WO1987004166A1

WO1987004166A1 - Nor-cholanic acid derivatives, a process for their preparation, their use and pharmaceutical compositions containing them

Info

Publication number: WO1987004166A1
Application number: PCT/EP1987/000006
Authority: WO
Inventors: Laura Chiodini; Mauro Gobbini; Sergio Mantegani; Daniela Ruggieri; Aldemio Temperilli; Gabriella Traquandi; Patrizia Ferrari
Original assignee: Farmitalia Carlo Erba SRL
Current assignee: Pfizer Italia SRL
Priority date: 1986-01-09
Filing date: 1987-01-07
Publication date: 1987-07-16
Anticipated expiration: 1988-07-09
Also published as: EP0253859A1; GR870012B; US4895836A; HUT46928A; DE3760176D1; FI873800A0; DK468887D0; DK468887A; KR880700813A; IL81163A0; EP0253859B1; PT84074A; PT84074B; JPS63502189A; NZ218854A; AU588668B2; ATE43350T1; IL81163A; AU6890687A; GB8600491D0

Abstract

Compounds (I) (either R1 = OH, C1-C4 alkoxy and R2 = H, C1-C4 alkyl, or R1 + R2 = a chemical bond; R3 = an aminodeoxy or aminodideoxy or aminotrideoxy sugar residue of the D and L series, the glycosidic linkage being alpha or beta: and either R4 = OH and R5 = H or R4 + R5 = a chemical bond) and their pharmaceutically acceptable salts are useful as antihypertensive agents. Their preparation and use as well as pharmaceutical compositions containing them are also described.

Description

Nor-cholanic acid derivatives, a process for their preparation, their use and pharmaceutical compositions containing them

The invention provides aminoglycoside steroids having the general formula I

wherein either R₁ represents a hydroxy group or an alkoxy group having from 1 to 4 carbon atoms and R₂ represents a hydrogen atom or an alkyl group having from 1 to 4 carbon atoms, or R₁ and R₂ taken together represent a chemical bond; R₃ represents an optionally alkyl-substituted aminodeoxy or aminodideoxy or aminotrideoxy sugar residue of the D or L series; and either R₄ represents a hydroxy group and R₅ represents a hydrogen atom or R₄ and R₅ taken together represent a chemical bond. Such sugar residues are, for example, 2-amino or 2-alkylamino-2-deoxy-hexopyranosyl, 3-amino or 3-alkylamino-3-deoxy-hexopyranosyl, 3-amino or 3-alkylamino-3,6-dideoxy-hexopyranosyl, 3-amino or 3-alkyl-amino-2,3,6-trideoxy-hexopyranosyl and 4-amino or 4-alkylamino-2,4,6-trideoxy-hexopyranosyl residues of the D and L series. The optional alkyl substituents of the sugar residue are preferably lower alkyl with C₁-C₄ atoms, e.g. methyl, ethyl, propyl and butyl. Advantageously the alkyl radicals may substitute the amino group of the sugar residue. The wavy lines in the formulae indicate that the hydrogen atom or the substituent may be above or under the plane of the ring system providing optically active isomer forms having different absolute configurations. These optically active forms either in the form of the racemate or as pure optical antipodes are encompassed by this invention. The racemates can be separated in accordance with methods known per se. Preferably the racemic mixture is reacted with an optically active separting agent to form diastereomers. As separating agents e.g. optically active acids such as the D- and L-form of tartaric acid, diacetyl-tartaric acid, dibenzoyl-tartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphersulfonic acids like β-camphersulfonic acid may be mentioned.

Of course it is also possible to obtain optically active compounds of formula I in utilizing starting materials being optically active.

The glycosidic linkage can be a or β. An alkoxy group may be methoxy, ethoxy, propoxy or butoxy group, an alkyl group may be methyl, ethyl, propyl or butyl group.

Pharmaceutically acceptable salts of these aminoglycoside steroids are included in the invention.

"Pharmaceutically acceptable salts" refers to those salts which retain the biological effectiveness and properties of the free basis and which are not biologically or otherwise undesirable. Such salts are formed with inorganic acids such as hydrochloric, hydrobromic, sulfuric, nitric or phosphoric acid and organic acids such as acetic, propionic, glycolic, pyruvic, oxalic, malic, malonic, succinic, maleic, fumaric, tartaric, citric, benzoic, cinnamic, mandelic, methanesulfonic, ethanesulfonic, p-toluene sulfonic or salicylic acid. The invention further provides a process for the preparation of the aminoglycoside steroids of the general formula I as herein defined, which process comprises condensing a nor-cholane derivative having the general formula II

wherein R₄ and R₅ have the above meanings with a protected 1-halo derivative of an optionally alkyl substituted aminodeoxy, aminodideoxy or aminotrideoxy sugar of the D or L series and removing the protecting groups from the resultant compound. The removal of the protecting group is advantageously obtained with a base.

The condensation is desirably carried out in a suitable organic solvent, such as chloroform, methylene dichloride, benzene, toluene, acetonitrile or nitromethane, in the presence of a catalyst. The catalyst may preferably by silver oxide or carbonate or trifluoromethanesulphonate, Fetizon's reagent, mercuric cyanide or bromide. In the process it may be desirable to use drierite or a molecular sieve as drying agents or the azeotropic distillation method during reaction, at a temperature of -5°C to 100°C. The reaction time may vary broadly and often be from one to eight hours. The base used to remove the protecting groups may be a hydroxide or methoxide of sodium or potassium or barium or may be triethylamine, and the reaction may be performed at room temperature over a period of some hours to several days.

This operation generally gives compounds of the general formula I wherein: R₁ is hydroxy or alkoxy group and R₂ is a hydrogen atom or alkyl group. These compounds may be converted by cyclization into the other compounds of the general formula I wherein R₂ and R₂ together represent a chemical bond, forming a lactone ring. The cyclization may be carried out in hydrochloric acid at room temperature and pH 1-2 for from three to five hours.

The nor-cholane derivatives of the formula II and the aminodeoxy, aminodideoxy and aminotrideoxy sugars, possibly in alkylated form respectively, used a starting materials are well known compounds or they may be prepared by procedures familiar to those skilled in the art.

The aminoglycoside nor-cholanic acid derivatives according to the invention and their pharmaceutically acceptable salts are capable of inhibiting specific ouabain binding without inhibiting Na⁺, K⁺-ATPase activity and thus they may form useful pharmaceutical compositions, particularly for the treatment of hypertension. Thus the inventive compounds may also be used for the making of medicaments effective against hypertension.

'In vitro' assays to test the ability of aminoglycoside steroids of formula I to displace specific ouabain binding to the (Na⁺ -K⁺)-ATPase receptors without inhibiting the

(Na⁺ -K⁺ ) -ATPase enzymatic activity.

Radiochemical assay:

A microsomial fraction enriched in (Na⁺ -K⁺) -ATPase was prepared from dog kidney outer medulla, according to Jorgensen (BBA 356: 36-52, 1974).

The partially purified enzyme (0.5 μg of protein) was incubated in 3 mM MgCl₂, 3 mM EGTA, 80 mM Hepes buffer

(pH 7.4) and 2 mM y² - P³²-ATP, final volume 110 μl, at

37ºC for 15 minutes with increasing concentrations of ouabain (as reference compound) or aminoglycoside steroids.

The reaction was stopped by the addition of 0.1 mM of cold perchloric acid (10% final concentration) and 0.5 ml of charcoal suspension (20% w/v). The suspension was centrifuged and the content of ³²P in the supernatant was measured by liquid scintillation counting. (ref. Mall F. et al.; Biochem. Pharm. 33: N.1, 47-53. 1984).

The effects of various concentrations of aminoglycoside steroids and ouabain were expressed as a percentage of inhibition of the total (Na⁺-K⁺)-ATPase activity and IC₅₀ values were calculated. The compounds of the formula I are inactive in this test.

Displacement of ouabain (H³) binding from human red blood cells

The procedure has been described elsewhere (Erdmann E. et al.; Arzneim. Forsh 34(11). no.10: 1314, 1984).

Washed erythrocytes (about 1-1.8 x 10 ⁹ /ml) were incubated in 130 mM NaCl, 1 mM MgCl₂, 10 mM glucose, 10 mM sucrose, 10 mM Tris/HCl buffer (pH 7.4) 2x10^-9M ³H ouabain and increasing concentration of the unlabelled aminoglycoside steroids, at 37°C for 5 hours. Bound ouabain was quantitated by a rapid filtration technique (Whatman GF/C glass filter membranes ; 'Whatman' is a Trade Mark) to separate free from membrane-bound ouabain. The radioactivity in the filters was determined by liquid scintillation counting Non specific binding was defined as the binding in the presence of 10^-3M unlabelled ouabain.

This dissociation constant (K_D value) was calculated from the concentration of unlabelled aminoglycoside steroids which inhibit ³H-ouabain bindung by 50% at equilibrium, by the method of Erdmann et al. (Schmiedeberg's Arch. Pharmacol. 283: 335, 1973). The compounds of the formula I are effective in this test with a K_D value range of from 10^-9 to 10^-6.

Inhibition of Na⁺ efflux mediated by the (Na⁺ -K⁺)--ATPase in human red blood cells

The procedure has been described elsewhere (Garay et al. , Biochem. Pharmacol. 33:2013-2020, 1984). Washed red blood cells were suspended to a hematocrit of 20-25% in 74 mM

MgCl₂, 2 mM KCl, 84 mM sucrose, 10 mM MOPS/Tris buffer (pH 7.4 at 37°C) and 10 mM glucose.

Red cell suspensions were added in the cold to tubes containing Mg⁺⁺ sucrose-K⁺ medium with increasing concentration of ouabain and fixed concentrations of aminoglycoside steroids. The tubes were incubated at 37ºC and aliquots of the suspensions were transferred to the cold and spun down at different times (0 - 10 - 20 - 30 minutes). External Na concentrations were measured in the superna tents by atomic absorption. A kinetic analysis of the inhibition of ouabain sensitive Na⁺ efflux as a function of different aminoglycoside concentrations was done and the IC₅₀ for each compound was calculated. The compounds of the formula I are effective in a concentration range from 10^-9 to 10^-6 M.

'In vivo' assays to test the hypotensive activities of aminoglycoside steroids of formula I

Indirect measurements of systolic blood pressure was carried out in groups of 4 spontaneously hypertensive rats (SHR, Kyoto). 8 to 10 weeks of age, supplied by Charles Rives, Italy. The animals were maintained in an environment of 36°C for 10 to 15 minutes to allow pulse pressure to be recorded and then systolic blood pressure and heart rate were measured by the indirect tail cuff method using a W+W, BP recorder, model 8005. The compounds were given orally, suspended in 5% arabic gum, once a day for 4 consecutive days and measurements were carried out before beginning the treatment and 1 and 5 hours after dosing in both the first and fourth day of treatment. Control animals received the vehicle only (0.2 ml/100 g body weight). Drug induced changes in systolic blood pressure were calculated as differences from the pretreatment values.

The formulation of the compounds of the invention as pharmaceutical composition may include solid formulations such as capsules, tablets and powders, or liquid formulations such as elixirs, syrups and suspensions for oral administration. Alternatively, the inventive compounds (I) may be formulated as injections or suppositories.

A carrier and diluent may be included in the pharmaceutical composition which is selected from pulverulent solid carriers such as lactose, saccharose, dextrose, mannitol, sorbitol, cellulose, and glycine etc.

The composition may further contain a lubricant, a binder or a disintegrator. Examples of suitable lubricants are silicon dioxide, talc, magnesium stearate and polyethylene glycol. Examples of suitable binders are starch, gelatin, tragacanth, methyl cellulose and polyvinyl pyrrolidone. Examples of suitable disintegrators are starch and agar etc. The following Examples illustrate preferred embodiments of the invention.

EXAMPLE 1

3- [(3-amino-3-deoxy-β-D-glucopyranosyl) oxy]-1 4 , 21 -dihydroxy- - ( 3 β , 5 β , 14 β , 2OR) -24-nor-cholanic acid .

376 mg of 3,14,21-trihydroxy-(3β,5β,14β,2OR)-24-nor-cholanic acid lactone were dissolved in 100 ml of anhydrous benzene, 3.4 g of Fetizon reagent were added, the mixture was brought to reflux and, with azeotropic distillation, a solution of 928 mg of 3-deoxy-2,4,6-tri-O-acetyl-3-trifluoroacetamido-α¬-D-glucopyranosyl bromide in 100 ml of anhydrous benzene was added dropwise over 3 hours. The mixture was cooled to r.t., filtered, the organic solution washed with a saturated sodium hydrogen carbonate solution, with a saturated sodium chloride solution, dried over sodium sulfate and evaporated to dryness. The crude residue was chromatographed on silica gel with cyclohexane-ethyl acetate 1:1 to afford 531 mg of 3-[(3-deoxy- 2,4,6-tri-O-acetyl-3-trifluoroacetamido-α-D-glucopyranosyl)oxy]- -14,21-dihydroxy-(3β,5β,14β,2OR)-24-nor-cholanic acid lactone. 531 mg of the aforementioned protected glycoside were dissolved in 28 ml of 0.1 N potassium hydroxide in methanol-water 99:1. After 24 hours at r.t., the solution was evaporated to dryness, the residue dissolved in water and acidified at pH 5 by addition of acetic acid. The resulting precipitate was filtered, washed with water and dried to afford 300 mg of the title compound. EXAMPLE 2

3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy--(3β,5β,14β,2OR)-24-nor-cholanic acid lactone.

540 mg of 3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-14,21--dihydroxy- (3β,5β,14β,2OR)-24-nor-cholanic acid were suspended in 150ml of distilled water and brought into solution acidifying at pH 1 with 1 N hydrochloric acid.

After 1 hour at r.t. the solution was brought at pH 8 with a saturated sodium hydrogen carbonate solution, extracted with chloroform-methanol 8:2, the organic layer washed with a saturated sodium chloride solution, dried over sodium sulfate, evaporated to dryness and crystallized from absolute ethyl alcohol to afford 430 mg of the title compound, m.p. 278-282 C.

EXAMPLE 3

3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy¬-(3β,5β,14β,20S)-24-nor-cholanic acid.

Operating as in Example 1 but employing 3,14,21-trihydroxy-(3β,5β,14β,2OS)-24-nor-cholanic acid lactone, the title compound was obtained in 58% yield.

EXAMPLE 4

3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,2OS)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3--deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid, the title compound was obtained in 78% yield.

EXAMPLE 5

3-[( 3-amino-3-deoxy-6-D-glucopyranosyl) oxy] -21 -hydroxy- ( 3 β , 5 β , 20R) -Δ ¹⁴-24-nor-cholenic acid.

Operating as in Example 1 but employing 3,21-dihydroxy- (3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone, the title compound was obtained in 72% yield.

EXAMPLE 6

3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy-(3 ,5 ,20R)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 90% yield, m.p. 188-190°C. EXAMPLE 7

3-[(3-amino-3-deoxy-β-D-glucoρyranosyl)oxy]-21-hydroxy- (3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 1 but employing 3,21-dihydroxy¬-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone, the title compound was obtained in 69% yield.

EXAMPLE 8

3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy¬-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy-(3β,5β,2OS)-Δ¹⁴-24-nor- -cholenic acid, the title compound was obtained in 79% yield, m.p.190-192°C.

EXAMPLE 9

3-[(2-amino-2-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy¬-(3β,5β,14β,20R)-24-nor-cholanic acid.

Operating as in Example 1 but employing 2-deoxy-3,4,6-tri-O¬-acetyl-2-trifluoroacetamido-α-D-glucopyranosyl bromide, the title compound was obtained in 52% yield.

EXAMPLE 10

3-[(2-amino-2-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(2-amino-2-deoxy-β-D-glucopyranosyl)oxy]-14,21-dihydroxy-(3β,5β, 14β,20R)-24-nor-cholanic acid, the title compound was obtained in 78% yield.

EXAMPLE 11

3-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]- -14,21-dihydroxy-(3β,5β,14β,2OR)-24-nor-cholanic acid.

Operating as in Example 1 but employing 2,3,6-trideoxy¬-3-trifluoroacetamido-4-O-trifluoroacetyl-α-L-lyxo-hexopyranosyl chloride, the title compound was obtained in 63% yield.

EXAMPLE 12

3-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]- -14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-2,3,6¬-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-14,21-dihydroxy-(3β, 5β,14β,20R)-24-nor-cholanic acid, the title compound was obtained in 75% yield. EXAMPLE 1 3

3-[(3-amino-2,3,6,trideoxy-α-L-lyxo-hexopyranosyl)oxy]¬-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid.

Operating as in Example 3 but employing 2,3,6-trideoxy¬-3-trifluoroacetamido-4-O-trifluoroacetyl-α-lyxo-hexopyranosyl chloride, the title compound was obtained in 60% yield.

EXAMPLE 14

3-[(3-amino-2,3,6-trideoxy-a-L-lyxo-hexopyranosyl)oxy]¬-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-2,3,6¬-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-14,21-dihydroxy-(3β, 5β,14β,20S)-24-nor-cholanic acid, the title compound was obtained in 74% yield.

EXAMPLE 15

3-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 5 but employing 2,3,6-trideoxy-3¬-trifluoroacetamido-4-O-trifluoroacetyl-α-L-lyxo-hexopyranosyl chloride, the title compound was obtained in 70% yield. EXAMPLE 16

3-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-2,3,6¬-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-21-hydroxy-(3β,5β, 20R)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 80% yield, m.p. 182-186°C.

EXAMPLE 17

3-[_(3-amino-2,3,6-trideoxy-α-L-lyxo-hexoρyranosyl)oxy]-21¬-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example- 7 but employing 2,3,6-trideoxy-3-tri-fluoroacetamido-4-O-trifluoroacetyl-α-L-lyxo-hexopyranosyl chloride, a mixture of α and β-glycosides was obtained in

70% yield.

By crystallization, the title compound was obtaiend in 40% yield, m.p.161-163°C.

EXAMPLE 18

3-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxyl-21¬-hydroxy-(3β,5β,2OS)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-2,3,6¬trideoxy-α-L-lyxo-hexopyranosyl)oxy]-21-hydroxy-(3β,5β ,20S)¬-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in

75% yield, m.p. 220-224°C. EXAMPLE 19

3-[(3-amino-2,3,6-trideoxy-β-L-lyxo-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20S)-Δ¹⁴-nor-cholenic acid.

The residue of the mother liquors obtained in Example 17 was crystallized giving the title compound in 25% yield.

EXAMPLE 20

3-[(3-amino-2,3,6-trideoxy-β-L-lyxo-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20S)-Δ¹⁴-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-2,3,6-trideoxy-β-L-lyxo-hexopyranosyl)oxy]-21-hydroxy-(3β,5β,2OS)-Δ¹⁴-nor-cholenic acid, the title compound was obtained in

76% yield.

EXAMPLE 21

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)-oxy]-14,21¬-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid.

Operating as in Example 1 but employing 2,4-di-O-acetyl-3,6-dideoxy-3-trifluoroacetamido-α-L-mannopyranosyl bromide, the title compound was obtained in 75% yield.

EXAMPLE 22

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-14,21¬-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3,6¬-dideoxy-α-L-mannopyranosyl)oxy]-14,21-dihydroxy-(3β,5β, 14β,20R)-24-nor-cholanic acid, the title compound was obtained in 80% yield.

EXAMPLE 23

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-14,21-dihydroxy¬-(3β,5β,14β,20S)-24-nor-cholanic acid.

Operating as in Example 3 but employing 2,4-di-O-acetyl-3,6¬-dideoxy-3-trifluoroacetamido-α-L-mannopyranosyl bromide, the title compound was obtained in 60% yield.

EXAMPLE 24

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-14,21-dihydroxy¬-(3β,5β,14β,20S)-24-nor-cholanic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3,6-dideoxy¬-α-L-mannopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor¬cholanic acid, the title compound was obtained in 95% yield.

EXAMPLE 25

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-21-hydroxy- (3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 5 but employing 2,4-di-O-acetyl-3,6¬-dideoxy-3-trifluoroacetamido-α-L-mannopyranosyl bromide, the title compound was obtained in 60% yield.

EXAMPLE 26

3- [( 3-amino-3 , 6-dideoxy-α-L-mannopyranosyl) oxy] -21 -hydroxy(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2 but employing 3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-21-hydroxy-(3β,5β,2OR)¬-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 80% yield.

EXAMPLE 27

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)oxy]-21-hydroxy- (3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 7, but employing 2,4-di-O-acetyl-3,6-dideoxy-3-trifluoroacetamido-α-L-mannopyranosyl bromide, the title compound was obtained in 66% yield. EXAMPLE 28

3-[(3-amino-3,6-dideoxy-α-L-mannopyranosyl)-oxy]-21--hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino¬-3,6-dideoxy-α-L-mannopyranosyl)oxy]-21-hydroxy-(3β,5β, 20S)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 74% yield.

EXAMPLE 29

3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid.

Operating as in Example 1, but employing 3,6-dideoxy-2,4¬-di-O-acetyl-3-trifluoroacetamido-α-D-altropyranosyl bromide, the title compound was obtained in 70% yield.

EXAMPLE 30

3-[(3-aminp-3,6-dideoxy-α-D-altropyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,2OR)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-3,6¬-dideoxy-α-D-altropyranosyl)oxy]-14,21-dihydroxy-(3β,5β, 14β,2O0)-24-nor-cholanic acid, the title compound was obtained in 75% yield. EXAMPLE 31

3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-14,21¬-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid.

Operating as in Example 3, but employing 3,6-dideoxy-2,4¬-di-O-acetyl-3-trifluoroacetamido-α-D-altropyranosyl bromide, the title compound was obtained in 55% yield.

EXAMPLE 32

3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-14,21¬-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[-(3-amino-3,6¬-dideoxy-α-D-altropyranosyl)oxy]-14,21-dihydroxy-(3β,5β, 14β,20S)-24-nor-cholanic acid, the title compound was obtained in 73% yield.

EXAMPLE 33

3-[ ( 3-amino-3 , 6-dideoxy-α-D-altropyranosyl) oxy] -21 -hydroxy¬(3β,5β, ,20R)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 5, but employing 3,6-dideoxy-2,4-di¬-O-acetyl-3-trifluoroacetamido-α-D-altropyranosyl bromide, the title compound was obtaiend in 75% yield.

EXAMPLE 34

3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-21-hydroxy- (3β,5β, 20R)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-21-hydroxy-(3β,5β,14β,20R)¬-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 77% yield.

EXAMPLE 35

3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-21-hydroxy- -(3β,5β, 20S)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 7, but employing 3,6-dideoxy-2,4-O¬-di-acetyl-3-trifluoroacetamido-α-D-altropyranosyl bromide, the title compound was obtained in 80% yield.

EXAMPLE 36

3-[_(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-21-hydroxy¬-(3β,5β, 20S)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-3,6-dideoxy-α-D-altropyranosyl)oxy]-21-hydroxy-(3β,5β,14β,2OS)-Δ¹⁴¬-24-nor-cholenic acid, the title compound was obtained in 73% yield. EXAMPLE 37

3-[(3,6-dideoxy-3-dimethylamino-α-D-altropyranosyl)oxy]¬-14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid.

Operating as in Example 1, but employing 3,6-dideoxy-2,4¬-di-O-acetyl-3-dimethylamino-α-D-altropyranosyl bromide hydrobromide, the title compound was obtaiend in 77% yield.

EXAMPLE 38

3-[(3,6-dideoxy-3-dimethylamino-α-D-altropyranosyl)oxy]-14,21¬-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[(3,6-dideoxy-3-dimethylamino-α-D-altropyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β, 20R)-24-nor-cholanic acid, the ttile compound was obtained in 80% yield.

EXAMPLE 39

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-14,21¬-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid.

Operating as in Example 1, but employing 2,3,6-trideoxy-3-trifluoroacetamido-4-O-trifluotoacetyl-α-D-arabino-hexopyranosyl chloride, a mixture of α and β-glycosides was obtained in 63% yield. The title compound was obtained by crystallization in 30% yield. EXAMPLE 40

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]- 14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-2,3,6¬-trideoxy-α-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy- (3β,5β,14β,20R)-24-nor-cholanic acid, the title compound was obtained in 67% yield.

EXAMPLE 41

3-[(3-amino-2,3,6-trideoxy-6-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid.

The residue of the mother liquors obtained in Example 39 was crystallized giving the title compound in 28% yield.

EXAMPLE 42

3-[(3-amino-2,3,6-trideoxy-6-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20R)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-2,3,6¬-trideoxy-β-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β, 5β,14β,20R)-24-nor-cholanic acid, the title compound was obtained in.73% yield. EXAMPLE 43

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid.

Operating as in Example 3, but employing 2,3, 6-trideoxy-3-trifluoroacetamido-4-O-trifluoroacetyl-α-D-arabino-hexopyranosyl chloride, the title compound was obtained in 55% yield.

EXAMPLE 44

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β,5β,14β,20S)-24-nor-cholanic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-2,3,6¬-trideoxy-α-D-arabino-hexopyranosyl)oxy]-14,21-dihydroxy-(3β,5β, 14β,20S)-24-nor-cholanic acid, the title compound was obtained in 78% yield.

EXAMPLE 45

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 5, but employing 2,3,6-trideoxy-3-trifluoroacetamido-4-O-trifluoroacetyl-α-D-arabino-hexopyranosyl chloride, the title compound was obtained in 58% yield. EXAMPLE 46

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β, 2OR)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 75% yield.

EXAMPLE 47

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid.

Operating as in Example 7, but employing 2,3,6-trideoxy-3¬-trifluoroacetamido-4-O-trifluoroacetyl-α-D-arabino-hexopyranosyl chloride, the title compound was obtained in 60% yield.

EXAMPLE 48

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21¬-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone.

Operating as in Example 2, but employing 3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β, 20S)-Δ ¹⁴-24-nor-cholenic acid, the title compound was obtained in 75% yield. EXAMPLE 49

3-[(2-amino-2-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 7, but employing 2-deoxy-2-diphenyl¬oxyphosphorylamino-3,4,6-O-acetyl-α-D-glucopyranosyl bromide, the title compound was obtained in 55% yield.

EXAMPLE 50

3-[(2-amino-2-deoxy-β-D-glucopyranosyl)oxy]-21-hydroxy¬-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone Operating as in Example 2, but employing 3-[(2-amino-2-deoxy¬-β-D-glucopyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor¬-cholenic acid, the title compound was obtained in 70% yield, m.p. 192-194°C.

EXAMPLE 51

3-[(3-amino-3,6-dideoxy-α-D-mannopyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 7, but employing 2,4-di-O-acetyl-3,6¬dideoxy-3-trifluoroacetamido-α-D-mannopyranosylbromide, the title compound was obtained in 65% yield.

EXAMPLE 52

3-[(3-amino-3,6-dideoxy-α-D-mannopyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20S)-Δ¹⁴-24-nor-chofenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-3,6-dideoxy-α-D-mannopyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴¬-nor-cholenic acid, the title compound was obtained in 70% yield, m.p. 200-202°C.

EXAMPLE 53

3-[ (3-amino-3-deoxy-α-D-mannopyranosyl ) oxy]-21-hydroxy¬

- ( 3β, 5β, 20S ) -Δ¹⁴-24-nor-cholenic acid

Operating as in Example 7, but employing 2,4,6-tri-O-acetil¬-3-deoxy-3-trifluoroacstamido-α-D-mannopyranosyl bromide, the title compound was obtained in 75% yield.

EXAMPLE 54

3-[(3-amino-3-deoxy-α-D-mannopyranosyl)oxy]-21-hydroxy¬-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-3¬-deoxy-o(-D-mannopyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴¬-nor-cholenic acid, the title compound was obtained in 85% yield, m.p. 251-253°C.

EXAMPLE 55

3-[(3-amino-3-deoxy-α-D-mannopyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 5, but employing 2,4,6-tri-O-acetyl¬-3-deoxy-3-trifluoroacetamido-α-D-mannopyranosyl bromide, the title compound was obtained in 50% yield.

EXAMPLE 56

3-[(3-amino-3-deoxy-α-D-mannopyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-3¬-deoxy-α-D-mannopyranosyl)oxy]-21-hydroxy-(3β,5β,20R) - Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 70% yield, m.p. 230-232°C.

EXAMPLE 57

3-[(3-amino-3,6-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 7, but employing 3,6-dideoxy-2,4-O-di-acetyl-3-trifluoro-acetamido-α-L-altropyranosylbromide, the title compound was obtained in 75% yield.

EXAMPLE 58

3-[(3-amino-3,6-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-3,6¬-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 72% yield, m.p. 192-194°C.

EXAMPLE 59

3-[(3-amino-3,6-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy¬

-(3β,5β,20R)-Δ¹⁴- 24-nor-cholenic acid

Operating as in Example 5, but employing 3,6-dideoxy-2,4-O-di-acetyl-3-trifluoroacetamido-α-L-altropyranosyl bromide, the title compound was obtained in 65% yield.

EXAMPLE 60

3-[(3-amino-3,6-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid lactone Operating as in Example 2, but employing 3-/(3-amino-3,6-dideoxy-α-L-altropyranosyl)oxy]-21-hydroxy-(3β,5β,20R)¬-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 83% yield, m.p. 152-155°C.

EXAMPLE 61

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)-oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 7, but employing 2,3,6-trideoxy-3¬-trifluoroacetamido-4-O-trifluoroacetyl-α-D-arabino-hexopyranosyl chloride, the title compound was obtained in 62% yield.

EXAMPLE 62

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-,hydroxy-(3β,5β,20S)-Δ¹⁴-24-nor-cholenic acid, the title compound was obtained in 72% yield, m.p. 222-226°C.

EXAMPLE 63

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]¬-21-hydroxy-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid

Operating as in Example 5, but employing 2,3,6-trideoxy-3¬-trifluoroacetamido-4-O-trifluoroacetyl-α-D-arabino-hexopyranosylchloride, the title compound was obtained in 68% yield. EXAMPLE 64

3-[(3-amino-2,3,6-trideoxy-α-D-arabino-hexopyranosyl)oxy]-21-hydroxy-(3β,5β,2OR)-Δ¹⁴-nor-cholenic acid lactone

Operating as in Example 2, but employing 3-[(3-amino-2,3,6-trideoxy-α -D-arabino-hexopyranosyl)-oxy[-21-hydroxy¬-(3β,5β,20R)-Δ¹⁴-24-nor-cholenic acid, the title componnd was obtained in 78% yield, m.p. 192-195°C.

The invention is illustrated further in the below formulation examples:

Formulation 1

Ingredient Part compound of the formula (I) 45 starch 10 lactose 45

The ingredients are mixed thoroughly, and tablets or capsules are formulated from the mixture.

Formulation 2 Ingredient Part compound of the formula (I) 10 lactose 75 magnesium oxide (MgO > 96%) 15 The above ingredients are mixed thoroughly, and powders or fine granules are formed from the mixture. Formulation 3

Ingredient Part compound of the formula (I) 1 surface active agent 1 physiological saline 98 The above ingredients are mixed under warming, and dispensed under sterile conditions into ampoules for use as injections.

Claims

CLAIMS :

A compound having the general formula I

wherein either R₁ represents a hydroxy group or an alkoxy group having from 1 to 4 carbon atoms and R₂ represents a hydrogen atom or an alkyl group having from 1 to 4 carbon atoms, or R₁ and R₂ taken together represent a chemical bond; R₃ represents an aminodeoxy or aminodideoxy or aminotrideoxy sugar residue of the D or L series; and either R₄ represents a hydroxy group and R₅ represents a hydrogen atom; or R₄ and R₅ taken together represent a chemical bond; or a pharmaceutically acceptable salt thereof.

2. A compound according to formula I wherein the sugar residue is alkyl substituted.

3. A compound according to claim 1 wherein said sugar residue is a 2-amino or 2-alkylamino-2-deoxyhexopyranosyl, 3-amino or 3-alkylamino-3-deoxy-hexopyranosyl, 3-amino or 3-alkylamino-3,6-dideoxy-hexopyranosyl, 3-amino or 3-alkylamino-2,3,6-trideoxy-hexopyranosyl or 4-amino or 4-alkylamino-2,4,6-trideoxy-hexopyranosyl residue of the D or L series.

4. A compound according to claim 1 wherein R₁ represents hydroxy, methoxy, ethoxy, propoxy or butoxy group and R₂ represents methyl, ethyl, propyl or butyl group or hydrogen atom.

5. A process for the preparation of a compound having the general formula I as defined in claim 1, which process comprises condensing a steroid having the general formula

wherein R₄ and R₅ are as defined in claim 1 with a protected 1-halo derivative of an aminodeoxy, aminodideoxy or aminotrideoxy sugar of the D or L series and removing the protecting groups from the resultant compound.

6. A process according to claim 5 wherein the protecting group is removed with a base.

7. A process according to claim 6 wherein the condensation is carried out in an organic solvent in presence of a catalyst and of a drying agent.

8. A process according to claim 7 wherein the condensation is carried out in chloroform, methylene dichloride, benzene, toluene, acetonitrile or nitromethane, in the presence of silver oxide or carbonate or trifluormethane-sulphonate, Fetizon's reagent or mercuric cyanide or bromide, and of drierite or a molecular sieve, or using the azeotropic distillation method, at a temperature of from -5°C to 100°C.

9. A process according to any of claims 5 to 8 wherein the base is a hydroxide or methoxide of potassium or of sodium or of barium or is triethylamine, and the removal of the protecting groups is effected at room temperature.

10. A process according to any of claims 5 to 9 further comprising cyclizing the resultant compound to form the corresponding lactone.

11. A process according to claim 10 wherein the cyclization is carried out in hydrochloric acid at room temperature and at pH 1-2 for a period of from three to five hours.

12. A pharmaceutical composition comprising a compound having the general formula I as defined in claim 1 or a pharmaceutically acceptable salt of such a compound in admixture with a pharmaceutically acceptable diluent or carrier.

13. A pharmaceutical composition according to claim 12 for use in the treatment of hypertension.

14. The use of compounds having the general formula I or the salts thereof for the making of medicaments against hypertension.