SI9210064A - NEW ANTI-INFLAMMATORY AND ANTI-ALLERGY ACTIVE COMPOUNDS, GLUCOCORTICOSTEROIDS AND PROCEDURES FOR THEIR PREPARATION - Google Patents

Abstract

A compound of the general formula 1*2 or a stereoisomeric component thereof, in which the 1,2 position is saturated or a double bond is present, Ri is hydrogen or a branched or unbranched hydrocarbon having a chain of 1-4 carbon atoms, R2 is acyl having a branched or unbranched, saturated or unsaturated hydrocarbon chain of 1-20 carbon atoms, X1 is hydrogen or halogen, X2 is hydrogen or halogen and it is determined that Ri and R2 are not simultaneously hydrogen, X1 and X2 are not simultaneously hydrogen when there is a double bond at the 1,2 position, Ri and R2 are not simultaneously methyl groups when there is a double bond at the 1,2 position, Ri is a hydrogen atom and R2 is a branched or unbranched hydrocarbon having a chain of 1-10 carbon atoms; R3 is acyl having 11-20 carbon atoms, processes for their preparation, pharmaceutical compositions containing them, and use of the compounds in the treatment of inflammatory and allergic conditions.

Description

NEW ANTI-INFLAMMATORY AND ANTI-ALLERGIC ACTIVE GLUCOCORTICOSTEROID COMPOUNDS AND PROCESSES FOR THEIR PREPARATION

Field of the invention

The present invention relates to novel anti-inflammatory and antiallergic active compounds and to processes for their preparation. The invention also relates to pharmaceutical compositions containing the compounds and to methods for the pharmacological use of the compositions.

The aim of the invention is to provide an anti-inflammatory, immunosuppressive and anti-allergic glucocorticosteroid or its pharmaceutical composition with high activity at the site of application, e.g. in the respiratory tract, on the skin, in the intestinal tract, in the joints or in the eye, by directing the drug to a limited target area, in this way inducing low glucocorticoid systemic effects.

A further object of the invention is to provide a pharmaceutical composition comprising liposomes containing a pharmacologically active steroidal fatty acid ester of the invention, in order to improve drug delivery and minimize side effects of therapy.

Scientific basis

Glucocorticosteroids (GCS) are the most valuable drugs for the relief of asthma and rhinitis. It is generally accepted that GCS exert their therapeutic efficacy by anti-inflammatory and anti-anaphylactic action within the airways and lung tissue. Long-term oral use of GCS is greatly hampered by strong side effects outside the pulmonary region. Thus, currently only a small proportion of patients with asthma or rhinitis undergo GCS therapy. Greater safety can be achieved by delivering GCS by inhalation. However, even the two predominant inhaled GCS currently in wide clinical use (beclomethasone 17a,21-dipropionate and budesonide) have a rather low margin of safety, both of which have been reported to have adverse effects within the general circulation at the highest recommended inhalation doses.

Liposomes are membrane-like vesicles consisting of a series of concentric lipid bilayers with alternating hydrophilic compartments.

Liposomes have been used as carriers for various types of pharmaceutically active compounds with the aim of improving drug delivery and minimizing the side effects of therapy.

Glucocorticosteroids are incorporated into liposomes only at low concentrations and are poorly retained in the vesicles. Esterification of GCS at the 21-position with fatty acids increases the degree of incorporation and retention of the steroid in the vesicles. It has been shown that the fatty acid chain acts as a hydrophobic anchor, holding the steroid core in the hydrated polar head groups of the phospholipids, thereby improving the interaction between the glucocorticosteroid and the liposome.

Glucocorticosteroids encapsulated in liposomes have been described (M. De Silva et al., Lancet 8130 (1979), 1320), US patent specification No. 4,693,999 describes liposomal formulations of glucocorticosteroids for inhalation.

Discovery of an invention

One object of the present invention is to provide novel GCS compounds. The novel compounds are characterized by anti-inflammatory, immunosuppressive and anti-anaphylactic potency at the site of application and, in particular, have a markedly improved ratio between this potency and the ability to elicit GCS action outside the treated regions. The most preferred form of administration of the novel compounds is by inhalation, when the site of application is within the airways.

Another object of the invention is to provide an anti-inflammatory and anti-allergic pharmaceutical composition comprising liposomes with steroid esters for topical administration primarily to the respiratory tract. Such a composition allows for the enhancement of the therapeutic properties of steroid esters by prolonging local retention in the airways and by targeting the drug to specific target cells.

The compounds of the invention are represented by formula 1

CH3COOH

C2H5COOH

C3H7COOH

C4H9COOH

C ₅ HjjCOOH

_{C6H13COOH ​}

C ₇ H ₅ COOH

C ₈ H ₁₇ COOH or their stereoisomeric component, in whose formula the 1,2 position is saturated or there is a double bond there,

Rj is hydrogen or a branched or unbranched hydrocarbon having 1-4 carbon atoms, R2 is hydrogen or a branched or unbranched hydrocarbon having 1-10 carbon atoms,

R3 is acyl having a branched or unbranched, saturated or unsaturated hydrocarbon chain having 1-20 carbon atoms,

Χ-j is hydrogen or halogen

Χ2 is hydrogen or halogen and it is determined that

1) Rj and R2 are not both hydrogen,

2) Xj and Χ2 are not simultaneously hydrogens,

3) when there is a double bond at the 1,2 position, R j and R2 are not both methyl groups,

4) when there is a double bond at the 1,2 position, R j is a hydrogen atom and R 2 is a branched or unbranched hydrocarbon having 1-10 carbon atoms, R 3 is an acyl having 1120 carbon atoms.

Acyl is derived from:

acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid,

CgH-jgCOOH

CioH-|gCOOH

C-j and H23COOH

C-12H25COOH

C-13H27COOH

C-14H29COOH

C ₁₅ H ₃₁ COOH c ₁₆ h ₃₃ cooh c ₁₇ h ₃₅ cooh

Ci7H33COOH

_{C17H31COOH ​}

C17H29COOH

C18H37COOH

C19H39COOH decanoic acid, capric acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, nonadecanoic acid, eicosanoic acid.

C13H27COOH

_{C15H31COOH ​}

_{C17H35COOH ​}

C-J7H33COOH

C-17H31COOH

C17H29COOH

The desired acyl groups are derived from:

C11H23COOH lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid and especially from palmitic acid.

The branched or unbranched hydrocarbon chain having 1-4 carbon atoms is preferably an alkyl group having 1-4 carbon atoms, especially a methyl group.

The branched or unbranched hydrocarbon chain having 1-10 carbon atoms is preferably an alkyl group having 1-10 carbon atoms, more preferably 1-4 carbon atoms, especially a methyl or propyl group.

Halogen in this specification is fluorine, chlorine or bromine. The preferred halogen is fluorine.

Preferred compounds of the invention are those where in formula 1 the 1,2 position is saturated,

R-| is hydrogen or a branched or unbranched hydrocarbon chain having 1-4 carbon atoms,

R2 is hydrogen or a branched or unbranched hydrocarbon chain having 1-10 carbon atoms,

R3 is acyl having a branched or unbranched, saturated or unsaturated hydrocarbon chain having 1-20 carbon atoms,

Xj is hydrogen or halogen,

Xp is hydrogen or halogen, it is determined that:

1) R-j and R2 are not both hydrogen,

2) Xj and Χ2 are not simultaneously hydrogens.

Particularly preferred compounds of the invention are those where in formula 1 the 1,2 position is saturated,

Rj is hydrogen,

R2 is a propyl group,

R3 is acyl having 11-20 carbon atoms,

Xj is fluorine,

X2 is fluorine.

A further preferred compound of the invention is that of formula 1, wherein in place of

1,2 double bond,

Rj is hydrogen,

R2 is a propyl group,

R3 is a palmitoyl group,

Xj is fluorine,

X2 is fluorine.

The most preferred compound of the invention has the formula

_{CH2OC ( CH2} ) _14CH3

C=O

CH.

H'

CH.

F

A preferred embodiment of the invention is a composition comprising a preferred compound of the invention in combination with liposomes.

In cases where the aim of the invention is to provide a pharmaceutical composition containing liposomes, the active compound of the composition should be a compound of formula 1, wherein R3 is acyl having 11-20 carbon atoms.

In cases where the aim of the invention is to provide a liposome-free pharmaceutical composition, the active compound should be a compound of formula 1, wherein R3 is acyl having 1-10 carbon atoms, preferably 5-10 carbon atoms.

The individual stereoisomeric components present in a mixture of a steroid having the above formula (1) can be explained as follows due to the chirality at the carbon atom at position 22 and with respect to the R2 substituent:

f ^H 2 ^OR 3

About

(epimer 22S)

The desired stereoisomeric components have the 22R configuration.

PREPARATION METHODS

Steroid not ester

About

St-OCR. 4 where St is

and Xj, X2, R-1, R2 have the meaning given above, R4 is a branched or unbranched, saturated or unsaturated alkyl group having 1-19 carbon atoms and the 1,2 position is saturated or there is a double bond, prepared by any of the alternative methods that follow.

A. Reaction of a compound with the formula

St-OH, where St is defined above, with a compound of the formula

About

R.COH where R4 is as defined above.

Esterification of the 21-hydroxy compound can be effected in a known manner, i.e. by reacting the parent 21-hydroxy steroid with a suitable carboxylic acid, preferably in the presence of trifluoroacetic anhydride and desirably in the presence of an acid catalyst, i.e. p-toluenesultonic acid.

The reaction is preferably carried out in an organic solvent such as benzene or methylene chloride; the reaction is conveniently carried out at a temperature of 20 to 100°C.

B. Reaction of a compound with the formula

St-OH where St is defined above, with a compound of the formula

About

R.CX where R4 is as defined above, X is halogen, such as chlorine, bromine, iodine and fluorine; or with the group

About

-O-C-R where R4 is as defined above.

The parent 21-hydroxy compound can be treated with a suitable carboxylic acid halide or anhydride, preferably in a solvent such as a halogenated hydrocarbon, e.g. methylene chloride or ethers, e.g. dioxane in the presence of a base such as triethylamine or pyridine, preferably at low temperature, i.e. -5°C to +30°C.

C. Reaction of a compound with the formula

St-Y where St is defined above and Y is selected from halogens, i.e. Cl, Br and iodine or from mesylate or p-toluenesulfonate, with a compound of formula

About

R.CO A ⁺ where R4 is as defined above and A+ is a cation.

An alkali metal salt of a suitable carboxylic acid, e.g. lithium, sodium or potassium, or a triethylammonium or tributylammonium salt, may be reacted with a suitable alkylating agent of formula St-Y. The reaction is preferably carried out in a polar solvent such as acetone, methyl ethyl ketone, dimethylformamide or dimethylsulfoxide, conveniently at a temperature in the range of 25-100°C.

If a pure epimer is desired, each method from A to C may require a final reaction step to resolve the epimeric mixture into its components.

PHARMACEUTICAL COMPOSITIONS

The compounds of the invention can be used for various local administration routes depending on the site of inflammation, i.e. percutaneously, parenterally or for local administration to the respiratory tract by inhalation. In formulating the formulation, an important goal is to achieve optimal bioavailability of the active steroid ingredient. For percutaneous formulations, this is advantageously achieved if the steroid is dissolved with high thermodynamic activity in a carrier. This is achieved by using a suitable system or solvents containing suitable glycols, such as propylene glycol or 1,3-butanediol as such or in combination with water.

The steroid ester can also be completely or partially dissolved in the lipophilic phase by the addition of a surfactant as a solubilizing agent. The percutaneous compositions may be an ointment, an oil-in-water cream, a water-in-oil cream or a lotion. The system containing the dissolved active ingredient may constitute the dispersed phase as well as the continuous phase in emulsion carriers. In the above compositions, the steroid may also exist as a micronized solid.

Pressurized steroid aerosols are intended for oral or nasal inhalation. The aerosol system is designed so that each delivered dose contains 10-1000 pg, preferably 20-250 pg, of active steroid. The most active steroids are administered in the lower end of the dosage range. Micronized steroid consists of particles significantly smaller than 5 pm, which are suspended in a propellant mixture by means of a dispersant (such as sorbitan trioleate, oleic acid, lecithin, or sodium dioctylsulfosuccinic acid).

The steroid can also be administered using a dry powder inhaler.

One option is to mix the micronized steroid with a carrier substance such as lactose or glucose. The powdered mixture is distributed into hard gelatin capsules, each containing the desired dose of steroid. The capsule is then placed in a powder inhaler and the patient inhales the dose into their airways.

Another option is to put the micronized powder into spheres that break during the dosing process. This micronized powder is filled into the drug reservoirs of a multidose inhaler, i.e. Turbuhaler. The dosing unit measures the desired dose, which the patient then inhales. In this way, the steroid is delivered to the patient with or without a carrier substance.

The steroid can also be included in formulations intended for the treatment of inflammatory bowel diseases, both orally and rectally.

Formulations for the oral route must be designed to deliver the steroid to the inflamed parts of the intestine. This can be achieved by various combinations of enteric and/or gradual or controlled release principles. For the rectal route, an enema-type formulation is suitable.

PREPARATION OF LIPOSOME COMPOSITIONS

The lecithins used in this invention have different fatty acid chain lengths and thus have different phase transition temperatures. Examples of lecithins used include those derived from eggs and soybeans, and synthetic lecithins such as dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), and distearoyl phosphatidylcholine (DSPC). Lecithin-based stable carriers with different biodegradation properties could be prepared by manipulating the structure. This would allow for a prolonged release of the entrapped steroid ester.

The extent of interaction of a steroid ester with vesicles, e.g. dipalmitoyl phosphatidylcholine (DPPC), depends on the length of the ester chain and increases with increasing chain length.

The incorporation of cholesterol or cholesterol derivatives into liposomal formulations has become very common due to its properties to increase the stability of liposomes.

For the initial stages of preparation of liposomes according to the present invention, it may be convenient to follow the procedures described in the literature, i.e. the components are dissolved in a solvent (e.g. ethanol or chloroform), which is then evaporated. The resulting lipid layer is then dispersed in the selected aqueous medium, after which the solution is stirred or sonicated. The liposomes of the present invention preferably have a diameter between 0.1 and 10 pm.

To modify the structure of the liposome membrane, other lipids (e.g., cholesterol or cholesterol stearate) can be added to the main lipids that make up the liposome (usually phospholipids) in amounts of 0-40 weight percent (w/w) of the total lipids. To optimize the uptake of liposomes, a third component can be included to provide a negative charge (e.g., dipalmitoyl phosphatidyl glycerol) or a positive charge (e.g., stearylamine acetate or cetylpyridine chloride).

During formation, a wide range of spheroid ester to lipid ratios can be used, depending on the conditions and the lipid used. Liposome drying (freeze drying or spray drying) in the presence of lactose can be used, with lactose content ranging from 0 to 95% of the final composition.

A particularly preferred composition of the invention comprises liposomes and (22R)-16a,17a-butylidenedioxy-6a.,9a-difluoro-11 B-hydroxy-21 -palmitoyloxypregn-4-ene-3,20-dione. Routes of administration include powder aerosols, instillation, nebulization, and pressurized aerosols.

WORKING EXAMPLES

The invention will be further illustrated by the following non-limiting examples. In the examples, a flow rate of 2.5 ml/cm2 χ h ¹ was used for the preparative chromatographic runs. Molecular weights were determined in all cases by chemical ionization mass spectrometry (CH4 as reagent gas) and melting points by a Leitz VVetzlar microscope with a hot stage. HPLC (High Performance Liquid Chromatography) analyses were performed on a pBondapak C-)g column (300 x 3.9 mm id) with a flow rate of 1.0 ml/min and with ethanol and water in a ratio of 40:60 to 60:40 as the mobile phase, unless otherwise specified.

EXAMPLE 1 (22R)-16a..17a-butylidenedioxy-6a.9a-difluoro-11B-hydroxy-21-palmitoyloxyprean-4-ene-3,20-dione

A solution of palmitoyl chloride (1.2 g) in 10 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B,21dihydroxypregn-4-ene-3,20-dione (200 mg) in 5 ml of pyridine. The reaction mixture was stirred for 16 hours at room temperature. Methylene chloride (150 ml) was added and the solution was washed with 1M hydrochloric acid, 5% aqueous potassium carbonate and water and dried. The crude product after evaporation was purified by chromatography on a Sephadex LH-20 column (87 x 2.5 cm) using chloroform as the mobile phase. Fractions between 210 and 255 ml were collected and evaporated, yielding 203 mg of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11 B-hydroxy-21 palmitoyl-oxypregn-4-ene-3,20-dione. Melting point 87-90°C; mol. wt. 706 (calcd. 707.0). Purity: 96% (HPLC analysis).

EXAMPLE 2 (22R)-16α,17α-butylidenedioxy-6α,9α-difluoro-11β-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione

To a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-113,21dihydroxypregn-4-ene-3,20-dione (50 mg) and palmitoyl chloride (35 mg) in 10 ml of methylene chloride was added dropwise a solution of triethylamine (13 mg) in 2 ml of methylene chloride. The reaction mixture was stirred for 2 hours at room temperature. A further 50 ml of methylene chloride was added and the reaction mixture was treated as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (87 x

2.5 cm) and chloroform was used as the mobile phase. Fractions between 210 and 255 ml were collected and evaporated, yielding 34 mg of (22R)-16a,17a-butylidenedioxy-6a,9adifluoro-11 B-hydroxy-21-palmitoyl-oxypregn-4-ene-3,20-dione. Mol. mass 706 (calcd. 707.0). Purity: 95% (HPLC analysis).

EXAMPLE 1 (22S)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione

A solution of palmitoyl chloride (0.4 ml) in 10 ml of dioxane was added dropwise to a solution of (22S)-16a,17a-butylidenedioxy-6a,9a-difluoro-113,21dihydroxypregn-4-ene-3,20-dione (70 mg) in 25 ml of pyridine. The reaction mixture was stirred for 16 hours at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (87 x

2.5 cm) and chloroform was used as the mobile phase. Fractions between 225 and 265 ml were collected and evaporated, yielding 92 mg of (22S)-16a,17a-butylidenedioxy-6a,9adifluoro-11 B-hydroxy-21-palmitoyl-oxypregn-4-ene-3,20-dione as an oil. Mol. wt. 706 (calcd. 707.0). Purity: 97% (HPLC analysis).

EXAMPLE 4 (22R)-16q,17a-butylidenedioxy-6a,9a-difluoro-11B-hydroxy-21-myristoyloxypreqn-4-ene-3,20-dione

Myristoyl chloride was synthesized by refluxing myristic acid (7.0 g) and thionyl chloride (9 ml) in trichloroethylene (100 ml) for 3 hours. The solvent was then evaporated.

To a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B,21 dihydroxypregn-4-ene-3,20-dione (51 mg) in 10 ml of methylene chloride was added myristoyl chloride (32 mg), followed by triethylamine (13 mg) dissolved in 5 ml of methylene chloride. The reaction mixture was stirred for 4 hours at room temperature. More methylene chloride was added and the mixture was washed successively with 0.1 M hydrochloric acid and water (3 x 50 ml). After drying and evaporation, the residue was purified by chromatography on Merck Kieselgel 60 using heptane-acetone, 6:4 as mobile phase, yielding 27 mg of (22R)-16a,17abutylidenedioxy-6a,9a,-difluoro-11B-hydroxy-21-myristoyloxypregn-4-ene-3,20dione. Mol. wt. 678 (calcd. 678.9). Purity: 96.8% (HPLC analysis).

EXAMPLE 5 (22R)-16a,17a-butylidenedioxy-6oi,9(x-difluoro-11B-hydroxy-21-lauroyloxypreqn4-ene-3,2Q-dione

To a solution of (22R)-16a,17a-butylidenedioxy-6a,9oc-difluoro-113,21 dihydroxypregn-4-ene-3,20-dione (51 mg) in 5 ml of methylene chloride was added lauroyl chloride (28 mg), followed by triethylamine (13 mg) dissolved in 2 ml of methylene chloride. The reaction mixture was stirred for 3 hours at room temperature, further methylene chloride was added and the organic phase was washed successively with 0.1 M hydrochloric acid and water (3 x 30 ml). After drying and evaporation, the residue was purified by chromatography on Merck Kieselgel 60 using heptane-acetone, 6:4 as the mobile phase. The obtained product was further purified in a second chromatographic step using petroleum ether:ethyl acetate, 3:2 as the mobile phase, which gave 33 mg of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro11B-hydroxy-21-lauroyloxypregn-4-ene-3,20-dione. Mol. mass 650 (calc. 650.8). Purity: 96.9% (HPLC analysis).

EXAMPLE 6 (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione

A solution of palmitoyl chloride (2.3 ml) in 15 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B,21dihydroxypregna-1,4-diene-3,20-dione (700 mg) in 30 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first example. The crude product was purified by chromatography on a Sephadex LH-20 column (76 x 6.3 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 1020 and 1350 ml were collected and evaporated, yielding 752 mg of (22R)-16a,17ct-butylidenedioxy-6a,9a-difluoro-11 B-hydroxy-21 palmitoyloxypregna-1,4-diene-3,20-dione. Melting point 141-145°C; [α] _D ²⁵ = +71.6° (c = 0.204; CH2Cl2); mol. wt. 704 (calcd. 704.9). Purity: 97.7% (HPLC analysis).

EXAMPLE 7 (22S)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione

A solution of palmitoyl chloride (0.5 ml) in 5 ml of dioxane was added dropwise to a solution of (22S)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B,21dihydroxypregna-1,4-diene-3,20-dione (150 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first example. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x 2.5 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 215 and 315 ml were collected and evaporated, yielding 132 mg of (22S)-16a,17a-butylidenedioxy-6a,9a-difluoro-11 B-hydroxy-21 palmitoyloxypregna-1,4-diene-3,20-dione. Melting point 176-180°C; [α] _D ²⁵ = +47.5° (c = 0.198; CH2Cl2); mol. wt. 704 (calcd. 704.9). Purity: 99% (HPLC analysis).

EXAMPLE 8 (22R)-21-acetoxy-16a,17a-butylidenedioxy-6a,9a-difluoro-11B-hydroxy-pregn-4-ene-3,20-dione

A solution of acetyl chloride (38 mg) in 5 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11 B,21 dihydroxypregn-4-ene-3,20-dione (75 mg) in 5 ml of pyridine. The reaction mixture was stirred for 16 hours at room temperature. After evaporation, methylene chloride (75 ml) was added and the solution was washed with cold 5% aqueous potassium carbonate and saturated sodium chloride solution. The crude product was purified by chromatography on a Sephadex LH-20 column (85 x 2.5 cm) using chloroform as the mobile phase. Fractions between 365 and 420 ml were collected and evaporated, yielding 57 mg of (22R)-21-acetoxy-16a,17a-butylidenedioxy-6a,9 a-difluoro-11B-hydroxy-pregn-4-ene-3,20-dione. Melting point 182-189°C; [α] _D 25 ₌ +112.0° (c = 0.225; CH ₂ Cl ₂ ); mol. wt. 510 (calcd. 510.6). Purity: 99.0% (HPLC analysis).

EXAMPLE 9 (22R)-iea.17a-butylidenedioxy-6a.9a-difluoro-11B-hydroxy-21-valeroyloxypregn-4-ene-3,20-dione

A solution of valeroyl chloride (60 mg) in 5 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a.,9a.-difluoro-11B,21dihydroxypregn-4-ene-3,20-dione (75 mg) in 5 ml of pyridine. The reaction mixture was stirred for 16 hours at room temperature and worked up as in the first example. After evaporation, methylene chloride (75 ml) was added and the solution was washed with cold 5% aqueous potassium carbonate and saturated sodium chloride solution. The crude product was purified by chromatography on a Sephadex LH20 column (85 x 2.5 cm) using chloroform as the mobile phase. Fractions between 265 and 325 ml were collected and evaporated to give 50 mg of (22R)-16a,17abutylidenedioxy-6a,9a-difluoro-11B-hydroxy-21-valeroyloxypregn-4-ene-3,20dione. Melting point 181-185°C; [α] _D ²⁵ = +109.4° (c = 0.212; CH ₂ Cl ₂ ); mol. wt. 552 (calcd. 552.7). Purity; 99.8% (HPLC analysis).

EXAMPLE 10 (22R)-16α,17α-butylidenedioxy-6α,9α-difluoro-11β-hydroxy-21-capryloxypregna-1,4-diene-3,20-dione

A solution of decanoyl chloride (0.2 ml) in 3 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a,9a-difluoro-11B,21dihydroxypregna-1,4-diene-3,20-dione (100 mg) in 6 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified after evaporation by chromatography on a Sephadex LH-20 column (71 x 6.3 cm) using chloroform as the mobile phase. Fractions between 1470-1725 were collected and evaporated, yielding 113 mg of (22R)-16 a,17a-butylidenedioxy-6a,9a-difluoro-11 B-hydroxy-21-capryloxypregna-1,4-diene3,20-dione. Melting point: 182-184°C; [α] _D ²⁵ = +71.5° (c = 0.186; CH ₂ Cl ₂ ); mol. mass: 620 (calc. 620.9). Purity: 97.7% (HPLC analysis).

EXAMPLE 11

6a,9a-difluoro-116,21-dihydroxy-16a,17a-f(1-methylethylidene)bis(oxy)lpreqn-4-ene-3,20-dione

A suspension of 0.9 g of tris(triphenylphosphine)rhodium chloride in 250 ml of deaerated toluene was hydrogenated for 45 min at room temperature and atmospheric pressure. A solution of 1.0 g of fluocinolone 16α,17α-acetonide in 100 ml of absolute ethanol was added and hydrogenation was continued for a further 40 h. The reaction product was evaporated and the residue was purified by flash chromatography on silica gel using acetone-petroleum ether as the mobile phase to remove the major part of the catalyst. The eluate was evaporated on a Sephadex LH-20 column (72.5 x 6.3 cm) using chloroform as the mobile phase. Fractions between 3555-4125 were collected and evaporated, yielding 0.61 g of 6a,9a-difluoro-110,21dihydroxy-16α,17a-[(1 -methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione. Melting point 146151°C; [α] _{D =} +124.5° (c = 0.220; CH2Cl2); mol. wt. 454 (calcd. 454.6). Purity: 98.5% (HPLC analysis).

EXAMPLE 12

6a,.9oc-difluoro-110-hydroxy-16a,17a-[(1-methylethylidene)bis(oxy)1-21-palmitoyloxyoreqn-4-ene-3,20-dione

A solution of palmitoyl chloride (2.1 ml) in 15 ml of dioxane was added dropwise to a solution of 6a,9a-difluoro-110,21 -dihydroxy-16a, 17a-[(1methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione (310 mg) in 30 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first example. The crude product was purified by chromatography on a Sephadex LH-20 column (76 x 6.3 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 1035-1260 were collected and evaporated, yielding 158 mg of 6a,9a-difluoro-110-hydroxy-16a,17a-[(1-methylethylidene)bis(oxy)]-21palmitoyloxypregn-4-ene-3,20-dione. Melting point 82-86°C; [α] _D ²⁵ = +85.3° (c = 0.232; CH2Cl2); mol. wt. 692 (calcd. 692.9). Purity: 98.6% (HPLC analysis).

EXAMPLE 13 (22R)- and (22S)-21-acetoxy-16α,17a-butylidenedioxy-6(x-fluoro-11B-hydroxypregn-4-ene-3,20-dione (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxypregn-4-ene-3,20-dione (68 mg) was dissolved in 1 ml of pyridine. Acetic anhydride (1 ml) was added and the reaction mixture was stirred at room temperature for 1 hour, poured into ice water and extracted with 3 x 25 ml of methylene chloride. The extract was dried and evaporated. The remaining 22R,S mixture was separated by chromatography on a Sephadex LH-20 column (89 x 2.5 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 380-400 (A) and 420-440 (B) were collected and evaporated.

After precipitation from methylene chloride-petroleum ether, fraction A yielded 14 mg of (22S)21-acetoxy-16a,17a-butylidenedioxy-6a-fluoro-11 B-hydroxypregn-4-ene-3,20dione. Melting point: 179-186°C; [a] _D ²⁵ = +86.2° (c = 0.188; CH ₂ CI ₂ ); mol. mass: 492 (calcd. 492.6). Purity: 97.5% (HPLC analysis).

After precipitation from methylene chloride-petroleum ether, fraction B yielded 20 mg of (22R)21-acetoxy-16a,17oc-butylidenedioxy-6a-fluoro-11 B-hydroxypregn-4-ene-3,20dione. Melting point: 169-172°C; [a] _D ²⁵ = +139.0° (c = 0.200; CH ₂ Cl ₂ ); mol. mass: 492 (calcd. 492.6). Purity: 97.9% (HPLC analysis).

EXAMPLE (22R.S)-16a,17a-butylidenedioxy-6".-fluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione

To a suspension of 1.4 g of tris(triphenylphosphine)rhodium chloride in 300 ml of deaerated toluene was added a solution of 1170 mg of 6a-fluoro-11B,16a,17cc,21-tetrahydroxypregna1,4-diene-3,20-dione in 250 ml of absolute ethanol. The mixture was hydrogenated for 22 hours at room temperature and atmospheric pressure and evaporated. The residue was precipitated from acetone-chloroform, yielding 661 mg of 6a-fluoro-11B,16a,17a,21tetrahydroxypregn-4-ene-3,20-dione. Mol. mass 396 (calcd. 396.5). Purity: 96.6% (HPLC analysis).

6a-Fluoro-11 B, 16α,17a,21 -tetrahydroxypregn-4-ene-3,20-dione (308 mg) was added portionwise to a solution of butanal (115 mg) and 70% perchloric acid (0.2 ml) in 50 ml dioxane. The reaction mixture was stirred at room temperature for 6 hours. Methylene chloride (200 ml) was added and the solution was washed with 10% aqueous potassium carbonate and water and dried. The residue after evaporation was purified on a Sephadex LH-20 column (87 x 2.5 cm) using chloroform as the mobile phase. Fractions between 420-500 ml were collected and evaporated, yielding 248 mg of (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-11 β,21 -dihydroxypregn-4-ene3,20-dione. Melting point 85-96°C; [ct] _D ²⁵ = +119.8° (c = 0.192; CH ₂ Cl ₂ ); mol. mass 450 (calcd. 450.6). Purity: 96.1% (HPLC analysis). The ratio of the 22R- and 22-S epimers was 59/41 (HPLC analysis).

A solution of palmitoyl chloride (0.21 ml) in 3 ml of dioxane was added dropwise to a solution of (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-116,21dihydroxypregn-4-ene-3,20-dione (50 mg) in 6 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 185-230 were collected and evaporated, yielding 42 mg of (22R,S)-16a,17abutylidenedioxy-6a-fluoro-116-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 99% (HPLC analysis). The ratio of the 22R- and 22-S epimers was 15/85 (HPLC analysis).

EXAMPLE 15 (22R)-16a,17a-butylidenedioxy-6a-fluoro-116-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-116,21-dihydroxypregn-4-ene-3,20-dione (225 mg) was separated by preparative HPLC in portions on a μBondapak Cjg column (150 x 19 mm) using 40:60 ethane/water as the mobile phase. Fractions centered at 265 ml (A) and 310 ml (B) were collected and evaporated. After precipitation from methylene chloride-petroleum ether, fraction A yielded 68 mg of (22R)-16a,17abutylidenedioxy-6a-fluoro-116,21-dihydroxypregn-4-ene-3,20-dione. Melting point: 180-192°C; [a] _D ²⁵ = +138.9° (c = 0.144; CH ₂ Cl ₂ ); mol. wt. 450 (calcd. 450.6). Purity: 99.4% (HPLC analysis).

After precipitation from methylene chloride-petroleum ether, fraction B yielded 62 mg of (22S)16a,17a-butylidenedioxy-6a-fluoro-116,21-dihydroxypregn-4-ene-3,20-dione. Mp 168-175°C; [a] _D ²⁵ = +103.7° (c = 0.216; CH ₂ Cl ₂ ); mol. wt 450 (calcd 450.6). Purity: 99.5% (HPLC analysis).

A solution of palmitoyl chloride (0.22 ml) in 5 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a-fluoro-11B,21dihydroxypregn-4-ene-3,20-dione (32 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (87 x

2.5 cm) and chloroform was used as the mobile phase. Fractions between 215-250 were collected and evaporated, yielding 38 mg of (22R)-16a,17a-butylidenedioxy-6a-fluoro11 B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 96.0% (HPLC analysis).

EXAMPLE 16 (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-11B,21-dihydroxypregn-4-ene-3,20-dione (68 mg) was dissolved in 1 ml of pyridine. Acetic anhydride was added and the reaction mixture was stirred at room temperature for 1 hour, poured into ice water and extracted with 3 x 25 ml of methylene chloride. The extract was dried and evaporated. The remaining 22R,S epimeric mixture was separated by chromatography on a Sephadex LH20 column (89 x 2.5 cm) using heptane:chloroform-.ethanol, 20:20:1 as the mobile phase. Fractions between 380-400 (A) and 420-440 (B) were collected and evaporated.

After precipitation from methylene chloride-petroleum ether, fraction A yielded 14 mg of (22S)21-acetoxy-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxypregn-4-ene-3,20dione. Melting point: 179-186°C; [a] _D ²⁵ = +86.2° (c = 0.188; CH ₂ Cl ₂ ); mol. mass: 492 (calcd. 492.6). Purity: 97.5% (HPLC analysis).

After precipitation from methylene chloride-petroleum ether, fraction B yielded 20 mg of (22 R)21-acetoxy-16a,17cc-butylidenedioxy-6a-fluoro-11 B-hydroxypregn-4-ene-3,20dione. Melting point 169-172°C; [<x] _D ²⁵ = +139.0° (c = 0.200; CH ₂ Cl ₂ ); mol. mass 492 (calcd. 492.6). Purity: 97.9% (HPLC analysis).

To a solution of 14 mg (22S)-21-acetoxy-16a,17a-butylidenedioxy-6a-fluoro11 B-hydroxypregn-4-ene-3,20-dione in 2 ml ethanol was added 2 ml 2M hydrochloric acid. After stirring for 5 hours at 60°C, the reaction mixture was neutralized with saturated aqueous sodium hydrogen carbonate and extracted with 3 x 25 ml methylene chloride. The combined extracts were washed with water, dried and evaporated. The residue was purified by chromatography on a Sephadex LH-20 column (87 χ 2.5 cm) using chloroform as the mobile phase. Fractions between 455510 were collected and evaporated to give 7 mg of (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B-21-dihydroxypregn-4-ene-3,20-dione. Mol. wt. 450 (calcd. 450.6). Purity: 96.6% (HPLC analysis).

A solution of palmitoyl chloride (195 mg) in 5 ml of dioxane was added dropwise to a solution of (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B,21dihydroxypregn-4-ene-3,20-dione (32 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 205-245 ml were collected and evaporated, yielding 38 mg of (22S)-16α,17αbutylidenedioxy-6α-fluoro-11β-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 96.4% (HPLC analysis).

EXAMPLE 17 (22RS)-16oc.17a-butylidenedioxy-6a-fluoro-11B-hydroxy-21-lauroyloxypregn-4-ene-3,20-dione

To a solution of (22RS)-16a,17a-butylidenedioxy-6a-fluoro-11B,21dihydroxypregn-4-ene-3,20-dione (50 mg) in 6 ml of pyridine was added dropwise a solution of lauroyl chloride (0.4 ml) in 3 ml of dioxane. The reaction mixture was stirred overnight at room temperature and worked up as in the first example. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x 2.5 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 215-255 ml were collected and evaporated, yielding 15 mg of (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxy-21-lauroyloxypregn-4-ene-3,20-dione. M.p. 125-143°C; [mp] _D ²⁵ = +92.8° (c = 0.208; CH ₂ CI ₂ ); mol. mass 632 (calcd. 632.9). Purity: 96.2% (HPLC analysis). The ratio of the 22R- and 22-S epimers was 58/42 (HPLC analysis).

EXAMPLE 18 (22R)-16α·17tx-butylidenedioxy-6α-fluoro-11β-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione

6a-Fluoro-11 B,16a,17a,21 -tetrahydroxypregna-1,4-diene-3,20-dione (400 mg) was added portionwise to a solution of butanal (0.18 ml) and 70% perchloric acid (0.2 ml) in 50 ml dioxane. The reaction mixture was stirred at room temperature for 16 hours. Methylene chloride (200 ml) was added and the solution was washed with 10% aqueous potassium carbonate and water and dried. The residue after evaporation was purified on a Sephadex LH-20 column (75 x 6.3 cm) using chloroform as the mobile phase. Fractions between 2880-3300 ml were collected and evaporated, yielding 1209 mg of (22R,S)-16a,17a-butylidenedioxy-6a-fluoro-11 B,21-dihydroxypregna1,4-diene-3,20-dione. Mol. wt. 448 (calc. 448.5). Purity: 95.7% (HPLC analysis). The ratio of 22R- and 22-S epimers was 55/45 (HPLC analysis).

(22R,S)-16a,17a-butylidenedioxy-6a-fluoro-113,21-dihydroxypregna-1,4diene-3,20-dione (36 mg) was chromatographed on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 1720-1800 ml (A) and 1960-2025 (B) were collected and evaporated. Two products were precipitated from methylene chloride-petroleum ether. The product from fraction A (12 mg) was determined by ¹ H-NMR and mass spectroscopy to be (22S)-16a,17a-butylidenedioxy-6afluoro-113,21-dihydroxypregna-1,4-diene-3,20-dione, and the product from fraction B (10 mg) to be the 22R-epimer.

The two epimers had the following properties. Epimer 22S: mp 172-180°C; [bp] _D 25 _ ₊ G2.3 ^O (c = 0.132; CH2CI2); mol. mass 448 (calcd. 448.5). Epimer 22R: mp 95-106°C; [a]D ²⁵ = +105.9° (c = 0.152; _CH2CI2 ); mol. mass 448 (calcd. 448.5). The purity of the epimers was determined by HPLC analysis and was 98.9% for 22Sin and 97.7% for 22R-epimer.

A solution of palmitoyl chloride (172 mg) in 5 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-6a-fluoro-113,21dihydroxypregna-1,4-diene-3,20-dione (56 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x 2.5 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 225-285 ml were collected and evaporated, yielding 31 mg of (22R)-16a,17a-butylidenedioxy-6a-fluoro-11 B-hydroxy-21 -palmitoyloxypregna1,4-diene-3,20-dione. Melting point: 95-100°C; [b.p.] _D ²⁵ = +68.0° (c = 0.200; CH ₂ Cl ₂ ); mol. mass: 686 (calc. 686.95). Purity: 97.7% (HPLC analysis).

EXAMPLE 19 (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione

A solution of palmitoyl chloride (110 mg) in 5 ml of dioxane was added dropwise to a solution of (22S)-16a,17a-butylidenedioxy-6a-fluoro-113,21 dihydroxypregna-1,4-diene-3,20-dione (46 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x 2.5 cm) using heptane:chloroform:ethanol, 20:20:1 as the mobile phase. Fractions between 185-225 ml were collected and evaporated, yielding 37 mg of (22S)-16a,17a-butylidenedioxy-6a-fluoro-11B-hydroxy-21-palmitoyloxypregna1,4-diene-3,20-dione. Melting point 65-68°C; [α] _D ²⁵ = +53.0° (c = 0.200; CH ₂ Cl ₂ ); mol. wt. 686 (calcd. 686.95). Purity: 95.9% (HPLC analysis).

EXAMPLE 20

6a-Fluoro-11B,21-dihydroxy-16a.,17a.-i(1-methylethylidene)bis(oxy)lpregn-4-ene-3,20-dione

A suspension of 2.1 g of tris(triphenylphosphine)rhodium chloride in 500 ml of toluene was hydrogenated for 45 min at room temperature and atmospheric pressure while the catalyst was in solution. A solution of 6a-fluoro-11B,21-dihydroxy-16a,17 a-[(1-methylethylidene)bis(oxy)]pregna-1,4-diene-3,20-dione in 1000 ml of absolute ethanol was added and the hydrogenation was continued for a further 65 h. The reaction mixture was evaporated and the residue was purified on a Sephadex LH-20 column (71 x 6.3 cm) using chloroform as the mobile phase. Fractions between 2010-2445 ml were collected and evaporated, yielding 1.51 g of 6a-fluoro-11B,21-dihydroxy-16a,17a-[(1methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione. Melting point 209-219°C; [a] _D ² θ = +133.5° (c = 0.230; CH ₂ CI ₂ ); mol. wt. 436 (calcd. 436.5). Purity: 99.6% (HPLC analysis).

EXAMPLE 21

6a-fluoro-11B-hydroxy-16a,17a-[(1-methylethylidene)bis(oxy)T21-palmitoyloxypregn-4-ene-3,20-dione

A solution of palmitoyl chloride (0.21 ml) in 3 ml of dioxane was added dropwise to a solution of 6a-fluoro-11B,21-dihydroxy-16a,17a-[(1methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione in 6 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (76 x

6.3 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 1035-1230 ml were collected and evaporated, yielding 63 mg of 6a-fluoro-11Bhydroxy-16a,17a-[(1-methylethylidene)bis(oxy)]-21-palmitoyloxypregn-4-ene-3,20dione. Melting point 99-101 °C; [<x] _D ²⁵ = +89.8° (c = 0.206; CH ₂ Cl ₂ ); mol. wt. 674 (calcd. 674.94). Purity: 97.9% (HPLC analysis).

EXAMPLE 22

9g-fluoro-113,21-dihydroxy-16g,17g-[(1-methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione

A suspension of 675 mg of tris(triphenylphosphine)rhodium chloride in 250 ml of toluene was hydrogenated for 45 min at room temperature and atmospheric pressure. A solution of 1 g of triamcinolone 16g,17g-acetonide in 100 ml of absolute ethanol was added and the hydrogenation was continued for a further 40 h. The reaction mixture was evaporated and the main part of the catalyst was removed by flash chromatography with acetone:petroleum ether (b.p. 40-60°C), 40:60 as the mobile phase. The crude product was further purified on a Sephadex LH-20 column (72.5 x 6.3 cm) using chloroform as the mobile phase. Fractions between 2746-3195 ml were collected and evaporated, yielding 404 mg of 9a-fluoro-113,21 -dihydroxy-16a,17a-[(1-methylethylidene)bis(oxy)]pregn-4ene-3,20-dione. Melting point 238-241 °C; [g] _D ²⁵ = +145.2° (c = 0.288; CH ₂ Cl ₂ ); mol. mass 436 (calcd. 436.5). Purity: 99.% (HPLC analysis).

EXAMPLE 23

9g-Fluoro-11B-hydroxy-16g,17g-f (1-methylethylidene)bis(oxy)]-21-palmitoyloxypregn-4-ene-3,20-dione

A solution of palmitoyl chloride (0.69 ml) in 10 ml of dioxane was added dropwise to a solution of 9g-fluoro-113,21-dihydroxy-16g,17g-[(1methylethylidene)bis(oxy)]pregn-4-ene-3,20-dione in 20 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 240-305 ml were collected and evaporated, yielding 102 mg of 9g-fluoro-11325 hydroxy-16g,17g-[(1-methylethylidene)bis(oxy)]-21-patmitoyloxypregn-4-ene-3,20dione as an oil. Mol. wt. 674 (calcd. 674.94). Purity: 98% (HPLC analysis).

EXAMPLE 24 (22RS)-16g,17g-butylidenedioxy-9g-fluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione

9g-Fluoro-11 β, 16a,17a,21 -tetrahydroxypregn-4-ene-3,20-dione (340 mg) was added portionwise over 20 minutes with stirring to a freshly distilled solution of butanal (100 mg) and 70% perchloric acid (0.2 ml) in 50 ml of purified and dried dioxane. The reaction mixture was stirred at room temperature for a further 5 hours. Methylene chloride (200 ml) was added and the solution was washed with aqueous potassium carbonate and water and dried with anhydrous magnesium sulfate. After evaporation, the crude product obtained was purified on a Sephadex LH-20 column (72.5 x 6.3 cm) using chloroform as the mobile phase. Fractions between 2760-3195 ml were collected and evaporated, yielding 215 mg of (22R,S)-16g,17g-butylidenedioxy-9g-fluoro-11B-21-dihydroxypregn-4-ene-3,20dione. Mol. mass 450 (calc. 450.6). Purity: 97.4% (HPLC analysis).

A solution of palmitoyl chloride (0.13 ml) in 2.5 ml of dioxane was added dropwise to a solution of (22R,S)-16g,17g-butylidenedioxy-9g-fluoro-11B,21dihydroxypregn-4-ene-3,20-dione (40 mg) in 5 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (87 x

2.5 cm) and chloroform was used as the mobile phase. Fractions between 220-300 ml were collected and evaporated, yielding 42 mg of (22R,S)-16g,17g-butylidenedioxy-9gfluoro-11 B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 99% (HPLC analysis). The ratio of the 22R- and 22-S epimers was 61/39 (HPLC analysis).

EXAMPLE 25 (22R)-16g,17g-butylidenedioxy-9g-fluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione (22R,S)-16g,17g-butylidenedioxy-9g-fluoro-11B-21-dihydroxypregn-4-ene3,20-dione (200 mg) was separated by chromatography on Sephadex LH-20 columns (76 x

6.3 cm) and used a mixture of heptane:chloroform:ethanol, 20:20:1 as the mobile phase.

Fractions between 7560-8835 ml (A) and 8836-9360 (B) were collected and evaporated. The product from fraction A (128 mg) was determined by ¹ H-NMR and mass spectroscopy to be (22S)-16a,17a-butylidenedioxy-9a-fluoro-11B,21-dihydroxypregn-4-ene-3,20dione, and the product from fraction B (50 mg) to be the 22R-epimer.

The epimers had the following properties. Epimer 22S: mp 180-190°C; [a] _D ²⁵ = +105.6° (c = 0.214; CH2CI2); mol. mass 450 (calcd. 450.6). Epimer 22R: mp 147-151°C; [a] _D ²⁵ = +133.7° (c = 0.196; CH2CI2); mol. mass 450 (calcd. 450.6). The purity of the epimers was determined by HPLC analysis and was 95.6% for 22Sin and 98.2% for the 22R-epimer.

A solution of palmitoyl chloride (0.34 ml) in 5 ml of dioxane was added dropwise to a solution of (22R)-16α,17α-butylidenedioxy-9α-fluoro-11β,21dihydroxypregn-4-ene-3,20-dione (50 mg) in 10 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 180-205 ml were collected and evaporated, yielding 36 mg of (22R)-16a,17abutylidenedioxy-9a-fluoro-11 B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 97.7% (HPLC analysis).

EXAMPLE 26 (22S)-16a,17a-butylidenedioxy-9g-fluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione

A solution of palmitoyl chloride (0.14 ml) in 15 ml of dioxane was added dropwise to a solution of (22S)-16a,17a-butylidenedioxy-9a-fluoro-11B,21dihydroxypregn-4-ene-3,20-dione (41 mg) in 3 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (89 x

2.5 cm) and heptane:chloroform:ethanol, 20:20:1 was used as the mobile phase. Fractions between 215-260 ml were collected and evaporated, yielding 26 mg of (22S)-16a,17abutylidenedioxy-9a-fluoro-11 B-hydroxy-21 -palmitoyloxypregn-4-ene-3,20-dione as an oil. Mol. wt. 688 (calcd. 688.97). Purity: 91.4% (HPLC analysis).

EXAMPLE 27 (22R)-16α,17α-butylidenedioxy-9α-fluoro-113-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione

A solution of palmitoyl chloride (75 mg) in 2.5 ml of dioxane was added dropwise to a solution of (22R)-16a,17a-butylidenedioxy-9a-fluoro-11B,21dihydroxypregna-1,4-diene-3,20-dione (25 mg) in 5 ml of pyridine. The reaction mixture was stirred overnight at room temperature and worked up as in the first case. The crude product was purified by chromatography on a Sephadex LH-20 column (85 x

2.5 cm) and chloroform was used as the mobile phase. Fractions between 235-285 ml were collected and evaporated, yielding 27 mg of (22R)-16a,17a-butylidenedioxy-9a-fluoro11B-hydroxy-21-palmitoyloxypregna-1,4-diene-3,20-dione. Melting point 116-121°C; [a] _D 25 _ +67.40 ( _{c =} 0.172; CH2Cl2); mol. wt. 686 (calcd. 687.0). Purity 96.5% (HPLC analysis).

EXAMPLE 28

PHARMACEUTICAL PREPARATIONS

The non-limiting examples that follow illustrate formulations that are intended for various targeted forms of administration. The amounts of active steroid in percutaneous formulations are typically 0.001-0.2% (w/w), preferably 0.01-0.1% (w/w).

Formulation 1, ointment [g]

Steroid, micronized 0.025

Liquid paraffin 10.0

White soft paraffin up to 100.0

Formulation 2, ointment[gj

Steroids 0.025

Propylene glycol 5.0

Sorbitan sesquioleate 5.0

Liquid paraffin 10.0

White soft paraffin up to 100.0

Formulation 3, oil-in-water cream[g]

Steroid 0.025 Cetanol 5.0 Glyceryl monostearate 5.0 Liquid paraffin 10.0 Cetomacrogol 1000 2.0 Citric acid 0.1 Sodium citrate 0.2 Propylene glycol 35.0 Water up to 100.0

Formulation 4, oil-in-water cream[gj

Steroid, micronized 0.025 White soft paraffin 15.0 Liquid paraffin 5.0 Cetanol 5.0 Sorbimacrogol strearate 2.0 Sorbitan monostearate 0.5 Sorbic acid 0.2 Citric acid 0.1 Sodium citrate 0.2 Water up to 100.0

Formulation 5, water-in-oil cream[g]

Steroid 0.025 White soft paraffin 35.0 Liquid paraffin 5.0 Sorbitan sesquioleate 5.0 Sorbic acid 0.2 Citric acid 0.1 Sodium citrate 0.2 Water up to 100.0

Formulation 6, lotion [mg]

Steroids 0.25

Isopropanol 0.5

Carboxyvinylpolymer 3

NaOH to top up

Water up to 1.0

Formulation 7, suspension for injection

Steroid, micronized 0.05-10 mg

Sodium carboxymethyl cellulose 7 mg

NaCl 7 mg

Polyoxyethylene (20) sorbitan monooleate 0.5 mg

Phenyl carbinol 8 mg

Water, sterile up to 1.0 ml

Formulation 8, aerosol for oral and nasal inhalation [% w/w]

Steroid, micronized 0.1 Sorbitan trioleate 0.7 Trichlorofluoromethane 24.8 Dichlorotetrafluoromethane 24.8 Dichlorodifluoromethane 49.6

Formulation 9, atomizing solution

Steroid 7.0 mg Propylene glycol 5.0 grams Water to 10.0 grams

Formulation 10, inhalation powder_

Gelatin capsule filled with micronized steroid 0.1 mg

Lactose 20 mg

The powder is inhaled using an inhalation device.

Formulation 11, inhalation powder

Powder in the form of pellets, filled into a multi-dose powder inhaler. Each dose contains micronized steroid. 0.1 mg

Formulation 12, inhalation powder

Powder in the form of pellets, filled into a multidose powder inhaler. Each dose contains micronised steroid. 0.1 mg Lactose, micronised 1 mg

Formulation 13, capsules for the treatment of the small intestine [mg]

Steroids 1.0

Sugar balls 321

Aquacoat EC D 30 6.6

Acetyltributyl citrate 0.5

Polysorbate 80 0.1

Eudragit L100-55 17.5

Triethyl citrate 1.8

Talc 8.8

Antifoaming agent MMS 0.01

Formulation 14, capsules for the treatment of colon [mg]

Steroids 2.0

Sugar balls 305

Aquacoat ECD 30 5.0

Acetyltributyl citrate 0.4

Polysorbate 80 0.14

Eudragit N E30 D 12.6

Eudragit S100 12.6

Talc 12.6

Formulation 15. Rectal enema

Steroid

Sodium carboxymethyl cellulose Disodium acetate Methyl parahydroxybenzoate Propyl parahydroxybenzoate Sodium chloride Citric acid anhydride Polysorbate 80 Purified water

0.02 mg 25 mg 0.5 mg 0.8 mg 0.2 mg 7.0 mg 1.8 mg 0.01 mg 1.0 ml

Formulation 16, a formulation containing a steroid bound in liposomes

A. Preparation of the instillation formulation

Synthetic dipalmitoylphosphatidylcholine (45 mg), dimyristoylphosphatidylcholine (7 mg), dipalmitoylphosphatidylglycerol (1 mg) and (22R)-16a,17a-butylidenedioxy-6a,9adifluoro-11B-hydroxy-21-palmitoyloxypregn-4-ene-3,20-dione (5 mg) were mixed in a glass tube. All components were dissolved in chloroform. Most of the solvent was evaporated using N2 and then under reduced pressure, forming a thin film of lipid components on the surface of the glass tube. An aqueous solution of 0.9% NaCl was added to the lipids. Liposome formation was carried out at a temperature above the phase transition temperature of the lipids. Liposomes were formed by shaking or sonication with a sonicator probe. The resulting suspension contained liposomes ranging in size from very small bubbles to 2 pm.

B. Preparation of the inhalation formulation

The preparation of liposomes was carried out as in Example A, where the aqueous solution contained 10% lactose. The ratio of lactose to lipid was 7:3.

The liposome suspension was frozen on dry ice and lyophilized. The dried product was micronized to yield particles with a mass median aerodynamic diameter (MMAD) of about 2 pm.

PHARMACOLOGY

Selectivity for local anti-inflammatory activity can be illustrated with the following airway model.

A significant portion of inhaled GCS is deposited in the pharynx and is later swallowed and ends up in the intestine. This portion contributes to the undesirable side effects of the steroid because it acts outside the area intended to be treated (the lung). Thus, it is advantageous to use GCS with high local anti-inflammatory activity but with low GCS-induced effects after oral administration. Thus, we conducted studies with the aim of determining the GCS-induced effects after local application to the lung as well as after oral administration and the differences between the action of the glucocorticosteroid in the treated region of the lung and outside this area, which was tested in the following manner.

TEST MODELS

A) Test model for desired lacal anti-inflammatory activity on mucous airways (left lung lobe)

Sprague Dawley rats (250 g) were lightly anesthetized with Ephrane and a glucocorticosteroid test preparation (in liposomes suspended in saline) was instilled directly into the left lung lobe at a volume of 0.5 ml/kg. Two hours later, a Sephadex suspension (5 mg/kg in a volume of 1 ml/kg) was instilled into the trachea sufficiently above the bifurcation to reach both the left and right lung lobes. Twenty hours later, the rats were sacrificed, the left lung lobes dissected, and weighed. Control groups received vehicle instead of glucocorticosteroid and saline instead of Sephadex suspension to measure the weight of normal lungs and Sephadex edema that was not treated with the drug.

B) Test model for adverse systemic effects of orally absorbed glucocorticosteroids

Sprague Dawley rats (250 g) were lightly anesthetized with Ephrane and given a glucocorticosteroid test preparation orally, volume 1.0 ml/kg. Two hours later, a Sephadex suspension (5 mg/kg in a volume of 1 ml/kg) was instilled into the trachea sufficiently above the bifurcation to reach both the left and right lung lobes. Twenty hours later, the rats were sacrificed, the lung lobes dissected, and weighed. Control groups received vehicle instead of glucocorticosteroid and saline instead of Sephadex suspension to measure the weight of normal lungs and Sephadex edema that was not treated with the drug.

The results of the comparative study are given in Table 1. The pharmacological profile of the compounds of the invention was compared with the profiles of budesonide-21-palmitate and flumethasone-21-palmitate in liposomes. All steroids of the invention showed a higher local anti-inflammatory potency in the lungs after topical administration than budesonide-21-palmitate in liposomes. Moreover, the results also show a higher pulmonary selectivity of the tested compounds of the invention compared to selected, previously known compounds, since the dose of the above-mentioned compounds required to inhibit pulmonary edema (Εϋςθ) by oral administration is 158 (Example 3), 247 (Example 7) and 559 (Example 1) times higher and that of budesonide-21-palmitate is 66 times higher and that of flumethasone-21-palmitate is 8 times higher than the dose required to inhibit pulmonary edema by topical administration of the drugs to the lungs.

Thus, it can be concluded that the compounds of the invention are well suited for the topical treatment of inflammatory disorders on the skin and in various body cavities (i.e. lungs, nose, intestines and joints).

Effects of the tested glucocorticosteroids in liposomes in a Sephadex-induced rat pulmonary edema model. Results are given relative to the corresponding Sephadex-treated control group.

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Claims (15)

PATENT APPLICATIONS 1) R-j and R2 are not hydrogen at one time, 1) R-, and R2 are not hydrogen at one time, A compound characterized in that it has the general formula 1 or a stereoisomeric component thereof in which the formula 1,2 is saturated or has a double bond therein, R-j is hydrogen or branched or unbranched hydrocarbon having a chain of 1-4 carbon atoms, R2 is hydrogen or branched or unbranched hydrocarbon having a chain of 1-10 carbon atoms, R3 is an acyl having a branched or unbranched, saturated or unsaturated hydrocarbon chain of 1-20 carbon atoms, X- | is hydrogen or halogen Χ2 is hydrogen or halogen and it is specified that 2) X- | and Χ2 are not hydrogen at one time. A compound according to the first claim, characterized in that, in general formula 1, the site 1,2 is saturated, R-l is hydrogen or a branched or unbranched hydrocarbon chain having 1-4 carbon atoms, R2 is hydrogen or a branched or unbranched hydrocarbon chain having 1-10 carbon atoms, R3 is an acyl having a branched or unbranched, saturated or unsaturated hydrocarbon chain having 1-20 carbon atoms, X- | is hydrogen or halogen, Χ2 is hydrogen or halogen, it is specified that: 2) Χ-j and Χ2 are not hydrogen at one time, A compound according to any one of claims 1-2, characterized in that R 3 is acyl having 11-20 carbon atoms. 3) when there is a double bond at the 1,2 position, R- | and R2 are not at one time methyl groups, A compound according to any one of claims 1-2, wherein R 3 is acyl having 1-10 carbon atoms. 4) when there is a double bond at the 1,2 position, R- | hydrogen atom and R2 is a branched or unbranched hydrocarbon having a chain of 1-10 carbon atoms, R3 is acyl having 11-20 carbon atoms. A compound according to the third claim, characterized in that the site 1,2 is saturated, R 2 is hydrogen, R 2 is a propyl group, X- | is fluorine and Χ2 is fluorine. A compound according to the first claim, characterized in that at the site 1,2 is a double bond, R 1 is hydrogen, R 2 is a propyl group, R 3 is a palmitoyl group, X- | is fluorine, Χ2 is fluorine. A compound according to the first claim, characterized in that the formula f f ^H 2 ^{OC (CH} 2> 14 ^CH 3 A process for the preparation of a compound of general formula 1, as defined in the first claim, which is designated stem to comprise a) reacting a compound of formula wherein R1, R2, Χ1 and Χ2 are defined as in the first claim, with a compound of formula R4COOH wherein R4 is a branched or unbranched, saturated or unsaturated alkyl group having 1-19 carbon atoms, or b) reacting a compound of formula wherein R 1, R ₂ , X 1 and X ₂ are as defined in the first claim, with a compound of formula R _{4 is} COX where R 4 is as defined above and X is a halogen or aliphatic moiety -OOCR4, or c) reacting a compound of formula O · where R1, Rp, Xj and X ₂ are as defined in the first claim and Y is halogen, mesylate or p-toluenesulfonate with a compound of formula R4COO-A + where R4 is defined above and A ⁺ is a cation, then, if the compound thus obtained is an epimeric mixture and a pure epimer is desired, the epimeric mixture is separated into its stereoisomeric components. The process according to claim 8, wherein the compound of any one of claims 2-7 is prepared. A pharmaceutical composition comprising as active ingredient a compound according to any one of claims 1-7. Pharmaceutical composition according to claim 10, characterized in that it contains liposomes containing a pharmacologically active compound according to claim 3. Pharmaceutical preparation according to claims 10-11, characterized in that it is in the form of a dosage unit. A pharmaceutical composition according to claims 10-12, characterized in that it contains an active ingredient coupled to a pharmaceutically acceptable carrier. A compound according to any one of claims 1-7, 'characterized in that it is useful as a therapeutically active substance. Use of a compound according to any one of claims 1-7 for the preparation of medicaments having anti-inflammatory and anti-allergic activity.