Pharmaceutical formulations

This invention relates to pharmaceutical formulations containing rapamycin. This invention particularly relates to galenic formulations which are in the form of microemulsions, microemulsion preconcentrates, emulsion, or emulsion preconcentrate.

Rapamycin is a macrolide antibiotic made from Streptomyces hyqroscopicus. It has been found to be pharmaceutically useless in a variety of applications, particularly as an immunosuppressant for the treatment and prevention of organ transplant rejection and autoimmune diseases. Rapamycin has the following structure:

<img file="AT408520B_D0001.tif" />

(Kesseler, H. et al., Helv. Chim. Acta (1993) 76: 117; US 3,929,992 A). A large number of rapamycin derivatives have been synthesized, including for example those described in US 5221670 A and US 5221740 A, certain acyl and aminoacyl rapamycins (see for example US 4316885 A, and US 4650803 A, and US 5151413 A), and carbonates and amide esters (see, for example, EP 509795 A2 and 515140 A1), 27-desmethylrapamycin (see, for example, WO 9214737 A1), 26-dihydrorapamycin (see, for example, US 5138051 A), Alkoxy ester derivatives (see for example US 5233036 A) and certain pyrazole derivatives (US 5164399 A).

Rapamycin class compounds are very powerful immunosuppressants and also have antitumoral and antifungal activity. Their use as pharmaceuticals, particularly when administered orally, has been limited by their low solubility, low and variable bioavailability, and high toxicity. Little is known about the causes of these properties and the point of absorption. Therefore, it can be assumed that poor bioavailability is due to extensive metabolism of the macrolide ring and cannot be resolved by a galenic formulation. What is needed, therefore, is an acceptable pharmaceutical composition containing compounds of the rapamycin class.

It has now surprisingly been found that stable compositions which contain rapamycin and which offer high absorption efficiency can be obtained by formulating rapamycin with certain carriers.

In one aspect, the invention relates to an oral pharmaceutical composition containing rapamycin and a carrier which

i) a hydrophilic phase, ii) a lipophilic phase, iii) a surfactant, wherein the composition is an emulsion or microemulsion pre-concentrate.

In another aspect, the invention relates to emulsion or microemulsion preconcentrate carriers for orally administrable rapamycin comprising the following

i) a reaction product of a castor oil and ethylene oxide, ii) a transesterification product of a vegetable oil and glycerol, which is mainly mono-, di- and

Triglycerides of linoleic or oleic acid, or a polyoxyalkylated vegetable oil,

AT 408 520 B iii) 1,2-propylene glycol, and iv) ethanol.

The pharmaceutical composition is stable and gives an amazingly high and constant rate of absorption when administered orally. Therefore, rapamycin can be administered in lower doses, which reduces toxicity problems. For example, animal studies in which the pharmaceutical compositions were administered orally have shown high bioavailabilities. The pharmaceutical compositions therefore have very surprising properties which offer great advantages.

The composition is preferably in the form of a microemulsion preconcentrate or emulsion concentrate, particularly in the form which provides O / W (oil-in-water) microemulsions or emulsions. The composition can, however, be in the form of a microemulsion or an emulsion which additionally contains an aqueous phase; preferably water.

A microemulsion pre-concentrate is defined in this specification as a formulation which forms a microemulsion spontaneously in an aqueous medium, for example in water or in gastric juice after oral administration.

A microemulsion is a non-opaque or substantially non-opaque colloidal dispersion that is formed spontaneously or substantially spontaneously when its components are brought into contact with one another. A microemulsion is thermodynamically stable and contains dispersed particles of a size less than about 200 nm. Microemulsions generally comprise droplets or particles with a diameter less than about 150 nm, typically from 3 to 100 nm. Further features can be found in GB 2 222 770 A.

An emulsion pre-concentrate ”is described in this specification as a formulation which spontaneously forms an emulsion in an aqueous medium, for example in water or in gastric juice after oral administration. The emulsion formed is opaque, thermodynamically stable and contains disperse droplets of a size larger than about 100 nm, usually larger than about 200 nm. Bimodal size range distributions are often obtained. The emulsion pre-concentrates are preferably of the type that they provide O / W (oil-in-water) emulsions.

A pharmaceutical composition means a composition in which the individual components or ingredients are themselves pharmaceutically acceptable and, if a particular form of administration is intended, is suitable or acceptable for that mode of administration.

The lipophilic phase can comprise 10 to 85% by weight of the carrier; preferably 15 to 70% by weight, more preferably 20 to 60% by weight, and even more preferably about 5% by weight.

The surfactant can comprise 5 to 80% by weight of the carrier; preferably 10 to 70% by weight, more preferably 20 to 60% by weight, and even more preferably about 40% by weight.

The hydrophilic phase can comprise 10 to 50% by weight of the carrier; preferably 15 to 40% by weight, more preferably 20 to 35% by weight, and even more preferably about 30% by weight.

Rapamycin is present in about 1 to 15% by weight of the composition, more preferably about 2 to 10%.

The hydrophilic phase can be selected from a compound of formula C₂H₅- [O- (CH₂)₂]₂-OH glycofurol (also known as tetrahydrofurfuryl alcohol polyethylene glycol ether) and 1,2-propylene glycol or mixtures thereof, and is preferably 1,2-propylene glycol. The hydrophilic phase can include further hydrophilic components, for example lower alcohols such as ethanol. These components will generally be present as partial substitutes for other components of the hydrophilic phase. The use of ethanol, although not essential in these compositions, has been found to be advantageous when the compositions are formulated in soft gelatin capsules. This is because the shelf life is improved, in particular because the risk of rapamycin precipitation after the encapsulation process is reduced. Therefore, the storage stability can be extended if ethanol or another similar component as an additional component of the hydrophilic

AT 408 520 B

Phase is used. Ethanol can comprise 0 to 60% by weight of the hydrophilic phase, preferably 20 to 55% by weight, and more preferably about 40 to 50% by weight. Small amounts of liquid polyethylene glycols can also be included in the hydrophilic phase.

Preferred lipophilic phase components are mixed mono-, di-, triglycerides and interesterified ethoxylated vegetable oils. Suitable fatty acid glycerides are known and available commercially under the trade names Miglyol, Captex, Myritol, Capmul, Captex, Neobee and Mazol; with Miglyol 812 being most preferred. These triglycerides are found in Fiedler, HP Lexicon of auxiliaries for pharmacy, cosmetics and related areas, Edition Cantor, D-7960 Aulendorf, third improved and expanded edition (1989).

The mixed mono-, di- and triglycerides preferably comprise mixtures of Ci2-2o fatty acid mono-, di- and triglycerides, in particular mixed Ci6-18-fatty acid mono-, di- and triglycerides. The fatty acid component of the mixed mono-, di- and triglycerides can comprise both saturated and unsaturated fatty acid residues. However, they preferably comprise mainly unsaturated fatty acid residues; especially unsaturated C₁₈-Fatty acid residues. Suitably the mixed mono-, di- and triglycerides comprise at least 60% by weight, preferably at least 75% by weight, more preferably at least 85% by weight of an unsaturated C₁₈-Fatty acids (for example linolenic, linoleic and oleic acid) -mono-, di- and triglyceride. The mixed mono-, di- and triglycerides comprise less than 20% by weight, for example 15% by weight or 10% by weight or less of saturated fatty acids (for example palmitic and stearic acid) mono-, di- and triglycerides.

The mixed mono-, di- and triglycerides preferably mainly comprise mono- and diglycerides; for example, mono- and diglycerides comprise at least 50%, more preferably at least 70%, based on the total weight of the lipophilic phase. The mono- and diglycerides more preferably comprise at least 75% by weight (e.g. about 80% or 85% by weight) of the lipophilic phase.

Preferably, the monoglycerides comprise from about 25 to about 50% of the mixed mono-, di- and triglycerides, based on the total weight of the lipophilic phase. More preferably from about 30 to about 40% (e.g. 35 to 40%) monoglycerides are present.

The diglycerides preferably comprise from about 30 to about 60% of the mixed mono-, di- and triglycerides, based on the total weight of the lipophilic phase. More preferably from about 40 to about 55% (e.g. 48 to 50%) diglycerides are present.

The triglycerides suitably comprise at least 5%, but less than about 25%, of the mixed mono-, di- and triglycerides, based on the total weight of the lipophilic phase. More preferably from about 7.5 to about 15% (e.g. 9 to 12%) triglycerides are present.

The mixed mono-, di- and triglycerides can be prepared by admixing individual mono-, di- and triglycerides in appropriate relative proportions. However, they conveniently include transesterification products of vegetable oils, for example almond oil, ground nut oil, olive oil, palm oil or, preferably, corn oil, sunflower oil or safflower oil and in particular corn oil with glycerol.

Such transesterification products are generally obtained by heating the selected vegetable oil with glycerol at a high temperature in the presence of a suitable catalyst in an inert atmosphere with continuous stirring (for example in a stainless steel reactor). The transesterification products generally also comprise small amounts of free glycerol in addition to their mono-, di- and triglyceride components. The amount of free glycerol present is preferably less than 10%, more preferably less than 5%, most preferably about 1 or 2% by weight based on the total weight of free glycerol plus mono-, di- and triglycerides.

First, some of the glycerol is preferably removed to give an essentially glycerol-free set when soft gelatin capsules are to be made.

Transesterification products of corn oil and glycerol provide particularly suitable mixed mono-, di- and triglycerides. An example of a suitably blended glyceride product is the transesterification product which is commercially available under the trade name MAISINE. This product mainly comprises linoleic and oleic acid mono-, di- and triglycerides along with small amounts of palmitic and stearic acid mono-, di- and triglycerides (corn oil itself includes

AT 408 520 B

Ingredients of about 56% by weight linoleic acid, 30% oleic acid, about 10% palmitic acid and about 3% stearic acid). The physical characteristics of MAISINE [available from Etablissement Gattefosso, 36, Chemin de Genas, PO Box 603, 69804 Saint-Priest, Cedex (France)] are as follows: up to 10% (typically 3.9 to 4.9% or, in substantially glycerol-free rates, approximately 0.2%) free glycerol; about 35% (typically 30 to 40%, or, in substantially glycerol-free sets, about 32 to 36%, for example about 36%) monoglycerides; about 50% (or, in substantially glycerol-free sets, about 46 to 48%) diglycerides; about 10% (or, in substantially glycerol-free sets, about 12 to 15%) triglycerides; and about 1% free oleic acid.

Further physical characteristics for MAISINE are as follows: a maximum acid value of approximately 2, an iodine number of approximately 85 to 105, a saponification number of approximately 150 to 175 (Fiedler Lexikon der auxiliaries, third improved and expanded edition (1989) Vol. 2, p. 768). The fatty acid content for MAISINE is typically as follows: about 11% palmitic acid, about 2.5% stearic acid; about 29% oleic acid; about 56% linoleic acid; and 1.5% other acids.

It is particularly preferred that the mixed mono-, di- and triglycerides be clear and remain clear for more than 20 days during storage at temperatures of 20 ° C to 25 ° C. A sample of the mixed mono-, di-, and triglycerides that has been stored in a refrigerator at approximately 2 and 8 ° C for 24 hours and then stored at room temperature for 1 hour should also be clear.

The mono-, di- and triglyceride es preferably have a low content of saturated fatty acids. Mixed mono-, di- and triglycerides meeting these requirements can be obtained from commercially available products by separation methods known in the art (e.g. freezing methods associated with separation methods such as centrifugation) to remove the saturated fatty acid components , and to improve the unsaturated fatty acid component content. Typically the total saturated fatty acid component content will be less than 15% by weight (e.g. &lt;10% by weight or &lt;5% by weight) based on the total weight of the lipophilic phase. A decrease in the content of saturated fatty acid components can be observed after it has been subjected to the separation process. A suitable method is described in WO 9309211 A1.

The mixed mono-, di- and triglycerides therefore preferably contain smaller amounts of saturated fatty acids (eg palmitic and stearic acids) and relatively high amounts of unsaturated fatty acids (eg oleic and linoleic acids) than the starting material.

A suitable example of a mixed mono-, di- and triglyceride product containing minor amounts of saturated fatty acids is the following: 32 to 36% by weight monoglycerides, 45 to 55% by weight diglycerides, and 12 to 20% by weight triglycerides, based on total weight the lipophilic phase. Other characteristics include the following:

<td>Fatty acid content (by chromatographic Determination of the methyl ester)</td><td>Methyl linoleate 53 to 63% Methyl oleate 24 to 34% methyl linolenate 0 to 3% methyl arachate 0 to 3% methyl palmitate 6 to 12% methyl stearate 1 to 3%</td>

<td>relativ density</td><td>0.94 to 0.96</td>

<td>Hydroxyl number</td><td>140 to 210</td>

<td>Iodine number</td><td>110 to 20</td>

<td>Peroxide number</td><td> &lt;4,0</td>

<td>Free glycerol</td><td> &lt; 1,0</td>

<td>Saponification number</td><td>about 150 to 185</td>

<td>Acid value</td><td>max. about 2</td>

AT 408 520 B

Mixed mono-, di- and triglycerides that correspond to these characteristics are used in this

Description referred to as refined glycerol transesterified corn oils. The refined glycerol transesterified corn oils have the advantage that they remain stable for a long time.

The lipophilic phase can, on the other hand, comprise suitable interesterified ethoxylated vegetable oils, such as those obtained by adding various natural vegetable oils (for example corn oil, kernel oil, almond oil, ground nut oil, olive oil, soybean oil, sunflower oil, safflower oil and palm oil, or mixtures thereof) with Polyethylene glycols, which have an average molecular weight of 200 to 800, are reacted in the presence of a suitable catalyst. These methods are known and an example is described in US 3,288,824 A. Interesterified ethoxylated corn oil is particularly preferred.

Transesterified ethoxylated vegetable oils are known and are commercially available under the trade name LABRAFIL (H. Fiedler, ioc sit, Vol. 2, page 707). Examples are LABRAFIL M 2125 CS (which is obtained from corn oil and has an acid number less than about 2, a saponification number from 155 to 175, an HBL value from 3 to 4, and an iodine number from 90 to 110), and LABRAFIL M 1944 CS (obtained from corn oil and having an acid number of about 2, a saponification number of 145 to 175, and an iodine number of 60 to 90). LABRAFIL M 2130 CS (which is a transesterification product of a C₁₂.i_ß-Glyceride and polyethylene glycol and which has a melting point of about 35 to 40 ° C, an acid number that is less than about 2, a saponification number of 185 to 200, and an iodine number that is less than about 3) can also be used will. The preferred interesterified ethoxylated vegetable oil is LABRAFIL M 2125 CS, available for example from Gattefosse, Saint-Priest Cedex, France.

Examples of suitable surfactants are as follows:

i) reaction products of a natural or hydrogenated castor oil and ethylene oxide. The natural or hydrogenated castor oil can be reacted with ethylene oxide in a molar ratio of from about 1:35 to about 1:60, with the optional removal of the polyethylene glycol component from the products. Numerous similar surfactants are commercially available. The polyethylene glycol hydrogenated castor oils available under the trade name CREMOPHOR are particularly suitable. CREMOPHOR RH 40, which has a saponification number of approximately 50 to 60, an acid number which is less than approximately 1, a water content (Fischer) which is less than approximately 2%, an n are particularly suitable_D.⁶⁰ from about 1.453 to 1.457 and an HLB value of about 14 to 16; and CREMOPHOR RH 60, which has a saponification number of about 40 to 50, an acid number that is less than about 1, an iodine number that is less than about 1, a water content (Fischer) of about 4.5 to 5.5% , an nD²⁵ from about 1.453 to 1.457 and an HLB value of about 15 to 17. A particularly preferred product of this class is CREMOPHOR RH40. Polyethylene glycol castor oils available under the trade name CREMOPHOR EL are also suitable, CREMOPHOR EL having a molecular weight (by steam osmometry) of about 1630, a saponification number of about 65 to 70, an acid number of about 2, an iodine number of about 28 to 32 and a nD²⁵ of about 1.471. Similar or identical products that can also be used are available under the trade names NIKKOL (e.g. NIKKOL HCO40 and NIKKOL HCO-60), MAPEG (e.g. MAPEG CO-40h), INCROCAS (e.g. INCROCAS 40), and TAGAT (e.g. TAGAT RH 40 ) available. These surface-active substances are further in Fiedler Ioc, cit. described.

ii) Polyoxyethylene sorbitan fatty acid esters, for example mono- and trilauryl-, -palmityl, -stearyl and -oleyl esters of the kind which are known and available commercially under the trade name TWEEN (Fiedler, loc. cit. p. 13001304), including the products TWEEN

20 [polyoxyethylene (20) sorbitan monolaurate],

21 [polyoxyethylene (4) sorbitan monolaurate],

40 [polyoxyethylene (20) sorbitan monopalmitate],

60iPolyoxyethylene (20) sorbitan monostearatej,

65 [polyoxyethylene (20) sorbitan tristearate],

80 [polyoxyethylene (20) sorbitan monooleate],

81 [polyoxyethylene (5) sorbitan monooleate],

85 [polyoxyethylene (20) sorbitan trioleate j.

Particularly preferred products in this class are TWEEN 40 and TWEEN 80.

AT 408 520 B iii) polyoxyethylene fatty acid esters, for example polyoxyethylene stearic acid esters of the type which are im

Trade under the trade name MYRJ (Fiedler, loc. Eit., 2. pp. 834-835) are known and available. A particularly preferred product in this class is MYJR 52 with a D²⁵ of about 1.1, a melting point of about 40 to 44 ° C, an HLB value of about 16.9, an acid value of about 0 to 1, and a saponification number of about 25 to 35.

iv) Polyoxyethylene-polyoxypropylene copolymers and block copolymers, for example of the type known and available under the trade names PLURONIC, EMKALYX and POLOXAMER (Fiedler, loc. eit., 2, p. 959). A particularly preferred product of this class is PLURONIC F68, with a melting point of about 52 ° C and a molecular weight of about 6800 to 8975. Another preferred product of this class is POLOXAMER 188.

v) Dioctyl sulfosuccinate or di- [2-ethylhexyl] succinate (Fiedler, loc. eit., 1, pp. 107-108).

vi) Phospholipids, especially lecithins (Fiedler, loc. cit., 2. pp. 943-944). Suitable lecithins include, in particular, soya lecithins.

vii) Propylene glycol mono- and difatty acid esters, such as propylene glycol dicaprylate (also known and available commercially under the trade name MIGLYOL 840), propylene glycol dilaurate, propylene glycol hydroxystearate, propylene glycol isostearate, propylene glycol isostearate, propylene glycol 8, propylene glycol laurate, propylene glycol and so on -809).

The components of the carrier can contain unreacted starting materials, for example polyethylene glycol.

The surfactant selected preferably has an HLB of at least 10.

The relative proportions of the hydrophilic phase component (s), the lipophilic phase and the surfactant lie within the microemulsion area on a normal three-way diagram. The compositions obtained in this way are microemulsion preconcentrates with high stability which, after the addition of water, are capable of producing microemulsions which have an average particle size of &lt;150 nm and which are stable for 24 hours.

The microemulsion pre-concentrate compositions show good stability characteristics, as shown by normal stability tests, for example with a storage stability of up to three years and even longer.

Alternatively, the components can be selected to provide an emulsion pre-concentrate. The emulsion pre-concentrate compositions also show good stability characteristics, as shown by normal stability tests, for example long-term stability of up to three years and even longer.

The pharmaceutical composition can also include other additives or ingredients, for example antioxidants (such as ascorbyl palmitate, butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT) and tocopherols) and / or preservatives. These additions or ingredients can comprise about 0.05 to 1% by weight of the total weight of the composition. The pharmaceutical composition can also include sweetening or flavoring agents in an amount of up to 2.5 or 5% by weight based on the total weight of the composition. The antioxidant is preferably α-tocopherol (vitamin E).

The composition can also include an FK506 compound. FK506 is a macrolide immunosuppressant made from Streptomvces tsukubaensis No. 9993. The structure of FK506 is given in the Appendix of the Merck Index as point A5. A large number of related compounds are also known that retain the basic structure and immunological properties of FK506. These compounds are described in a large number of publications, e.g. EP 184162 A2, EP 315973 A2, EP 323042 A1, EP 423714 A2, EP 427680 A1, EP 465426 A1, EP 474126 A1, WO 9113889 A1, WO 91 19495A1, EP 484936 A1, EP 532088 A1, EP 532089 A1, WO 93 5059 A1, and the like. Little is known about the biopharmaceutical properties of such components. These components are collectively referred to as FK506 compounds in this description.

The pharmaceutical composition exhibits particularly advantageous properties when administered orally; for example as to persistence and a high level of bioavailability obtained in normal bioavailability tests, e.g. 2 to 4 times higher than emulsions. These experiments are carried out on animals or healthy volunteers using HPLC or a specific or non-specific monoclonal test system to determine the level of the rapamycin class compound in the blood. For example be

AT 408 520 B in the test described in Example 3, 10 mg rapamycin orally administered to rats and the surprisingly high C_MaxValues ​​between 2670 and 3400 ng / ml are found by ELISA using a specific monoclonal antibody. It is also found that an emulsion pre-concentrate and a microemulsion pre-concentrate composition have much better pharmacokinetic properties than a normal solvent system.

In a further aspect the invention provides the use of the emulsion or microemulsion pre-concentrate according to the present invention for the manufacture of a medicament for oral administration.

Pharmacokinetic parameters such as absorption and blood levels also become surprisingly more predictable and administration problems with erratic absorption can be eliminated or reduced. In addition, the pharmaceutical composition is effective with surfactant materials, for example bile salts, which are present in the gastrointestinal tract. That is, the pharmaceutical composition is completely soluble in aqueous systems comprising such natural surfactants, and therefore can provide microemulsion systems in situ that are stable and show no precipitation of the active ingredient or other fine particle structure disturbance. The function of the pharmaceutical composition after oral administration remains essentially independent and / or unaffected by the relative presence or absence of bile salts at any particular time or for any particular person.

The pharmaceutical composition is preferably in single-dose containers, for example by filling it into orally administrable capsules. The capsules can be soft or hard gelatin capsules. Where the pharmaceutical composition is in a unit dose drug form, each unit dose will suitably be between 10 and 100 mg rapamycin, more preferably between 10 and 50 mg; for example 15, 20, 25 or 50 mg rapamycin. Such single-dose containers are suitable for administration from 1 to 5 times a day, depending on the particular therapeutic purpose, the therapeutic phase and the like.

However, if desired, the pharmaceutical composition may be in the form of beverage concentrates and may include water or some other aqueous system to provide microemulsion systems suitable for drinking.

The utility of the pharmaceutical composition can be demonstrated in normal clinical tests, for example, by observing the dose / blood level curve of compounds of the rapamycin class; to be determined. For example, dosages ranging from 2.5 mg to 1000 mg of rapamycin class compounds per day for an adult weighing 75 kilograms and in normal animal models are used. The increased bioavailability of a compound of the rapamycin class provided by the compositions can be observed in animal studies and in clinical trials. When an FK506 compound is included in the pharmaceutical composition, this usefulness can also be seen in normal clinical tests and animal models. The rapamycin dosages to be used in clinical trials are as indicated above; while those for an FK 506 compound can be 2.5 mg to 1000 mg per day for an adult weighing 75 kg.

The attending physician must carefully consider the optimal dosage of rapamycin, and optionally FK506 derivatives, to be administered to a particular patient as an individual reaction to the metabolism of rapamycin. It may be advisable to monitor serum levels of the rapamycin compound by radioimmunoassay, monoclonal antibody assay, or other suitable conventional means. The rapamycin doses are generally 2.5 mg to 1000 mg per day for an adult weighing 75 kg, preferably 25 mg to 500 mg, the optimal dosage being 50 to 100 mg per day. Satisfactory results have been obtained by administering approximately 75 mg per day, for example in the form of two capsules, one containing 50 mg and the other containing 25 mg; or 3 capsules, each containing 25 mg. When an FK506 compound is included in the pharmaceutical composition, the dose of FK-506 can be 2.5 mg to 1000 mg per day, preferably 10 mg to 250 mg.

Rapamycin compounds are particularly useful for the following conditions:

a) Treatment and prevention of organ transplant rejection for example for the treatment of recipients of heart, lung, combined heart-lung, liver, kidney,

AT 408 520 B

Pancreas, skin or corneal grafts. The pharmaceutical compositions are also indicated for the prevention of graft versus recipient disease, which sometimes occurs after bone marrow transplants.

b) Treatment and prevention of autoimmune diseases and inflammatory conditions, particularly inflammatory conditions with an etiology that includes an autoimmune component such as arthritis (e.g. rheumatoid arthritis, chronica progressive arthritis and arthritis deformans) and rheumatic diseases. Specific autoimmune diseases for which the pharmaceutical compositions can be used include autoimmune hematological diseases (including e.g. haematological anemia, aplastic anemia, red blood cell anemia and idiopathic thrombocytopenia), systemic lupus erythematosus, polychondritis, sclerodoma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven-Johnson syndrome, autoimmune inflammatory diseases ulcerative colitis and Crohn's disease), endocrine ophthalmopathy, Greaves disease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis, juvenile diabetes (diabetes mellitus type I), uveitis (anterior and posterior), keratoconjunctivitis sicca and vernal keratoconjunctivitis, pulmonary fibrosis, psoriasis interstitial Glomerulonephritis (with and without nephrotic syndrome, e.g. including idiopathic nephrotic syndrome or minimal change nephropathy and adolescent dermatomyositis.

c) Treatment and prevention of asthma.

d) Treatment of multi-drug resistance (MDR = insensitivity to a variety of drugs.) Rapamycin compounds suppress P-glucoproteins (Pgp), which are membrane transport molecules that play a role in MDR. MDR is particularly problematic in cancer patients and AIDS patients who do not respond to conventional chemotherapy because the medication is pumped out of the cells by Pgp. Thus, pharmaceutical compounds that increase the effectiveness of other chemotherapeutic agents used in the treatment and control of conditions resistant to many drugs, such as cancer, or AIDS, are useful.

Since the rapamycin compounds also show antitumoral and antifungal effects, these pharmaceutical compositions can also be used as antitumoral and antifungal agents.

In a further aspect, the invention also provides a method of making a pharmaceutical composition as defined above, the method comprising (1) a hydrophilic phase; (2) a lipophilic phase; and (3) mix a surfactant and add rapamycin, and optionally an FK-506 compound. If desired, the composition can be formulated in unit dose containers; for example by filling the composition in gelatin capsules.

Optionally, further components or additions, in particular a component of the hydrophilic phase, for example ethanol, can be mixed with components (1), (2) and (3), with or after the addition of rapamycin, and optionally an FK-506 compound .

The composition can be combined with enough water or a sufficient amount of an aqueous solvent that a microemulsion is obtained.

The following examples illustrate compositions in single dose containers which, when administered from 1 to 5 unit doses / day, are useful, for example, to prevent graft rejection or to treat autoimmune diseases.

EXAMPLE 1: Refined giycerol interesterified corn oil is prepared as follows:

Corn oil transesterified with glycerol, which is essentially free of glycerol, is slowly cooled to a temperature of + 20 ° C and stored at this temperature for one night. The corn oil is centrifuged at an acceleration of 12,000 G and a flow rate of 103 kg / h in a continuous flow centrifuge to give a liquid phase (62 kg / h) and a sediment-containing phase (41 kg / h). The liquid phase is slowly cooled to + 8 ° C and kept at this temperature for one night. The liquid phase is then centrifuged at an acceleration of 12,000 G and a flow rate of 112 kg / h to give a liquid phase (76.2 kg / h) and a sediment-containing phase (35.8 kg / h). the

AT 408 520 B liquid phase is refined, glycerol-transesterified corn oil. On the other hand, an improved product can be obtained by effecting centrifugation in three steps, for example at + 20 ° C, + 10 ° C and + 5 ° C.

The process is characterized by a small percentage reduction in the monoglyceride component in the refined glycerol interesterified corn oil compared to the starting material (e.g. 35.6% compared to 38.3%).

EXAMPLE 2:

The refined corn oil obtained as described in Example 1 and transesterified with glycerol is used in the preparation of the following oral unit dose:

<td>COMPONENT</td><td>QUANTITY (mg / capsule)</td>

<td>Rapamycin</td><td> 20,0</td>

<td>1) ethanol</td><td> 75,0</td>

<td>2) 1,2 propylene glycol</td><td> 81,0</td>

<td>3) refined oil</td><td> 121,5</td>

<td>4) Cremophor RH40</td><td> 202,5</td>

<td>Total</td><td> 500,0</td>

The rapamycin is suspended in (1) with stirring at room temperature, and (2), (3) and (4) are added to the resulting solution while stirring. The mixture obtained is filled into size 0 hard gelatin capsules and sealed using the quasi-seal method.

EXAMPLE 3: Pharmokinetics

Two formulations prepared as in Example 2 are used:

<td>formulation</td><td>component</td><td>Lot %</td>

<td>A.</td><td>Tween 80</td><td> 41,5%</td>

<td></td><td>Maisine</td><td> 24,9%</td>

<td></td><td>Propylene glycol</td><td> 16,6%</td>

<td></td><td>Ethanol</td><td> 15,0%</td>

<td></td><td>Rapamycin</td><td> 2,0%</td>

<td>B.</td><td>Cremophor RH40</td><td> 41,5%</td>

<td></td><td>Maisine</td><td> 24,9%</td>

<td></td><td>Propylene glycol</td><td> 16,6%</td>

<td></td><td>Ethanol</td><td> 15,0%</td>

<td></td><td>Rapamycin</td><td> 2,0%</td>

Formulation A is an emulsion pre-concentrate and Formulation B is a microemulsion pre-concentrate. 6 male Wistar rats with an average body weight of 300 g are used per formulation. One day before treatment, the rats are deprived of food, but the rats are allowed free access to water. The rats are then anesthetized by injecting 2 x 1 ml of 20% urethane intraperitoneally and a permanent catheter is inserted into the right jugular vein to allow blood to be drawn. 500 ml / animal of the formulation is administered through a nasogastric tube 20 hours after the operation. A total dose of 10 mg of the drug per animal is administered. Blood samples with a size of 0.7 m are taken! removed from the jugular catheter of each animal 15 minutes before drug administration and then 0.17, 0.5, 1, 1.5, 2, 3, 5 and 8 hours after drug administration. The samples are taken in heparinized tubes and subjected to ELISA

AT 408 520 Β

Using microtitration plates coated with specific antibodies from rapamycin, analyzed. The animals are killed immediately after the last blood sample has been taken. The results are given in the following table:

<td>For m</td><td>AUC (0-8 hours) [ng.h / ml]</td><td>CV [%1</td><td>ömax [ng / ml]</td><td>CV [%]</td><td>tfnax [Hours]</td><td>CV [%]</td>

<td>A.</td><td> 11951</td><td> 44</td><td> 2671</td><td> 42</td><td> 3,8</td><td> 29*</td>

<td>B.</td><td> 13826</td><td> 13</td><td> 3405</td><td> 30</td><td> 4,0</td><td> 35+</td>

*) n = 5

+) n = 2 due to difficulties in drawing blood

The results indicate that rapamycin is well absorbed.

EXAMPLE 4: Comparison

Formulations A and B are compared to a formulation comprising 38.6% corn oil, 41.6% Labrafil M21 / 25C, 17.8% ethanol and 2% rapamycin (Formulation C). The same procedure is used as in Example 3, except that the animals each receive a total dosage of 0.5 mg of the formulation.

The results are given in the following table:

<td>For m</td><td>AUC (0-8 hours) [ng.h / ml]</td><td>CV [%]</td><td>Dmax [ng / ml]</td><td>CV [%]</td><td>tfnax [Hours]</td><td>CV [%]</td>

<td>A.</td><td> 105,8</td><td> 28</td><td> 31,22</td><td> 35</td><td> 1,6</td><td> 51*</td>

<td>B.</td><td> 96,6</td><td> 32</td><td> 36,13</td><td> 60</td><td> 0,4</td><td> 30</td>

<td>C.</td><td> 36,2</td><td> 31</td><td> 7,83</td><td> 27</td><td> 3,0</td><td> 78</td>

*) n = 4

The results indicate that Formulations A and B provide much better pharmacokinetic properties than Formulation C.