HK1139954A - Fatty acid esters of glucocorticoids as anti-inflammatory and anti-cancer agents - Google Patents

Description

Fatty acid esters of glucocorticoids as anti-inflammatory and anti-cancer agents

Technical Field

The present invention relates to certain unsaturated fatty acid derivatives of therapeutically active glucocorticoids and pharmaceutical formulations comprising the same. Said derivatives are referred to in the present description and claims as "compounds of formula (I)". The compounds of formula (I) are useful in the treatment of cancerous and inflammatory diseases. Including hematologic and solid cancers as well as steroid-resistant cancers, asthma and steroid-resistant inflammation and the treatment of COPD.

Background

Glucocorticoids are widely used in the treatment of inflammatory diseases such as asthma, rheumatoid arthritis, inflammatory bowel disease and autoimmune diseases. Glucocorticoids are also key drugs in the treatment of Acute Lymphoblastic Leukemia (ALL) in childhood (1). Although glucocorticoids are commonly used in the treatment of chronic inflammatory diseases and cancer, optimal efficacy is limited by inherent or acquired resistance to the drug. Steroid resistance is also a major problem in the treatment of patients suffering from inflammatory bowel disease (2). A few asthmatic patients do not respond adequately or at all to corticosteroid treatment (3). Although the problem of glucocorticoid resistance may not be adequately addressed in, for example, rheumatoid arthritis as in hematological cancers, the latter two share a common mechanism of resistance, and thus strategies to overcome resistance after disease treatment would have a common benefit (7).

The beneficial effects of glucocorticoids are thought to be mediated by the inhibition of inflammatory gene expression. They act by binding to a single antibody (glucocorticoid receptor, GR) that is concentrated within the cytosol. When activated, the GR translocates into the nucleus where it turns on (transactivation) or off (transrepression) the inflammatory gene (4). Gene induction requires GR dimerization and DNA binding to specific GR Response Elements (GREs) located within the promoter region of the response gene. Glucocorticoids increase transcription of genes encoding anti-inflammatory proteins such as lipocortin-1 and interleukin-10. The most striking effect of glucocorticoids is to inhibit the expression of a variety of inflammatory genes (cytokines, enzymes, receptors, and adhesion molecules). The proinflammatory cytokine granulocyte-macrophage colony stimulating factor (GM-CSF) is an example. Glucocorticoids mediate this inhibitory effect through an interaction between activated GR and activated transcription factors, such as nuclear factor- κ B (nfkb) and activin-1 (AP-1), that regulate the expression of inflammatory genes. GM-CSF plays a key role in inflammatory and autoimmune diseases, and GM-CSF depletion has been identified as a sole drug target for suppression of disease symptoms (8).

The use of glucocorticoids is limited by serious side effects due to their endocrine and metabolic effects (5), presumably mediated by inducible transcriptional regulation of target genes by direct GR regulation. Possible side effects reported include osteoporosis and depression of the hypothalamic pituitary-adrenal cortical axis after systemic use, a reduced growth rate in children after intake of corticosteroids, loss of bone mineral, ocular symptoms and skin changes (6). Modulation of transactivation and transrepression in a tissue and pathway specific manner, due to reduced transcriptional regulation of GR regulatory genes important for side effects, can provide an improvement in therapeutic index and exhibit highly selective neocorticosteroids with higher therapeutic effect against inflammation or cancer and lower side effects (6).

In addition, the mechanistic basis for the anti-cancer effects of glucocorticoids does also involve the interaction of GR with its target genes that control the expression of cell death/apoptosis proteins.

Cell lines U937 and THP1 are human monocytes/macrophages and represent a model system of cell lines for testing the anti-inflammatory and anti-cancer activity of the products, which are also associated with glucocorticoid resistance, as they are resistant to growth inhibition by dexamethasone and prednisolone (7).

Commonly assigned publication WO98/32718 describes adrenal steroids that can be derivatized with lipophilic groups. Among the compounds mentioned are betamethasone, dexamethasone and beclomethasone. These derivatized compounds are useful for treating inflammation.

Budesonide fatty acid conjugates have been described, more specifically budesonide oleate, budesonide palmitate, budesonide linoleate, budesonide palmitoleate, and budesonide arachidonic acid. These conjugates are formed during the treatment of inflammation, more particularly asthma, but the fatty acid conjugates of the steroids mentioned are said to be pharmacologically inactive lipid conjugates (12).

Disclosure of Invention

The present invention surprisingly shows that certain unsaturated fatty acid derivatives of glucocorticoids show new and surprising therapeutic activity. A large number of these derivatives constitute novel compounds which have not been described previously. The derivatives of the invention show increased activity and fewer side effects than the previously described glucocorticoid derivatives, and they show activity in steroid resistant systems.

Glucocorticoids are known to have a site in the treatment of cancerous diseases and inflammation. Intrinsic or acquired resistance to glucocorticoid therapy is well known. Human U937 and THP1 monocytes/macrophages have been reported to represent a cell line model system characterized by intrinsic resistance to the glucocorticoids dexamethasone and prednisolone (7). The present inventors have confirmed these findings and additionally demonstrated that cells were cross-resistant to the high potency steroid fluticasone propionate, until no activity was found at 100 μ M.

As demonstrated by the examples, the inventors have surprisingly found that glucocorticoid fatty acid derivatives have high antiproliferative activity in these drug-resistant cell lines. This finding enables the treatment of diseases that are resistant to treatment with known glucocorticoids.

The present inventors have surprisingly found that the anti-inflammatory activity is increased when using glucocorticoid fatty acid derivatives in combination with a significant reduction of activation of genes associated with side effects. In activated lung cells, 22-fold stronger inhibition of the release of the cytokine GM-CSF by the fluticasone elaidic acid derivative was observed than by fluticasone propionate. At the same time, a 7-fold reduction in side effects was found. Side effects are manifested by activation of GRE elements. In this system, where activation is taken as a measure of side effects, the gene activation of steroid derivatives is reduced by 85%. These results increased the relative therapeutic index by 154-fold.

The invention relates to compounds of formula (I)

Wherein:

X₁and X₂Independently represent H or F;

R₁represents OH, OCH₂CH₃，OCH₂Cl，SCH₂F，CH₂OCOCH(CH₃)₂，CH₂-O-R₄Or O-R₄；

R₂Denotes OCOEt, OCOCHCl₂，OCO₂CH₂CH₃Or O-R₄；

R₃Represents H, CH₃Or OH; or

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Independently represent H, C₁-C₆Alkyl or C₃-C₆A cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the following conditions:

when X is₁When is F, X₂Is not H;

-at least one acyl group of formula (II) is present in the compound; and

-the compound of formula (I) is not one of the following compounds: budesonide oleate, budesonide palmitate, budesonide linoleate, budesonide palmitoleate, and budesonide arachidonic acid;

or a pharmaceutically acceptable salt thereof.

In a preferred embodiment of the invention, the compound of formula (I) is a compound wherein

X₁And X₂Both represent F;

R₁indicating SCH₂F；

R₂Represents OCOEt or O-R₄；

R₃Represents CH₃；

R₄H or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the proviso that at least one acyl group of the formula (II) is present in the compound,

or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention are the compounds of formula (I) wherein

X₁And X₂All represent H;

R₁represents CH₂-O-R₄；

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Represents H, C₁-C₆-alkyl or C₃-C₆-a cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the proviso that at least one acyl group of the formula (II) is present in the compound,

or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention are the compounds of formula (I) wherein

X₁And X₂Both represent F;

R₁represents O-R₄；

R₂Represents O-R₄；

R₃Represents CH₃；

R₄H or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the proviso that at least one acyl group of the formula (II) is present in the compound,

or a pharmaceutically acceptable salt thereof.

Preferred compounds of the invention are, for example, but not limited to, the following:

the compounds of formula (I) are useful in the treatment of diseases. More specifically, the compounds of formula (I) are useful for the treatment of cancer. In this regard, cancers include hematologic cancers, solid cancers and steroid-resistant cancers. The compounds are also useful for treating inflammation. More specifically, the compounds are well suited for the treatment of steroid-resistant inflammation. The compounds of formula (I) may also be useful in the treatment of COPD (chronic obstructive pulmonary disease).

The compounds of the present invention cause less side effects than the base steroid and have more specific anti-inflammatory activity.

The compounds of the present invention are typically, but not necessarily, formulated into a pharmaceutical composition prior to administration to a patient. Thus, in another aspect the invention relates to a pharmaceutical composition comprising a compound of the invention and one or more pharmaceutically acceptable excipients.

The excipients are selected according to the desired pharmaceutical form and the desired mode of administration. The excipients are typically formulated with the compounds of formula (I) of the present invention in a dosage form suitable for administration to a patient by a predetermined route of administration. Thus, dosage forms include forms suitable for the following routes: (a) inhalation administration, such as aerosols, solutions, and dry powders; (b) topical administration, such as creams, lotions, pastes, ointments, solutions, sprays, and gels; (c) intravenous administration, such as sterile solutions, suspensions, and powders for reconstitution; (d) oral administration, such as tablets, capsules, pills, powders, syrups, elixirs, suspensions, solutions, emulsions and sachets. The compounds of formula (I) may also be administered as eye drops.

Suitable pharmaceutically acceptable excipients may vary depending on the particular dosage form. Pharmaceutically acceptable excipients include, inter alia, the following: diluents, lubricants, fillers, disintegrants, solvents, wetting agents, suspending agents, emulsifiers, granulating agents, coating agents, binders, flavoring agents, taste masking agents, sweeteners, plasticizers, tackifiers, antioxidants, stabilizers, surfactants, and buffers. Certain excipients may perform more than one function.

Suitable diluents and fillers include lactose, sucrose, glucose, mannitol, sorbitol, potato starch, corn starch, pregelatinized starch, cellulose and derivatives thereof. Suitable binders include starch, pectin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives. Suitable disintegrants include sodium starch glycolate, alginic acid, crospovidone and sodium carboxymethyl cellulose.

The compounds of formula (I) may also be combined with biodegradable polymers for controlled release of drugs.

The pharmaceutical compositions of the present invention may be prepared using methods and techniques known to those skilled in the art.

Accordingly, one aspect of the present invention is a compound of formula (I) according to claims 9-12 for use in the treatment of cancer.

Another aspect of the invention is the use of a compound of formula (I) according to claims 14-21 for the preparation of a pharmaceutical composition for the treatment of said diseases.

The preparation of the compounds of formula (I) is illustrated below.

11 beta-elaidic acid ester of fluticasone propionate

To a solution of fluticasone propionate (1.02g, 2.04mmol) in dichloromethane (35ml) was added elaidic acid (576mg, 2.04mmol) followed by 4- (dimethylamino) pyridine (249mg, 2.04mmol) and 1, 3-dicyclohexylcarbodiimide (421mg, 2.04 mmol). The resulting solution was stirred for 24 hours at which time TLC showed a significant amount of unreacted starting material. The mixture was concentrated in vacuo (-10 ml solution) and stirred for an additional 140 hours. The mixture was further concentrated in vacuo and flash chromatographed on silica eluting with hexane/EtOAc (3: 1) to give 1.23g (79%) of the desired compound as a colorless oil.

6 alpha, 9 alpha-difluoro-11 beta, 17 alpha-dihydroxy-16 alpha-methyl-3-oxoandrostane-1, 4- Diene-17 beta-carboxylic acid

To a suspension of difluoromethylenepine (1.19g, 2.9mmol) in tetrahydrofuran (11ml) was added a solution of periodic acid (2.25g, 9.9mmol) in water (5.3 ml). The resulting solution was stirred at room temperature for 1 hour. The tetrahydrofuran was removed in vacuo leaving an aqueous suspension which was filtered, the solid washed with water and dried to give 1.17g (100%) of the title compound as a colourless solid.

6 alpha, 9 alpha-difluoro-16 alpha-elaidic acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxo Androstane-1, 4-diene-17 beta-carboxylic acid

To a solution of 6 α, 9 α -difluoro-11 β, 17 α -dihydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene-17 β -carboxylic acid (1.22g, 3.1mmol) and triethylamine (0.72g, 7.1mmol) in acetone (25ml) was added elaidic acid chloride in dichloromethane (10 ml). Elaidic acid chloride was prepared from elaidic acid (2.0g, 7.1mmol), oxalyl chloride (2.6ml, 30mmol) and DMF (catalytic amount) in toluene (45ml) at ambient temperatureStirred at room temperature for 17 hours and then evaporated to dryness. The resulting mixture was stirred at room temperature for 2 hours, treated with diethylamine (0.97ml, 9.2mmol) and the resulting solution stirred at room temperature for an additional 1.5 hours. 1M HCl was then added and the mixture was extracted with dichloromethane (3X 50 ml). The combined organic extracts were dried (Na)₂SO₄) Filtered and evaporated in vacuo. The crude product was dissolved in EtOAc, hexanes were added and the freezer was put overnight. The off-white solid obtained (2.07g, containing some N, N-diethylamide) was collected by filtration, dried and used directly in the next step. Some of the title compound (170mg) was flash chromatographed on silica gel eluting with hexane/EtOAc (4: 1) then hexane/EtOAc/AcOH (100: 1) to give 123mg (72%) as a white solid.

6 alpha, 9 alpha-difluoro-17 beta- (N, N-dimethylcarbamoylthio) carboxy-16 alpha-trans-oil Acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxoandrosta-1, 4-diene

A solution of 6 α, 9 α -difluoro-16 α -elaidoyloxy-11 β -hydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene-17 β -carboxylic acid (1.9g, 2.9mmol) and N, N-dimethylthiocarbamoyl chloride (0.71g, 5.8mmol) in acetone (40ml) at room temperature was treated sequentially with triethylamine (0.58g, 5.8mmol), anhydrous sodium iodide (0.43g, 2.9mmol) and water (0.19ml, 10% w/w steroid). The solution was stirred for 18 h, water (100ml) and EtOAc (100ml) were added and the phases were separated. The organic phase was washed with 1M HCl, 5% aqueous sodium bicarbonate, water and dried (Na)₂SO₄) Filtered and evaporated in vacuo. Flash chromatography on silica eluting with hexane/EtOAc (3: 2) afforded 1.55g (. about.70%, two steps) of the title compound as a yellow solid.

6 alpha, 9 alpha-difluoro-16 alpha-elaidic acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxo Androstane-1, 4-diene-17 beta-carbothioic acid

A solution of 6 α, 9 α -difluoro-17 β - (N, N-dimethylcarbamoylthio) carboxy-16 α -elaidoyloxy-11 β -hydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene (1.55g, 2.1mmol) in diethylamine (16ml) was refluxed for 3 hours. The solution was cooled to ambient temperature, poured into cold 3M HCl (150ml) and extracted with EtOAc (2 × 150 ml). The combined organic extracts were washed with water and brine and then dried (Na)₂SO₄) Filtered and evaporated in vacuo. The crude product (1.35g, off-white solid) was used directly in the next step.

6 alpha, 9 alpha-difluoro-16 alpha-elaidic acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxo Nondrostan-1, 4-diene-17 beta-carbothioic acid S-chloromethyl ester

To a stirred solution of 6 α, 9 α -difluoro-16 α -elaidoyloxy-11 β -hydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene-17 β -carbothioic acid (1.35g, 2mmol) in dichloromethane (40ml) was added triethylamine (0.28ml, 2mmol) followed by chloroiodomethane (0.56ml, 8 mmol). The resulting solution was stirred at ambient temperature for 70 hours, a saturated aqueous solution of ammonium chloride was added and the phases were separated. The aqueous phase was extracted with dichloromethane (2 times) and the combined organic phases were dried (Na)₂SO₄) Filtered and evaporated in vacuo. Flash chromatography on silica eluting with hexane/EtOAc (3: 1) afforded 1.04g (72%) of the title compound as a colorless solid.

6 alpha, 9 alpha-difluoro-16 alpha-elaidic acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxo O-methyl androst-1, 4-diene-17 beta-carbothioic acid S-iodomethyl ester

To a solution of 6 α, 9 α -difluoro-16 α -elaidoyloxy-11 β -hydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene-17 β -carbothioic acid S-chloromethyl ester (1.04g, 1.4mmol) in acetone (100ml) was added sodium iodide (0.86g, 5.7mmol) and refluxed for 22 hours. The solvent was evaporated in vacuo and the residue was dissolved in EtOAc, washed with water, 10% aqueous sodium bicarbonate, water and dried (Na)₂SO₄) Filtered and evaporated in vacuo. The crude product (1.1g) was used directly in the next step.

6 alpha, 9 alpha-difluoro-16 alpha-elaidic acyloxy-11 beta-hydroxy-16 alpha-methyl-3-oxo O-fluoro-1, 4-diene-17 beta-carbothioic acid S-fluoromethyl ester

To a solution of 6 α, 9 α -difluoro-16 α -elaidoyloxy-11 β -hydroxy-16 α -methyl-3-oxoandrosta-1, 4-diene-17 β -carbothioic acid S-iodomethyl ester (1.1g, 1.35mmol) in acetonitrile (65ml) was added silver fluoride (1.7g, 13.5mmol) and stirred at room temperature in the dark for 40 h. The mixture was diluted with EtOAc and filtered through a short celite and silica gel. The filtrate was washed with water and dried (Na)₂SO₄) Filtered and evaporated in vacuo. Of the crude product¹HNMR showed some unreacted chloride present and the last two steps were repeated to completely convert the chloride to fluoride. The product was then purified by flash chromatography on silica gel eluting with hexane/EtOAc (3: 1) to give 0.65g (46%, two steps) of the title compoundThe title compound was a colorless solid.

Examples

The following examples illustrate the invention. However, these examples are not to be construed as limiting the scope of the invention.

Example 1:

u937 and THP-1 cell lines were seeded in 96-well plates at 20000 cells per well. 50 μ l of cell culture medium was added to each well. At the same time, test compounds at 5 different concentrations were added and incubated for 48 hours. Use ofThe non-radioactive cell proliferation assay (Promega) investigated the cytotoxicity of test compounds in these cells. The assay is a colorimetric method for determining the number of viable cells in a proliferation or chemosensitivity assay. It consists of a solution of the novel tetrazolium compound (3- (4, 5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium, inner salt; MTS) and an electron binding reagent (phenazine methosulfate; PMS). MTS is bioreduced by cells into formazan soluble in tissue culture mediumAnd (3) obtaining the product. Nail can be measured directly from a 96-well assay plate without additional processingAbsorbance of the product at 490 nm. The amount of the formazan product and the number of viable cells in the culture (IC) measured by absorbance at 490nm₅₀Value) is proportional. The test compounds were fluticasone propionate and fluticasone elaeate derivatives. In this glucocorticoid-resistant cell line, fluticasone elaidic acid has a high potency, IC₅₀Values in the micromolar range, U937 cells were most sensitive.

TABLE 1

Compound (I)	U937 cells, IC50μM±SD	THP1 cells, IC50μM±SD
			Fluticasone propionate	＞100μM±0,03	＞100μM±0,02
Fluticasone elaeate derivative	1μM±0,02	10μM±0,04

Example 2:

monkey kidney COS-1 cells (ATCC CRL 1650) were grown in Dulbecco's modified Eagle Medium (Gibco BRL, Grand Island, NY) as described elsewhere (9). Transient transfection of COS-1 cells was performed as described elsewhere (10). Cells were plated at 2X 10⁵The density of individual cells/well was plated. Each well received 5. mu.g of the test plasmid, 5. mu.g of the beta-galactosidase control plasmid as an internal control and 2. mu.g of the expression plasmid, pMT-hGR or pGL3-basic as a vector. Plasmid details: the mouse PPAR α gene is described elsewhere (11). Vectors expressing the LUC reporter under the control of the PPAR α 5' -flanking region and the promoter region were constructed in pGL3-LUC vector (Promega). PPAR alpha 5' -flanking sequence between-2800 bp to +100bpClones were cloned into NheI digested pGL3-LUC to generate PPAR (-2800/+100) LUC plasmids. Each transfection was performed in triplicate. After plating with fresh medium containing ligand, cells were transfected for 24 hours. Cells were harvested after 72 hours, cytosolic extracts were prepared and LUC activity was measured according to the Promega protocol. The results were normalized to the measured beta-galactosidase activity, which was measured as follows: mu.l of the extract was incubated with 0.28mg of o-nitrophenyl-D-galactoside (ONPG) in 50mM phosphate buffer (pH 7.0), 10mM KCl, 1mM MgCl2 at 30 ℃ for 30 minutes and the absorbance read at 420 nm.

The test compounds were fluticasone propionate, fluticasone elaeate derivatives and diflumetosone-elaidic acid ester derivatives. The steroid fluticasone propionate was able to induce reporter activity at 100%, whereas the fluticasone elaidic acid ester derivative and the diflumetosone elaidic acid ester derivative surprisingly induced reporter activity only at 15% and 2%, respectively. Steroid analogues are less effective in gene activation of genes causing side effects.

TABLE 2

Compound (I)	Luciferase Activity/beta-galactosidase (percentage of Fluticasone propionate (%))
		Fluticasone propionate	100％
Fluticasone elaidic acid ester derivative	15％
		Difluorometasone elaidic acid ester derivative	2％

Example 3:

in A549TRE (AP-1-regulated) cells, IL-1 β -stimulation caused an increase in granulocyte-macrophage colony-stimulating factor (GM-CSF) release after 24 hours. Inhibition of GM-CSF release was measured after 24 hours of exposure to test compounds. Fluticasone elaidic acid ester derivative in A549TRE cells as IC₅₀＝1.4×10^-11M produces a concentration-dependent inhibition of GM-CSF release stimulated by IL-1 β. Fluticasone propionate in IC in A549TRE cells₅₀Is 3.1 × 10^-10M produced a concentration-dependent inhibition of GM-CSF release (Table 3). The fluticasone elaidic acid ester derivative inhibited GM-CSF release 22-fold more strongly, and its anti-inflammatory effect was 22-fold more strongly in this assay.

Induction of GRE-luciferase activity was observed in stably transfected a549 GRE-luciferase cells. Compared to concentration-dependent induction of GRE-luciferase reporter gene activity in A549 GRE-luciferase cells exposed to fluticasone propionate (where fluticasone propionate is in EC₅₀＝5.1×10^-10M is the maximum stimulus that elicits luciferase activity), no effect on GRE-luciferase activity was observed in a549 GRE-luciferase cells.

Surprisingly, the inhibition of GM-CSF by the fluticasone elaidic acid ester derivative was increased by more than 20-fold compared to the absence of effect on GRE-luciferase by the fluticasone elaidic acid ester derivative. It also shows a higher therapeutic index in this test system.

TABLE 3

Steroid compound	A549 TRE(+IL-1β)IC50
		Dexamethasone	8.5×10-10M
Fluticasone propionate	3.1×10-10M
		Fluticasone elaidic acid ester derivative	1.4×10-11M

Reference to the literature

1.Genome-wide identification of prednisolone-responsive genes in acutelymphoblastic leukaemia cells.，Tissing W.J.et al.，Blood，2007，jan.11

2.Identification of budesonide and prednisolone as substrates of theintestinal drug efflux pump P-glycoprotein.，Dilger K.et al.Inflamm.BowelDis.2004，Sept，；10(5)：578-83.

3.Mechanisms of steroid action and resistance in inflammation.，I.M.Adcock and S.J.Lane，J.of Endocrinology(2003)178，347-355.

4.Anti-inflammatory actions of glucocorticoids：Molecular mechanism.，Barnes et al.(1998)J.Clinical Science 94，557-572.

5.Inhaled corticosteroids：past lessons and future issues，Allen D.B.etal.，J.Allergy Clinic.Immunol.2003；112，p.1-40.

6.Designing corticosteroid drugs for pulmonary selectivity.；K.Biggadike，I.Uings and S.N.Farrow；Proceedings of the American Thoracic Society，Vol1，p.352-355，2004.

7.Sulfasalazine sensitizes human monocytic/macrophage cells forglucocorticoids by up-regulation of receptor(alpha)andglucocorticoid-induced apoptosis.，Oerlemans R.et al.，Ann.Rheum.Dis.，2007Jan 31.

8.GM-CSF in inflammation and autoimmunity，Hamilton J.A.，Trends Immunol.，2002，Aug.，23(8)；403-8.

9.Establishment of a clonal strain of hetatoma cells which maintain inculture the five enzymes of the urea cycle.Richardson U.I.，Snodgrass P.J.，Nuzum C.T.，Tashjian A.H.，Jr.(1974)J.Cell Physiol.83：141-149.

10.A new technique for the assay of infectivity of human adenovirus 5 DNA，Graham et al.1973)Virology 52：456-467.

11.Structure of the mouse peroxisome proliferator activated reporter gene.Gearing et al.(1994)BBRC 199，255-

12.Reversible formation of fatty acid esters of budesonide，anantiasthma glucocorticoid，in human lung and liver microsomes.Tunej，A.Sjodin，K.and Hallstrom，G.，Drug Metabolism and Disposistion，25(11)；1311-7(1997)

Claims (27)

1. A compound of the general formula (I)

Wherein:

X₁and X₂Independently represent H or F;

R₁represents OH, OCH₂CH₃，OCH₂Cl，SCH₂F，CH₂OCOCH(CH₃)₂，CH₂-O-R₄Or O-R₄；

R₂Denotes OCOEt, OCOCHCl₂，OCO₂CH₂CH₃Or O-R₄；

R₃Represents H, CH₃Or OH; or

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Independently represent H, C₁-C₆Alkyl or C₃-C₆A cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

provided that

When X is₁When is F, X₂Is other than H, and

-at least one acyl group of formula (II) is present in the compound; and

-the compound of formula (I) is not one of the following compounds: budesonide oleate, budesonide palmitate, budesonide linoleate, budesonide palmitoleate, and budesonide arachidonic acid;

or a pharmaceutically acceptable salt thereof.

2. A compound of formula (I) according to claim 1,

wherein

X₁And X₂Both represent F;

R₁indicating SCH₂F；

R₂Represents OCOEt or O-R₄；

R₃Represents CH₃；

R₄H or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

or a pharmaceutically acceptable salt thereof.

3. A compound of formula (I) according to claim 1,

wherein

X₁And X₂All represent H;

R₁represents CH₂-O-R₄；

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Represents H, C₁-C₆-alkyl or C₃-C₆-a cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

or a pharmaceutically acceptable salt thereof.

4. A compound of formula (I) according to claim 1,

wherein

X₁And X₂Both represent F;

R₁represents O-R₄；

R₂Represents O-R₄；

R₃Represents CH₃；

R₄H or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

or a pharmaceutically acceptable salt thereof.

5. A compound of formula (I) according to claim 1,

wherein

X₁And X₂All represent H;

R₁represents OCH₂Cl or O-R₄；

R₂Represents OCO₂CH₂CH₃Or O-R₄；

R₃Represents H;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the proviso that at least one acyl group of the formula (II) is present in the compound,

or a pharmaceutically acceptable salt thereof.

6. A compound of formula (I) according to claim 1,

wherein

X₁And X₂All represent H;

R₁represents OCH₂CH₃Or O-R₄；

R₂Represents OCOCOCHCl₂Or O-R₄；

R₃Represents H;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

with the proviso that at least one acyl group of the formula (II) is present in the compound,

or a pharmaceutically acceptable salt thereof.

7. A compound of formula (I) as claimed in any one of claims 1 to 4 wherein the acyl group of formula (II) is a elaidic acid residue.

8. The compound of claim 1, 2, 3, 4, 5, or 6 which is:

9. a compound of formula (I)

Wherein:

X₁and X₂Independently represent H or F;

R₁represents OH, OCH₂CH₃，OCH₂Cl，SCH₂F，CH₂OCOCH(CH₃)₂，CH₂-O-R₄Or O-R₄；

R₂Denotes OCOEt, OCOCHCl₂，OCO₂CH₂CH₃Or O-R₄；

R₃Represents H, CH₃Or OH; or

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Independently represent H, C₁-C₆Alkyl or C₃-C₆A cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

provided that

When X is₁When is F, X₂Is not H; and

-at least one acyl group of formula (II) is present in the compound;

or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

10. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 9 for use in the treatment of hematological cancer.

11. A compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 9, for use in the treatment of steroid-resistant cancer.

12. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 9 for use in adjuvant and/or palliative treatment.

13. A compound of formula (I)

Wherein:

X₁and X₂Independently represent H or F;

R₁represents OH, OCH₂CH₃，OCH₂Cl，SCH₂F，CH₂OCOCH(CH₃)₂，CH₂-O-R₄Or O-R₄；

R₂Denotes OCOEt, OCOCHCl₂，OCO₂CH₂CH₃Or O-R₄；

R₃Represents H, CH₃Or OH; or

R₂And R₃Together with the carbon atom to which they are attached, form 2-R₅，R₆-1, 3-dioxolane ring, wherein R is₅And R₆Independently represent H, C₁-C₆Alkyl or C₃-C₆A cycloalkyl group;

R₄h or C of the formula₈-C₂₄Acyl radical

CH₃-(CH₂)_k-(CH＝CH-CH₂)_l-(CH＝CH)_m-(CH₂)_n-CO (II)

Wherein k is an integer of 0 to 10, l is an integer of 0 to 6, m is an integer of 0 to 1, and n is an integer of 2 to 7;

provided that

-at least one acyl group of formula (II) is present in the compound;

or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical composition for the treatment of cancer.

14. The use according to claim 13 for the preparation of a pharmaceutical composition for the treatment of hematological cancer.

15. The use according to claim 13 for the preparation of a pharmaceutical composition for the treatment of steroid resistant cancers.

16. The use of claim 13 for the preparation of a pharmaceutical composition for adjuvant and/or palliative treatment.

17. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, for use in the treatment of inflammation.

18. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 for use in the treatment of steroid-resistant inflammation.

19. Use of a compound of formula (I) according to claim 1 for the preparation of a pharmaceutical composition for the treatment of inflammation.

20. The use according to claim 19 for the preparation of a pharmaceutical composition for the treatment of steroid resistant inflammation.

21. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 for the manufacture of a pharmaceutical composition for the treatment of chronic obstructive pulmonary disease, COPD.

22. A method of treating cancer in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a compound of formula (I) of claim 1, or a pharmaceutically acceptable salt thereof.

23. The method of claim 22, wherein the condition to be treated is a hematological cancer.

24. The method of claim 22 wherein the condition to be treated is a steroid-resistant cancer.

25. A method of treating inflammation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) of claim 1, or a pharmaceutically acceptable salt thereof.

26. The method of claim 25 wherein the condition to be treated is steroid-resistant inflammation.

27. A pharmaceutical composition comprising a compound of formula (I) according to claim 1 and a pharmaceutically acceptable excipient, carrier and/or diluent.