GB2102003A - New 11 beta ,18-epoxy steroids having anti-aldosterone activity - Google Patents

Abstract

Compounds of the formula: <IMAGE> wherein X is a hydrogen atom or a halogen atom, Y<1> is a hydrogen atom, an acetylthio group, a carboxy or esterified carboxy group, a lower alkyl group or a halogen atom, Y<2> is a hydrogen atom or a halogen atom Z<1> is a hydroxy group and Z<2> is a group of the formula -CH2CH2COOM (in which M is potassium or sodium); or Z<1> and Z<2> when taken together with the adjacent carbon atom to which they are attached represent a gamma -lactone ring of the formula: <IMAGE> and the dotted lines between 1- and 2-positions and between the 6- and 7-positions represent double bonds which can optionally be present. Activity: antialdosterone, with reduced side-effects.

Description

SPECIFICATION Steroids having anti-aldosterone activity and their production The present invention relates to new steroid compounds having anti-aldosterone activity and to their production. More particularly, it is concerned with iip, 18 - oxido - steroids having improved antialdosterone activity and their production.

Anti-aldosterone agents are competitive antagonists to aldosterone and inhibit both resorption of sodium ions and excretion of potassium ions at the distal tubule. Thus, they produce a diuretic effect without causing a loss of potassium ions from body fluids and therefore can be used for the diagnosis and treatment of primary hyper-aldosteronism.

Further, they are extensively employed for the treatment of primary or renal hypertension as well as cardiac or renal edema in combination with other therapeutic agents.

Examples of typical anti-aldosterone agents which have heretofore been used are as follows: 3- (7a- Acetylthio - 17,t- hydroxy - 3- oxo - 4- androsten - 17a- yl) propionic acid- y- lactone (i.e.

spirono- lactone) (The Merck Index, 9th Ed., p. 8537)

Potassium 3 - (17ss- hydroxy - 3 - oxo - 4,6 - androstadien - 17a- yl) propionate (i.e. potassium canrenoate) (The Pharmaceuticals Monthly, 22, p. 1316

Potassium 3 - (i7 - hydroxy - 7 - methoxycarbonyl - 3 - oxo - 4- androsten - 17a- yl) propionate (i.e.

mexrenoate potassium) (J. Pharm. Exper. Ther., 209,

These known anti-aldosterone agents have a strong androgen receptor affinity and produce an anti-androgen-like side effect. Thus, their continuous use causes libido-degradation, impotence, gynecomastia, etc. in men, and paramenia, mastauxe, etc. in women.

The present invention is based upon the discovery of certain steroids which exhibit excellent antialdosterone activity with a low androgen receptor affinity.

The present invention provides a compound ofthe formula:

wherein X is a hydrogen atom or a halogen atom, Y' is an acetylthio group, a carboxy or esterified carboxy group, a lower alkyl group or a halogen atom, V2 iS a hydrogen atom or a halogen atom, Z' is a hydroxy group and Z2 is a group of the formula -CH2CH2COOM [in which M is one equivalent of a metal (especially an alkali metal or an alkaline earth metal) or an ammonium or amine cation]; orZ1 and Z2 considered together with the carbon atom to which they are attached represent a lactone ring of the formula:

and the dotted lines between the 1- and 2-positions and between the 6- and 7-positions represent the optional presence of double bonds.

In the above definitions of substituents, the term "halogen" includes fluorine, chlorine, bromine and iodine, fluorine being the most preferred. The "esterified carboxy" group may be, for instance, lower alkoxycarbonyl. The term "lower" is intended to mean any group having not more than 5 carbon atoms. Thus, "lower alkyl" may be straight or branched alkyl having 1 to 5 carbon atoms, of which examples are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl and isopentyl.

The 1 1p, 18 - oxido - steroids (A) include the following 4 kinds of compounds:

wherein X, Vi andY2 are each as defined above; which compounds can be produced from 3,3 ethylenedioxy - lip, 18-oxido-5-androsten- 17 - one or its 9os- halogenated derivatives according to the following: Preparation Scheme (I)

o HO CZECH yi̇st step o.

0 (II) &verbar; 2nd step 0t1 a r 0 HO CeCCO H 0- b I (IV1 step t 4th step (A1) 15th ste I (A3) 6th step (A4) wherein X is as defined above.

The first step in the above scheme is concerned with an acetylene condensation with the carbonyl group at the 17-position of a compound (I) using a nucleophilic reagent. As the nucleophilic reagent, there may be used, for example, an alkali metal acetylide or Grignard reagent type acetylene derivative. The reaction is usually carried out in an inert solvent at room temperature or under cooling. When desired, heating may be applied. Examples of solvents which may be used are aprotic solvents such as ether, tetrahydrofuran, glyme, diglyme, benzene and toluene. The reaction is normally accomplished within a period of from several minutes to several hours.

In the second step, a carboxy group is introduced into a compound (II). The introduction may be achieved, for example, by reacting a compound (II) with an alkyl alkali metal (e.g. n-butyl lithium) to give an alkali metal acetylide (e.g. lithium acetylide) and reacting the latter with carbon dioxide at a temperature from cooling temperature to room temperature.

These reactions are normally carried out in an inert solvent such as an aprotic solvent (e.g. ether, tetrahydrofuran, glyme, diglyme or benzene).

In the third step, the triple bond of a compound (III) is reduced simultaneously with formation of a lactone ring. As a catalyst for the reduction, there may be used, for example, Lindlar catalyst, palladiumcarbon catalyst or rhodium catalyst, whereby the triple bond is reduced to a saturated bond in a singie step. Preferably, however, the triple bond is first reduced by the use of a catalyst such as Lindlar catalyst to a cis-type double bond and then the resulting unsaturated lactone is catalytically reduced to the corresponding saturated lactone. The reaction is usually carried out in an inert solvent under a hydrogen stream at room temperature for a period of from several minutes to several hours. Examples of the solvent which may be used are ethers (e.g. ether, tetrahydrofuran or dioxane), alcohols (e.g. methanol or ethanol) or esters (e.g. ethyl acetate).

In the fourth step, a compound (IV) is hydrolyzed, preferably in the presence of an acid catalyst, to give the corresponding ketone. Examples ofthe acid catalyst are inorganic acids such as, for example, perchloric acid, hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid. The reaction is usually effected in an inert solvent at room temperature or with heating for a period of from several minutes to several hours. As the solvent, there may, for example, be used aqueous methanol, aqueous ethanol, aqueous tetrahydrofuran, aqueous dioxane or aqueous acetone. In this step, a compound (A1) is produced, i.e. a compound (A) wherein V is hydrogen, V2 is hydrogen and there is no double bond present between the 1- and 2-positions and between the 6and 7-positions.

In the fifth step, a double bond is formed in the ring A or B by the use of a dehydrogenating agent, usually under strongly acidic conditions. As the dehydrogenating agent, there may, for example, be used chloranil or 2,3 - dichloro - 5,6 - dicyanobenzoquinone. When chloranil is used as the dehydrogenating agent, a compound (A3) is produced.

When 2,3 - dichloro - 5,6 - dicyanobenzoquinone is used as the dehydrogenating agent, a compound (A2) is produced under kinetically controlled conditions and a compound (A3) is produced underthermodynamically controlled conditions. In order to keep the conditions strongly acidic, hydrochloric acid or conc. sulfuric acid may, for example, be used.

The reaction is normally effected in an inert solvent at a temperature from cooling temperature to room temperature for a period of from several minutes to several hours. Examples of the solvent which may be used are alcohols (e.g. methanol, ethanol or propanol), ethers (e.g. dioxane, ether or tetrahydrofuran) or aromatic hydrocarbons (e.g. benzene or toluene).

In the sixth step, a group selected from acetylthio, carboxy or esterified carboxy, lower alkyl or halogen is introduced into the 7a-position of a compound (A3). In the case of acetylthio, the compound (A3) may be heated under reflux in thioacetic acid for a period of from several minutes to several hours. In the case of carboxy or esterified carboxy, the compound (A3) may be reacted with an alkali metal cyanide (e.g. potassium cyanide or sodium cyanide) in an alcohol (e.g. methanol). Alternatively, the compound (A3) may be reacted with dialkyt aluminium cyanide in an aprotic solvent (e.g. benzene, toluene ortetrahydrofuran) to give a cyanide compound, which may then be heated in the presence of an acid or alkaline catalyst in an inert solvent (e.g. water, methanol or ethanol).In the case of lower alkyl, the compound (A3) may be reacted with a Grignard reagent, preferably n-propyl magnesium bromide, in an ether (e.g. tetrahydrofuran or diethyl ether).

In the seventh step, a halogen atom is introduced into the 6a-position of a compound (A1). For exam ple, the compound (A1) may first be converted into its enol ether or enol acetate according to a per se conventional procedure, and the resulting product then reacted with a fluorinating agent (e.g. perc hloryl fluoride) in a pyridine solvent at a temperature of from cooling temperature to room temperature for several minutes.

The thus-produced compounds (A1), (A2), (A3) and (A4) all have a lactone ring. The lactone ring can be cleaved by contact with an aqueous alkaline solution to give the corresponding hydroxy acid salts (e.g.

sodium salts or potassium salts).

The starting compound (I) in the above Prepration Scheme (I) can be produced from 4 - pregnene 11per 21 - diol - 3, 20 - dione 21 - acetate or its 9a - fluoro derivative which is known. Thus, the 21- acetyl group in the said known substance may be hydrolyzed with an alkali to form a hydroxyl group, and the resulting product oxidised at the 18-position by microbiological oxidation (Kondo et al.: J. Am.

Chem. Soc., 87,4665(1965)) to give an 11 ss - 18 - oxido group. The resulting compound may then be oxidised with periodic acid, followed by esterification with diazomethane to give the 17a-methoxy carbonyl derivative. This compound may then be subjected to conversion of the carbonyl group at the 3-position to a ketal according to a conventional procedure, followed by conversion of the methoxy carbonyl group at the 17-position into an aldehyde group and oxidation ofthe corresponding enamine derivative with sodium bichromate to give the 17keto derivative (I). These conversions are shown in the following scheme: Preparation Scheme

wherein X is as defined above.

The 11p, 18 - oxido - steroids (A) of the invention show excellent anti-aldosterone activity with low side effects, particularly having a low affinity to the androgen receptor. Comparative test results, using 3 (9a-fluoro- 17ss-hydroxy-3-oxo- 11ss, 18-oxido- 4- androsten - 17a- yl) propionic acid - y- lactone as a representative compound of the invention and spironolactone as a comparison on both antialdosterone activity and affinity to the aldosterone receptor and the androgen receptor are given below.

(1) Anti-aldosterone activity Both adrenal glands were excised from male SLC-Wister rats of 180 to 200 g in body weight. The animals were given normal food and 0.45% sodium chloride solution on the first day and sugar instead of normal food on the second day. Eighteen hours before the start ofthe collection of urine, 0.45% sodium chloride solution was intraperitoneally injected into the animals at a dose of 6ml per 100 g of body weight. On the third day, 0.45% sodium chloride solution was intraperitoneally injected into them at a dose of 6 ml per 100 g of body weight.

Aldosterone with or without the test compound or spironolactone was subcutaneously administered to the animals using sesame oil as a vehicle. Two hours after administration, the collection of urine was started, and the amount of urine collected for 4 hours was measured. The concentrations of sodium and potassium were determined by atomic absorption analysis, and their ratio was calculated therefrom. Results are shown in Table 1.

Table 1

Amount of urine Dose (mllloogofbody NalK (;gl1g) weight) ratio Control - 2.0 4.1 Aldosterone 5 16 0.8 Aldosterone 5 Test compound 250 1.9 1.3 Aldosterone 1.9 2.1 Test compound 2500 Aldosterone 5 Spironolactone 250 1.9 0.9 Aldosterone 5 Spironolactone 2500 2.0 15 (2) Affinity to aldosterone receptor Rat kidney cytosol was incubated with about 1 nM 3H-aldosterone and various concentrations of the test compound or spironolactone at O for 18 to 24 hours or at 250C for 60 to 100 minutes. Bound type and non-bound type 3H-aldosterone were separated by the Dextran coated charcoal method, and the amount of bound type was measured. The relative binding affinity (hereinafter referred to as "RBA value") taking aldosterone as 100 was determined when using the test compound and spironolactone.

The average RBA value was 9 in the case of spironolactone and 87 in the case of the test compound when incubated at 0 C. The value was 16 in the case of spironolactone and 40 in the case of the test compound when incubated at 25"C.

(3) Affinity to androgen receptor The prostate lobes obtained from 12 week old castrated male rats were used for Experiment (a), and the kineys obtained from 4 week old castrated male rats were used for Experiment (b). The cytosol was incubated with 1.1 nM 3H-dihydroteststerone (androgen) and the test compound or spironolactone at OOC overnight in Experiment (a) and with 5.0 nM 3H-dihydroteststerone and the test compound or spironolactone at 0 C overnight in Experiment (b).

The bound type and the non-bound type 3H dihydroteststerone were separated by the Dextran coated charcoal method, and the amount of the bound type was measured. Taking androgen as 100, the relative binding affinity was determined when using the test compound and spironolactone.

The RBA values of spironolactone and of the test compound were, respectively, 1.6 and less than 0.1 in Experiment (a) and, respectively, 7.0 and 0.02 in Experiment (b).

As may be appreciated from the above, the results in the in vivo test (1) are coincident with those in the in vitro test (2), and the tested compound of the invention shows higher anti-aldosterone activity than spiconolactone. The in vitro test (3) shows that the tested compound ofthe invention is lower than spironolactone in its affinity to the androgen receptor. Thus, the 11 p- 8 - oxide - steroids (A) ofthe invention have significant anti-aldosterone activity with low affinity to the androgen receptor. In other words, they are useful as anti-aldosterone agents having fewer side effects.

For the oral or parentheral administration of the llp,18 - oxido - steroids (A), they may be formulated into various types of preparation depending upon the mode of administration. For instance, the steroids (A) may be admixed with solid or liquid carriers or diluents to make preparations such as, for example, tablets, capsules, pills, granules, fine granules, solutions or emulsions. Examples of solid diluents are lactose, sucrose, starch, cellulose, talc, magnesium stearate, magnesium oxide, calcium sulfate, arabic gum, gelatine, sodium arginate, sodium benzoate and stearic acid. Examples of liquid diluents are distilled water, physiological saline solution and Ringer's liquid.

Thus, the present invention provides a pharmaceutical or veterinary formulation comprising a compound of the invention formulated for pharmaceutical or veterinary use, respectively. Such formulations may be in unit dosage form and/or may inciude a pharmaceutically acceptable or veterinarily acceptable diluent, carrier or excipient.

Further included in the invention is a steroid (A) of the invention for use in producing an antialdosterone effect in an animal, i.e. when intended for such use by any person in possession thereof, or when made, sold, kept, disposed of, or otherwise dealt with intent for such use.

For oral administration, the compounds of the invention may be used for each adult at a dose of 10 to 200 mg per day. For intravenous injection, they may be administered in an amount of about 0.5 to 50 mg per adult.

The invention will now be further illustrated and described by way of Examples. Reference Examples describing the preparation of starting materials are also given below.

Examples Example 1 Preparation of 3 - (17p - hydroxy - 3 - oxo - 11p,18 oxido - 4 - androsten - 17a - yl) propionic acid -y- lactone (6): (a) Preparation of 3,3 - ethylenedioxy - 17a ethynyl -17ss - hydroxy - 1 1p,18 - oxido - 5 - androstene (2):

Into a mixture of tetrahydrofuran (20 ml) and diethyl ether (10 ml) cooled at -40 C, acetylene gas was introduced to saturation. To the resultant solution, a solution of 1.50 M n-butyl lithium in hexane (3.53 ml; 5.3 mmol) was dropwise added in 10 minutes.To the milky suspension, a solution of 3,3 ethylenedioxy - 1 1p,18 - oxido - 5 - androsten - 17 - one (1) (183 mg; 0.53 mmol) in tetrahydrofuran (8 ml) was dropwise added. The resultant mixture was stirred at -40"C for 15 minutes and at 0 C for 30 minutes. The reaction mixture was poured into saturated ammonium chloride solution and extracted with ethyl acetate. The ethyl acetate extract was washed with saturated sodium chloride solution, dried over magnesium sulfate and concentrated. The residue was subjected to silica gel chromatography using a mixture of ethyl acetate and benzene (1:2) as an eluting solvent. The product was recrystallized from a mixture of dichloromethane and diethyl ether to give the said product (2) (158 mg).M.P., 220-224"C. Yield, 81%.

(b) Preparation of 3 - (3,3 - ethylenedioxy - 17P hydroxy - 11p,18 - oxido - 5 - androsten - 17a - yl) propiolic acid (3):

To a solution of the compound (2) obtained in Example 1 (a) (200 mg; 0.54 mmol) in tetrahydrofuran (5 ml) cooled at -75 C, a solution of 1.50 M n-butyl lithium in hexane (0.87 ml; 1.30 mmol) was dropwise added. The resultant mixture was stirred at -78 C for30 minutes and at -600C for 15 minutes.

Dry ice was added thereto. The temperature of the reaction mixture was elevated up to room temperature. Then, excess of ammonium chloride solution was added thereto, followed by extraction with ethyl acetate. The ethyl acetate layer was washed with water twice, dried over magnesium sulfate and concentrated. The product was recrystallized from ethyl acetate to give the said compound (3) (164 mg). M.P., 188-191 C (decomp.). Yield, 72%.

(c) Preparation of 3 - (3,3 - ethylenedioxy - 17ss hydroxy - 11p,18 - oxido - 5 - androsten -17sr - yl) ) propenic acid -y- lactone (4):

Into a reaction vessel wherein the atmosphere was replaced by nitrogen, 5% palFadium-barium sulfate (50 mg), dioxane (1.6 ml) and puridine (160 iLl) were charged, and the atmosphere was replaced by hydrogen. To the suspension, a solution of the compound (3) obtained in Example î (b) (160 mg; 0.39 mmol) in dioxane (2 ml) was added, the reaction vessel was flushed by hydrogen, and stirring was continued in a hydrogen stream for 1.6 hours. The catalyst was eliminated by filtration. The filtrate was concentrated under reduced pressure.The residue was purified by silica gel chromatography using a mixture of ethyl acetate and benzene (2:1) as an eluting solvent and recrystallized from a mixture of dichloromethane and diethyl ether to give the said compound (4) (91 mg). M.P., 288-291"C. Yield, 59.1%.

(d) Preparation of3 - (3,3 - ethylenedioxy -17ss - hydroxy - llp,18 - oxido - 5 - androsten - 17a -yl) propionic acid -y - lactone (5):

To a solution ofthe compound (4) obtained in Example 1 (c) (300 mg; 0.753 mmol) in a mixture of dioxane (20 ml) and ethanol (4 ml), 5% palladium carbon (60 mg) was added, and stirring was carried out for 20 minutes in a hydrogen stream. The catal yst was eliminated by filtration. The filtrate was con centrated under reduced pressure to give the said compound (5) as crude crystals, which were recrys tallized from a mixture of dichloromethane and diethyl ether to give the pure product. M.P., 285-286"C.

(e) Preparation of 3 - (17,B - hydroxy -3 - oxo 1 lp,18 - oxido - 4 - androsten - 17a - yl) propionic acid -e - lactone (6):

To a solution of the crude product (5) obtained in Example 1 (d) in a mixture of dioxane (35 ml) and water (12.5 ml), 60% perchloric acid (1.12 ml) was added, and the resultant mixture was stirred at room temperature for 15 hours. The reaction mixture was poured onto ice water and extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography using a mixture of benzene and ethyl acetate (4:1) as an eluting solvent and recrystallized from a mixture of dichloromethane and diethyl ether to give the said compound (6) (233 mg).M.P., 186-187"C. Yield, 73.3%.

Example 2 Preparation of 3 - (7 - acetylthio - 17ss - hydroxy - 3 -oxo-11ss,18-oxido-4-androsten-17a-yl)prop- ionic acid -y- lactone (8): (a) Preparation of 3 - (17p - hydroxy - 3 - oxo ?lp,18-oxido-4,6- androstadien- 17a!-yl)prop- ionic acid -y- lactone (7):

To a solution ofthe compound (6) obtained in Example 1 (310 mg; 0.87 mmol) in a mixture of acetic acid (8 ml) and toluene (2 ml), chloranil (227 mg; 0.91 mmol) was added, and the resultant mixture was heated under reflux for 1.5 hours. The reaction mixture was cooled and extracted with chloroform.The extract was washed with water twice, saturated sodium chloride solution once, 10% NaOH solution twice and water twice in order, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was recrystallized from a mixture of dichloromethane and diethyl etherto give the said compound (7) (204 mg). M.P., 205-208"C. Yield, 66%.

(b) Preparation of 3 - (7 - acetylthio - 17p - hyd rnxy-3-oxo-11p,18-oxido-4-andrnsten - 1 7cz - yl ) propionic acid -y- lactone (8):

A mixture ofthe compound (7) obtained in Example 2(a) (250 mg; 0.71 mmol) and thioacetic acid (1 ml; 14 mmol) was heated under reflux for 1 hour.

The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography using a mixture of benzene and ethyl acetate (4:1) as an eluting solvent to give the said compound (8) (180 mg). M.P., 259-262"C. Yield, 59%.

Example 3 Preparation of 3 - (9a - fluoro -17ss - hydroxy - 3 - oxo - 1 1p,18 - oxido -4 - androsten - 17a - yl) propionic acid -y - lactone (14):

In the same manner as in Example 1, the said compound (14) was prepared as shown in the foregoing scheme. Physical data for the reaction intermediates and products of Examples 1 to 3 are shown in Table 2.

Table2

M.P., Molecular formula, Compound Molecular weigh IR NMR No. Elementary analysis (CHCl3 cm-1) (CDCl3#) 1 M.P., 222-224 C 1730 1.14 (3H,s), 3.40 (1H. d, J = 8Hz), C21H28O4(344.46) 4.01 (1H. d. J = 8Hz), 3.93 (4H, s).

4.58 (1H, d,J = 6Hz), 5.37 (1H, m) 2 M.P., 220-224 C 3590,3295 1.14 (3H. s). 2.53 (1H, s), 3.68 C23H30O4(370.47) (1H, d, J = 8Hz), 3.88 (1H, d, J = 8Hz), 3.92 (4H. s), 4.45 (1H, d.

J = 6Hz), 5,32 (1H, m) J = 6Hz), 5.32 (1H, m) 3 M.P., 188-191 C (Nujol)1719, 1.13(3H,s),3.71 (1H,d,J=8Hz), C24H20O6 (424.52) 2215, 2500-3500 3.90 (1H, d, J = 8Hz), 3.96 (4H, s), 4.50 (1 H, d, J = 6Hz), 5.33 (1 H, m) 4 M.P.,288-291 C 1760 1.13 (3H, s), 3.93 (6H, s), 4.45 C24H2aOs (398.48) (1 H, d, J = 6Hz), 5.33 (1 H, m), 5.98 (1H, d,J = 5Hz), 7.48 (1H, d.

J = 5Hz) 5 M.P., 285-286 C 1765 1.13 (3H,s),3.79(2H, s),3.93 C24H300s (398.48) (4H, s), 4.47 (1 H, d, J = 6Hz), 5.33(1K, m) 6 M.P., 186-187 C 1605, 1660, 1.27 (3H, s), 3.83 (2H, s), 4.47 C22H2404 (352.41) 1762 (1 H, d, J = 6Hz), 5.70 (H, br s) 7 M.P., 205-208"C 1610,1650, 1.23 (3H, s), 3.88 (2H, s), 4.52 C22H2204 (354.44) 1660, 1765 (1 H, d, J = 6Hz), 5.63 (1 H, s) 6.11 (2K, s) 8 M.P., 259-262 C 1615, 1675, 1.32 (3H, s), 2.33 (3H, s) 3.85 C24H3005 (430.48) 1693,1775 (2H, s),4.10 (1H, m),4.45 (1 H, d, J = 6Hz), 5.68 (1 H, br s) 9 M.P., 229-231 C 1738 1.23 (3H, s), 4.25 (4H, s), 4.62 C21H27O4F (362.43) (1H, m), 5.37(1H,m) Anal. C H Calcd: 69.59; 7.51 Found: 69.67; 7.68 10 M.P., 239-240 C 3580, 3300 1.23 (3H, s), 2.33 (1H, s), 3.68 C23H2904F (388.46) (1 H, d, J = 8Hz), 3.90 (1 H, d, Anal. C H J=8Hz), 3.93 (4H, s), 4.52 (1H, Calcd: 71.11; 7.52 m), 5.35 (1 H, m) Found: 71.25; 7.51 12 C24H29O5F (416.33) 1760 1.23 (3H, s), 3.92 (6H, s), 4.49 (lH, m), 5.37(1K, m), 5.98 (1H, d, J = 6Hz), 7.50 (1 H, d, J = 6Hz) 13 M.P., 239-241 C 1767 1.23 (3H, s), 3.84 (2H), 3.93 C24H3,OsF (418.49) (4H, s), 4.55 (1 H, m), 5.38 (LH, m) Anal. C H Calcd: 68.88; 7.47 Found: 69.03; 7.64 14 M.P., 252-253 C 1620, 1670, 1.40 (3H, s), 3.87 (2H, s), 4.51 C22H2704F (374.44) 1767 (1 H, m), 5.78 (1 H, br s) Anal. C H F Calcd: 70.56; 7.27; 5.07 Found: 70.32; 7.48; 5.27 Reference Example 1 Preparation of 3,3 - ethylenedioxy - 9a - fluoro 11 ss,18 - oxido - 5 - androsten - 17 - one (9): (a) Preparation of 9a - fluoro -4 - pregnene lip,21 - diol -3,20 - dione (16):

To a solution of 9a - fluoro - 4 - pregnene - 1 lp,21 diol -3,20 - dione 21 - acetate (15) (J. Am. Chem.

Soc., 77, 1068 (1955)) (22.35 g; 55.0 mmol) in a mixture of methanol (450 ml) and dioxane (150 ml) cooled with ice, a solution of potassium hydroxide (3.59 g; 55 mmol) in methanol (56 ml) was dropwise added, and the resultant mixture was stirred under cooling with ice for 10 minutes. The reaction mixture was neutralized with 10% hydrochloric acid and extracted with dichloromethane. The dichloromethane layer was washed with water, sodium bicarbonate solution and sodium chloride solution in order, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give a crystalline residue. The residue was recrystallized from a mixture of dichloroethane and diethyl ether to give the said compound (16) (18.84 g). M.P., 182.5484'C.

Yield, 94% (b) Preparation of9a - fluoro - 1 lop,18 - oxido - 4 pregnen -21 - ol -3,20 - dione (17):

The compound (16) obtained in Reference Example 1 (a) (20.93 g) was subjected to microbiological oxidation in the following manner. Corynespora cas siicola (IMI 56007) was cultured on a slant medium comprising 20% of potato juice, 1% of sugar and 20% of agar media at 28"C for 8 to 10 days. The seed culture was inoculated into a 500 ml volume Sakaguchi flask accommodating a liquid medium (100 ml) comprising glucose (3.5%), polypeptone (2.0%) and corn steep liquor (0.3%; pH, 7.0) and incubated at 280C while shaking for 4 days.After incubation, the fermentation broth was filtered, and the collected bacterial cells were washed with deion ized water to give wet cells in a yield of 10 to 11 g per 100 ml of the medium. The wet cells (10 g) were suspended in deionized water (100 ml) charged in a 500 ml volume Sakaguchi flask, and the fine powder (less than 100 mesh) ofthe compound (16) (70 mg) was added thereto. Using 299 Sakaguchi flasks, reaction was effected while shaking at 28"C for 2 days.

The reaction mixture was filtered through a filtrating cloth to separate the bacterial cells and the filtrate, which were respectively extracted with ethyl acetate.

The extracts were combined together and concentrated at40 C under reduced pressure. The residue (22.5 g) was suspended in an appropriate amount of a mixture of ethyl acetate and acetone, and the insoluble material was collected by filtration to give the crude 18 - hydroxyl product (11.6 g). The crude product (11.5 g) was suspended in a mixture of acetic acid (700 ml) and water (300 ml) and heated under reflux for 1.5 hours. After removal of acetic acid and water by distillation under reduced pressure, the residue was admixed with acetone, and the acetone insoluble material was eliminated by filtration. The acetone soluble material was subjected to silica gel chromatography using a mixture of benzene and ethyl acetate (2:1) as an eluting solvent.The polar fraction was recrystallized from a mixture of dichloromethane and diethyl ether to give the said compound (17) (2.114 g). M.P., 180-181"C. Yield, 10.2%.

(c) Preparation of 9a - fluoro - 17P - methoxy carbonyl - 11 ss,18 - oxido - 4 - androsten - 3 - one (t8):

The compound (17) obtained in Reference Example 1(b) (1.10 g; 3.0 mmol) was dissolved in tetrahydrofuran (12 ml), a solution of periodic acid dihydrate (1.40 g; 6.2 mmol) in water (3 ml) was added thereto, and the resultant mixture was stirred at room temperature for 15 hours. The reaction mixture was diluted with ethyl acetate and washed with saturated sodium chloride solution three times. The organic layer was separated, dried over anhydrous magnesium sulfate and filtered. To the filtrate, an ethereal solution of diazomethane was added for esterification, followed by removal of the solvent by distillation. The residue was recrystallized from a mixture of dichloromethane and diethyl ether to give the compound (18) (996 mg). M.P., 194-196"C. Yield, 90.8%.

(d) Preparation of 3,3 - ethylenedioxy - 9a- fluoro 17p- methoxycarbonyl - 1 1p,18 - oxido - 5 - androstene (19):

The compound (18) obtained in Reference Example 1 (c) (965 mg; 2.66 mmol), ethylene glycol (0.75 ml), p-toluenesulfonic acid monohydrate (20 mg) and dry benzene (15 ml) were charged in a reactor equipped with a Dean Stark type dehydration apparatus comprising molecular sieve 4A and heated under reflux for 5.5 hours. After cooling with ice, anhydrous sodium carbonate (100 mg) was added to the reaction mixture, and stirring was effected for 5 minutes. The resultant mixture was extracted with ethyl acetate.The extract was washed with saturated sodium chloride solution, dried over magnesium sulfate and concentrated under reduced pressure. The residue was recrystallized from a mixture of dichloromethane and methanol to give the said compound (19) (500 mg). The mother liquor was subjected to silica jel chromatography using a mixture of ethyl acetate and benzene (1:1) as an eluting solvent to give an additional amount ofthe said compound (19) (458 mg). Total amount, 958 mg.

M.P., 151-153"C. Yield, 88.5%.

(e) Preparation of 3,3 - ethylenedioxy - 9o- fluoro 17p- formyl - 1 ss,18 - oxido - 5 - androstene (20):-

The compound (19) obtained in Referenc Example 1(d) (951 mg; 2.34 mmol) was dissolved in dry toluene (10 ml), and the resultant solution was cooled to -78 C in a dry ice-acetone bath. To this solution, a solution of 1.43 M diisobutyl aluminum hydride in toluene (2.1 ml; 2.93 mmol) was dropwise added in 15 minutes, and the resultant mixture was stirred at -78 C for 45 minutes. To the reaction mixture, saturated ammonium chloride solution and ethyl acetate were added while stirring.The insolu ble material was eliminated by filtration through a celite layer, and the filtrate was washed with saturated sodium chloride solution, dried over magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel chromatography using a mixture of ethyl acetate and benzene (1:4) as an eluting solvent to give the said compound (20) (619 mg). Recrystallization from a mixture of dichloroethane and diethyl ether gave the pure product. M.P., 155-1 59 C. Yield, 70.3%.

(f) Preparation of 3,3 - ethylenedioxy - 9u'- fluoro 11P18-oxido-5- androsten-17-one (9):-

Into a reactor equipped with a Dean Stark type dehydration apparatus comprising molecular sieve 4A, the compound (20) obtained in Reference Example 1(e) (619 mg; 1.64 mmol), morpholine (0.29 ml; 3.28 mmol) and dry toluene (5 ml) were charged, and the resulting mixture was heated under reflux for 3.5 hours. The solvent and excess of the morpholine were eliminated by distillation under reduced pressure.The residue was dissolved in dry benzene, and the resultant solution was added to a solution of sodium bichromate dihydrate (976 mg; 3.28 mmol) in anhydrous acetic acid (commercially available acetic acid 5 ml + acetic anhydride 0.05 ml) (5 ml) kept at 0"C. The resulting mixture was stirred with ice cooling for 2 hours. To the resultant mixture, methanol (2 ml) was added, and stirring was continued for additional 0.5 hour. The reaction mixture was extracted with ethyl acetate. The extract was washed with 7% sodium hydroxide solution (20 ml) and saturated sodium chloride solution in order, dried over magnesium sulfate and concentrated. The residue was purified by silica gel chromatography using a mixture of ethyl acetate and benzene (1:2) as an eluting solvent to give the said compound (9) (315 mg). M.P., 229-231"C. Yield, 52.9%.

The said compound (9) can be used as the starting material in Example 3.

Reference Example 2 Preparation of 3,3 - ethylenedioxy - 1 1p,18 - oxido 5 - androsten - 17 - one (1):

In the same manner as in Reference Example 1,4 pregnene - 1 ss, 21 - diol - 3,20 - dione - 21 - acetate (21) (synthesized by the process as described in Helv. Chim. Acta., 20, 953 (1937)) was subjected to reaction as shown in the above scheme to give the said compound (1).

The said compound (1 ) can be used as the starting material in Example 1.

The physical data ofthe major intermediates in Reference Examples 1 and 2 are shown in Table 3. Table 3

M.P., Molecular formula, Compound Molecular weight, IR NMR No. Elementary analysis (CHCl3 cm-1) (CDCl3 #) 15 M.P., 213-214 C 1623, 1660 (sh), 0.93 (3H, s), 1.53 (3H, s), 2.17 C23H31O05 (406.47) 1665, 1725, (3H, s), 4.33 (1H, m), 4.67 (1H, d, 1747, 3250-3600, J = 17Hz), 4.82 (1 H, d, J = 17Hz), 3600 5.83(1H, br s) 16 M.P., 182.5-184 C 1625, 1660 (sh), 0.93 (3H, s), 1.55 (3H, s), 4.20 C21H2gO4F (364.45) 1670, 1710, (2H, br s), 4.33 (1H, m), 5.77 3200-3600, (1H, brs) 3625 17 M.P., 180-181 C 1618, 1655 (sh), 1.35 (3H, sl, 3.03 (1H, d, J = 8Hz), C2,H2704F (362.43) 1665, 1710, 3.35(1K, d, J = 8Hz), 3.38(1K, d, Anal. C H 3490 J = 8Hz), 3.73 (1H, d, J =8Hz), Calcd: 69.59; 7.51 4.23 (2H, s), 4.45 (1H,m), Found: 69.02; 7.49 5.78(1H,brs) 18 M.P., 194-196 C 1627, 1675, 1.38 (3H, s), 3.71 (3H, s), 3.32 C2, H2704F (362.43) 1735 (1K, dd, J = 8 & 2Hz), 3.75 (1H, d, Anal. C H J = 8Hz)4.43(1H, d, m), 5.80 Calcd: 69.59; 7.51 brs) Found: 69.14; 7.37 19 M.P., 151-153 C 1722 1.22 (3H, s), 3.67 (3H, s), 3.92 C23H3105F (406.48) (4H, s), 4.43 (1 H, m), 5.37 Anal. C H (1H, m) Calcd: 67.96; 7.69 Found: 67.66; 7.71 20 M.P., 155-159 C 1718, 2700, 1.23 (3H, s), 3.48(1K, dd, J = 8 & BR< C22H2904F (376.45) 2825 2Hz), 3.72 (1 H, d, J = 8Hz), 3.93 Anal. C H (4H, s), 4.48 (1H, m),5.38 Calcd: 70.18; 7.76 (1H, m), 9.73 (1H, d, J = 2Hz) Found: 70.13; 7.78 24 M.P., 193-194 C 1608, 1660, 1.27 (3H, s), 3.30 (1H, d, J = 8Hz), C21H28O4(344.43) 1722 3.43 (1H, d, J = 8Hz), 3.31 (3H, s), 4.43 (1H, d, J = 6Hz), 5.37 (1H, brs) 25 M.P., 171-175 C 1725 1.12 (3H, s), 3.25 (1H, d, J = 8Hz), C23H3205 (388.49) 3.72 (1 H, d, J = 8Hz), 3.68 (3H, s), 7.79 (4H, s), 4.42 (1 H, d, J = 6Hz), 5.38(1K, m) 26 M.P., 142-144 C 1720, 2720, 1.13 (3H, s), 3.48 (1H, d, J = 8Hz), C22H3004 (358.46) 2830 3.75 (1H, d, J = 8Hz), 3.95 (4H, s), 4.43 (1H, d, J = 6Hz), 5.35 (1H, m), 9.73(1H, d,J = 1Hz)

Claims (29)

1. Acompound oftheformula:

wherein X is a hydrogen atom or a halogen atom, Y' is an acetylthio group, a carboxy or esterified carboxy group, a lower alkyl group or a halogen atom, V2 is a hydrogen atom or a halogen atom, Z' is a hydroxy group and Z2 is a group of the formula -CH2CH2COOM (in which M is potassium or sodium); orZ1 and Z2 considered together with the carbon atom to which they are attached represent a lactone ring of the formula:

and the dotted lines between the 1- and 2-positions and between the 6- and 7-positions represent the optional presence of double bonds.

2. A compound as claimed in claim 1, wherein X is hydrogen, Y' is hydrogen, V2 is hydrogen, Z' and Z2 together with the carbon atom to which they are attached represent a group of formula (a) as defined in claim 1 and there are no double bonds between the 1- and 2-positions and between the 6- and 7-positions.

3. A compound as claimed in claim 1, wherein X is hydrogen, Y' is acetylthio, V2 is hydrogen, Z' and Z2 together with the carbon atom to which they are attached represent a group of formula (a) as defined in claim 1 and there are no double bonds between the 1- and 2-positions and between the 6- and 7-positions.

4. A compound as claimed in claim 1, wherein X is fluorine, Y' is hydrogen, V2 is hydrogen, Z' and Z2 together with the carbon atom to which they are attached represent a group of formula (a) as defined in claim 1 and there are no double bonds between the 1- and 2-positions and between the 6- and 7-positions.

5. A compound as claimed in claim 1 and of formula A1, A2, A3 orA4 as defined hereinbefore.

6. A compound as claimed in claim 1 wherein X, Y', and/or V2 is/are (a) substituent(s) referred to hereinbefore in exemplification of substituents X, Y', and/or V2.

7. A process for preparing a compound as claimed in claim 1, which process comprises hydrolysing a compound of the formula:

so as to produce a compound of the formula:

8. A process as claimed in claim 7, wherein the hydrolysis is effected substantially in accordance with the foregoing description of the 4th step in the foregoing Preparation Scheme (1).

9. A process as claimed in claim 7 or claim 8 followed by reacting the resulting compound of formula (A,) (as hereinbefore defined) with a dehydrogenating agent so as to produce a compound as claimed in claim 1 having a double bond between the 1- and 2-positions and/or between the 6- and 7-positions.

10. A process as claimed in claim 9, wherein the reaction with a dehydrogenating agent is effected substantially in accordance with the foregoing description of the 5th step in the foregoing Preparation Scheme (I).

11. A process as claimed in claim 9 or claim 10, wherein a compound ofthe formula (A3) (as hereinbefore defined) is produced and said compound of formula (A3) is thereafter subjected to introduction of an acetylthio group, a carboxy or esterified carboxy group, a lower alkyl group or halogen at the 7a-position.

12. A process as claimed in claim 11, wherein the introduction at the 7a-position is effected substantially in accordance with the foregoing description of the 6th step in the foregoing Preparation Scheme (I).

13. A process as claimed in claim 7 or claim 8 followed by subjecting the resulting compound of formula (A1) (as hereinbefore defined) to 6ahalogenation so asto produce a compound offormula (A4) (as hereinbefore defined).

14. A process as claimed in claim 13, wherein the 6a-halogenation is effected substantially in accordance with the foregoing description of the 7th step in the foregoing Preparation Scheme (I).

15. A process as claimed in any one of claims 7 to 14, wherein the compound of formula (IV) used to produce the compound of formula (A,) has been prepared by reduction of and lactone ring formation from a compound ofthe formula:

16. A process as claimed in claim 15, wherein the reduction and lactone ring formation are effected substantially in accordance with the foregoing description of the 3rd step in the foregoing Preparation Scheme (I).

17. A process as claimed in claim 15 or claim 16, wherein the compound of formula (III) has been prepared by introducing a carboxy group into a compound of the formula:

18. A process as claimed in claim 17, wherein the carboxy group introduction is effected substantially in accordance with the foregoing description of the 2nd step in the foregoing Preparation Scheme (I).

19. A process as claimed in claim 17 or claim 18, wherein the compound of formula (II) has been prepared by an acetylene condensation with the 17carbonyl group of a compound of the formula:

20. A process as claimed in claim 19, wherein the condensation is effected substantially in accordance with the foregoing description of the 1 sot step in the foregoing Preparation Scheme (I).

21. A process as claimed in claim 19 or claim 20, wherein the compound of formula (I) has been pre pared from 4- pregnene - 11 lp, 21 - diol - 3, 20 - dione 21 - acetate or its 9a- fluoro derivative.

22. A process as claimed in claim 21, wherein said preparation is effected substantially in accordance with the foregoing Preparation Scheme (II).

23. A process as claimed in any one of claims 7 to 22, wherein the resulting lactone ring-containing compound as claimed in claim 1 is subjected to lactone ring cleavage using an aqueous alkali to give a compound as claimed in claim 1 which is a hydroxy acid salt.

24. A process for preparing a compound as claimed in claim 1 and substantially as hereinbefore described in any one ofthe foregoing Examples.

25. A process for preparing a compound as claimed in claim 1, wherein 3,3 - ethylenedioxy - 9a fluoro - 1 1p,18 - oxido - 5 - androsten - 17 - one or 3,3 ethylenedioxy 1 1p,18- oxido - 5 - androsten - 17- one is used as an intermediate, which intermediate has been prepared by a procedure substantially as hereinbefore described in Reference Example 1 or Reference Example 2, respectively.

26. A pharmaceutical or veterinary formulation comprising a compound as claimed in claim 1 formulated for pharmaceutical or veterinary use, respectively.

27. A formulation as claimed in claim 26 and in unit dosage form.

28. Aformulation as claimed in claim 26 or claim 27 also comprising a pharmaceutically acceptably or veterinarily acceptable, respectively, diluent, carrier or excipient.

29. A compound as defined in claim 1 for use in producing an anti-aldosterone effect in an animal.