of tetrahydrofuran, stirred for 1 hour in a bath of ice

  this, added 296 mg of (Z) 5o-19 nor-pregn 17 (20) -en-3o-amine in solution in 3 ml of tetrahydrofuran, stirred for 1 hour at room temperature, cooled in an ice bath, added

  slowly 2 ml of 6N sodium hydroxide, stir for 5 minutes at

  room temperature, decanted, extracted the tetrahydro-

17 2494698

  furan, wash the organic phase, add 4 ml of 5N sodium hydroxide, 2 ml of hydrogen peroxide at 110 volumes, stir for 45 minutes, extract

  with ethyl acetate, wash, dry and dry distil the gold phase.

  under reduced pressure, the dry extract is taken up in 10 ml of methanol with 5 ml of N hydrochloric acid, the mixture is heated for 30 minutes in a bath at 50 ° C., poured into a saturated solution of sodium bicarbonate and extracted with methylene chloride. , washing,

  dry, evaporate under reduced pressure and obtain 257 mg of

  expected product rate. PF - 190 C.

  Example 11: 2-Amino N- [17α-hydroxy-20-oxo] hydrochloride

  pregnan-3-ol / acetamide By following a procedure analogous to that described in Example 1 (variant 2) from BOC-Glycine and 3-amino

  17β-hydroxy-5-pregnan-20-one, the expected product is obtained.

PF> 300 C.

/ "/ D = + 50 + 1 (c = 1% ethanol 95).

  The starting 3'-amino-17α-hydroxy-5-pregnan-20-one may be prepared as follows: Perchromic oxidation of the product obtained in Step 5 of

  Example 9 is prepared N- (17c-hydroxy-20-oxo-5-pregnan-3 <-

  yl) trifluoroacetamide (mp = 178 ° C then 186 ° C.), which is treated

  as shown in Step 6 of Example 9 to obtain the 34-

  amino 17α-hydroxy 5oc-pregnan-20-one. PF = 216 C by crystal

in water.

  Example 12: 2-Amino N - (20R) -20-hydroxy-5-hydroxychloride

  Preqnan-3 "-yl / acetamide Using a method analogous to that described in Example 1 (variant 1) from BOC-Glycine and (20R)

  3o-amino-5-pregnan-20-ol, the expected product is obtained.

PF = 2100C then 260 C.

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

  Example 13: N - / (20S) -20-hydroxy (5c) -preanan-3-yl / ethyl carbamate 5 g of (20S) 3c-amino are added dropwise in the course of 10 minutes under an inert atmosphere. , -pregnan-20-ol in solution in 500 cm3 of methylene chloride to 15 cm3 of ethyl chloroformate and 45 cm3 of methylene chloride, stirred for 30 minutes.

  minutes, add 20 cm3 of N sodium hydroxide, stir again for 30 minutes.

  Naked, decanted, extracted with methylene chloride, washed with water, dried, filtered, evaporated to dryness under reduced pressure, resumed at

  the ethyl ether, wrings out and obtains 5.7 g of the expected product.

PFo 198oC.

  /> / D = +25 + 1.5 (c = 1% methylene chloride).

  Example 14: 2-amino N - [(20S) -20-hydroxy-5H-hydrochloride

  By reduction of the product of Example 13, (20S) 3o-methylamino 5x-pregnan-20-ol (mp = 170.degree. C.) described by Vetter et al in Bull was prepared by reduction of the product of Example 13. . Soc. (1963) 1324, which is reacted with BOC-Glycine according to a method analogous to that described in

  Example 1 (variant 2) to obtain the expected hydrochloride.

PFo "2500C.

Example 15

  Tablets having the formula:

  2-amino hydrochloride N - / (20S) -20-hydroxy-5-pregnanediol

  3 "-yl / propanamide .................................... 20 mg;

  excipient q. s. for a tablet finished at ............ 100 mg.

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

Example 16

  Tablets having the formula:

  2-amino hydrochloride N - / (20S) -20-hydroxy-5-pregnane

  3a-yl / N-methyl acetamide ............................. 15 mg;

  - Excipient q. s. for a tablet finished at ............ 100 mg.

  (Details of the excipient: lactose, starch, talc, stearate

magnesium).

  Pharmacological study 1. - Adjuvant of anaphylactic shock Principle: The administration to animals of a product capable of stimulating the activity of the immune systems results in an increase of the shock in response to the administration of an

  antigen to which the animal was sensitized.

  Male mice weighing 30 to 35 g are sensitized intraplantarly with serum albumin of beef. 8 days

  later, they receive this antigen intravenously.

  In conditions of minimum sensitization the animals

  less do not make a fatal shock during the latter administration.

  tration. The product to be tested is injected intra-plantar, mixed with the antigen: if this product is an adjuvant, it goes

19 2494698

  increase awareness and it will result in a deadly shock

  during intravenous administration.

  As the active dose, the dose which causes a

  mortality equal to or greater than 50% of the animals.

  Results: 2. - Rosette test with sheep red blood cells Principle: The administration to animals of a product capable of stimulating the activity of the immune systems results in an increase in their reaction capacity to the injection

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 spleno-

  cytes are put in contact with sheep erythrocytes: the percentage of leucocytes around which the

  erythrocytes formed rosettes.

  The product to be studied is administered per os daily

from day -1 to day 1.

  An immunostimulatory dose is considered to be the dose of

  duit that multiplies by about 2 the percentage of rosettes

observed in control animals.

  Product of the example Dose per animal in mg

1 0.5

2 g 1

4 5

5

6 2

7 1

8 5

9 1

0.5

13 5

14 0.5

  Results: 3. Study of the Toxicity Assay LD0 lethal doses of the various compounds were evaluated

  tested after oral administration in mice.

  DLO is the maximum dose that does not cause any

after 8 days.

  The results are as follows: Product of the example Dose per animal in mg / kg per os

1 1

2

6> 5

8 5

5

he 5

14 10

Product of the example DLO in mg / kg

1,200

2> 400

3> 400

4> 400

> 1000

6> 800

7> 400

9> 600

> 400

he> 1000

12> 400

13> 400

14> 200

Claims (7)

  1. - New 3-amino substituted steroid derivatives, as well as their addition salts with mineral or organic acids, characterized in that they correspond to the general formula I: x R <W I   R2 R3 wherein W represents a hydrogen atom or a hydroxyl radical or together with X, represents an ethylidene radical, IH3 CH3   X represents an ethyl radical or a group or CH " O NOH   or, together with W, an ethylidene radical, the wavy lines signify that the corresponding radical is in one or other of the configurations i and (, R1 represents an atom   of hydrogen or a methyl radical, R2 represents a hydrogen atom   or 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, an alkylene radical,   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 a caprent peptide 2 or 3 e-amino acids being understood that:   - on the one hand at least one of R2, and R3 does not represent a   hydrogen volume, - on the other hand: i) when both R1 represents a methyl radical, W and R2 represent a hydrogen atom and X represents the radical CH3   % O, in this case, R3 can not represent a   dical methyl, or acetyl, or a residue derived from an amino carboxylic acid,   ii) when both R 1 represents a methyl radical, W represents   a hydrogen atom, R2 represents a methyl radical and CH X represents the radical / C <, in which case R3 can not represent a methyl or acetyl radical, 22 2494698   iii) when, at the same time, R 1 represents a methyl radical, R and W represent a hydrogen atom, X represents a CH 3 CH group whose hydroxyl is of (S) configuration, and the OH amino radical is in position 30; in this case, R3 can not represent a methyl radical,   iiii) when both R2 and W represent an atom of hy = -   CH, X represents a CH group and the OH radical   amino is in position 34, in this case at least one of the   substituents R1, R2 and R3 do not represent a methyl radical and,   iiiii) when both R2 and W represent an atom of hy-   R1 represents a methyl radical, X represents a group of   CH3 element 6 whose hydroxyl is of (R) and OH configuration   the amino group is in position 3c, in which case R3 does not represent   does not feel an ethoxycarbonyl radical.   2. - Novel steroid 3-substituted derivatives according to the   claim 1 and their addition salts with   inorganic or organic, characterized in that X represents an OH 3 group. - Novel 3-amino steroid derivatives substituted according to the   claim 2 and their addition salts with   inorganic or organic, characterized in that X represents CH3 a CH group and R2 represents a hydrogen atom OH or a methyl radical.   4. - Any one of the derivatives of formula I whose names follow:   2-amino N - [(20S) -20-hydroxy (5 ") - pregnan-3" -yl / propan] formamide,   (2S) 2-Amino N - [(20S) -20-hydroxy (5c) -pregnan-3c-yl] -1H- indole 3-propanamide,   2-Amino N - [(20S) -20-hydroxy-19No (50) -pregnan-3-yl] acetoacetate   tamide, 2-amino N - [(20S) -20-hydroxy (5) -pregnan-3-yl] N-methylacetamide,   as well as their addition salts with the mineral acids or   ganic. 5. - Process for the preparation of the derivatives of formula I, such as   defined in claim 1, as well as their salts,   characterized in that an amine of formula II: B3 R is reacted 1 X II   H2 in which the wavy line, W, X and R1 have the meaning already indicated, or with a halogenide of formula III Hal-R'3 III in which Hal 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 R 2 represents a hydrogen atom, R 3 represents an acyl or alkoxycarbonyl radical containing from 2 to 8 carbon atoms, and W, X, R1 and the wavy line have the   meaning already indicated, a product of formula I which is isolated   and salt if desired, - either with an amino acid or a peptide comprising 2 or 3   amino acids whose amine function is protected by a group   easily cleavable especially by acid hydrolysis, then   treated in order to eliminate the protective group, to ob-   to contain a product of formula I in which R 2 represents a hydrogen atom, R 3 represents a radical derived from an amino acid or a peptide comprising 2 or 3 amino acids, and W, X, R 1 and corrugated line have the meaning already indicated, which is isolated and salified if desired,   - with either a C1-C5 alkyl halide or hydroxyalkyl containing   mantle of 2 to 5 carbon atoms and whose halogen is an atom   chlorine, bromine or iodine to obtain a product of   mule I, wherein 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,   R1 and the wavy line have the meaning already indicated,   formula I which either is isolated and salified if desired, or in the case where R 3 represents a hydrogen atom, said product of formula I is reacted with a halide of formula IV: R "3 IV wherein R" 3 represents an alkyl radical containing 1   with 5 carbon atoms or hydroxyalkyl containing from 2 to 5 atoms   carbon monoxide or acyl or alkoxycarbonyl containing from 2 to 8 carbon atoms and Hal has the meaning already indicated, to obtain a product of formula I wherein R2 represents a C1-C5 alkyl or hydroxyalkyl radical containing from 2 to 5 carbon atoms, R3 represents an alkyl radical containing from 1 to 5 carbon atoms or hydroxyalkyl containing from 2 to 5 carbon atoms, or acyl or acyloxy containing from 2 to 8 carbon atoms and W, X, R1 and the corrugated line have the meaning   already indicated, whether it is insulated and salified if desired, or   in the case where R 3 represents a hydrogen atom, said product of formula I is reacted with an amino acid or a peptide comprising 2 or 3 amino acids whose function is   amine is protected by an easily cleavable group,   by acid hydrolysis and then treated to remove the protecting group, to obtain a product of formula I wherein R 2 is an alkyl radical containing 1   with 5 carbon atoms or hydroxyalkyl containing from 2 to 5 atoms   carbon monoxide, R3 represents a radical derived from an o acid (-   amino acid or a peptide comprising 2 or 3 o-amino acids and W, X, R1 and the wavy line have the meaning already indicated, that it is isolated and salified if desired.   6. - Drugs, characterized in that they consist of   the new 3-amino substituted steroid derivatives, as de-   finished by formula I of renvedication 1, as well as by   their addition salts with the pharmaceutically acceptable acids tables.   7. - Medicines, characterized in that they consist of   the new 3-amino substituted steroid derivatives, as de-   finished in one of claims 2 or 3, and by their   addition salts with pharmaceutically acceptable acids.   8. - Drugs, characterized in that they consist of steroid-substituted 3-amino derivatives, as defined in 2494698   claim 4, as well as by their addition salts with the   pharmaceutically acceptable acids.   9. Pharmaceutical compositions, characterized in that   contain, as an active ingredient, at least one of the   cameras, as defined in one of claims 6, 7 or 8.