CA1098897A

CA1098897A - D-homosteroids

Info

Publication number: CA1098897A
Application number: CA285,723A
Authority: CA
Inventors: Leo Alig; Marcel Muller; Andor Furst; Ulrich Kerb; Klaus Kieslich; Rudolf Wiechert
Original assignee: F Hoffmann La Roche AG
Current assignee: F Hoffmann La Roche AG
Priority date: 1976-09-03
Filing date: 1977-08-30
Publication date: 1981-04-07
Also published as: ES469573A1; NL182314C; GR73039B; FI57600C; IE45671L; GB1586504A; ATA656076A; PH14434A; DK393277A; IL52831A; JPS5331655A; NL7709585A; DK140947C; MY8500293A; ES469574A1; FR2363580A1; ES462076A1; AU516174B2; AR220320A1; HK26484A

Abstract

Abstract of the Disclosure The specification discloses novel D-homosteroids, having inflammation-inhibiting activity, and having the formula

Description

5a7 The present invention relates to novel D-homosteroids of the fo.rmula fH3 H~,F~17 a ~ ~ J

wherein the broken line in the 1,2-position denotPs an optional carbon-carbon bond; R6 represents a hydrogen, fluorine or chlorine atom or a methyl group; R9 represents a hydrogen, fluorine or chlorine atom; and R17a represents a hydroxy or acyloxy group, with the proviso that, where R6 represents a hydrogen atom in a 11,17a-dihydroxy compound or where R represents a fluorine atom in a 11,17a-dihydroxy-4-ene compound, R9 must represent a fluorine or chlorine atom.
An acyloxy group can be derived from a saturated or unsaturated aliphatic carboxylic acid, a cycloaliphatic, araliphatic or an aromatic carboxylic acid pxe~erably containing up to 15 carbon atoms. Examples of such acids are formic . .

`, ', '. ~
. ~ ~

acid, acetic acid, trifluoroacetic acid/ pivalic acid, propionic acid, butyric acid, caproic acid, oenanthic acid, undecylenic acid, oleic acid, cyclopentylpropionic acid, cyclohexylpropionic acid, phenylacetic acid and benzoic acid.
Especially preferred acyloxy groups are alkanoyloxy groups containing from 1 to 7 carbon atoms.
D-Homosteroids of formula I containing a double bond in the l,~-position are preferred.
Examples of D-homosteroids for formula I provided by the present invention are:
17a-butyryloxy~ hydroxy-D-homopregna-1,4-diene-3,20--dione, 6~-fluoro~ ,17a-dihydroxy-D-homopregna-1,4-diene-3,20--dione, 6~-chloro-11~,17a-dihydroxy-D-homopregna-1,4-diene-3,20--dione, 17a-butyryloxy-6a-fluoro~ -hydroxy-D-homopregna-1,4--diene-3,20-dione, 17a-valeroyloxy-6~-chloro-11~ hydroxy-D-homopregna-1,4---diene-3,20-dione, 9-fluoro~ ,17a-dihydroxy-D-homopregna-1,4-diene-3,20--dione, 9-chloro-11~,17a-dihydroxy-D-homopregna-1,4-diene-~,20--dione, 17a-butyryloxy-9-fluoro~ hydroxy-D-homopregna-1,4--diene-3,20-dione, ~,.-~. . . .

~.
.

~.~g~ 7 9-chloro~ -hydroxy-17a-propionyloxy-D-homopregna-1,4--diene-3,20-dione, 11~,17a-dihydroxy-6a-methyl-D homopregn-4-ene-3,20-dione, 17a-butyryloxy-11~-hydroxy-6a-methyl-D-homopregn-4-ene--3,20-dione, 11~,17a-dihydroxy-6a-methyl-D-homopregna-1,4-diene~3,20--dione, 17a-butyryloxy-11~-hydroxy-6a-methyl-D-homopregna-1,4--diene-3,20-dione, 109-1uoro~ ,17a-dihydroxy-6a~methyl-D-homopregn-4-ene--3,20-dione, 17a-butyryloxy-9-fluoro~ -hydroxy-6a-methyl-D-homopregn--4-ene-3,20-dione, 9-fluoro-11~,17a-dihydroxy-6a-methyl-D-homopregna-1,4-15-diene-3,20-dione, 9-fluoro-11~-hydroxy-6a-methyl-17a-valeroyloxy-D-homo-pregna-1,4-diene-3,20-dione, 9-chloro-6a~fluoro-11~,17a-dihydroxy-D-homopregna-1,4---diene-3,20-dione, 206a,9-difluoro-11~,17a-dihydroxy-D-homopregna-1,4-diene--3,20-dione, 6a,9-difluoro-11~-hydroxy-17a-propi.onyloxy-D-homopregna--1,4-diene-3,20-dione.
The D-homosteroids of formula I are prepared in 25accordance with the present invention by ~a) hydroxylating a D-homosteroid of the general formula ,, : ',~ .

. ~ .

, . . , ~

fH3 ~0 ~J~ R I7 a (lIj ~ ''~
R~
in the ll-position by means of microorganisms or enz~nnes producPd there~rom, or (b~ replaclng the iodine atom in a D-homo~3teroid of the gerleral forrnula E~O~,~.R17 a by a hydrogen atom, or ~c) dehydrogenating a 1,2-saturated D-homosteroid of formula I in the 1, 2~position, or - ' :.~ :

g8~

(d) adding hyp~chlorous acid to the 9,11-double bond of a D-homosteroid of the general formula fH3 CO
~I~ R 17 a (IV), or (e) treating a D-homosteroid of the general formula f~3 c~
H3C ~.R17 a (v) with hydrogen fluoride or chloride, or -~ , ~.c ~, ... .

, ~9~7 ( f ) sapon~ fying a 17ad-acyloxy group in a D-homosteroid of ~ormula I, or (g) isomerising a 6~-(fluoro, chloro or methyl)oD-homo steroid corre~ponding to formula I to the 6a-isomer, or (h~ ~luorlnating or chlorina~-ing a D-homosteroid of the general formula fH3 C~
H~ R 17 a )\J
(VI ) in the 6-position, or (i) acylating a 17~-hydroxy group in a D-homo-steroid of foxmula I, ..

~: ;

.:

or ( j ) oxidising the 3-hydroxy-~5~grouping in a D-homostexoid of the general formula C~O
H ~ ~ R17a ~ (VIII) to the 3-keto-~4~gxouping, - or (k) reducing the ll-keto group in a ~-homosteroid of the general formula fH3 ~o ~:
0~.F~ 17 a t~3 l \~ (IX) ~z b~

to the hydxoxy group with protection of the 3- and 20-keto group, or (1) oxidising the 17a(20)-double bond in a D-homosteroid .if~, o the general formula .~=^J -- 7 --.

~:, , , . :, .... .. .....

~ ~98~97 ``
fH3 HO ~
~ (X) ~6 to the hydroxyketone grouping, o~
(m) methylating a D-homosteroid o formula VI in the 6-posltion, or ~ n) converting the 17~-ethynyl group in a D-homosteroid of the general formula IllH

H~.R~7 a (XI) 19 into the acetyl group, or (o) dehydrogenating a D~homosteroid of the general formula , ,; , .:, : . ~., . . . -. , , , :, 7 ~
f~3 ~o H~a~,R17a H3Ç 1 1 1 (XII) in the 4-positlon;
in whlch formulae R6, R9, Rl7a and the broken lin~ in the l,2-positlon have thP
signlficance given earller and R6~
repre~ents a hydrogen atom or a methyl group.

The hydroxylation of a D-homosteroid of formula II in accordance w.ith embodiment (a) of the process can be carried out according to methods known per ~e for the microbial ll--hydroxylation of sterolds. For thi~ hydroxylation there can be used microorganisms of the taxonomic groups Fungi and Schizomycetes, especially of the sub-groups Ascomycekes, Phycomycetes, Basidiomycetes and Actinomycetales~ There can also be used mutants produced in a chemical manner (e.g. by treatment with nitrite) or in a physical manner (e.g~ by irradiation) as well as cell-free enzyme preparations obtained from the microorganismsO Especially suitable microorganisms for the ll~-hydroxylation are those of the genera Curvularia (e.g. C. lunata NRRL 2380 and NRRL 2178; ATCC 13633, l3432, 14678, IMI 77007, IF0 28ll), Absldia (e.g. A. coerula IF0 4435), g _ :

- ' ~ ~ ' ' ' ,, -'; ` "' ' ~ ' I

t,~ 7 -- 10 ~
Colletotrichum (e . g . C . pisl ATCC 12520~, Pelllcolarla (e . g ~
P. filamentosa IF0 6675) ~ Streptomyces (e.g. S. fradiae ATCC
10745), Cunninghamella (e.g. C. bainieri ATCC 9244, C.
verticellata ATCC 8983~ C. elegans NRR$ 1392 and ATCC 9245, C. blakesleeana ATCC 8688, 8688a, B688b, 8983 and C. echinulata ATCC 8984), Pycnosporium (e.g. sp. ATCC 12231), Verticillium (e.g~ V. theobromae CBS 39858), Aspergillus (e.g. A.
quadrilieatuæ JAM 2763), Trichothecium (e.g. T. roseum ATCC
12519) and Phoma (e.g. sp. ~TCC 13145).

The replacement of the lodine atom tn a D-homosteroid of formula III by a hydrogen atom in accordance with embodiment (b) of the process can be carr~ed out by treatment with a reduction agent such as sodlum hydrogen sulphite.

The 1,2-dehydrogenation of a 1,2-saturated D-homosteroid of formula I in accordance with embodiment ~c) o the process can be carried out in a manner known per se; for example, in a microbiological manner or using a dehydrogenating agent such as iodine pentoxide, periodic acid, selenium dioxide, 2,3-dichloro--5,6-dicyanobenzoquinone, chloranil or lead tetraacetate.
Suitable microorganisms for the 1,2-dehydrogenatlon are, or example, Schizomycetes, especially those of the genera Arthrobacter (e.g. A. slmplex ATCC 6946), Bacillus ~e.g. B.
lentus ATCC 13805 and B. sphaericus ATCC 7055), Pseudomonas (e.g. P. aeruginosa IF0 3505), Flavobacterium (e.g. F~
flavescens IF0 3058), hactobacillus (e.g~ L. brevis IF0 3345) and Nocardia (e.g. N. opaca ATCC 4276).

.; ~

:' :,:

..

In carrying out ~mbodiments (d) and (e) vf the process, a D-homosteroid of fonmula I~ or V is conveniently dissolved in a suitable solvent (e.g. an ether such as tetrahydrofuran or dioxan, a chlorinated hydrocarbon such a~ methylene chlorlde or S chloroform, or a ketone such as acetone) and le~t to react with the reagent which is added thereto. Hypochlorous acid is conveniently generated in situ; for example, from N-chloroamides or imides such as N-chlorosuccinimide and a strong aci~,.pr~.erably perchloric acid. Embodiment ~e) is preferred for ~he preparation o~ 9--fluoro ll~hydroxy-D-homosteroids of formul? I.

The saponification of an acyloxy group in a D-homosteroid of formula I in accordance with embodiment (f) of the process can be carried out in a manner known per se; for example, with aqueous-methanolic potassium carbonate solutlon or sodium hydrogen carbonate solution.

The isomerisation of a 6~-~fluoro, chloro or methyl)-D--homosteroid corresponding to formula I, especially a 6~-(fluoro or chloro)-D homosteroid, in accordance with embodiment ~g) of the process can be carried out by treatment with an acid, especially a mineral acid such as hydrochloric acld, in a solvent (e.g. dloxan or glacial acetic acid)~

~ he fluorination or chlorination of a D-homosteroid of formula VI in the 6-position in accordance with embodiment (h) of the process can be carried out in a manner known per se. A
6,7-saturated D-homosteroid of formula VI can be fluorinated or .~ , f ,, . '. , . ,' ' .. ,' .
'` ~, ' ' ' ' .
` ' ' ' . ' . . ' `
'' ,, ~ ~ "
' ' ' ,': ' . . :

8~7 ~`

chlorinated by reaction with a fluorinating or chlorinating agent such as a N-chloroamide or imide (e.g. N-chlorosuccinimide) or with elemental chlorine [see J. Am. Chem. Soc. 72, 4534 (1950)]. This embodiment of the process is preferably carried out by converting a 6,7-saturated D-homosteroid of formula VI
into a 3-enol ester or 3-enol ether (e.g. the 3-enol acetate) and reacting the 3-enol ester or 3-enol ether with chlorine [see J. Am. Chem. Soc. 82, 1230 ~1960)], with a N-chloro~mide [see J. Am. Chem. Soc. 82, 1230 (1960); 77, 3827 (1955)] or with perchloryl fluoride [see J. Am. Chem. Soc. 81, 5259 (1959);
Chem. and Ind. 1959, 1317]. Trifluoromethylhypofluorite can also be used as the fluorinating agent.

Insofar as the previously described fluorlnation or chlorination yields an isomer mixture (i.e. a mixture of 6a-and 6~-tfluoro or chloro)-D-homosteroids, the mixture can be separated into the pure isomers according to known methods such as chromatography.

The acylation of a 17a-hydroxy group in a .
D-homosteroid of formula I in accordance with ~mbodiment (i) of the process can be carried out in a manner known per se; for example, by treatment with an acylating agent such as an acyl chloride or anhydride in the presence of an acid binding agent (e.g. pyridine or triethylamine) and a suitable catalyst te.g.

p-dimethylaminopyridine) or in the presence of a strong acid catalyst (e.g. p-toluenesulphonic acid). As the solvent for the acylation there may be mentioned organic solvents which do not contain hydroxyl groups (e.g. chlorinated hydxocarbons such ~ I - 12 -, " . , .

. ",, ~-- . .

~- . . ` ,:

as methylene chloride or hydrocarbons such as benzene).

The oxidation of the 3-hydroxy-~5-grouping ln a D-homo-steroid of formula VIII in accordance with embodiment (j) of the process can be carried out according to the Oppenauer procedure 5e.g. using aluminium isopropyla~e) or by means OI oxidlsing agents such as chromium trioxide ~eOg~ Jones' reagent3 or according to the Pfitzner Mofatt procedure using dimethyl-sulphoxide/dicyclohexylcarbodiimide ~the initially obtained ~5-3-ketone requiring subsequent isomerisatio~ to the ~4-3--ketone) or ~y means of pyridine/sulphur trioxide.

In carrying out embodiment (k) of the process, the keto groups in the 3- and 20~positions of a D-homosteroid of formula IX are first protected (e.g. as the semicarbazone). Where a 1,2-double bond ls present, the 3-keto group can also be protscted by the formation of an enamine. The protecting gxoups can subsequently bP removed by acid hydrolysis. A ~1~4_ -3-ketone can also be converted lnto a al'3'5-3-enamine using a secondary amine in the presence of titanium tetrachloride. The reduction of the ll-keto group of a thus-protected D-homosteroid 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~teroid of formula X in accordance with embodiment (1) of the process ~ ~ 13 --~ ^ ' :.

. . ~ ., , : , : .

"

OE ~g~97 can be carri~d out, for example, with an oxidising agent such as a tertiary amine N-oxide peroxide in tert.butanol/pyridine ln the presence of ca~alytic amounts of osmium tetroxide. Exa~ples of tertiary amlne N~oxide peroxides which can be used in this embodiment are N-me~hylmorpholine N-oxide peroxide and triethyl-amine oxide peroxide. The 17 (20)-double bond can al~o be oxidised with an oxidising agen~ such as osmium tetroxide or permanganate to give a 17,20-glycol which can be fur~her oxidised to ~he hydroxyketone w~th an oxidising ag~nt such as chromic acid.

The methylation of a D-homos~eroid of formula VI in the 6-position in accordance with embodiment ~m) of the procPss can be carried out, for example, by conver~ing a D-homosteroid of fQrmula VI into a 3-enol eth~r (e.g~ by treatment with an orthoformic acid ester such as ethyl orthoformate in the presence of an acid such as p-toluenesulphonic acid, if desired, with addition Qf the corre~ponding alcohol; or by treatment with a dialkoxypropane such as 2,2-dimethoxypropane in methanol/
dimethylformamide in the presence of p-toluenesulphonic acid) and reacting the 3-enol ether with a tetrahalomethane (e.g.
CBr4, CC12Br2 or CC13Br) to give a trihalomethyl-~4-3-ketoneO
A trihalomethyl-~4-3-ketone can be dehydrohalogenated with a base such as collldine to give a dihalomethylene-a4-3-ketone which can be converted by catalytic hydrogenation under mild conditions (e.g. using a Pd/SrC03 catalyst) into a 6a-methyl -~ -3-ketone.

Another methylation procedure consists in converting a ~1,2-saturated D-homosteroid of formula VI into a 3-enol ether .
.. ~ .

. - 15 - ~

as described earliex afi~ reacting this 3-enol ether ln a manner known per se to give a corresponding 6-~ormyl derivative, reducing the formyl group with sodium borohydride to the hydroxymethyl group and finally dehydrating the product obtained 5 with cleavage of the enol ethex, there being obtalned a 6--methylene-D-homosteroid o~ the general formula ~3 CO
HO ~ ~17a (VII) ~`1~

wherein R9 and ~17a have the significance given earlier.

6 Methylene-D-homosteroids of formula VII can also be obtained by converting a 1,2-saturated D-homosteroid of formu}a VI into a 3-enamine (e.g. the 3 pyrrolidinium enamine), hydroxy-methylating the 3-enamine with formaldehyde and cleaving water from the hydroxymethylation product using an acid such as p-toluenesulphonic acid.

A 6-me'hylene-D-homosteroid of formula VII can he catalytically hydrogenated to give a corresponding 6-methyl-D--homosteroid of formula I in a manner known per se; for example, using a known hydrogenation catalyst.

~, - ~ . '' ~

9~7 ~r ~

The convexslon of the 17~-e hynyl group in a D~homosteroid of formula XI into the acetyl group in accordance wlth e~bodiment (n~ of the process can be carxied out in the presence of a suitable catalyst such as p-toluenesulphonamide-mercury or with ' acid ion exchangers activated with mercury salts.

In accordance with embodiment (o) of the process, a D-homostero1d of formula XII ca~ be dehydrogenated in the 4-position or 1,4-positlon by bromination and subsequent dehydrobromination.

The starting materials used ln the foregoing process, insofar as they are not known or insofar as their preparation is not descri~ed hereinafter, can be prepared in analogy to known methods or methods described ln the Examples hereinafter.

The D-homosteroids of formula I possess inflammation--inhibiting activity and can accordingly be used, for example, for the treatment of inflammatory conditions such as ecæemas.

In general, pharmaceutical preparations for internal administration can contain 0.01% to 5.0~ of a D-homosteroid of ~ormula I. The da~ly dosage can vary between 0.05 mg and lO.0 mg depending on the condition to b~ treated and the duration of the desired treatmentO The amount of D~homosteroid of fonmula I
in topical preparations lies, in general, in the range of from 0.0001 wt.% to 5 wt.%, advantageously in the range of from 0.001 wt.% to 0.5 wt.% and preferably in the range of from 0.01 w~.% to 0.25 wt.%.

: ,~ -: .

- :: : , ~: :

.: :

. '~
_ 17 -The D-homosteroids of formula I can be used as medicament~
in the f~rm of pharmaceutical preparations which contain them in association with a compatibla pharmaceutical carrier material.
This carrier material can be an organic or inorganic inert S carrier material suitable for ènteral t percutaneous ox parenteral administratlon such as, for example, wa~er, gelatin, gum arabic, lactose, starch, magne~um stearate, talc, vegetable olls, polyalkyleneglycols, petroleum ~elly etc. ~he pharmaceutical preparations can be made up, for example, as salves or as solutions, suspens~ons or emulsions. The pharmaceutlcal preparations may be sterilised and/or may contain adjuvants such as preserving, stabilising, wetting or emulslfying agents, salts fox the varlation of the osmotic pressure or buffers.
They can also contain still other therapeutically valuable materials.

~ 17 -8g7 . , 4.9 g of 9-fluoro-llB,17a-dihydroxy-21-lodo~D-homopregna--1,4-diene-3,20-dione, 80 ml of ether, 80 ml of benzene, 40 ml of water and 40 ml o saturated sodium hydrogen sulph$te solution were stirred a~ 25C for 30 hours. The mixtur~ was diluted with ethyl a¢etate. The aqueou~ phase was ~eparated and extracted twice wlth ethyl acetate. The ethyl acetate solutions ware washed twice w~th sodium chloride solution, dried over sodium sulphate and evaporated-. Filtration on silica gel and crystallisatlon from acetone/hexane gave 9-~luoro~ 17a-~dihydroxy-D-homopregna-1,4-diene-3,20-dione of melting point 268~-269C; W : E329 = 15200; [a]D ~ +67~ (c = 0.1~ ~n me~hanol).

-- The starting material can be prepared by reacting 9--fluoro-D-homopr~dnisolone r melting point 241-246C; []D ~
+101 ~c = 0.1% in dioxan); W : E23~ = 14540 7 with methane-sulphonyl chlorlde in pyridine to gi~e 9-fluoro~ ,17a--dihydroxy-21-methanesulphonyloxy-D-homopregna-.l,4 diene-3,20--dione and reacting this with sodium lodide in acetone to give 9~fluoro~ ,17a-dihydroxy-21-iodo-D-homopregna-1,4-diene-3,20-. . -~ : : -. - ~ - .. . . . .....
:, . ~. ,: : . .

g7 -;

-dione of melting point 190C ~decomposition); [a]~ = +118 (c = 0.1~ in dioxan)i W : 23~ = 15750-In an analogous manner, from 6a~luoro~ ,17a-dihydroxy 21-iodo-D-homopregna-1,4--diene~3,20-dione L mel~1ng polnt 175-177C; W : ~243 ~
15830; [a]D = +121 (c ~ 0.1% in dioxan) 7 there was obtained 6a-fluoro-11~,17a-dihydroxy-~ homopregna--1,4-diene-3,20-dione of melting point 183-184C; W : 242 = 14400; [a]D = +56 ~c = 0.1% in dioxan)i and from 6a,9-difluoro-11~,17a-dihydroxy 21-iodo-D-homopxegna-1,4--dien-3,20-dione ~ melting point 189-190C; W: E238 = 16750;
[a]D = +115 (c = 0.1% in dioxan) 7 there was obtained 6,9-~difluoro~ ,17a-dihydroxy-D-homopregna-1,4-diene-3,20-dione of m~lting point 230-231~C; W : E238 = 16000; ~a3D = +57 (c = 0.1~ in dioxan).

60 mg of 9,11~-epoxy-17a-hydroxy D-homo-9~ pregna-1,4--diene~3,20-dione were stirred at 25C for 15 minutes in 1.5 ml of glacial acet~c acid and 0.15 ml of 37% hydrochloric acid.
The mixture was poured into dilut~ sodium hydrogen carbonate solution and extracted three times with methylene chlorid~.
The methylene chloride solutions were washed twice with dilute sodium chloride solution, dried and evaporated. From acetone there was obtained 9-chloro~ ,17a-dihydroxy-D-homopregna-1,4-

2~ -diene-3,20-dione of melting point 265-270C (decomposition)i W . 240 - 15080p [~]D = ~94 (c = 0.1~ in dimethylsulphoxide).

.' - 19 _ .
.; . . ~ ~ -139~

Example 3 100 mg of 9,11~-epoxy~17a-hydroxy-D-homo-gB-pregna-1,4-dlene-3,20-dione were stirred at 25C for 1 hour ln 2 ml of a solutlon of 1 part of ur~a and 1.3 parts of hydrogen fluoride.
The mixture was poured into water and extracted with methylene chloxide as usual. Chromatography of the crude product on silica gel gave 9-fluoro-11~,17a-dihydroxy-~homopregna-1,4--diene-3,20-dione of melting polnt 268-269C; UV:
15~00; [a]D = ~67~ (c - 0.1~ in methanol).

10 The starting material, 9,ll~epoxy-17a-hydroxy-D-homo-9~--pregna-1,4-diene-3,20-dione r melting point 179-180C;
W: E248 = 16060; [a]D = -15 ~c - 0.1~ in dloxan) 7, is obtained from 9 bromo~ ,17a-dlhydroxy-D-homopregna-1,4-diene-

-3,20-dione and potassium acetate in alcohol after heating to . : 15 reflux for several hours.

1 g of 6~-fluoroT11~,17a-dlhydroxy-D-homopregn-4-ene-3,20--dione a~d 660 mg of selenium dioxlde were stirred at reflux for 24 hours under argon ~n 50 ml of tert.butanol and 0.5 ml of glacial acetic ac~d. ~ The mixture was filtered and evaporated.
The resldue was di~solved ln e~hyl ace~ate and washed ~uccessively with sodium hydxogen carbonate solu~ion, water, lce~cold ammon$um sulphide ~olution, dilute ammonia, water, ~ 20 -.~
. ~1 .. .... ... . . ,~ .

, , , , . , .-. :. . ......
., .. . ,:

dilute hydrochloric acid and water. The ethyl acetate solution was dried over sodium sulphate and evaporated in vacuo.
Chromatography on silica gel gave 6a-fluoro~ ,17a-dihydroxy--D-homopregna-1,4-diene-3,20-dione of melting point 183~-184C;
UV: ~242 = 14400; [a]D = ~56 (c = 0.1~ in dioxan).

Exam~le 5 Preparation of The Starting Material 1.1 g o 11~,17a-d~hydroxy-D-homopregna-1,4-diene-3,20--dione were dissolved at -10C in 6.2 ml of pyridine and 0.474 ml of trifluoroacetic acld anhdyrlde and stirred at 0C for 50 minutes under argon. The mixtu.re was poured into dilute hydrochloric acid and extracted three times wlth methylene chloride. The methylene chloride solut~ons were washed neutral with sodium hydrogen carbonate solution and sodium :~
chloride solution, dried and evaporated. Chro~tography on silica gel gave pure non-crystalline 17a-hydroxy~ trifluoro-acetoxy-D-homopregna-1,4-diene-3,20-dione; W : ~239 = 14200;
ta]D = +84 (c = 0.1% in dioxan~.

1o2 g of 17a-hydroxy llB-trifluoroaceto~y-D-homopregna--1,4-diene-3,20-dlone were dissolved in a mixture of 12 ml of butyric acid and 4.8 ml of trifluoroacetic acid anhydride a~d stlrred at 50C for 4 hours. The mixture was poured into aqueous pyridine, stirred for 10 minutes, acidified with 2-N

~. .

~' ~

. . ~ - . - -: . ;

22 ~

hydrochloric acid and extracted three times with methylene chloride. ~he methylene chlorlde solutions wPre washed neutral with ~odium hydrogen carbonate solut~on and sodium chloride solution, dried over sodium sulphate and ~vaporated.
Chromatography of the residue on silica gel gave puxe 17a--butyryloxy~ trifluoroacetoxy-D-homopregna-1,4-dlene-3,20--dione as a foam. W: 240 = 13900; t~ D - +41 (c = 0.1% in dioxan).

The Process 1.1 g of 17a-butyryloxy~ rifluoroacetoxy-D-homopregna--1,4;~diene-3,20-dione were treated in 55 ml of methanol and 4~2 ml of wa~er with 4.2 ml of saturated sodlum hydrogen carbonate solution and stirred at 25C for 48 hours. The methanol was evaporated and the re~idue taken up in methylene chloride and water. The methylene chloride solution was washed with dilute sodium chloride solution, dried and evaporated. There was obtalned 17a-butyryloxy~ hydroxy-D-homopregna~1,4-diene-3,20--dione as a foam which was pure according to thin-layer chromatography. W : ~244 = 13940; [a]D - +22 (c = 0.13 in dioxan).

~

In an analogous manner to Example 5, from 9-fluoro~ ,17a-d~hydroxy-D~homopregna-1,4-diene-3,~0--dione there was obtained 17a-~utyrylo~y-9-fluoro-llB-hydroxy-- 22 ~
,, j ~- . .. ~. , .. . , - , ~--D-homopregna-1,4-diene-3,20-dione of melting point 187-188C;
W: E~4~ = 14000; ~a3D = +13 ~c = 1% in dioxan); and from 6a,9-difluoro-11~,17a-dihydroxy-D-homopregna-1,4~dlene--3,20 dione there was obtained 17a-butyryloxy-6~9-difluoxo-~ -hydro~y-D-homopregna-1,4-diene-3,20-dione of melting point 224-2~5C; W: 238 = 16400; [a]D ~ ~14 (c = 0.1~ ln dioxan).

~e~

In analogy to Example 5, 1~from 6a-fluvro~llB,17a-dlhydroxy-D-homopregna-1,4-diene 3,20- :
-dione ther~ was obtained 17a-butyryloxy 6a-fluoro-11~-hydroxy--D-homopregna-1,4-diene-3,20 dlone of melting point 168-16gC;
W : ~24~ = 166~0; [a]D = +13 (c 5 O. 1% in dioxan).

15If, ln Example 5, there is used acetic acid, propionic acid or valeric acid in p}ace of butyrlc acid, then there are obtained from 9-fluoro-11~,17a-dihydroxy-D-homopxegna 1,4-dlene--3,20-d~one in analogy to ~xample 5 17a-acetoxy-9~fluoro~ -hydroxy-D-homopregna-1,4-diene-3,20--dione of melting point 232-233C; W : ~239 = 13900; [a]D =
~29 (c ~ 0.1~ in dioxan);

, . , , ~ ~ ~ . - , . ~. . .

9-fluoro~ hydroxy-17a-propionyloxy D-homopregna-1,4-diene--3,20-dione of melting point 204-205C; W : ~238 = 15100;
~a]D = +23 (c = 0.1% in dioxan); and 9-fluoro~ -hydroxy-17a-valeroyloxy-D-homopregna-1,4-diene~
-3,20-dione ~f melting point 144-146C; W : ~23g = i5400;
[~]D = ~17 (c = 0.1% in dioxan).

300 mg of llB,17a-dihydroxy-6-methylene-D-homopregn-4--ene-3,20-dione, 150 mg of 5~ palladium/carbon, 1.5 ml of cyclohexene and 15 ml of ethanol were boiled at reflux for 8.5 hours under argon. The mixture was cooled to 25C, treated with 0.75 ml of 25% hydrochloric acid and stirred for 1 houx.
The cataly~t was ~iltered off and the filtrat2 evaporated.
Chromatography ~n silica gel gav~ ,17a-dihydroxy-6a-methyl~ .
-D-homopregn-4-ene-3,20-dione of ~eltlng point 223 Z25C;
W : ~242 = 14100j ~aJD = +46 (c 2 0.1% in dioxan).

The ~tarting material can be prepared as follow~:

11~,17a-~ihydroxy-D-homopregn-4-ene-3,?0-dlone ls reac~ed ln boiling methanol with pyrrolidine to give 11~,17a-dihydro~y--3~ pyrrolidinyl)-D-homopregna-3,5-dien-20-one. This is reacted with formalin in benzene and methanol to give 11~,17a-. . ~

.

- . . , .,: ::
., . . :: .

-dihydroxy~6~-hydroxymethyl-D~homopregn-4-ene-3,20-dione.
Treatment with hydrochloric acid ln dioxan gives llB,17a--dihydroxy 6-methylene-D-homopregn-4-ene 3,20-dione.

A 2 litre Erlenmeyer flask containlng 500 ml of nutrient solut~on (ster~lised for 30 minutes at 120C in an autoclave) comprising 1~ cornsteep liquor, 1% soya powder and 0.005% soya oil, adjusted to pH 6.2, is inoculat~d with a lyophilised culture of Cur w larla lunata (NRRh 2380) and shaken on a rotary shaker at 30C for 72 hours. A 20 litre stainless steel fermenter containing 15 litres o~ a medium (sterilised at 121C
and 1.1 atmospheres) comprising 1% cornsteep llquor, 0.5% starch sugar and 0.005% soya oil, adjusted to pH 6.2, is then inoculated with the aforementioned pre-culture. Cultlvation 1~ is carried out for 24 hour~ at 29C wlth aeratlon (10 litres/
minute), at 0.7 atmospheres and while stirring (220 revolutions/
minute) with the addition of a silicon oll (Silicon SH) a~ an , . ~ .

anti-foam agent. 1 litre of the culture broth is tran~ferred under sterile conditions into 14 litres of a medium (sterilised as described earlier) romprising 1~ cornsteep l~quor, 1.25~ soya powder and Q.005~ soya oil and cuitivated under the same conditions. ~fter 12 hours, there is added a solutivn of 4 g of 17a~-aceto~y-D-homo-4 pregnene-3,20-dione in 100 ml of dimethylformamide. After 52 houxs, the conten~ of the fermenter is extracted twice by stirring with 10 litres of methyl i~obutyl ketone each time and the extract evaporated at 50C (bath tempera~ure) in vacuo. In order to remove the silicon oll, the residu~ is washed several times with hexane and separated from unreacted starting material by column chromatography on silica gel tgxadient elution: hexan~ + hexane/ethyl acetate 1 ~ (1/1)]. The 17aa-aceto~y~ hydroxy-D-homo-4-pregnene--3,20-dione is recrystallised from i~opropyl ether; melting point 234~/235~237C; ~24~ ' 16700.

A 2 litre Erlenmeyer flask containing 500 ml of a nutrient solution (sterilised for 30 minutes at 120C in an autoclave~
comprising 1.5~ peptone, 1.~ cornsteep and 0.2% magnesium sulphate, adjusted to pH 6.5, ls inoculated with a lyophilised culture of Bacillus lentus (ATCC 13805) and shaken at 30C for 24 hours~ A 20 litre stainless steel fermenter con~aining 15 litres of a liquid nutrient medium (sterilised at 121C and 1.1 atmospheres) comprising 0.2~ yeast extract, 1% cornsteep liquor and 0.1% starch sugar, adjusted to pH 7.0, is then inoculated with the aforementioned pre-culture. Cultivation is carri~d ~ ~ ~6 _ ., .~i :: . :, .. :
i ` . ~ . -~ , .
:: :: :

... : .

out at 29C wlth aeration and ~tirring and with the additlon of a sllicon oll (Silicon SH) as an an~i-foam agent. ~fter a growth-phase of 6 hours, there i~ added a solutlon of 1. 6 g of 17aa-acetoxy~ hydro~y-~-homo-4-pregnene-3,20-dione in 50 ml of dimethylformamide. After 15 hours, the content of ~he fermenter is extracted twice wlth 10 litres of methyl ~sobu~yl ketone each time and the extract evaporated in vacuo. In order to remove the silicon oll, the residue i~ washed wlth hexane and recrystallis~d from acetona/dii~opropyl ether ln the pre~ance of ac~ive carbon, thex~ being obtalned 17aa-acetoxy~ hydroxy-D~
-homo~ pregnadiene-3,20-dione of meltlng poin~ 218/219-220C
and 2~ = 15100, The ~ollowing Example illustrates a pharmaceutical preparatlon prov1ded by th~ present invention and the manufacture thereof:

Example A

The D-homosteroids of formula I' can be made up, for example, in the form of salves as follows:

D-Homosteroid 0.01-1 wt.
Liquid paraffin 10.0 wt.%
White soft paraffin ad loO parts by weight The D~homosteroid is ground with a portion of the liquid paraffin in a ball mill until a particle size of less than 5 ~
is achieved. The paste is diluted and ~he mill washed out with ~ - 27 . ~ _ .:

l~B9~72 8 the remaining l~quid parafin. The ~u~penslon 18 added to the molten colourless soft paraffin at 50C ~nd stirred until the mass is cold, there being obtained a homogeneous salve.

,, .
:

, . .

Claims

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1) A process for the preparation of the D-homosteroids of the general formula (I) wherein the broken line in the 1,2-position denotes an optional carbon-carbon bond; R6 represents a hydrogen, fluorine or chlorine atom or a methyl group; R9 represents a hydrogen, fluorine or chlorine atom; and R17a represents a hydroxy or acyloxy group, with the proviso that, where R6 represents a hydrogen atom in a 11,17a-dihydroxy compound or R6 represents a fluorine atom in a 11,17a-dihydroxy-4-ene compound, R9 must represent a fluorine or chlorine atom, which process comprises (a) hydroxylating a D-homosteroid of the general formula (II) in the 11-position by means of microorganisms or enzymes produced therefrom, or (b) replacing the iodine atom in a D-homosteroid of the general formula (III) by a hydrogen atom, or (c) dehydrogenating a 1,2-saturated D-homosteroid of formula I in the 1,2-position, or (d) adding hypochlorous acid to the 9,11-double bond of a D-homosteroid of the general formula (IV), or (e) treating a D-homosteroid of the general formula (V) with hydrogen fluoride or hydrogen chloride, or (f) saponifying a 17a?-acyloxy group in a D-homosteroid of formula I, or (g) isomerising a 6.beta.-(fluoro, chloro or methyl)-D-homo-steroid corresponding to formula I to the 6.alpha.-isomer, or (h) fluorinating or chlorinating a D-homosteroid of the general formula (VI) in the 6-position, or (i) acylating a 17a-hydroxy group in a D-homo-steroid of formula I, or (j) oxidising the 3-hydroxy-.DELTA.5-grouping in a D-homosteroid of the general formula (VIII) to the 3-keto-.DELTA.4-grouping, or (k) reducing the 11-keto group in a D-homosteroid of the general formula (IX) to the hydroxy group with protection of the 3- and 20-keto group, or (1) oxidising the 17a(20)-double bond in a D-homosteroid of the general formula (X) to the hydroxyketone grouping, or (m) methylating a D-homosteriod steroid of formula VI in the 6-position, or (n) converting the 17.beta.-ethynyl group in a D-homosteroid of the general formula (XI) into the acetyl group, or (o) dehydrogenating a D-homosteroid of the general formula (XII) in the 4-position;
in which formulae R6, R9, R17a and the broken line in the 1,2-position have the significance given in claim 7 and R611 represents a hydrogen atom or a methyl group.

2) A process according to claim 1, wherein in the starting material and in the product there is a double bond in the 1,2-position, and wherein a 1,2-unsaturated D-homosteroid of formula I is prepared.

3) D-Homosteroids of the general formula (I) wherein the broken line in the 1,2-position denotes an optional carbon-carbon bond; R6 represents a hydrogen, fluorine or chlorine atom or a methyl group; R9 represents a hydrogen, fluorine or chlorine atom; and R17a represents a hydroxy or acyloxy group, with the proviso that, where R6 represents a hydrogen atom in a 11,17a-dihydroxy compound or R6 represents a fluorine atom in a 11,17a-dihydroxy-4-ene compound, R9 must represent a fluorine or chlorine atom, whenever prepared by the process as claimed in claim 1 or by an obvious chemical equivalent thereof.

4) 1,2-Unsaturated D-homosteroids of formula I according to claim 3, whenever prepared by the process as claimed in claim 2 or by an obvious chemical equivalent thereof.

5) A process according to claim 1, wherein 17a-butyry-loxy-11.beta.-hydroxy-D-hompregna-1,4-diene-3,20-dione is prepared from 11.beta., 17a-dihydroxy-D-homopregna-1,4-diene-3,20-dione.

6) A process according to claim 1, wherein 17a-butyryloxy 9-fluoro-11.beta.-hydroxy-D-homopregna-1,4-diene-3,20-dione is prepared from 9-fluoro-11.beta., 17a-dihydroxy-D-homopregna-1,4-diene-3,20-dione.

7) 17a-butyryloxy-11.beta.-hydroxy-D-homopregna-1,4-diene-3,20-dione whenever prepared by the process as claimed in claim 5 or by an obvious chemical equivalent thereof.

8) 17-butyryloxy-9-fluoro-11.beta.-hydroxy-D-homopregna-

1,4-diene-3,20-dione whenever prepared by the process as claimed in claim 6 or by an obvious chemical equivalent thereof.