NZ230570A - The preparation of intermediate 16,17-acetal substituted androstang-17#b#-carboxylic acid esters - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number £30570 2305 70 r'io. 3aze\z}: U ; Ci_r: Specif cation Filed- '29 t a vj iggg P.O. Journal. !\!o: •5C^ } • ^ X..V Under the previsions cf Rccu-ic.:.-n 23 (1) ths ..CQrO.p.lCj.C Spccific&t.cn beer, ante-datca •u: B5...fB2r„Ch.. ig lb.

NEW ZEALAND Initials THE PATENTS ACT 1953 COMPLETE SPECIFICATION "PREPARATION OF INTERMEDIATES FOR 16,17-ACETALSUBSTITUTED ANDROSTANE-17B-CARBOXYLIC ACID ESTERS" We, AXTIEBOLAGET DRACO, a Swedish Company, of Box 31, S-221 00 Lund, Sweden, hereby declare the invention, for which we pray that a parent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statements: 230570 DESCRIPTION Field of the Invention ->"•* The present invention relates to a process for the preparation of intermediates for producing pharmacologically active 5 compounds that are described and claimed in New Zealand Patent Specification No. 215586. 3ackcround Art It is known that certain glucocorticosteroids (GCS) can be used for local therapy of inflammatory, allergic or immunologic 10 diseases in respiratory airways (e.g. asthma, rhinitis), in skin (eczema, psoriasis) or in bowel (ulcerative colitis, Morbus Crohn). With such local glucocorticoid therapy, clinical advantages over general therapy (with e.g. glucocorticoid tablets) are obtained, especially regarding 15 reduction of the unwanted glucocorticoid effects outside the diseased area. To reach such clinical advantages, in e.g. severe respiratory airway disease, GCS must have a suitable pharmacological profile. They should have high intrinsic glucocorticoid activity at the application site but also a — 20 rapid inactivation by e.g. hydrolysis in the target organ or after uptake into the general circulation.

Since binding of GCS to the glucocorticoid receptor is a pre-recuisite for their anti-inflammatory and allergic effects to occur, the ability of steroids to bind to their receptor(s) "*^25 can be used as an adequate method for determining the biological activity of GCS. A direct correlation between the affinity of GCS's to the receptor ana their antiinflammatory effects has been shown using ear edema test in the rat. {^Correlation between chemical structure, receptor binding, and 3 0 biological activity of some novel, highly active, 16a,17a- >*■»>> 2305 7 acetalsubstitutec glucocorticoids. E. Dahlberg, A. Thalen, R. Brattsand, J-A Gustafsson, U. Johansson, X. Roempke, and T. Saartok, Mol. Pharmacol. .25 (1984, 7 0.] New Zealand Patent Specification No. 215586, which is the 5 parent of the present divisional specification, discloses that certain 3-oxoandrosta-l,4-diene-173-carboxyiic acid esters possess high binding affinity to the glucocorticosteroid receptor- In particular, the compounds disclosed in New Zealand Patent Specification No. 215586, which can be used for 10 the treatment and control of inflammatory conditions, are characterized by the formula: CR-jRj wnerem the 1,2-position is saturated or is a double bond; 15 Xx is selected from hydrogen, fluorine, chlorine and bromine; X2 is selected from hydrogen, fluorine, chlorine and bromine; R2 is selected from hydrogen and a straight.or branched hydrocarbon chain having 1-4 carbon atoms; R2 is selected from hydrogen and straight or branched 20 hydrocarbon chains having 1-10 carbon atoms; and 230570 R3 is selected from CR4R5OCR6 and CR4R5OCYR6 "v Y is 0 or S; R4 is selected from hydrogen, straight or branched 5 hydrocarbon chains having 1-10 carbon atoms and phenyl; R5 is selected from hydrogen and methyl; and Rg is selected from hydrogen, straight or branched, saturated v or unsaturated hydrocarbon chains having 1-10 carbon atoms, an alkyl group substituted by at least one halogen 10 atom, a heterocyclic ring system containing 3-10 atoms in the ring system, -(CK2)m CK(CH2)n (m = 0,1,2: n = 2,3,4,5,6), and phenyl or benzyl groups which are unsubstituted or substituted by one or more alkyl, nitro, carboxy, alkoxy, halogen, cvano, carbalkoxy or 15 trifluoromethyl groups(s), provided that when R2 is hydrogen R^_ is methyl.

Disclosure of the Invention The present invention is directed to a process for the preparation of a compound of the formula D: ch5-or7 I 2 7 23 0 5 7 or a stereoisomeric comoound thereof, which formula: the 1,2-position is saturated or is a double bond; Xj_ is selected from hydrogen, fluorine, chlorine and bromine; X2 is selected from hydrogen, fluorine, chlorine and bromine; P2 is selected from straight and branched hydrocarbon chains having 1-10 carbon atoms; and Rj is hydrogen or an acvl group with 1-10 carbon atoms arranged in a straight or branched chain, characterized by reaction or a comoouna of :ne romuia F: ch,or7 l 2 7 c=o - °>c^CH3 — 0' *ch. with a compound of the formula 0=cCH 2 wherein Xt_, X2, R2, ^7 an°- have the meaning given above, in the presence of an acid catalyst.

New Zealand Patent Specification No. 215586 discloses that the individual stereoisomeric components present in a mixture of a steroid having the above formula (I) can be elucidated in the following way: - 4A - 230 5 70 (II; epirr.er S) (III; epimer R) The individual stereoisomeric components present, in a mixture of ster id 175-carboxylic acid esters having the formulas 0 i| 0 i| StCOCR-R^OCR. (IV) or 4 3 0 0 II 0 II StCOCR.R-OCYR, 4-0 b (v) - 4B - 230 5 70 where St is the steroid moiety, can be elucidated in the following way 0 R.O 0 R.O a I StCOCGCR, stcoCoca- : o | o - RS R5 VI VII and 0 R„0 0 R.O n l**:s StCC-COCYR, StCOCCCYR- ; o [o R- R- 3 2 VIII IX In diasteroisomers like II, III, VI, VII, VIII and IX, the ccnfiguraticn differs only at one out of several asyTmetric caroon atoms. Sucn dia-stereoisomers are denoted epimers.

Alkyl in the definitions aDove is a straignt or brar.c.neti nycrccaroon chain with 1-5 carbon atoms, preferably 1-4 C.

Alkoxy in the definition above is a group -0-alkyl wherein the alkyl moiety has the above given definition.

Halogen in the definition above is preferaoly a chlorine, bromine or fluorine atom.

Carbalkoxy in the definition above is a group -COO-alkyl wherein the 15 alkyl moiety has the above given definition.

Heterocyclic ring system is a ring system containing as hetero atoms N, 0 or S.

Preferred systems are pyrry1, pyrridyl, pyrimidyl, pyrazinyl, furyl, pyranyl, benzofuranyl, indolyl and thienyl.

Preferred compounds described and claimed in New Zealand Patent Specification No. 215586 are: 230570 1'-Ethoxycarbonyloxyethyl 6a,9a-dif1uoro-115-hydroxy-16a,17a-[(]-methylethyl idenejbi s(oxy) j-androsta-1 ,4-diene-3-one-175-carboxy1ate, the epi-meric mixture A - 3 and epimer 3. 1'-i sopropoxycarbony1oxyethy1 9a-f1uoro-11S-hydroxy-16a,17a-[(1-methyl -5 ethyliaene)bis(oxy)j-androsta-1,4-diene-3-one-17S-carboxylate, epimer 3. 1'-prcpoxycarbonyloxyethy! 6a,9a-dif1uoro-11S-hydroxy-16a,17a-[(1--methylethyl ide.ne)bis(oxy) ]androsta-l ,4-diene-3-one-l75-carboxylate, epimer 3. 1'-isopropoxycarbony1oxyethy1 6a,9a-di f1uoro-115-hydroxy-16a,17a-[ (1 -10 -methylethylidene)bis(oxy)]androsta-l,4-diene-3-one-175-carboxylate, epimeric mixture A B and epimer 3. 1'-Acetoxyethyl (20R)-9a-f1uoro-11 S-hydroxy-16a,17a-propy1 methylene-dioxyandrosta-1,4-diene-3-one-l75-carboxylate, epimer B. 1'-Ethoxycarbonyloxyethyl (22R)-9a-fluoro-115-hydroxy-16a,17a-propyl -15 methylenedioxyandrosta-1,4-diene-3-one-l7£-carboxylate, epimer B. 1'-i sopropoxycarbonyloxyethyl (20R)-9a-f1uoro-115-hydroxy-16a,17a--propylmethylenedioxyandrosta-1,4-diene-3-one-l7S-carboxylate, epimer 8. 1'-Ethoxycarbonyloxyethyl (20R)-6a,9a-difluoro-11S-hydroxy-16a,17a-propylmethylenedioxyandrosta-1,4-diene-3-one-175-carboxylate, epimeric 20 mixture A + B and epimer 3.

Methods of Preparation The compounds of formula I of New Zealand Patent Specification No. 215586 are prepared by the oxidation of a compound of the formulas X, XI ana XII to the corresponding 173-carboxylic acid: 2305 70 CH2-OR? ch£(- "2 CH2-OR? c=o XI CH2-OR7 XII wherein the solid and broken lines between C-1 and C-2 represent a single or double bond, ( 2305 70 , X^, R-j and have the meaning given above for Formula I, and R_ is hydrogen or an acyl group with 1-10 carbon atoms arranged in a straight or branched chairs.

The 176-carboxylic acids then are esterified to give compounds character-5 ized by the formula I-IX, wherein ="="= X^, X^, R,, R2 and R., have the meaning given above for Formula I.

The process to convert a compound of formulas X, XI or XII to the corresponding 17-carboxylic acids is carried out in a suitable oxygenated hydrocarbon solvent such as a lower alkanol. Methanol 10 and ethanol are preferred, particularly the former. The reaction medium is made slightly alkaline by the addition of a suitable weak inorganic base such as an alkali metal carbonate, for example sodium, lithium or potassium carbonate. The latter is preferred. The conversion of a compound of formula X, XI or XII to a 175-carboxylic acid of formula 15 I, II or III (R^H) takes place at ambient temperatures, i.e. 20-253C.

The presence of oxygen is necessary for the reaction. Oxygen can be supplied by bubbling a stream of air or oxygen into the reaction mixture.

'The oxidative degradation of the 175 side-chain of compounds of formula X, XI and XII to the corresponding 175 carboxylic acids can also be 20 carried out with periodic acid, sodium h^'pobromate °r with sodium bis- muthate. The reaction is performed in a mixture of water and a suitable oxygenated hydrocarbon solvent such as a lower ether. Dioxane and tetra-hydrofurane are preferred, particularly the former.

/ The parent 175-carboxylic acids of compounds of formula I, II and III 25 (Rg=H) may be esterified in known manner to provide the desired 17f5 carboxylate esters. Tor example, the J7f5-carboxylic acid may be reacted with an appropriate alcuriul and a carboaiimiae, e.g. dicyclohexylcarbodiimide, in a suitable solvent such as diethylether, tetrahydrofurane, methylene chloride or pyridine advantageously at a 30 temperature of 25-100°C. Alternatively, a salt of the 175-carboxylic acid with an alkali metal, e.g. lithium, sodium or potassium, a salt 23 0 5 of a quaternary ammonium compound, such as a salt of triethyl-or tri-butylamine, or tetrabutylammonium, may be reacted with an appropriate alkylating agent, for example an acyloxyalkylnaliae or haloalkyl alkyl-carbonate preferably in a polar solvent medium such as acetone, methyl -5 ethylketone or dimethyl formamids, dimethyl sulpnoxide, methylenechlo-ride or chloroform, conven-iently at a temperature in the range 25-100°C. The reaction may also be performed in the presence of a crown ether.

The crude steroid ester derivatives formed are after isolation purified by chromatography on a suitable material, for instance cross-linked 10 dextran gels of Sepnadex® IH-type with suitable solvents as eluants, e.g. halogenated hydrocarbons, ethers, esters such as ethyl acetate or acetonitrile.

The individual epimers, which are formed at the acetal isation of the 16a,17a-hydroxy groups or at the esterification of the 175-carboxylic 15 acids, possess practically identical solubility characteristics. Accordingly, they have turned out to be impossible to separate and isolate from the epimeric mixture by conventional method for resolution of stereoisomers, e.g. fractionated crystallization. In order to obtain the individual epimers separately the stereoisomeric mixtures according 2 0 to the formulas I, IV and V above are subjected to column chromatography, thus separating the epimers II, III, VI, VII, VIII and IX in view of different mobility on the stationary phase. The chromatography may be carried out for instance on cross-linked dextran gels of the type Sepnadex3 IH, e.g. Sepnadex3 LH-20 in combination with a suitable organic 25 solvent as eluting agent. Sepnadex® LH-20, prepared by Pharmacia Fine Chemicals AB, Uppsala, Sweden, is a beadformed hydroxypropylated dextran gel wherein the dextran chains are cross-linked to give a three-dimen-sional polysaccharide network. As eluting agent, halogenated hydrocarbons e.g. chloroform or a mixture of heptane-chloroform-ethanol in the 30 proportions 0-50:50-100:10-1 has successfully been used, preferably a 20:20:1 mixture.

Compounds of the formulas X, XI and XII are used as starting materials for compounds of formula I. They are prepared by reaction of comDOunds with the formula 230570 CH OR, I 2 / C = 0 wherei n the solid and broken lines between C-1 and C-2 represent a single or double bond, and X^, X2 and have the meaning given above, with an aldehyde of the formula 0=C' / H wherein R^ has the meaning given above.

The aldehyde is preferably acetaldehyde, propansl, butanal, isobutanal, pentanal, 3-methylbutanal, 2,2-dimethylpropanal, hexanal, heptanal, 10 octane!, nonanal and aodecanal. The reaction is carried out by adding the steroid to a solution of the aldehyde together with an acid catalyst, e.g. perchloric acid, p-toluene sulphonic acid, hydrochloric acid in an ether, preferably dioxane, or halogenated hydrocarbons, preferably methylene chloride or chloroform.

Compounds X, XI and XII are also prepared by transacetalisation of the corresponding 16a,17a-acetonictes CH^ORy 2305 70 wherein the solid and broken lines between and represent a single or double bond and X.j, and R^ have the meaning given above with an aldehyde of the formula 0 = C wherein R^ has the meaning given above.

The aldehyde is preferably acetaldehyde, propanal, butanal, isobutanal, pentanal, 3-methylbutanal, 2,2-dimethylpropanal, hexanal, heptanal, octanal, nonanal and doaecanal. The reaction is carried out by adding the steroid to a solution of the aldehyde together with a strong 10 inorganic acid as catalyst, preferably perchloric or hydrochloric acid, in an ether, preferably dioxane or tetrahydrofurane, a halogenated hydrocarbon, preferably methylene chloride or chloroform, an aromatic hydrocarbon, preferably toluene, an alicyclic hydrocarbon, preferably cyclonexane or an aliphatic hydrocarbon, preferably heptane or isooctane, 15 under the latter conditions eliminating the chromatographic step for preparation of the epimers III and XII.

Pharmaceutical Preparations The compounds of formula I may be used for different modes of local administration dependent on the site of inflammation, e.g. percutaneous-20 ly, parenterally or for local administration in the respiratory tract by inhalation. An important aim of the formulation design is to reach optimal bioavailability of the active steroid ingredient. For percutaneous formulations this is advantageously achieved if the steroid is dissolved with a high thermodynamic activity in the vehicle. This is 25 attained by using a suitable system of solvents comprising suitable glycols, such as propylene glycol or 1,3-butandiol either as such or in combination with water.

It is also possible to dissolve the steroid either completely or partially in a lipophilic phase with the aid of a surfactant as a 30 solubilizer. The percutaneous compositions can be an ointment, an oil in water cream, a water in oil cream or a lotion. In the emulsion vehicles the system comprising the dissolved active component can make up the disperse phase as well as the continuous one. The steroid can also exist in the abov? compositions as a micronized, solid substance. 230570 Pressurized aerosols for steroids are intended for oral or nasal inhalation. The aerosol system is designed in sucn a way that each delivered dose contains 10-1000 pg, preferably 20-250 pg of the active steroid. The most active steroids are administered in the lower part of the dose 5 range. The micronized steroid consists of particles substantially smaller than 5 pm, which are suspended in a propellent mixture with the assistance of a dispersant, such as sorbitan trioleate, oleic acid, lecithin or sodium salt of dioctylsulpnosuccinic acid.

Working Examples The invention will be further illustrated by the following non-limita- 2 -1 tive examples. In the examples a flow-rate of 2.5 ml/cm • h is used at the preparative chromatographic runs. Molecular weights are in all examples determined with electron impact mass spectrometry and the melting points on a Leitz Wetzlar hot stage microscope. All HPLC analyses 15 (HPLC = High Performance Liquid Chromatography) were performed on a Waters pBondapak C-jg column (300x3.9 mm internal diameter) with a flow-rate of 1.0 ml/min and with ethanol-water in ratios between 50:50 and 60:40 as mobile phase, if not otherwise stated.

Example 1. This example sets forth a process for preparing (22RS)-, 20 (22R)- and (22S)-115,16c:,17a,21-tetrariydroxypregna-l ,4-diene-3,20-dione 16a,17ct-acetals.

Preparation of (22RS)-, (22R)- and (22S)-117a-butylidenedioxy-6a,9a-dif1uoro-115,21-aihydroxypregna-1,4-diene-3,20-aione.

A. To a suspension of 1.0 a of 6a,9a-difluoro-ll£,16oL,17a,21-tetra-25 hydroxypregna-1 -4-aiene-3,20-dione in 500 ml of methylene chloride 0.32 ml of freshly distilled n-butanal and 2 ml of 72% perchloric acid were added. The reaction mixture was allowed to stand for 24 h at room temperature under stirring. The reaction mixture was washed with 10% aqueous potassium carbonate solution and water, dried over sodium 30 sulphate ana evaporated. The residue was dissolved in ethyl acetate and precipitated with petroleum ether leaving 883 mg of (22RS)-16a,17a-butyli denedi oxy-5ci,9a-aif1uoro-11£,21-dihydroxypregna-1,4-di ene-3,20-dione. HPLC-analysis showed 99% purity ana the ratio 15:84 between the 22S- ana 22R-epimers. Molecular weight: 456 (calculated 466.5). 230570 The (22RS) epimeric mixture was chromatograpned on Sephadex LH-20 column (76x6.3 cm) using heptane:chloroform:ethanol, 20:20:1 , as mobile phase. The fractions 12315-13425 ml (A) and 13740-15590 ml (B) were collected and evaporated and the residue dissolved in methylene chloride and 5 precipitated with petr.-ether. Fraction A gave 62 mg of (22S)- and fraction B 687 mg of (22R)-l 6«, 17cx-butyl idenedi0x^-6^,9^-difl uoro-ll£,21-dihydroxypregna-l ,4-diene-3,20-dione. The (22S)-epimer: Molecular weight 466 (calculated 466.5), m.p. 196-200°C.

The (22R)-epimer: Molecular weight 466 (calculated 466.5), m.p. 10 169-72°C. _B. To a solution of 1.0 g of 6«,9o-difl uoro-115,21-dihydroxy-!6a, 17c<-[(1-methylethylidene)bis(oxy)j pregna-1,4-diene-3,20-dione in 500 ml of methylene chloride was added 0.30 ml freshly distilled £-butanal and 2 ml of 72% perchloric acid. The reaction mixture was allowed to stand 15 for 24 h at 33°C under stirring, extracted with aqueous potassium carbonate and water, dried over sodium sulphate and evaporated. The residue was dissolved in methylene chloride ana precipitated with petr.--ether yielding 848 mg of (22RS)-16^s IT^-butylidenetiioxy-6cs9<*-dif luoro-llp ,21-cihydroxyprecna-20 1,4-diene-3,20-dione. HPLC-analysis showed 93% purity ana the ratio "12/88 between the 22S- and 22R-epimers.

Bl. To a suspension of 4.0g of 6a, 9a-difluoro-115, 21-di-hydroxv-16a, 17a- [(1-methylethylidene) bis (oxv)] pregna-1, 4—aiene-3,20-dione in 100 ml of heptane was added 1.2 ml of 25 freshly distilled normal-butanal and 3.8 ml of perchloric acid (72%). The reaction mixture was allowed to stand for 5h at room temperature under vigorous stirring, extracted with aqueous potassium carbonate and water, dried over sodium sulphate ana evaporated, yielding 4.0g of (22RS)-16a, 17cx-30 butylideneaioxy-6a, 9cx-dif luoro-116, 21-aihydroxy-pregna-l, 4-diene-3, 20-dione. HPLC analysis showed 98.5% purity and the ratio 3:97 between the 22S- and 22R-epimers. After two recrystallisations from chloroform-petroleum ether, 3.1g of 22R-epimer was obtained, which contained only 1.1% of the 35 22S-epimer and 1.3?, of other impurities. c3 0 5 ^Similarly, jy following the procedure set forth in the example by substituti ng 6a, 9«.-dif 1 uoro-1lp , 1 6°s 17cs 21 -tetrahydroxypregn a-1,4-diene-3,20-dione for 112,16=^ 17<*.,21-tetrahydroxypregna-! ,4-diene-3,20-dione, 9oc-f 1 uoro- and 6c*-f 1 uoro-ll£>, 16°"-, 17c*, 21 -tetrahydroxypregna-1,4-di ene-3,20-dione or the corresponding 16^, 17ct-acetonides non-fluorinated and fluorinated non-symmetric (22RS)-, (22R)- and (22S)-113,16»,17<x,21--tetrahydroxypregna-1,4-diene-3,20-dione 16csl7^-acetals from acetaldehyde, propanal, butanal, isobutanal, pentanal, 3-methy!butanal, 2,2-dimethylpropanal, hexanal, heptanal, octanal, nonanal and dodecanal are prepared.

Example 2 A. Prednacino!on 1 &n,17«-acetonide (250 mg;0,6 mmol) was dissolved in 75 ml of C^C^- n-Butanal (130 mg; 1,8 mmol and 70% perchloric acid (0,025ml) were added. The solution was stirred at 33°C for 15 hours. The yellow solution was washed with 2x10 ml of 10% l^CO^ and 4x10 ml of H^O, dried and evaporated. Yield: 257 mg (97,7%). HPLC gave 91,1% purity. Unreacted acetonide consists of 7,4% of the impurities. Epimer ratio 14,5/85,4.

B_. Triamcinolon 16cs 17=*-acetonide (0,5 g; 1,1 mmol) was dissolved in 150 ml of CH^C^- n-Butanal (260 mg; 3,5 mmol) and 70% perchloric acid (0,22 ml) were added. The mixture way stirred at 33°C for 16 hours. CK^C^ was taker, over into a separation funnel and the reaction flask was washed several times with 10 ml f^CO^ and Ch^C^* respectively. The solution was then washed with 2x10 ml of 10% K-C0-, and 4x10 ml of H,0, dried and 2 j 2 evaporated. Yield: 438 mg (84,9%). HPLC gave 80,2% purity. Epimer ratio 19/81.

C. Fluocinolon 16<*, 17»-acetonide (0,5 g; 1,1 mmol) was dissolved in 150 ml of CH2C12. n-Butanal (260 mg; 3,6 mmol) and 70% perchloric acid (0,22 ml )were added. The mixture was stirred at 33°C for 24 hours. The CH^Cl^ phase was taken over into a separation funnel. The reaction flask was washed several times with 15 ml of 10% ^2^3 anc* CH^Cl^, respectively. The solution was washed with 2x15 ml of 10% ^CO^ ana 4x15 ml of H^O, dried and evaporated. Yield: 513 mg (100%). HPLC gave 97,4% purity. Epimeric ratio 8.6/91,4. 2 3 0 5 7 0 j/grn^ Example 3. This example sets forth a process for preparing 11^-hydroxy-16<*,17o^- [(1-methyl ethyl i dene) bis (oxy)] - ana (20RS)-, (20R)- ana (20S) -112-hydroxy-16**, 17«*- al kylmethyl eneai oxyanarosta-1,4-di ene-3-one-175-car-boxylic and -4-ene-3-one-l7^-carboxylic acids.

Preparation of 6a,9a-difluoro-n = -hydroxy-16a,17a-[ (1-methyl ethylidene)-bis(oxy)]androsta-l,4-diene-3-one-17S-carboxylic acid.

A. To a solution of 1.99 —g of fluocinolone 16a,17a-acetonide in 120 ml of methanol 40 ml of 20% aqueous potassium carbonate was added. A stream of air was bubbled through this solution for about 20 h under stirring 10 at room temperature. The methanol was evaporated and 200 ml of water was added to the residue. The solution was extracted with methylene chloride. The aqueous phase was acidified with diluted hydrochloric acid. The precipitate formed was collected by filtration and dried to yield 1.34 g of 6a,9a-difluoro-llS-hydroxy-16a,17a-[ (1-methylethylidene)-15 bis(oxy)]anarosta-l,4-diene-3-one-17£-carboxylic acid, melting point 254-68°C, molecular weight 43S. The purity determined by HPLC was 94.0%. The aqueous phase was extracted with ethyl acetate. After drying the solvent was evaporated leaving another 0.26 g portion of acid.

Purity: 93.7%.

B. Periodic acid (15.1 g) in 16.5 ml of water was added to a solution of fluocinolone 16a,17a-acetonide (5.0 g) in 55 ml dioxane. The reaction mixture was stirred at room temperature for 20 h, neutralized with saturated aqueous sodium hydrogen carbonate and evaporated. The residue was dissolved in 200 ml of methylene chloride and washed with 8 x 100 ml 25 10% aqueous potassium carbonate. The aqueous phase was acidified with conc. hydrochloric acid and extracted with 6 x 100 ml of ethyl acetate.

After drying the solvent was evaporated. The residue was dissolvea in 400 ml of ethyl acetate and precipitated with petroleum ether yielding 3.96 g of 6a,9a-difluoro-115-hydroxy-16a,17a-f (1-methyl ethyl i dene) bis-30 (oxy)jandrosta-1,4-diene-3-one-l75-carboxylic acid. The purity determined by HPLC was 99.5%.

C. Similarly, by following the procedure set forth in the example by substituting fluocinolone 16a,17a-acetonide for 115,16a,17a,21-tetra-hydroxypregna-1,4-diene-3,20-dione, 6a-f1uoro-115,16a,17a,21 -tetranyd-35 roxypregna-1,4-diene-3,20-dione, ana triamcinolone 16a,17a-ecetoniae 11S-hydroxy-16a,17a-[(1-methylethyli dene)bi s(oxy) ]androsta-l,4-diene- . : 0 5 7 0 3-one-17£-carboxylic acids are prepared. By substituting the 16a,17a-acetonide group for 16a, 17cs-acetals between 16a-hydroxyprednisolor.e 6ct-fluor-16a-hydroxyprednisolone, triamcinolone and fluocinolone and acetaldehyde, propanal, butanal, isobutanal, pentanal, 3-methylbutanal, 5 2,2-dimethylpropanal, hexanal, heptanal, octanal, nonanal and dodecanal and their 21-esters (20RS)- (20R)- and (20S)-115-hydroxy-16a,17a-alkyl-methylenedioxyandrosta-1,4-diene- and 4-ene-3-one-l75-carboxylic acids are prepared.

Example 4. 1'-Ethoxycarbonyloxyethyl 5a,9a-difluoro-11S-hydroxy-16a,17q-10 I(1-methyl ethylidene)bis(oxy)}androsta-l,4-diene-3-one-17£-carboxylate.

A. 6a,9a-Difluoro-ll£-hydroxy-15a,17a-i (1-methylethyl idene)bis (oxy) ]-androsta-1,4-diene-3-one-l7£-carboxylic acid (600 mg) and potassium hydrogen carbonate (684 mg) were dissolved in 45 ml of dimethyl form-amide. 1-Bromoethyl ethyl carbonate (2 ml) was added and the reaction 15 mixture stirred at room temperature overnight. Water (200 ml) was added and the mixture was extracted with methylene chloride. The combined extracts were washed with 5% aqueous sodium hydrogen carbonate and water, and the residue purified by chromatography on a Sephadex LH-20 column (72x6.3 cm) using chloroform as mobil phase. The fraction 1515-2250 20 ml was collected and evaporated yielding 480 mg of 1'-ethoxycarbonyloxyethyl 6a,9a-difluoro-ll£-hydroxy-16a,17a-[ (1-methylethyl idene )bi s (oxy) ]-androsta-1,4-diene-3-one-17£-carboxylate. The purity determined by HPLC was 98.1% and the ratio epimer A/B, 48/52. Melting point: 218-27°C. pc [o]q = +53.2° (c=0.214; CH2C12). The molecular weight was 554.

The 1'-ethoxycarbonyloxyethyl 5a,9a-dif1uoro-11 S-hydroxy-15a,17a-(l-methylethylidene)bi s( oxy)]androsta-1,4-diene-3-one-l 75-carboxylate (480 mg) was chromatographed on a Sephadex LH-20 column (75x6.3 cm) using heptane:chloroform:ethanol, 20:20:1, as mobile phase. The fraction 2325-2715 ml was collected, evaporated and the residue dissolved in 3 0 methylene chloride and precipitated by petroleum ether giving 200 mg of a compound (A) of purity 97.3% (determined by HPLC analysis). Melting point: 246-50°C. [a]^ = +100.5° (c=0.214; ^C^)- The molecular weight was 554.

The fraction 4140-5100 ml yielded 250 mg of a compound (B) with purity 35 99.OS. Melting point: 250-55cC. [ct]p^ = +28.5° (c=0.246; Cr^C^L The molecular weight was 554. The methine signal from the ester group is *3 0 5 shifted 0.13 ppm downfield in ^H-NMR spectrum of B compared to A, while the rest of the spectra are nearly identical. The electron impact mass spectra of A and B are identical apart from the intensities of the mass peaks. These spectroscopic differences and similarities indicate that A 5 and B are epimers due to the chiral centre in the ester group.

B_. 6cx,Sx-Difluoro-1 IB-hydroxy-l6cs 17a- f( 1 -methylethylidene)bis(oxy)J androsta-1,4-diene-3-one-l73-carboxylic acid (200 mg) was dissolved in 25 ml of dimethylformamide. 1-Chloroethyl ethyl carbonate (100 mg), potassium hydrogen carbonate (70 mg) ana 18-crown-6-ether were added. 10 The reaction mixture was stirred at 80°C for 3 h, cooled, extracted with methylene chloride after addition of 150 ml of water, dried and evaporated. The crude.product was purified in the same way as in procedure A leaving 207 mg of 1'-ethoxycarbonyloxyethyl 6«,9^-difluoro--116-hydroxy-16«,17»- [(1 -methylethylidene)bis(oxy)1 androsta-1,4-diene-15 -3-one-178-carboxylate. The purity (HPLC) was 98.4% and the ratio epimer A/B, 54/46.

C_. 6cs,9«.-Dif 1 uoro-118-hydroxy-16oc, 17c.- j( 1 -methylethylidene)bis(oxy)j androsta-1,4-diene-3-one-17B-carboxylic acid (200 mg) ana 1,5-diazebicyclo [5.4.o] undecene-5 (140 mg) were suspended in 25 ml of 20 benzene and warmed to reflux. A solution of 1-bromoethyl ethyl carbonate (175 mg) in 5 ml of benzene was added and the mixture was refluxed for 2 1/2 h. After cooling 50 ml of methylene chloride was added and the solution was washed with water, dried and evaporated. The crude product was purified in the same way as in procedure A, yielding 207 mg of 1'-25 -ethoxycarbonyloxyethyl 6=S 9«-dif luoro-HS-hydroxy-16% 17s- [(1 -methyl -ethylidene)bi s(oxy)1 androsta-1,4-diene-3-one-17S-carboxylate. The purity (HPLC) was 96.4% and the ratio epimer A/B, 44/56.

JD. To a solution of 6«,9^-difluoro-nB-hydroxy-16«, 17=\- [(1 -methylethyl -idene)bis(oxy)J androsta-1,4-diene-3-one-17S-carboxylic acid (100 mg) in 30 25 ml of acetone 175 mg of <X-bromodiethylcarbonate and 45 mg of anhydrous potassium carbonate were added. The mixture was heated for 6 h at reflux. The cooled reaction mixture was poured into 150 ml of water and extracted with methylene chloride. The extract was washed with water, dried over sodium sulphate and evaporated yielding 65 mg of solid 23 0 5 7 1 '-ethoxycarbonyloxyethyl 6a,9c<-difluoro-11 S-hycroxy-l5^17^, £( 1-methylethylidene)bis(oxy)1 androsta-1,4-di ene-3-one-176-carboxylate.

The purity determined by HPLC was 97.5% and the ratio eoimer A/S, 49/51.

E. 5«, 9a-Difluoro-11B-hydroxy-1 ccn, 17:x- £( 1-methyl ethyl idene)bi s(oxy)( 5 androsta-l,4-diene-3-one-17B-carboxylic acid (500 mg) and tetrabutylammonium hydrogen sulphate (577 mg) were added to 3 ml of 1M sodium hydroxide.A solution of 435 mg of 1-bromoethyl ethyl carbonate in 50 ml of methylene chloride was added. The mixture was refluxed with stirring overnight. The two layers were separated. The organic layer was Xo washed with 2x10 ml of water, dried and evaporated. The crude product was purified by chromatography on a Sephadex LH-20 column (72x5.3 cm) using*chloroform as' mobile phase. The fraction 1545-1950 ml was collected and evaporated and the residue precipitated from methylene chloride - petroleum ether leaving 341 mg of 1'-ethoxycarbonyl-15 oxyethyl 6«,9c*.-difl uoro-11 S-hydroxy-16«, 17oc- [j( 1-methyl ethyl i dene)--bis(oxy)] androsta-1,4-diene-3-one-17B-carboxyl ate. The purity determined with HPLC was 99.2% and the ratio epimer A/B, 55/^. £. 6®-,9«-Difluoro-116-hydroxy-l6<*,17w.- f( 1-methylethylidene)bis--(oxy)l androsta-1,4-diene-3-one-17B-carboxylic acid (200 mg) ana 20 tricaprylmethylammonium chloride (200 mg) were added to 5 ml of saturated aqueous NaHCO^. A solution of 100 mg of 1-bromoethyl ethyl carbonate in 10 ml of methylene chloride was added. The mixture was stirred at 45=c for 20 h, diluted with 10 ml of methylene chloride and isolated and purified in the same way as in procedure E yielding 25 254 mg of 1'-ethoxycarbonyloxyethyl oosS^-difluoro-llB-hydroxy- -1517^x- ["(1-methylethv 1 idene)bis(oxy)j - androsta-1,4-diene-3-one--17B-carboxylate. The purity (HPLC) was 97.4% and the ratio epimer A/B, 60M0.

G. 6a, 9o!-Difl uoro-11 S-hydroxy-16oc, 17 <x- [(1 -methyl ethyl i dene)bi s (oxy) j 30 androsta-1,4-diene-3-one-17B-carboxylic acid (200 mg), 1-bromoethyl ethyl carbonate (135 mg) and triethyl amine (275 mg) were dissolved in 20 ml of dimethylformamide. The mixture was stirred at 80°C for 3 h, dilutee with 200 ml of methylene chloride, washed with water, dried and evaporated. The crude product was purified in the same way as in - IS - . 0 5 i_ v/ U yj procedure A yielding 69 mg of 1'-ethoxycarbonyloxyethyl 6*,9^-dif 1 uoro--nB-hydroxy-16a, 17«- |_( l -methyl ethylidene)bis(oxy)~ ancrosta-1,4--diene-3-one-178-carboxylate. The purity (HPLC) was 97.3% and the ratio epimer A/B, 48/52.

Example 5. 1'-Acetoxyethyl 6a,9ot-dif 1 uoro-1 IS-hydroxy-16=<, 17c<- 1 -methyl ethyl idene) bi s (cxy) J androsta-1,4-di ene-3-one-175-carboxyl ate. 6«, 9».-Oifl uoro-1 IB-hycroxy-l 6<X. 17«.- }_{1 -methyl ethyl i cene)bi s{ oxy) 1 androsta-1,4-diene-3-one-173-carboxylic acid (500 mg) and potassium hydrogen carbonate (575 mg) were dissolved in 40 ml of 10 dimethyl foruiamide. 1-chloroethyl acetate (1 ml) was added and the reaction" mixture was stirred at room temperature for 40 h. The reaction • mixture was poured into 50 ml of water and extracted with metnylene chloride. The extract was washed with aqueous socium hydrogen carbonate and water, dried and evaporated. The residue was chromatocraphed on 15 Sephadex LH-20 column (72x6.3 cm) using chloroform as mobile phase. The fractions 1755-2025 and 2026-2325 ml were collected and evaporated.

The solid product from fraction 1755-2025 ml was further purified by chromatography on a sephadex LH-20 column (76x6.3 cm i.d.) using a mixture of heptane-chloroform-ethanol, 20:20:1,.as mobile phase. The 2v fraction 2505-2880 ml was collected and evaporated, the residue was dissolved in methylene chloride and precipitated with petroleum ether leaving 167 nig of solid product (A). Tne purity determine by HPLC was 99.1%. Melting point 23S-59°C. = -94° (c=0.192; CH2C12). The molecular weight was 524. z5 The solid product from fraction 2025-2325 ml above was further purified by chromatography in the same way as above. The fraction 5100-5670 ml was collected and evaporated. The residue was dissolved in methylene chloride and precipitated with petroleum ether yielding 165 mg of solid product (3). The purity determined with HPLC was 99.4%. Melting point 30 261-65°C. = +34° (c=0.262; Ch^C^). The molecular weight was 524.

The "'h-NMR spectra of a and 3 are nearly identical with the exception of the methine quartet from the ester group which is shifted 0.16 ppm 23 0 5 7 downfield in compound B compared to A. The fragmentation patterns of A and Bin electron impact mass spectra are identical apart from the intensities of the mass peaks. These spectroscopic properties of A and B indicate that they are epimers due to the chiral centre in the ester 5 group.

Example 6-88. The substance given in Table 1-3 below were prepared, isolated and purified in a manner analogous to that described in Examples 4 and 5.

Table 1.

M OOCOCRr CH3ils. -0>CfC"3 - O^ CH.

Example X^ X,, R^ Rg ^6/'^R6 no. 6 H H fenyl 1 Cll3 7 H H fenyl ch3 8 F H CH(CH3)2 1 cii3 9 F II CH(CH3)2 1 cii3 F II fenyl 1 ch3 11 F H fenyl 1 ch3 12 F F CH3 1 C(CH3) 13 F F CI13 1 C(CH3) ') ' ) C00C0CR6 CH^-5- - 0nc^CH3 Epimer A CM ?4o COOfOCR ' Rc X -J 0\, -CM- ■Oy ^ CM.

Epimer B i-i?b Epimer Mp °C (c=0.2 1n Molecular weight Retention CH2C}2> calc, found volume (ml) A 242 (dec) + 79° 550.7 550 1665-1890' B 221 (dec) +89° 550.7 550 1891-21751 A - +102° 534.6 534 2325-2500 ^ B - +40° 534.6 534 3165-3555 ^ A 249 (dec) +73° 568.6 568 2040-23551 B 238 (dec) +75° 568.6 568 2895-3285' A 262-70 +87° 566.6 566 2190-2505 ^ B 268-77 +50° 566.6 566 3525-39901 230570 U ( ) Table 1. (continued) Example X^ X2 R^ Rg ^6^6 Epimer no. • l 14 F F CH3 H fenyl A F F CH3 H fenyl B 16 F F CH3 CH3 CH3 17 F H CM3 II 00(0113)2' A 18 F H CH3 II 0C(CH3)3' B 19 H F CH3 H OCH(CH3)2 A H F Cll3 H OCH(CH3)2 3 21 F F C»3 H OCH3 A 22 F F Cll3 II 0CII3 0 23 F1 F Cll3 H 0(CH2)2CII3 A 24 F F CH3 H 0(CII2)2CH3 B F F CH3 H OCH(CH3)2 A+B 26 F F CH3 H OCII(CII3)2 A 27 F F CH3 II 0CH(CH3)2 B 28 F F CH3 H 0CH(CH2CII3)2 A 29 F F CH3 H 0CH(CH2CH3)2 B ' F F CH3 II OCH2CH(CII2CH3)2 A 31 F F CH3 H 0CH2CH(CII2CH3)2 B < ) («)d5 Hp °C (c=0.2 1n Molecular weight Retention CH^Clg) calc. found volume (ml) ) 224-30 +96° 586.6 586 2325-26251} 259-67 +48° 586.6 586 4350-4B751' 130-42 461° 538.6 538 1965-22201' 184*87 +98° 564.7 564 235-280 3) >300 430° 564.7 564 525-630 2) 250-53 4109° 550.6 550 1530-177015 230-35 450° 550.6 550 2295-2Q501) 235-42 4)02° 540.6 540 590-690 225-33 431° 540.6 540 395-430 3) 224-31 4106° 568.6 568 410-495 2) 227-30 428° 568.6 568 690-900 2) 205-28 459° 568.6 568 1365-15605) 210-25 495° 568.6 568 400-475 2) 242-47 431° 568.6 568 625-780 2) 226-28 495° 596.7 596 1785-208511 183-97 430° 596.7 596 3150-36001* 217-21 489° 610.7 610 1725-19801* 207-10 430° 610.7 610 3120-348015 CJ1 Table 1. (continued) ( ) Example X^ no.

V™6 (a) Epimer Mp °C (c=0.2 1n ch?CI2) Molecular weight calc. found Retention volume (ml) 32 f f ch3 h 0c(ch3)3 a+b 170-70 +65° 502.6 582 33 f f ch3 h 0c(ch3)3 a 177-79 + 100° 582.6 502 34 f f cll3 h 0c(ch3)3 b 190-92 +27° 582.6 582 f f cii3 11 ocii2c(ch3)3 a+b 208-36 +60° 596.7 596 36 f f ch3 h 0ch2c(ch3)3 a 248-56 +90° 596.7 596 1 N> U» 1 37 f 38 f f cll3 f ch3 h 0ch2c(cii3)3 cii3 ocii2cii3 b 226-2b +20° 596.7 560.6 596 560 1290-1920 255-310 650-800 1605-1995 1845-2130 3270-3750 405-460 ) 3) 2) 1) 1) 1) 2) 1) On a Sephadex LH- column (76x6.3 cm) using chloroform-heptane-ethanol (20:20:1) as mobile phase. 2) On a Sephadex LH- •20 column (07.5x2.5 cm) using chloroform-heptane-ethanol (20:20:1) as mobile phase. 3) On a Sephadex LH- •20 column (85x2.5 cm) using chloroform as mobile phase. 4) On a Sephadex Lll- •20 column (72x6.3 cm) using chloroform as mobile phase.

) On a Sephadex LH- •20 column (71.5x6.3 cm) using chloroform as mobile phase. ro O cn C; ( i Table 2.

M COOWR, ? 6 CI,J H 3ic - - - O v r,^R -O^S..

R.O l4ll C00C0CR6 I ch-.I 11 oxr^^2 - \ Epimer A Ml COOCOCR.

I 6 !! 0^(/R2 - -0'%, Epimer B ro Example no.

V™6 Epimer Hp °C (a)0 (c=0.2 in ch2ci2) Molecular weight calc. found Retention volume (ml) 39 h h cll3 h c(cii3)3 40 ii ii (chz)2ch3 ii ci,3 41 h ii (cii2)2cii3 ii c(ch3)3 42 f h (ch2)2ch3 ii c(ch3)3 43 h ii (chz)2ch3 ii 0(chz)3ch 44 h h ich2)2ch3 11 0c(cii3)3 45 h ii (ch2)2ch3 ch.l 0ch2ch3 46 f h (ch2)2ch3 cii3 och2cii3 47 f f (ch2)2ch3 cii3 och2ch3 1) 2) a+b a+b afb 109-92 63-70 192-96 254-50 40-46 155-50 163-75 130-60 160-07 + 70° +79° +74° +64° +70° +67° +63° 502.6 480.6 530.7 548.7 546.7 546.7 532.6 550.6 56B.6 502 488 530 548 546 546 532 550 568 On a Sephadex LH-20 column (72x6.3 cm) using chloroform as mobile phase. On a Sephades LH-20 column (83x2.5 cm) using chloroform as mobile phase. 1290-1665 1110-1260 1245-1440 1485-1800 1200-1395 320-400 225-285 1410-1545 1620-2175 1) 1) 1) 1) 1) 2) 2) 1) 1) ro CM o CJl ^sj Table 3. o R.O Vll 00(j0CR6 . _ 0^>Q-''R2 1 Epimer A l4|| C00(j0CR6 1 ^_0.c.-R2 0y XR Epimer B Example X no. 48 h 11 ch3 ii ii 49 ii h h ch3 ii 50 f ii (ch2)2ch3 ii h 51 f 11 ii (ch2)2ch3 h ii 52 f h (ch2)2ch3 ch3 53 f ii (ch2)2ch3 11 ch3 54 f ii ii (ch2)2ch3 cll3 55 f h ii (ch2)2ch3 cll3 56 f h h (ch2)2ch3 cimch 57 f ii 11 (ch2)2ch3 ch (ch 2 3'2 VYR6 c(c,,3)3 C(CH3)3 C<CM3,3 C(CH3,3 CIL CIL CH CM, c"3 CIL i-i? Epimer Hp °C (c=0.2 In CH2C12) A 0 A D A 0 192-97 196-200 261-67 255-59 226-31 232-38 176-88 211-19 133-35 210-12 Holecular weight calc. found 467° 407° +69° 463° 4101° 435° 4104° 446° 4110° 444° 502.6 502.6 548.7 548.7 520.6 520.6 520.6 520.6 548.7 548.7 502 502 548 548 520 520 520 520 548 Retention volume (ml) 1650-1995 1305-1560 1950-2100 2145-2370 1905-2175 3300-3720 430-490 630-715 1) 3) 1) 1) 1) 1) 2) 2) 1) 2100-2400 .548 2850-3225*> 23 0 5 7 0 (, () Table 3. (continued) Example X1 X2 R1 R2 R^ Rg R6^R6 58 f h 59 f h 60 h h 61 h ii 62 f h 63 f h 64 f f 65 f f 66 h ii 67 h ii 68 f h 69 f h 70 f h 71 f h 72 f h 73 f ii 74 f h 75 f h h II (ch2)2ch3 (ch2)2ch3 (cm, (ch, (CH, (cil (ch, (ch, 2ch3 2c,,3 2ch3 -cil pcil 2 3 fenyl fenyl )2ch3 ii ch3 h (ch2 2ch3 cii3 11 (cll2 2ch3 ch3 h (cll2 2ch3 ch3 )2ch3 h ch3 )2ch3 h ch3 h (ch2 2cii3 ch3 ii (ch2 2ch3 ch3 h (ch2 2ch3 c»l3 h (ch2 2c,i3 ch3 ch, cil '3 :ci 0C(CH oc (ch3)3 3 3 oc(ch3)3 oc(ch3)3 och2ch3 och(ch3)2 och2ch3 och2ch3 och2cii3 och2ch3 oc(ch3)3 0C(CH3)3 ocikch3)2 0ch(ch3)2 oc(ch3)3 0c(ch3)3 la|» .

Epimor Mp °C (c=0.2 in Holecular weight Retention CHgC1g) calc. found volume (ml) a 235-10 +75° 502.7 582 2100-240011 b 157-82 +75° 582.7 i582 2760-30751) 140-42 +77° 546.7 546 1500-1665^ 160-65 *69° 546.7 546 1620-170511 171-73 +66° 564.7 564 250-295 4) 161-64 +72° 564.7 564 245-290 4) 203-11 +99° 554.6 554 325-370 41 196-209 +70° 568.6 568 2235-25501) a+b 138-52 4102° 532.6 532 300-370 21 a4b 158-91 +33° 532.6 532 400-460 z) a 196-90 4110° 550.7 550 405-475 2) b 212-14 436 550.7 550 585-670 a 154-57 492° 578.7 570 345-400 2) b 161-60 +27° 570.7 570 405-565 2) 2) a 221-24 +107° 564.7 564 355-425 b 212-15 +39° 564.7 564 535-635 a 168-71 4103° 570.7 570 405-570 2) b 174-79 431° 578.7 570 255-310 5) 230570 Table 3. (continued) ( > Example Xj no. x2 R1 r2 R4 R5 R6/YR6 Epimer Hp °C I°l (c=0. ch2c 76 F H C(CH3)3 II ch3 II och2ch3 A 220-22 + 95 77 F H C(C,13)3 II ch3 II 0CII2CII3 0 227-37 + 18 78 F II H C(CH3)3 ch3 H och2cii3 A 229-32 + 115 79 F h ii C(CH3)3 ch3 II 0CII2CII3 B 246-51 + 34 80 F F (ch2)2ch3 H ch3 II och2ch3 A 167-70 + 95 01 F F (ch2)2ch3 II cll3 H 0ch2ch3 D 100-90 + 26 02 F F H (ch2)2ch3 ch3 II och2ch3 A+B 170-96 +60 83 F F H (ch2)2ch3 cn3 II 0CII2CII3 A 217-21 + 105 04 F F II (cii2)2ch3 cii3 H 0CH2CH3 g 211-15 ♦32 85 F F H (ch2)2ch3 ch3 II 0cii(cii3)2 A+B 190-210 ♦ 67 06 F F II (ch2)2ch3 ch3 II ocikch3)2 A 232-37 + 96 07 F F H (ch2)2ch3 ch3 H 0cii(cii3)2 B 225-32 + 37 88 F F H (ch2)2cii3 ch3 cll3 ocii2ch • - - 0 Holecular weight calc, found Retention volume (ml) 1) 21 3) 4) ) 564.7 564.7 564.7 564.7 560.6 560.6 560.6 560.6 560.6 502.6 502.6 502.6 502.6 564 564 564 564 560 -560 560 560 560 582 502 502 502 On a Sephadex LH-20 column (76x6.3 cm) using heptane-chloroform-ethanol (20:20:1) as mobile phase On a Sephadex LH-20 column (07.5x2.5 cm) using heptane-chloroform-ethanol (20:20:1) as mobile phase On a Sephadex LH-20 column (72x6.3 cm) using chloroform as mobile phase On a Sephadex LH-20 column (00x2.5 cm) using chloroform as mobile phase On a Sephadex LH-20 column (01.5 x2.5 cm) using chloroform as mobile phase 300-430 540-630 305-455 565-695 300-330 365-395 3720-4155 290-340 341-395 2190-3900 2190-2355 3630-3900 305-440 2) 2) 2) 2) 5) 5) 1) 5) 5) 1) 1) 1) 2) ro Cf4 O cn 2 3 0 5 7 C Example 89 . Pharmaceutical Preparations The following non-limitative examples illustrate formulations intended for different topical forms of administration. The amount of active steroid in the percutaneous formulations are ordinarily 0.001-0.2% (w/w), 5 preferably 0.01-0.1% (w/w).

Formulation 1, Ointment Steroid, micronized 0.025 g Liquid paraffin .0 9 White soft paraffin ad 100.0 g Formulation 2, Ointment Steroid 0.025 9 Propylene glycol .0 9 Sorbitan sesquioleate .0 9 Liquid paraffin .0 g White soft paraffin ad 100.0 g Formulation 3, Oil in water cream , ,, Steroid 0.025 g Cetanol .0 g Glyceryl monostearate .0 g Liquid paraffin .0 g Cetomacrogol 1000 2.0 g w' Citric acid 0.1 g Sodium citrate 0.2 g Propylene glycol .0 g Water ad 100.0 g 230570 Formulation 4, Oil in water cream Steroid, micronized 0.025 9 White soft paraffin .0 g Liquid paraffin .0 g Cetanol .0 g Sorbimacrogol stearate 2.0 g Sorbitan monostearate 0.5 g Sorbic acid 0.2 g Citric acid 0.1 g Sodium citrate 0.2 g Water ad 100.0 g Formulation 5, Water in oil cream 0.025 g .0 g .0 g .0 g 0.2 g O.l g 0.2 g ad 100.0 g 0.25 mg 0.5 ml 3 mg q.s. ad 1.0 g Steroi d White soft paraffin 15 Liquid paraffin •Sorbitan sesquioleate Sorbic acid Citric acid Sodium citrate 20 Water Formulation 6, Lotion Steroid Isopropanol Carboxyvinylpolymer 2 5 NaOH Water 230570 Formulation 7, Suspension for injection Steroid, micronized 0.05-10 mg Sodium carboxymethylcellulose 7 mg NaCl 7 mg Polyoxyethylene (20) sorbitan monoleate 0.5 mg Phenyl carbinol 8 mg Water, sterile ad 1.0 ml Formulation 8, Aerosol for oral and nasal inhalation Steroid, micronized 0.1 % w/w Sorbitan trioleate 0.7 % w/w Trichlorofluoromethane 24.8 % w/w Dichlorotetrafluoromethane 24.8 % w/w Dichlorodifluoromethane 49.6 % w/w Formulation 9, Solution for atomization Steroid 7.0 mg propylene glycol 5.0 g Water ad 10.0 g Formulation 10, Powder for inhalation A gelatin capsule is filled with a mixture of Steroid, micronized 0.1 mg Lactose 20 mg The powder is inhaled by means of an inhalation device. 2 3 0 5 7 0 Pharmacology The affinity of the new androstane-176-carboxylic acid esters to the glucocorticoid receptor.

All steroids according to formula I are physiologically 5 active compounds. The affinity of the novel androstane-175-carboxylic acid esters to the glucocorticoid receptor has been used as a model for determination of the anti-inf1ammatory potency. Their receptor affinities have been compared to budesonide ([22R,S]-16c:,17a-butylidenedioxy-11£,21-dihydroxypregna-l,4-diene-3,20-dione) a highly active glucocorti-10 coid with a favourable ratio between local and systemic effects (Tnalen and Rrattsand, Arzneim.-Forsch. 29, 1687-90 (1979)).

Male Sprague-Dawley rats, one to two months of age, were used throughout the investigation. The thymus was removed and put into ice-cold saline. The tissue was homogenized in a Potter Elvehjem homogenizer in 10 -ml 15 of a buffer containing 20 mM Tris, pH 7.4, 10 % (w/v) glycerol, 1 mM EDTA, 20 mM NaMoO^, 10 mM mercaptoethanol. The homogenate was centri-fuged for 15 min at 20,000 x g. Portions of the 20,000 x g supernatant (230 pi) were incubated for-about-24 h at 0°C with 100 pi phenylmethyl -sulphonylfluoride (an esterase inhibitor, final conc. 0.5 mM), 20 pi 20 unlabelled competitor and 50 pi ^H-labelled dexamethasone (final conc. 3 nM). Bound and free steroid were separated by incubating the mixture with 60 pi 2.5 % (w/v) charcoa-T--and 0-25 % (w/v) dextran T70 suspension in 20 mM Tris, pH 7.4, 1 mM EDTA, and 20 mM NaMoO^, for 10 min at 0°C. Following a centrifugation at 500 x g for 10 min, 230 pi of the super-25 natant was counted in 10 ml Insta-Gel in a Packard scintillation spectro- 3 photometer. The supernatants were incubated with a) [ Hjdexamethasone alone, b) [ Hjdexamethasone plus 1000 fold excess of unlabelled dexa-methasone and c) [ Hjdexamethasone plus 0.03-300 fold "excess" of competitor. The nonspecific binding was determined when 1000 fold excess 30 of unlabelled dexamethasone was added to [ Hj-labelled dexamethasone.

The radioactivity bound to the receptor in the presence of competitor divided by the radioactivity bound to the receptor in the absence of competitor multiplied by 100 gives the percentage specific binding of labelled dexamethasone. For each concentration of a competitor the per-

Claims (2)

>30570 centage specifically bound radioactivity is plotted against the log of concentration of competitor. The curves are compared at the 50 % specific binding level and referenced to budesonide, which is assigned a relative binding affinity (RBA) of 1. 5 Table 4. Table summarizing relative binding affinities (RBA) to the glucocorticoid receptor of some of the investigated compounds. Compound according RBA to Ex. No. Budesonide 1 10 4 epimer B 0.30 5 epimer B 0.17 27 0.50 38 0.04 55 0.20 15 64 0.05 57 0.04 69 0.44 84 1.03 87 0.63 20 The matter contained in each of the following- claims is to be read as part of the general description of the present invention. - 32 - 2305 70 What we claim is:

1. A process for the preparation of a compound of the formula D: or a stereoisomeric compound thereof, in which formula: 10 the 1,2-position is saturated or is a double bond ^1 is selected from hydrogen, fluorine, chlorine and bromine; is selected from hydrogen, fluorine, chlorine and bromine; is selected from straight and branched hydrocarbon chains having 1-10 carbon atoms;and is hydrogen or an acyl group with 1-10 carbon atoms arranged in a straight or branched chain, characterized by reaction of a Com-Doun.d of the formula F: 0- — 0' -CH. "CH- - 33 - 230570 with a compound of the formula 0=c: 15 wherein X-j, X£, R^, and =-r-= have the meaning given above, in the presence of an acid catalyst.

2. A compound according to formula D of claim 1 when obtained by the process according to claim 1. DATED this day of SA.D. 19 89 AKTIEBOLAGET DRACO, By itc Patent Attorneyo-, By i«*?/7hs:r Attorneys. HEKRY K'JGHZ3 LIMITED, per A - 34 -