CZ234297A3 - Pharmaceutical preparation containing corticosteroid and the use thereof - Google Patents

Abstract

A pharmaceutical composition comprising, as active ingredient, a compound of formula (I) contained in liposomes or dehydrated liposomes.

Description

Technical field

The invention relates to pharmaceutical compositions, the active ingredient comprising a particular corticosteroid, use for the treatment of asthma by inhalation therapy, and the preparation of such pharmaceutical compositions.

BACKGROUND OF THE INVENTION

To a large extent, obstructive bronchopulmonary diseases such as bronchitis and bronchial asthma have become widespread as a result of environmental improvements. Their pathogenesis and severity vary between individuals. External allergic bronchial asthma, caused by environmental influences (such as waste gases, inverted atmospheric layers) and internal bronchial asthma, are often characterized by severe seizures with various respiratory disorders. The intensity of coughing and expectoration also varies.

Transient and mixed forms of asthma are common and should be taken into account during medical treatment.

Three groups of active substances with acceptable risks are available for the treatment of such disorders of varying intensity and origin, in addition to the combined formulations. These three groups are 3 £ ₂ adrenergic agents such as adrenaline, bamethan, clenbuterol, fenoterol, sulbutamol and terbutaline, xanthine derivatives such as theophylline and diprophylline and antichlolinergika containing the atropine derivatives ipatropium bromide and oxitropium bromide. If therapy with formulations based on these three classes of active substances is unsuccessful, the use of certain corticosteroids, such as beclomethasone or budesonide, is recommended and is administered orally or by inhalation. However, all corticosteroid formulations used in current inhalation therapy, in addition to the desired antiallergic, antiexsudative anti-inflammatory properties, have mild but undesirable systemic side effects due to absorption of the inhaled corticosteroid. Corticosteroid inhalation therapy is usually required for many years, significantly increasing the problem of systemic side effects.

SUMMARY OF THE INVENTION

It has now surprisingly been found that methyl-Sa-chloro-6α-fluoro-β-hydroxy-16α-methyl-3-oxo-17α-propionyloxyandrosta-1,4-diene-17β-carboxylate, previously proposed as an active ingredient for dermatological ointments, creams, gels and foams exhibit particularly good antiasthmatic properties when administered encapsulated in liposomes. It has also been found that a surprisingly low systemic absorption is observed with this compound.

Accordingly, in one embodiment, the invention provides a pharmaceutical composition comprising as an active ingredient a compound of formula I

F contained in liposomes or dehydrated liposomes.

The compound of formula I, methyl 9α-chloro-6α-fluoro-11β-hydroxy-16α-methyl-3-oxo-17α-propionyloxyandrosta-1,4-diene-17β-carboxylate, can be prepared as described in British patent no. For administration using an inhalation device, the liposomes may be in aqueous suspension or in dehydrated form, as a dry powder.

It has been found that liposomes containing a compound of formula I are readily taken up by alveolar macrophages and effectively inhibit the increase in eosinophils in the Brown-Norway model of allergen-induced eosinophilia in rats.

Suitable liposomes generally include those in which the lipid component comprises at least one synthetic phospholipid. Examples of synthetic phospholipids include synthetic phosphatidylcholines such as dimyristoyl-phosphatidylcholine, dipalmitoyl-phosphatidylcholine, distearoyl-phosphatidylcholine, dioleoyl-phosphatidylcholine, dilinoleoyl-phosphatidylcholine, dilauryloyl-phosphatidylcholine, 2-dilauryloyl-phosphatidylcholine, 2 phosphatidylcholine and 1-palmitoyl-2-myristoylphosphatidylcholine, synthetic phosphatidylglycerols such as dilauryloylphostatidylglycerol, dimyristoylphostatidylglycerol, dipalmitoylphostatidylglycerol, and dioleoylphostatidylglycerol acid such as synthetic phosphatidylphosphididylglycerol, and dioleoylphostatidyl glycerol phosphatidylethanolamine and dipalmitoyl phosphatidylethanolamine and synthetic phosphatidylserines such as dimyristoyl phosphatidylserine, dipalmitoyl phosphatidylserine and dioleoyl phosphatidylserine.

Preferably, the lipid component of the liposomes comprises a synthetic phosphatidylcholine such as the phosphatidylcholines listed above, optionally together with a synthetic phosphatidylserine or a synthetic phosphatidylglycerol such as the phosphatidylserines and phosphatidylglycerols mentioned above, wherein the weight ratio of phosphatidylcholine to phosphatidylserine or phosphatidylglycerol

it is preferably from 60: 40 to 95: 5, in particular from 70: 30 to: 10.

According to one preferred embodiment, the lipid component of the liposome comprises a synthetic, substantially pure phospholipid of formula IIi

CH ₂ - O - R

R ₂ -o- ² ch

CH ₂ in which

R _x is C 10 -C 20 alkanoyl, the number of carbon atoms being an even number,

R ₂ is alkanoyl group having 10 to 20 carbon atoms., The number of carbon atoms is even, R _a, R _b and R _c are hydrogen or alkyl having 1 to 4 carbon atoms, n is an integer from two to four.

In a phospholipid of formula II is an alkanoyl group having 10 to 20 carbon atoms in which the carbon number is an even number, in the meaning of the symbol R _{L R} is preferably n-dodecanoyl, n-tetradecanoyl, n-hexadecanoyl, n-octadecanoyl or n-eicosanoyl .

The C 10 -C 20 alkenoyl group in which the number of carbon atoms is an even number, in the meaning of the symbol R ₂ , is preferably 9-cis-dodecenoyl, 9-cis-tetradecenoyl, 9-cis-hexadecenoyl, 6-cis-octadecenoyl, 6-trans-octadecenoyl, 9-cis-octadecenoyl, 9-trans-octadecenoyl, 11-cis5

-octadecenoyl or 9-cis-eicosenoyl. The radicals R _a , R _b and R _c in the phospholipid of the general formula II preferably each represent a C 1 -C 4 alkyl group, in particular a methyl group.

The variable n in formula II is an integer from two to four, preferably n is two. The group of the formula - (C _n -H _2n ) - is a branched or unbranched alkylene group, for example 1,1-ethylene, 1,1-, 1,2- or 1,3-propylene or 1,2-, 1, 3- or 1,4-butylene. 1,2-ethylene (n = 2) is preferred.

Particularly preferred is a phospholipid of formula II wherein R 1 is n-dodecanoyl, n-tetradecanoyl, n-hexadecanoyl or n-octadecanoyl, R ₂ is 9-cis-dodecenoyl, 9-cis-tetradecenoyl, 9-cis-hexadecenoyl 9-cis-octadecenoyl or 9-cis-eicosenoyl, R _a , R _b and R _c are each methyl, and n is two.

A particularly preferred phospholipid of formula II is synthetic 1-n-hexadecanoyl-2- (9-cis-octadecenoyl) -3-sn-phosphatidylcholine.

In certain particularly preferred liposomes, the lipid component comprises a phospholipid of formula II in combination with a synthetic, substantially pure phospholipid of formula III

CH ₂ -or ₃

III)

R ₄ -OCH ₂ -o -P-O- (C _n H _2n ) (S)

- CH

OH in which R ₃ and R independently of one another are each alkanoyl group having 10 to 20 carbon atoms, wherein the number of carbon atoms is even, n is an integer from one to three and

Y ® 'represents a cation of a pharmaceutically acceptable base.

The C 10 -C 20 alkenoyl group in which the number of carbon atoms is an even number, in the meaning of R ₃ or R ₄ in formula III, is preferably 9-cis-dodecenoyl, 9-cis-tetradecenoyl, 9-cis-hexadecenoyl 6-cis-octadecenoyl, 6-trans-octadecenoyl, 9-cis-octadecenoyl, 9-trans-octadecenoyl, 11-cis-oct-adecenoyl or 9-cis-eicosenoyl.

The cation of the pharmaceutically acceptable base, indicated by the symbol Y ®, is, for example, an alkali metal ion such as lithium, sodium or potassium ion, ammonium ion, mono-, di- or trialkylammonium ion having 1 to 4 carbon atoms in each alkyl moiety, e.g. ethyl, diethyl or triethylammonium ion, tetramethylammonium ion, 2-hydroxyethyl-trialkylammonium ion having 1 to 4 carbon atoms in each alkyl t

or a 2-hydroxyethylammonium ion, or a basic amino acid cation such as lysine or arginine. The cation Y® is preferably sodium ion.

In a particularly preferred phospholipid of the formula III, R _{@ 3} and R _{@ 4 are the} same and are, for example, 9-cis-dodecenoyl, 9-cis-tetradecenoyl, 9-cis-hexadecenoyl, 9-cis-octadecenoyl or 9-cis-eicosenoyl, n is 1 and Y ® is sodium ion.

A particularly preferred phospholipid of formula III is synthetic sodium 1,2-di- (9-cis-octadecenoyl) -3-sn-phosphatidyl-S-serine.

According to another preferred embodiment, the lipid component of the liposomes comprises di (alkanoyl) phosphatidylcholine of 10 to 20 carbon atoms in each alkanoyl moiety, together with di (alkanoyl) phosphatidylglycerol of 10 to 20 carbon atoms in each alkanoyl moiety, wherein the number of carbon atoms in the alkanoyl groups is even, and preferred weight ratios are described above. The two alkanoyl groups present in each phospholipid may be the same or different and are preferably n-dodecanoyl (lauroyl), n-tetradecanoyl (myristoyl), n-hexadecanoyl (palmitoyl), n-octadecanoyl (stearoyl) or n-eicosanoyl. According to a particularly preferred embodiment, the di (alkanoyl) phosphatidylcholine having 10 to 20 carbon atoms in each alkanoyl moiety is distearoyl-phosphatidylcholine and the di (alkanoyl) phosphatidylglycerol having 10 to 20 carbon atoms in each alkanoyl moiety is dipalmitoyl-phosphatidylglycerol.

The lipid component of the liposomes may contain cholesterol in addition to the phospholipid (or phospholipids), the amount of cholesterol being, for example, from 20 to 60, preferably 30 to 50 mol% of the total lipid content. According to another preferred embodiment, the lipid component of the liposomes comprises a synthetic phosphatidylcholine as described above, a synthetic phosphatidylserine or a phosphatidylglycerol such as those described above, and cholesterol, wherein a preferred weight ratio of phosphatidylcholine to phosphatidylserine or phosphatidylglycerol is above and preferred weight ratio of cholesterol to total. The phospholipid is from 1: 1 to 1: 5. According to a particularly preferred embodiment, the lipid component comprises dimyristoyl-phosphatidylcholine, cholesterol and dioleoyl-phosphatidylserine.

In general, it is desirable that the weight ratio of active compound to lipid be as high as possible while maintaining the stability of the liposomes. The maximum weight ratio may vary depending on the nature and composition of the lipid component, but is generally about 1: 20. However, it has been found that good results can be obtained with liposomes in which the weight ratio is from 1: 100 to 1:50 .

Liposomes containing the active compound of the invention can be prepared using known methods for preparing drug-containing liposomes. For example, according to one method, a solution of a compound of formula I and one or more lipids in an organic solvent such as an alcohol, an ether or a halogenated hydrocarbon or a mixture thereof is gradually added, preferably dropwise, to a stirred aqueous medium such as phosphate buffered sodium chloride solution. thereby obtaining an aqueous liposome suspension. In another method, one or more lipids and the compound of formula I are dissolved in an organic solvent such as an alcohol, ether or halogenated hydrocarbon or a mixture thereof, the solvent is removed from the resulting solution, for example by freeze-drying or rotary evaporation, and the residue dispersed in aqueous a medium, such as a phosphate buffered saline solution or an aqueous sugar solution, for example lactose, to form an aqueous liposome suspension.

The aqueous liposome suspension can be treated by known methods to remove the solvent and reduce the size of the liposomes. For example, the aqueous liposome suspension prepared by the first method described above using a water-miscible organic solvent may be subjected to dialysis, optionally after further dilution with an aqueous medium, and the dialyzed suspension concentrated by ultrafiltration. The aqueous liposome suspension prepared by the first method, but using a water-immiscible organic solvent, can be evaporated to remove the solvent and then concentrated by ultrafiltration. The aqueous liposome suspension prepared by the second method described above, which typically forms multilamellar vesicles (MLVs), can be pushed through one or more membranes, for example polycarbonate membranes, of a specified pore size, to reduce the size of the liposomes. more than 1 μηα, more preferably 20-200 nm and in particular 50-100 nm.

Liposomes containing a compound of formula I can be dehydrated, preferably by freeze-drying to form a dry powder for administration by a dry powder inhaler in the treatment of asthma .The dehydrated liposomes are rehydrated with a liquid in the patient's airways. The antifreeze agent is preferably a sugar, for example a monosaccharide such as glucose, a polymeric sugar such as dextran, or preferably a disaccharide such as sucrose, lactose, maltose or trehalose. Lactose and trehalose are particularly preferred frost protection agents In accordance with a conventional freeze-drying procedure, the primers are preferably freeze-dried at a temperature below the phase transition temperature of the material to be lyophilized.

Dehydration of the liposomes in the presence of the antifreeze agent produces a dry powder containing a mixture of dehydrated liposomes and the antifreeze agent. When, as preferred, the frost-protecting agent is present in the aqueous medium in which the liposomes are formed, the frost-protecting agent is present on both the inner and outer surfaces of the liposome particles. By weight, the ratio of the antifreeze agent to the lipid in Liposomes is generally from 1: 1 to 4: 1, although lower or higher ratios may be used if desired.

If necessary, the product obtained by dehydrating liposomes containing the compound of formula I is ground, especially when dehydrating is carried out by lyophilization in the presence of a freeze-protecting agent to form particles of a size suitable for use in inhalation therapy, for example using an inhalation device. dry powders. A suitable size is generally less than 10 μπι, preferably 1 to μπι. Liposomes containing a compound of formula I can also be used in inhalation therapy in the form of a suspension in an aqueous medium, if necessary after the treatment described above to reduce the particle size of the liposomes to a suitable range. For use in this form, the liposomes may be prepared in an aqueous medium to be used as a carrier in inhalation therapy, or may be prepared in another medium and separated therefrom and optionally dehydrated as described above prior to incorporation into the aqueous medium to be administered. used as a carrier in inhalation therapy. The aqueous medium may be an aqueous medium that is commonly used as a carrier in inhalation therapy. Usually it is water, in. which is dissolved with one or more pharmaceutically acceptable excipients such as sodium chloride, buffering agents, antioxidants and surfactants. A suitable aqueous medium is a phosphate buffered sodium chloride solution; which may contain an antioxidant such as α-tocopherol. When used in inhalation therapy, the aqueous liposome suspension of the invention may be administered using a known nebulizer, for example a pneumatic nebulizer.

The dry powder of the invention containing the compound of formula I enclosed in dehydrated liposomes may be placed in capsules, for example gelatin or plastic, or in pads for use in a dry powder inhaler. The capsules or pads preferably contain dry powder dosage units, which may contain, for example, 10 to 100 µg., Preferably 50 to 400 µ%. Of a compound of Formula I, together with sufficient carrier to obtain 4 to 40 mg, preferably 20 to 30 mg. dry powder. Alternatively, the dry powder may be placed in a reservoir of a multi-dose dry powder inhaler device adapted to:. that each time it is started up, it applies, for example, 2 mg of dry powder.

The invention also relates, in a further embodiment, to a method of treating asthma which comprises inhaling an effective amount of a compound of formula I as defined above contained in liposomes or dehydrated liposomes as such. as defined above.

The daily dose of the compound of formula I may vary depending on the age and weight of the patient being treated and the severity of the disease. In general, daily doses may range from 50 to 2000 µg, more usually 100 to 1000 µ%.

The following Examples illustrate the invention in which parts are by weight unless otherwise indicated.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

700 mg of 1-n-hexadecanoyl-2- (9-cis-octadecenoyl) -3-sn-phosphatidylcholine and 300 mg of sodium 1,2-di- (9-cis-octadecenoyl) -3-sn-phosphatidyl- S-serine was dissolved in 20 ml of tert-butanol at 40 ° C. The solution obtained is mixed with a solution formed by dissolving 10 mg of methyl 9α-chloro-6α-fluoro-β-hydroxy-16α-methyl-3-oxo-17α-propionyloxyandrosta-1,4-diene-17β-carboxylate (compounds). 1) in 5 ml of tert-butanol at 40 ° C, and the temperature returns to 40 ° C. The resulting solution is added dropwise at room temperature to 200 ml of well stirred phosphate buffered saline (PBS) pH 7.4 . The obtained aqueous liposome suspension is diaysed against PBS using an AMICON YM 100 membrane under a nitrogen atmosphere, and concentrated to -20 ml. The thickened aqueous liposome suspension is successively filtered through filters with a pore size of 0.8 μπι and 0.2 μτη and distributed by 2 ml into sterile ampoules. The suspension obtained is suitable for nebuliser administration in the treatment of asthma by inhalation therapy.

Example 2

The preparation of Example 1 was repeated except that instead of the phosphate buffered sodium chloride solution used in Example 1, both an aqueous solution containing 94.4 g of lactose monohydrate per liter and 0.24 g of sodium chloride was used for both the liposome formation and the dialysis. liters to obtain a concentrated aqueous liposome suspension.

Example 3

The concentrated aqueous liposome suspension prepared in Example 2 was freeze-dried in a Lyovac GT4 lyophilizer. The obtained dry mass is micronized using a Pulverizer pulverized by a stream of air to give a dry powder having a mean particle size of 6-7 µm, which is suitable for administration by a dry powder inhaler device for the treatment of asthma by inhalation therapy.

Example 4

700 mg of distearoyl phosphatidylcholine, 300 mg of dipalmitoyl phosphatidylcholine and 20 mg of the compound of formula I are dissolved in 20 ml of a 2: 1 by volume mixture of chloroform and methanol. The solvent was removed on a rotary evaporator. The residue is dispersed in 40 ml of an aqueous lactose solution containing 94.4 g of lactose monohydrate per liter and 0.24 g of sodium chloride per liter to give an aqueous liposome suspension. This slurry was passed through two polycarbonate membranes with an aperture size of 200 nm and ten times through two polycarbonate membranes with an aperture size of 100 nm at 70 ° C under nitrogen pressure to reduce the particle size of the liposomes. The resulting suspension is lyophilized and micronized as in Example 3 to give a dry powder that is suitable for administration by a dry powder inhaler device for the treatment of asthma by inhalation therapy.

Example 5

678 mg of dimyristoyl-phosphatidylcholine, 193 mg of cholesterol, 81 mg of dioleoyl-phosphatidylserine and 20 mg of the compound of formula 1 are dissolved in 20 ml of tert-butanol. The tert-butanol is removed from the resulting solution by freeze-drying. The residue is dispersed in an aqueous lactose solution as described in Example 4 and the liposome suspension is forced through membranes as described in Example 4 but at a temperature of 35 ° C instead of 70 ° C to reduce the size of the liposomes to 100 nm. The resulting suspension is lyophilized and micronized as in Example 3 to give a dry powder that is suitable for administration by a dry powder inhaler device for the treatment of asthma by inhalation therapy.

Example 6

The effect of liposomes containing the compound of formula I enclosed therein on increasing the number of eosinophils is tested in the Brown-Norway model of allergen-induced eosinophilia in rats as described by Elwood et al., J. Allergy Clin. Immunol 1991, 88, 951-60. Four groups of inbred male rats weighing 180-220 g are examined:

Group 1: Animals are sensitized by intraperitoneal injection of ml 0.9% (w / v) suspension of 1 mg ovalbumin and

100 mg of Al (OH) ₃ and 21 days later exposed to aerosol of sodium chloride solution alone for 15 minutes.

Group 2: Animals were sensitized with ovalbumin as Group 1 animals and 21 days later exposed to 1% ovalbumin aerosol for 15 minutes.

Group 3: Animals are sensitized with ovalbumin as Group 1 animals and, 19 days later, 0.5 ml of the liposome suspension of Example 1 containing 3 μ $ of the compound of formula I is injected transcutaneously under ketamine anesthesia and 24 hours later transtracheal injection. 24 hours after the second injection, the animals are exposed to 1% ovalbumin aerosol for 15 minutes.

Example 4: Animals are treated in the same way as animals from the group

3, but instead of liposomes containing a compound of formula I, placebo-liposomes prepared using the same procedure but omitting the compound of formula I are used.

In each group of rats, bronchoalveolar lavage was performed 24 hours after aerosol challenge to determine the number of eosinophils. The results are as follows:

group number of eosinophils per rat 1 (treatment with solution sodium chloride) 0,25xlG ⁵ 2 (ovalbumin action) 19,0xl0 ⁵ Liposomes containing liposomes compound of formula I) 10,3xl0 ^s 4 (placebo-liposomes) 16,9xl0 ⁵

Claims (20)

What is claimed is: 1. A pharmaceutical composition comprising as active ingredient a compound contained in liposomes or dehydrated liposomes. 2. The composition of claim 1 wherein the lipid component of the liposomes or dehydrated liposomes comprises at least one synthetic phospholipid. 3. A composition according to claim 2 wherein the lipid component comprises synthetic phosphatidylcholine optionally together with synthetic phosphatidylserine or synthetic phosphatidylglycerol. A composition according to claim 3, wherein phosphatidylserine or phosphatidylglycerol is present and the weight ratio of phosphatidylcholine to phosphatidylserine or phosphatidylglycerol is from 60: 40 to 95: 5. 5. A composition according to claim 4 wherein said weight ratio is from 70: 30 to 90: 10. Composition according to any one of claims 3 to 5, characterized in that the lipid component comprises a synthetic phospholipid of the general formula II. CH ₂ -OR in which _t R _x is C 10 -C 20 alkanoyl, the number of carbon atoms being an even number, R ₂ is alkanoyl group having 10 to 20 carbon atoms, wherein the number of carbon atoms is even, R _a, R _b and R _c are hydrogen or alkyl having 1 to 4 carbon atoms, n is an integer from two to four. 7. A composition according to claim 6, wherein the phospholipid of formula II is 1-n-hexadecanoyl-2- (9-cis-octadecenoyl) -3-sn-phosphatidylcholine. Composition according to any one of claims 3 to 7, characterized in that the lipid component comprises a synthetic phospholipid of the formula II according to claim 6 together with a synthetic phospholipid of the formula III ch ₂ -o - r ₃ (S) CH ₂ -O -P - O- (CnH ^) - CH (III) OH NHn in which R ₃ and R _{4 are each} independently C 10 -C 20 alkenoyl, the number of carbon atoms being an even number, n being an integer from one to three, and Y 6 is a cation of a pharmaceutically acceptable base. Composition according to claim 8, characterized in that the phospholipid of the formula III is sodium-1,2-di- (9-cis-octadecenoyl) -3-sn-phosphatidyl-Ξ-serine. A composition according to any one of claims 3 to 5, wherein the lipid component comprises di (alkanoyl) phosphatidylcholine of 10 to 20 carbon atoms in each alkanoyl moiety, together with di (alkanoyl) phosphatidylglycerol of 10 to 20 carbon atoms in each alkanoyl moiety. 11. A composition according to claim 10, wherein the phosphatidylcholine is dithearoyl phosphatidylcholine and the phosphatidylglycerol is dipalmitoyl phosphatidylglycerol. Composition according to any one of claims 2 to 11, characterized in that the lipid component also contains cholesterol. 13. A composition according to claim 12, wherein the amount of cholesterol is from 20 to 60 mol% by weight. total lipid content. The composition according to claim 12 or 13, wherein the lipid component comprises dimyristoyl phosphatidylcholine, cholesterol and dioleoyl phosphatidylserine. A composition according to any one of claims 1 to 14, wherein the weight ratio of the compound of Formula I to the lipid is from 1: 100 to 1:50. Composition according to any one of claims 1 to 15, characterized in that the liposomes have a particle size of less than 1 μπι. Composition according to any one of claims 1 to 16, characterized in that the liposomes are in aqueous suspension. Composition according to any one of claims 1 to 16, characterized in that it is in the form of a dry powder comprising a mixture of (a) dehydrated liposomes comprising a compound of formula I, and (b) a frost-protecting agent. The composition of claim 18, wherein the weight ratio of the antifreeze agent to the lipid in the liposomes is from 1: 1 to 4: 1. Use of a composition according to any one of claims 1 to 19 for the preparation of a medicament for the treatment of asthma.