IL41752A - D-homosteroids of the pregnane series their manufacture and pharmaceutical compositions containing them - Google Patents

Description

oniR α·**3>»ίτ n*npn *·Ί*ι_ η .ρ¾ι*»· Hovel IMiomosteroida of the pregoane eerie8» their manufacture and pharmaceutical compositions containing them SPARA EDICA. A.G.

C. 41752/2 ' : " The present invention. relates to novel steroids of the pregnane series. More particularly, the invention- is . concerned with D-homosteroids of the pregnane series and vith a process for the manufacture thereof.

The D-homosteroids provided "by the present invention compounds of the general formula , wherein R represents a hydrogen, fluorine or chlorine atom or the methyl group, R 9 represents a hydrogen, fluorine, chlorine or bromine atom and R 17' and R21. each independently represent a alkenoyloxy, hydroxy or an alkanoyloxy ,/ aralkanoyloxy or aroyloxy group and 1,2-dehydro derivatives thereof.

The aforementioned acyloxy groups are preferably derived from saturated or unsaturated aliphatic araliphatic or aromatic carboxylic acids containing up to 20, preferably up to 12, carbon atoms. Examples of such acids butyric acid, caproic acid, oenanthic acid, oleic acid, palmitic acid, stearic acid, succinic acid, malonic acid, fumaric acid, citric acid, oyolohoxylpropionio aoid, phenyl-acetic acid and benzoic acid.

A preferred group of compounds of formula I are those 6 in which R and R7 re nt a h d en or fluorine atom and R group. Especially preferred are the 1,2-dehydro derivatives. Of the compounds which are substituted in the 6-position, the 6a-isomers are preferred.

According to the process provided by the present invention, the D-homosteroids aforesaid (i.e. the compounds of formula I and 1,2-dehydro derivatives thereof) are manufactured by a) hydroxylating in the 11-position a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof by meads of microorganisms or enzymes obtained therefrom, or b) treating a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof with an alkali acylate, or c) dehydrogenating a D-homosteroid of formula I in the 1,2--position, or d) adding hypochlorous or hypobromous acid to the 9ill-double bond of a D-homosteroid of the general formula or of a 1,2-dehydro derivative thereof, or e) treating a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof with hydrogen fluoride, hydrogen chloride or hydrogen bromide, or alkenoyloxy, \ f) saponifying an alkanoyloxy ,/aralkanoyloxy or aroyloxy group in a D-homosteroid of formula I or in a 1,2-dehydro derivative thereof in which at least one of R1^ and R21 represents an acyloxy group, or g) isomerising a 63-fluoro-, 63-chloro- or 6 -methyl-D--homosteroid of formula I or a 1,2-dehydro derivative thereof to a 6a-isomer, or h) treating a D-homosteroid of the general formula (VI) (Via) with a fluorinating or chlorinating agent, or alkenoylating, i) alkanoylating,/ aralkanoylating or aroylating a 17a- or 21-hydroxy group in a D-homosteroid of the general formula or in a 1,2-dehydro derivative thereof, or j) dehydrating a D-homosteroid of the general formula , or k) reducing the 11-keto group in a D-homosteroid of the general formula or in a 1,2-dehydro derivative thereof to the hydroxy group with protection of the 3- and 20-keto groups, or 1) oxidising the 17(20)-double bond in a D-homosteroid of the general formula to the hydroxy-ketone group, or m) dehydrohalogenating a D-homosteroid of the general formula ; in the foregoing formulae R , R , R 1 and 21 R have the significance given earlier, R 61 represents an acyloxy or alkoxy group, R represents a fluorine or chlorine atom or the methyl group and Hal represents a chlorine, bromine or iodine atom.

The hydroxylation according to embodiment (a) of the process can be carried out using methods which are known per for the microbiological introduction of an 11-hydroxy group into steroids. For the 11-hydroxylation there can be used microorganisms of the taxonomic unit Fungi and Schizomycetes, especially of the sub-units Ascomycetes, Phycomycetes, Basidiomycetes and Actinomycetales. Mutants produced by chemical means (e.g. by treatment with nitrite) or by physical means (e.g. by irradiation) as well as cell-free enzyme preparations obtained from the microorganisms can also be used. Suitable microorganisms for the llfl-hydroxylation are particularly those of the genus Curvularia, e.g., C.lunata NRRL 2380 and NRRL 2178, Absidia, e.g., A. orchidis ATCC A. 8990 and Λ. coerula IPO 4 35, Colletotrichum, e.g., C. pisi ATCC 12520, Pellicolaria, e.g., PT filamentosa IFO 6675, streptomyces, e.g., S. fradiae ATCC 107 , Cunninghamella, e.g., C. bainieri ATCC 9244, C. verticellata ATCC 8983, C. elegans ATCC 924 and C. echinulata ATCC 8984 , and Pycnosporium, e.g., sp. ATCC 12231.

The replacement of a 21-halogen atom in a D-homosteroid of formula III or in a 1 ,2-dehydro derivative thereof by an acyloxy group according to embodiment (b) of the process can be carried out by warming a D-homosteroid of formula III or a 1 , 2-dehydro derivative thereof with an appropriate alkali acylate in the presence of the acid corresponding to the acylate (e.g. with potassium acetate in glacial acetic acid).

The 1 , 2-dehydrogenation of a D-homosteroid of formula I according to embodiment (c) of the process can be carried out in a manner known per se; for example, by microbiological means or by means of dehydrogenating agents such as iodine pentoxide, periodic acid, selenium dioxide, 2 , 3-dichloro-5i 6--dicyanobenzoquinone, chloranil or lead tetraacetate.

Suitable microorganisms for the 1 , 2-dehydrogenation are, for instance, Schizomycetes, particularly those of the genus Arthrobacter, e.g. A. simplex ATCC 6946, Bacillus, e.g., B. lentus ATCC 13805 and B. sphaericus ATCC 7055, Pseudomonas, e.g., P. aeruginosa IFO 350 , Flavobacterium, e.g., P. flavescens IFO 3085* Lactobacillus, e.g., L. brevis IFO 3345 and Nocardia, e.g., N. opaca ATCC 4276.

In carrying out embodiment (d) of the process, a D-homosteroid of formula IV or a 1,2-dehydro derivative thereof is expediently dissolved in a suitable solvent (e.g. an ether such as tetrahydrofuran or dioxane, a chlorinated hydrocarbon such as methylene chloride or chloroform or a ketone such as acetone) and reacted with hypochlorous or hypobromous acid. The hypochlorous or hypobromous acid is itself expediently formed in situ in the reaction mixture; for example, from N-bromo or N-chloroamides or -imides such as N-chloro-succinimide or N-bromoacetamide and a strong acid, preferably-perchloric acid.

Embodiment (e) of the process is carried out in a manner known per se. If desired, the D-homosteroid of formula V or a 1,2-dehydro derivative thereof is dissolved in an inert solvent and the solution treated with the appropriate hydrogen halide. This embodiment of the process is the preferred embodiment for the manufacture of 9-fluoro-D-homosteroids of formula I and 1,2-dehydro derivatives thereof.

The saponification of an acyloxy group in a D-homosteroid of formula I or in a 1,2-dehydro derivative thereof according to embodiment (f) of the process can be carried out in a manner known per se; for example, using aqueous-methanolic potassium carbonate solution_ The isomerisation of a δβ-fluoro-, δβ-chloro- or 6β--methyl-D-homosteroid of formula I or of a 1,2-dehydro derivative thereof, especially of a 63-fluoro- or 63-chloro--D-homosteroid or of a 1,2-dehydro derivative thereof, according to embodiment (g) of the process can be carried out by treatment with an acid, especially a mineral acid such as hydrochloric acid, in a solvent such as dioxane or glacial acetic acid.

According to embodiment (h) of the process, the chlorination of a D-homosteroid of formula VI or Via can be carried out, for example, by treatment with a chlorination agent such as elemental chlorine or a N-chlorimide (e.g.

N-chlorosuccinimide) or a N-chloroamide . The fluorination, also according to embodiment (h) of the process, is expediently carried out by treatment with perchloryl fluoride. Mixtures of 6a- and δβ-isomers can be separated by chromato raphy.

The acylation of a 17a- or 21-hydroxy group in a D-homosteroid of formula VII or in a 1,2-dehydro derivative thereof according to embodiment (i) of the process can be carried out in a manner known per se by treatment with an acylating agent such as an acyl chloride (e.g. acetyl chloride) or an acid anhydride in the presence of an acid binding agent such as pyridine. The acylation of a 17a-hydroxy group is expediently carried out in the presence of an acid catalyst such as p-toluenesulphonic acid.

The dehydration of a D-homosteroid of formula VIII according to embodiment (J) of the process can be carried out by treatment with an acid (e.g. a mineral acid such as hydrochloric acid) .

In carrying out embodiment (k) of the process, the keto groups in the 3- and 20-positions of a D-homosteroid of formula IX or of a 1,2-dehydro derivative thereof are first 17 21 protected (e.g. by ketalisation) . Where R ' and R both represent hydroxy groups, a 20-keto group can also be protected by formation of the 17»20 ;21-bismethylenedioxy derivative.

The reduction of the 11-keto group of the protected D-homosteroid or 1,2-dehydro derivative thereof can be carried out using a complex metal hydride such as lithium aluminium hydride, sodium borohydride or diisobutyl aluminium hydride.

The oxidation of the 17(20)-double bond in a D-homo-steroid of formula jin accordance with embodiment (1 ) of the process can be carried out, for example, using oxidising agents such as a tertiary amine N-oxide peroxide in tertbutanol/ pyridine in the presence of catalytic amounts of osmium tetroxide. Examples of tertiary amine N-oxide peroxides are N-methylmorpholine N-oxide peroxide and triethylamine oxide peroxide. On the other hand, one can oxidise to a 17 ,20- -glycol using an oxidising agent such as osmium tetroxide or permanganate and then further oxidise the 17 , 20-glycol to the desired hydroxy ketone using an oxidising agent such as chromic acid.

The dehydrohalogenation of a D-homosteroid of formula XI in accordance with embodiment (m) of the process can be carried out using a base (e.g. an organic base such as pyridine).

The starting materials used in the foregoing process, insofar as they are unknown or are described hereinafter, can be prepared in analogy to known methods or to methods described hereinafter.

The D-homosteroids provided by this invention (i.e. the compounds of formula I and 1 ,2-dehydro derivatives thereof) possess endocrinal, especially anti-inflammatory, activity.

In the felt-pellet test in the rat a 4-0% inhibition of the granuloma formation was achieved with. the following dosages; 2 .7 mgAg of lip, 17aa , 21-trihydroxy-pregn- -ene-3» 20-dione; 0.9 mgAg of 113 , 17aa , 21-trihydroxypregna-l, -diene-3i20--dione; 0.35 mgAg of 113 , 17aoc , 21-trihydroxy-9a-fluoro--pregn- -ene-3, 20-dione and 0 .05 mgAg of 11 β , 17βα , 21--trihydroxy-9«-fluoro-pregna-l ,4—diene-3» 20-dione„ The D-homosteroids provided "by the present invention can be used as medicaments; for example, in the form of pharmaceutical preparations which contain them in association with a compatible pharmaceutical carrier. This carrier can be an inert organic or inorganic carrier material suitable for enteral, percutaneous or parenteral administration; for example, water, gelatine, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, petroleum jelly etc. The pharmaceutical preparations can be made up in solid form (e.g. as tablets, dragees, suppositories or capsules), in semi-solid form (e.g. as ointments) or in liquid form (e.g. as solutions, suspensions or emulsions).

If necessary, the pharmaceutical preparations may be sterilised and/or may contain adjuvants such as preservatives, stabilisers, wetting agents, emulsifiers, salts for varying the osmotic pressure or buffers. They can also contain other therapeutically valuable substances „ The following Examples illustrate the process provided by the present invention: Example 1 3.81 g of D-homo-113 , 17aa-dihydroxy-pregn-4-ene-3,20- 5 -dione in 20 ml of methanol are mixed with 4.7 ml of 10% methanolic calcium chloride solution and 2.3 g of ignited calcium oxide. Then a solution of 3.5 g of iodine and 2.2 g of calcium chloride in 22 ml of methanol is slowly added dropwise and the mixture is stirred for about a further 10 10 minutes. The mixture is poured on to ice-water and. extracted with methylene chloride. The methylene chloride extracts are washed with water, dried and evaporated. The crude iodide obtained is dissolved in 56 ml of acetone, the solution mixed with 0.56 ml of water, 0.56 ml of glacial acetic acid and 15 5·6 g of potassium acetate and boiled under reflux for 18 hours. The solution is concentrated and worked up with water/methylene chloride. Chromatography of the crude product on silicagel gives D-homo-21-acetoxy-ll , 17aa-dihydroxy-pregn-*~* -4-ene-3»20-dione of melting point 212°-213°C; [alp = +145° 20 (c ■ 0.104 in dioxane).

The starting material can be prepared as followss 3> 113-Diacetoxy-androsta-3» 5-dien-17-one is reacted in methylene chloride with ethylene glycol in the presence of orthoformic acid ester and p-toluenesulphonic acid at room 25 temperature to give 3ill3-acetoxy-17 » 17-ethylenedioxy-androsta- -3,5-diene of melting point 183°-186°C; [a3D - -112° The foregoing 17-ketal is reduced in tetrahydrofuran/ methanol with sodium borohydride to give 113-acetoxy-17,17--ethylenedioxy-33-hydroxy-androst-5-ene of melting point 125°-126°C; Ca]D = -66° (c - 0.102 in dioxane).

Cleavage of the thus-obtained ketal in aqueous acetone with p-toluenesulphonic acid gives lip-acetoxy-33-hydroxy--androst-5-en-17-one of melting point 193°-195°C; Ca3D = -4° (c = 0.102 in dioxane).

The resulting 17-ketosteroid is reacted with dimethyl-sulphoxonium methylide in dimethylformamide to give 21-nor--llβ-aceto y-17^20ξ-epo ~3β-hydro y-pregn-5-ene of melting point 155°-156°C; [a D = -52° (c = 0.103 in dioxane).

The foregoing epoxide is reacted in alcohol and concentrated ammonia in an autoclave to give Ιΐβ-acetoxy--174-aminomethyl-33 »175-clihydroxy-androst-4~ene . By reaction with sodium nitrite in glacial acetic acid and water there is obtained D-homo-113-acetoxy-33-hydroxy-androst-5-en-17a-one of melting point 230°-232°C; Ca3D - -121° (c ■= 0.103 in dioxane).

Saponification of the thus-obtained Ιΐβ-acetate in boiling methanolic potassium hydroxide gives D-homo-33 »11β--dihydroxy-androst-5-en-17a-one of melting point 234-°-236°C; [a]D = -1^3° (c = 0.107 in dioxane).

The resulting 3»ll-diol is reacted in dimethyl sulphoxide with sodium hydride and triphenylethylphosphonium bromide to give D-homo-3 >113-dihydroxy-pregna-5,17a(20)-diene of melting point 172°-173°C; Ca]D = -137° (c = 0.104 in dioxane).

Subsequent oxidation according to the Oppenauer method gives D-homo-113-hydroxy-pregna-4-,17a(20)-dien-3-one of melting point 160°-161°C; [a]D = +96° (c = 102 in dioxane); ε2 3 = ^OOO.

Oxidation of the 4-,17a(20)-diene thus obtained' ith osmiuin tetroxide and N-methylmorpholine oxide/hydrogen peroxide gives D-homo-lip,17aa-dihydroxy-pregn- -ene-3»20-dione of melting point 213°-215°C; [a]D - +10 ° (c = 0.102 in dioxane); ε2½ = 16,250.

Example 2 In an analogous manner to that described in Example 1, from D-homo-lip,17aa-dihydroxy-pregna-l,4-diene-3,20-dione there is obtained D-homo-21-acetoxy-110,17aoc-dihydroxy-pregna--l, -diene-3,20-dione of melting point 220°-222°C; [a]D = +108 (c = 0.105 in dioxane); E2-2 a ^'500.

The starting material can be prepared by the microbiological dehydrogenation of D-homo-llp,17aa-dihydroxy-pregn-- -ene-3>20-dione with Arthrobacter simplex to give D-homo--113,17aa-dihydroxy-pregna-l,4-diene-3i20-dione' of melting point 208°-212°C; [a]D - + 7° (c = 0.107 in dioxane); ε2 2 = 13,900.

Example 5 500 mg of D-homo.cortisol are stirred in 2 ml of pyridine and 2 ml of acetic acid anhydride for 2 hours at room temperature. The mixture is poured on to ice-cold dilute hydrochloric acid and extracted with methylene chloride „ The extracts are washed with water, dried and evaporated,, There is obtained pure D-homocortisol acetate which is identical with the product obtained according to Example 1.

Example I.25 g of D-homo-21i-acetoxy-17aa-hydroxy-pregna-4-, 9(ll)--diene-3 , 20-dione are dissolved in 53 ml of dioxane, mixed with IO.5 ml of water, 865 nig of N-bromoacetamide, » ml of 10% perchloric acid and stirred for 15 minutes at room temperature. Then „ 5 g of sodium sulphite and 90 ml of water are added. After stirring for a short time, the mixture is extracted with methylene chloride, the extracts are washed with water, dried over sodium sulphate and evaporated. There is obtained D-homo-21-acetoxy-113 , 17acx--dihydroxy-9a-bromo-pregn-4--ene-3 » 20-dione which is almost pure according to thin-layer chromatography.

The starting material can be prepared as follows; D-Homohydrocortisone acetate is dehydrated in dimethyl-formamide with methanesulphonyl chloride in the presence of pyridine at an elevated temperature. There is obtained D-homo-21-acetoxy-17aoc-hydroxy-pregna- , 9 ( 11 )-diene- 3 » 20- -dione of melting point 238°-24-0°C; [α]π = +71° (c = 0.10 in dioxane); e2,<} = 16, 750.

Example 5 In an analogous manner to that described in Example 4-, from D-homo-21-acetoxy-17aa-hydroxy-pregna-l , , 9(H)-triene--3 , 20-dione /"melting point 188°-190°C; [a]D = -1 (c = 0.084-in dioxane); ε2^β ~ 16, 700j7 there is obtained D-homo-21--acetoxy-lip, 17aa-dihydroxy-9a-bromo-pregna-l, -diene-3i 0--dione .

The starting material is prepared from D-homoprednisolone -21-acetate in an analogous manner to that described in the second paragraph of Example 4-„ Example 6 905 mg of D-homo-21-acetoxy-9 * 113-epoxy-17aa-hydroxy--pregn-A-ene-3i20-dione are stirred in 20 ml of a solution of l o25 parts of hydrogen fluoride in 1 part of urea for 20 minutes at room temperature. The mixture is poured on to a mixture of 70 ml of concentrated ammonia and 200 g of ice and extracted with methylene chloride. The extracts are washed with sodium chloride solution, dried and evaporated.

Chromatography on silicagel gives D-homo-21-acetoxy-113 , 17aa--dihydroxy-9a-fluoro-pregn- -ene-3 ,20-dione of melting point 24-2°-24A°C; [a]D = +137° (c = 0.102 in dioxane); ε∑^ = 16,220.

As byproduct, D-homocortisone acetate, was also isolated.

The starting material can be prepared as follows: D-Homo-21-acetoxy-9a-bromo-11 ,17aoc-dihydroxy-pregn- — -ene-3»20-dione is boiled under reflux in absolute alcohol in the presence of anhydrous potassium acetate for 24· hours.

There is obtained D-homo-21-acetoxy-9ill3-epoxy-17aa-hydroxy--pregn-4-ene-3i20-dione of melting point 226°-228°C; [a]D -+51° (c m 0.103 in dioxane); £3Ll ■ 14,100.

Example 7 In an analogous manner to that described in Example 6, from D-homo-21-acetoxy-9»113-epoxy-17aa-hydroxy-pregna-l ,4--diene-3,20-dione "melting point 225°-226°C; [a]D = +63° (c = 0.103 in dioxane); e2n& B ^-^,900 (prepared from D-homo--21-acetoxy-9a-bromo-11 ,17aa-dihydroxy-pregna-l, -diene-3,20--dione)_7 there is obtained D-homo-21-acetoxy-9a-fluoro- -113,17aoc-dihydroxy-pregna-l,4—diene-3»20-dione of melting point 24-0°-250°C; [a]D - +109° (c = 0.106 in dioxane); ε259 = 15,200.

Example 8 418 mg of D-homo-21-acetoxy-113,17aa-dihydroxy-pregn- - -ene-3,20-diohe and 250 mg of selenium dioxide are stirred under reflux in 20 ml of tertbutanol and 0.2 ml of glacial acetic acid under argon for 20 hours. The mixture is filtered and evaporated. The oil obtained is dissolved in ethyl acetate and washed successively with sodium bicarbonate solution, water, ice-cold ammonium sulphide solution, dilute ammonia, water, dilute hydrochloric acid and water. The ethyl acetate solution is dried over sodium sulphate and evaporated in a vacuum. Chromatography on silicagel gives D-homo-21-acetoxy-113 , 17aoc-dihydroxy-pregna-l ,4-diene-3 i 20--dione of melting point 220°-222°C; [o]D « +108° (c = 0.105 in dioxane); ^ h 53 1 , 500.

Example 9 A suspension of 955 mg of D-homoCortisol acetate in 16 ml of absolute methanol is flushed with argon. Then 250 mg of potassium carbonate in 3.5 ml of water are added dropwise and the mixture is stirred for 0.75 hour at room temperature. The mixture is then poured on to sodium chloride solution and extracted with methylene chloride. The extracts are washed, dried and evaporated. There is obtained pure D-homq.cortisol of melting point 2450-24-6°C ; [a]^ = +142° (c = 0.102 in dioxane); ε ,Ρ = 15*850.

In. a similar manner there are obtained: from D-homoprednisolone-21-acetate D-homoprednisolone of melting point 250°C (decomposition) ; Ca]D » +105° (c 0.101 in dioxane); = 14,400; from D-homo-9a-fluoro-cortisol-21-acetate D-homo-9a-fluoro- Cortisol of melting point 239°-2 1°C; [a]D = +151° (c = 0.102 in dioxane); ε239 05 16,820 ; and from D- omo-9a-fluoro-prednisolone-21-acetate D-homo-9<*- uoro-prednisolone of melting point 2 1°-2 6°C (decomposition); Ca]D = +101° (c = 0.097 in dioxane); e2^ " 1^'5^0. from βα-fluoro-D-homocortisol -21-acetate 6a-fluoro-D-homocortisol of melting point 233-235°C ; [a]D + 110°C (c,= O.IO3 in dioxane) = 1 . ^00 from 6a-fluoro-D-homoprednisolon-21-acetate 6a-fluoro-D-homoprednisolon of melting point 2bl-26 °C; ( ]D + 93° (c = 0. 100 in dioxane) C ^ = 15. 200.

Example 10 5OO ml of a sterile nutrient medium containing 5$ starch syrup, 0.5$ corn steep liquor, 0. 2$ sodium nitrate, 0. 1$ potassium dihydrogenphosphate, 0. 05$ potassium chloride, 0. 05$ magnesium sulfate^ 0.002$ ferrous sulfate is inoculated with Curvularia lunata NRRL 2380 from a two weeks old agar slant culture. The culture is grown for five days at 30°C with shaking. The so obtained culture serves to inoculate 15 liters of a sterile medium containing 5$ starch syrup, 0.25$ corn steep liquor, 0. 1$ sodium nitrate, 0. 05$ potassium dihydrogen phosphate, 0.025$ potassium chloride, 0. 025$ magnesium sulfate, 0.001$ ferrous sulfate. The culture is grown for 72 hours at 29°C with stirring (220 r.p.m.) and aeration (1¾ liters/min. ) . 900 ml of the so-obtained culture are transferred to a fermenter containing 14,1 liters of the nutrient medium described before. This culture is grown for 2 hours at 29°C with stirring and aeration. Thereafter, a solution of ),0 g of 21-acetoxy-17ao-hydroxy-D-homo-pregn- 4-ene-3,20-dione in 150 ml of dimethylformamide is filtered sterile and added. The culture is then incubated for another 40 hours, filtered and the filtrate and the mycelium is extracted with methyl isobutyl ketone. The extracts are combined and concentrated under reduced pressure. The residue is chromatographed on silicagel and the crude product recrystallized from acetone-hexane to yield D-homocortisol-21-acetate.

Exam-pie 11 A nutrient medium consisting of 0.15% corn steep, 0.5% peptone and 0.5% glucose in distilled water, pH 7.3, is inoculated with Arthrobacter simplex ATCC 694-6. The culture is grown for 24- hours at 28°C and then a solution of 25 mg of D-homocortisol in 1 ml of 80% aqueous ethanol is added.

After an incubation time of 48-72 hours, the mycelia of the substrate are separated, washed with water and the washings and the substrate extracted with methylene chloride. Working up of the extract yields D-homo^ll3,17aa,21-trihydroxypregna--l, -diene-3,20-dione (D-homoprednisolone) .

Example 12 690 mg of 6p-fluoro-D-homocortisol-21-acetate in 50 ml of acetic acid are mixed at room temperature with O.J ml of a 20 per cent solution of hydrogen bromide in acetic acid. After 30 minutes, 1.5 ml of pyridine are added. The mixture is evaporated and the residue is taken up in methylene chloride and washed with diluted sodium carbonate solution and water. The organic phase is dried over sodium sulfate and evaporated. Chromatography of the' residue on silicagel using methylene chloride 5 per cent acetone gives pure 6a-fluoro-D-homocortisol-21-acetate of melting point 137-139°C [a)D + 110°C (c = 0.09^ in dioxane), .14.200.

Example 13 600 mg of D-homocortisol-?21-acetate are dissolved in 6 ml of orthoethyl formate and 6 ml of absolute ethanol with heating. The solution is cooled to room temperature and a solution of 6 mg of p-toluene sulfonic acid in 6 ml of absolute ethanol is added. The mixture is left to stand at room temperature for 8 minutes. After one drop of pyridine is added the mixture is poured into water and extracted with methylene chloride. The organic phase is dried and evaporated and yields 700 mg of 21-acetoxy-3-ethoxy-ΙΙβ , 17aoc-dihydroxy-D-homopregna-3> 5-diene-20-one , which is dissolved in 25 ml of dimethylformamide and 2. 5 ml of water. Gaseous perchloryl fluoride is introduced at 0°C during 15 minutes. The reaction mixture is poured on ice water and extracted with methylene chloride. The organic phase is washed with water, dried and evaporated. There is obtained a mixture of 6a-fluoro-D-homocortisol-21-acetate and its 6 -isomer in a ratio of about 1 : 5.

Example 14 3. 9 g of D-homo-21-acetoxy-llp-hydroxypregna- , 17a (20)-dien-3-one are dissolved in 95 ml of t-butanol. 7 ml of pyridine and subsequently, 20 mg of osmium tjetroxide and 23 ml of 1 , 5 N N-methylmorpholine oxide/hydrogen peroxide solution in t-butanol are added. After 24 hours, - ψ peroxide are added. The reaction mixture is stirred for 24 hours, poured into water and thoroughly extracted with methylene chloride. The organic layers are washed with water, dried and evaporated. The crude oily product is chromatographed on silicagel and gives l.j5 g of D-homo-cortlsol -21-acetate, which is identical with the product of Example 1.

Example 15 In analogy to Example 6, there is obtained from D-homo-21-acetoxy-9, Ιΐβ-epoxy -6a-fluoro-17aa-hydroxypregna-l,4-diene-j5, 20-dione (prepared from D-homo-21-acetoxy-9a-bromo-6a-fluoro-ΙΙ , 17aa-dihydroxypregna-l,4-diene-j5,20-dione) the compound D-homo-21-acetoxy-6a , 9a-difluoro-llt3, 17aa-dlhydroxypregna-l,4-diene-3, 20-dione, melting point 24o-24l°C 16,100.

Example 16 A mixture of 1.0 g of 5a , 0a-dichloro-21-acetoxy-ll| - 17aa-dihydroxy-D-homo-pregnane-3, 20-dione, 1.0 g of anhydrous sodium acetate and 40 ml of 95 per cent ethanol is heated to reflux for JO minutes. The reaction mixture is then poured into 250 ml of ice water and extracted with methylene chloride. The extract is washed with water, dried and evaporated. The residue is recrystallized from acetone-hexane and yields pure 21-aceto y-6β-chloro-llβ, l?aa- dihydroxy-D-homopregn-½-ene-;5, 20-dione.

The starting material can be prepared as follows: 21-acetoxy-li -17aa-dihydroxy-D-homopregn- -ene-3, 20-dione is reacted with ethylene glycol and p-toluene sulfonic acid in benzene to yield the 3-monoketal. The ketal is dissolved in chloroform and the solution treated with 1.05 equivalents chlorine at 0°C. The reaction mixture is allowed to warm to room temperature where upon nitrogen is bubbled through the solution. After washing and drying, the solution is evaporated on the residue left to stand with acetone 2 N for 5 hours at room temperature. Usual work-up gives the desired compound.

Example 17 A mixture of 2. 5 g of 21-acetoxy-6j3-chloro-5a ,ΙΙβ , 17aa-trihydroxy-D-homo-pregna-3, 20-dione, 250 ml of glacial acetic acid and 10 ml of cone, hydrochloric acid is left to stand for 17 hours at 5°C, then poured on ice water and extracted with methylene chloride. The extract is washed with sodium hydrogen carbonate solution and water until neutral, dried and evaporated under reduced pressure. The residue gives after recrystallization from acetone/hexane pure 21-acetoxy-0a-chloro-lip, 17aa-dihydroxy-D-homo-pregri- -ene-j5, 20-dione.

The starting material can be prepared as follows: 3, 3-ethylenedioxy-21-acetoxy-llp, 17aa-dihydroxy-D-homopregn- - - - - yield a mixture of the 5 Example 18 3* 5 g of 21-acetoxy-17aa-hydroxy-D-homo-pregn-4-ene-3, 11 , 20-trione is suspended in 50 ml of methanol and 2 ml of water.Nitrogen is blown through the suspension to remove oxygen. Thereafter, 2. ? g of semicarbazide hydrochloride and 1. 5 g of sodium hydrogenacarbonate are added. The reaction mixture is heated to reflux for 3 l/2 hours and subsequently kept ot 5°C for 20 hours. 60 ml of water are slowly added and the mixture is cooled to 0°C. The crystalline semicarbazone is sucked off and dried at 70° under reduced pressure. The semicarbazone is dissolved in 300 ml of tetrahydrofuran and reacted with a solution of 6.0 g of sodiumborohydride in 100 ml of water. The reaction mixture is stirred at room temperature for 2 hours, cooled to 5¾ , cautiously adjusted to pH 5· 5 by the addition of acidic acid and concentrated. Water is then added and the mixture is filtered. The precipitate is dissolved in 500 ml of 2.5 N hydrochloric acid under nitrogen atmosphere. 50 ml of water is added within 10 minutes at 0-5°C. After stirring for 30 minutes, a solution of 0 g'of urea in 50 ml of water is .added within 5 minutes. The resulting solution is neutralized with 20% sodium hydroxide solution at a temperature not exceeding 15°C and extracted with chloroform several times. The extract is evaporated to dryness and the residue after the addition of 50 ml of acetic anhydride and 0 ml of pyridine left to stand at room temperature for 2 hours. Usual work-up affords 14 g of crude product which after chromatography on silicagel yields pure 21-acetoxy-l^ , 17aoc-dihydroxy-pregn-4-ene-3-20-dione of melting point 212-213°C.

Example 19 In analogy to the process of Example 1 there is obtained from 6cc-fluoro-lip, 17aa-dihydroxy-D-homo-pregn-4-ene-3> 20-dione the compound 21-acetoxy-6a-fluoro-lip, 17aa-dihydroxy-D-homo-pre,gna-l , 4-diene-3, 20-dione , m. . 229-232°C [ EXAMPLE 26 By a procedure analogous to that described in Example 28d - - Example 28 A solution of 375 mg of D-homo-prednisolone , 0.3 ml of pyridine and 0.96 ml of oleic chloride in 10 ml of methylene chloride was stirred at 25° C for 70 minutes. After the addition of 5 ml of water, the mixture was stirred for another 5 minutes, diluted with methylene chloride and washed ith 2 n hydrochloric acid, diluted sodium carbonate, solution and water and evaporated.

After chromatography on silica gel, there was obtained D-homo-prednisolone-21-oleate, m.p."118-120° C, [a]^4 = + 86° (c = 0,1 in dioxane ) .

Example 2,¾. mg of 17a,21-dihydroxy-D-homopregna- ,9(ll)-diene- -3,20-dione 21-hemisuccinate in 20 ml of dioxane and ml of water are treated with 270 mg of N-bromoacetamide and 1.85 ml of 10% perchloric acid and the mixture is stirred at room temperature for 15 minutes. 1.5 g of. sodium sulphite and 30 ml of water are then added. After stirring for a short time, the mixture is extracted with methylene chloride, washed with water dried and evaporated. There are obtained ^0 mg of 113,17a,21- -trihydroxy-9-bromo-I)-homopregn- -ene-3♦20-dione 21-hemisucc nate which is uniform according to thin-layer chromatography; UV: eO Q = 148 0.

The starting material can be prepared as follows: - 21-Acetoxy-17a-hydroxy-D-homopregna- , 9( 11 )-diene-3 , 20- -dione in methanolic solution is saponified with potassium carbonate, there being obtained 17a,21-dihydroxy-D-homopregna- - , 9(ll)-diene-3,20-dione; UV: *= 15800. This is converted in carbon tetrachloride with succinic acid anhydride in the presence of pyridine at an elevated temperature into 17a, 21- -dihydroxy -D-homopregna-4#-9(11 )-diene-3,20-dione 21 -hemisuccinate UV : £239 = 1 620° - ' Example 30 880 mg of 21-bromo-113i 17a-dihydroxy-D-homopregn- -ene--3,20-dione, 600 mg of succinic acid and 1.4 ml of triethylamine are boiled at reflux for 24 hours under argon in 16 ml of acetone. The mixture is poured on to dilute hydrochloric acid and extracted with methylene chloride. The methylene chloride extract is washed with water, dried over sodium sulphate and the solvent removed in vacuo. After crystallisation of the residue from acetone/hexane, there is obtained 11β, 17a,21-tri-hydroxy-D-homopregn-4-ene-3,20-dione 21-hemisuccinate ; UV: ^2 2 '' 15900·

Claims (3)

1. 41752/2 CLAIMS: 1) A process for the manufacture .of the D-homosteroids of the general formula , wherein R represents a hydrogen, fluorine or chlorine atom or the methyl group, R represents a hydrogen, fluorine, chlorine or 17 21 bromine atom and R ' and R each independently alkenoyloxy, represent a hydroxy ^ alkanoyloxy ,/ aralkanoyloxy or aroyloxy group and 1,2-dehydro derivatives thereof which process comprises a) hydroxylating in the 11-position a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof by means of microorganisms or enzymes obtained therefrom, or b) treating a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof with an alkali acylate, or c) dehydrogenating a D-homosteroid of formula I given in claim 1 in the 1 ,2-position, or d) adding hypochlorous or hydrobromous acid to the 9»H-double bond of a D-homosteroid of the general formula 41752/2 or of a 1,2-dehydro derivative thereof, or e) treating a D-homosteroid of the general formula or a 1,2-dehydro derivative thereof with hydrogen fluoride, hydrogen chloride or hydrogen bromide, or alkenoyloxy, f) saponifying an alkanoyloxy ,/ aralkanoyloxy or aroyloxy group in a D-homosteroid of formula I given in claim 1 or in a 1,2-dehydro derivative thereof in 17 21 which at least one of R and R represents an acyloxy group, or g) isomerising a 6p-fluoro-, 6 -chloro- or δβ-methyl-D--homosteroid of formula I given in claim 1 or a 1,2-dehydro derivative thereof to a 6a-isomer, or h) treating a D-homosteroid of the general formula (VI) (Via) with a fluorinating or chlorinating agent, or alkenoylating , i) alkanoylating,/aralkan°ylatini-T or aroylating a 17a- or 21-hydroxy group in a D-homosteroid of the general formula or in a 1,2-dehydro derivative thereof, or j) dehydrating a D-homosteroid of the general formula > or k) reducing the 11-keto group in a D-homosteroid of the general formula or in a 1,2-dehydro derivative thereof to the hydroxy group with protection of the 3- and 20-keto groups, or 1) oxidising the 17(20)-double bond in a D-homosteroid of the general formula to the hydroxy ketone group, or m) dehydrohalogenating a D-homosteroid of the general formul ; in the foregoing formulae R , R~, R ' and 21 R have the significance given in claim 1, R 61 represents an acyloxy or alkoxy group, R represents a fluorine or chlorine atom or the methyl group and Hal represents a chlorine, "bromine or iodine atom.

2. ) A process according to claim 1, wherein there is manufactured a compound of formula I given in claim 1 or a 1,2-dehydro derivative thereof in which R and R^ represent 17 21 a hydrogen or fluorine atom and R ' and R represent a hydroxy or C, .--aikanoyloxy group.

3. ) A process according to claim l or claim 2, wherein D-homo-9tt-fluoro-prednisolone-21-acetate is saponified. A process for the manufacture of the D-homosteroids set forth in claim 1, substantially as hereinbefore described with reference to the foregoing Examples. 5) Process for the manufacture of pharmaceutical preparations characterized in that a D-homosteroid as claimed in claim 1, is mixed, as active substance, wi nontoxic, inert, therapeutically compatible solid or liquid carriers, commonly used in such. preparations, and/or. excipienta. 6) A pharmaceutical preparation containing a D-jhomo-steroid as claimed, in claim 1 in association with a compatible pharmaceutical carrier. 7) D-Homosteroids of the general formula , wherein R represents a hydrogen, fluorine 9 or chlorine atom or the methyl group, R represents a hydrogen, fluorine, chlorine or 17 21 bromine atom and R 1 and R each independently alkenoyloxy, represent a hydroxy^ alkanoyloxy ,/ aralkanoyloxy or aroyloxy group and 1,2-dehydro derivatives thereof, whenever prepared according to any one of claims 1-4, 8 ) Compounds of formula I given in claim 1 , wherein RD and R 9 represent a hydrogen or fluorine atom and R171 and 21 R represent a hydroxy or C^_g-al'kanoyloxy group and 1 , 2-dehydro derivatives thereof, whenever prepared according to any one of claims 1-4 , or by an obvious chemical equivalent thereof. 9 ) 9