Pharmaceutical cyclosporin composition

(42) Start of patent duration: 15. 7.1997 ⁵⁴ (45) Date of issue: 25. 2.1998

<td>(30) Priority:</td><td>(73) Patent owner:</td>

<td>27. 6.1991 GB 9113872 claims.</td><td>NOVARTIS INVESTMENTS ADMINISTRATIVE COMPANY MBH</td>

<td>(56) Documents:</td><td>A-1235 VIENNA (AT).</td>

<td>GB 2228198A GB2222770A</td><td></td>

(54) PHARMACEUTICAL COMPOSITION (57) A pharmaceutical composition in the form of a microemulsion concentrate containing a cydosporin as an active ingredient in a carrier medium, characterized in that

1) as the hydrophilic phase 1,2-propylene glycol,

2) a mixture of mono-, di- and tri-glycerides and

3) contains a surfactant.

CQ

AT 403 436 eur a-raas

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The present invention relates to novel galenic formulations, in particular to novel galenic formulations in which the active ingredient comprises one or more compounds selected from the cyclic poly-N-methylated undecapeptides of the class of cyclosporins - see, for example, GB-A 2 222 770 and US Pat GB-A 2 228 198 and worldwide equivalents thereof.

As discussed in these GB patents, the cyclosporins represent highly specific problem cases with respect to the application in general and the galenic formulation in particular, including the particular problems of bioavailability of the drug and the variability of the patient's response to the dose administered.

In order to address these and related difficulties, GB-A-2 222 770 has disclosed galenic formulations containing a cyclosporin active agent which, inter alia, take the form of a microemulsion or a microemulsion preconcentrate. Such compositions typically contain: 1) a hydrophilic phase, 2) a lipophilic phase, and 3) a surfactant. Specifically cited hydrophilic phase components are both commercially available products under the trade names Transcutol and Glycofurol, as well as 1,2-propylene glycol. Preferred components of the lipophilic phase are tri-glycerides of medium chain fatty acids known and commercially available under the trade names Miglyol, Captex, Myritol, Capmul, Captex, Neobee and Mazol, with Miglyol 812 being the most preferred. Suitable surfactant components include, in particular, reaction products of natural and hydrogenated vegetable oils and ethylene glycol, as known and commercially available under the trade names Cremophor and Nikkol, the products Cremophor RH40 and Nikkol HCO-40 being found to be particularly preferred.

GB-A-2 228 198 proposes another way to overcome the difficulties with cyclosporin administration. In particular, it discloses formulations based on oils in which the oily ingredient comprises a combination of a tri-glyceride and (i) a glycerol partial ester or (ii) a 1,2-propylene glycol full or partial ester or (iii) a sorbitol full or partial ester contains. The products known and available under the trade name Maisine are proposed as suitable tris and partial glyceride components. The disclosed agents additionally contain a surface-active component, for example Cremophor RH40, but are preferably free from any hydrophilic components such as ethanol. The described and exemplified means are free of such components.

According to the present invention it has now surprisingly been found that particularly stable galenic cyclosporin formulations are obtainable which have particularly interesting bioavailability characteristics and a reduced variability of the inter- and intra-individual bioavailability parameters. Since such agents are new, the present invention provides, in its broadest aspect:

A pharmaceutical composition in the form of a microemulsion preconcentrate containing a cyclosporin active agent in a carrier material, the

1) as the hydrophilic phase 1,2-propylene glycol;

2) a mixture of mono-, di- and tri-glycerides; and

3) contains a surfactant.

The term pharmaceutical composition as used herein and in the appended claims should be understood to define agents whose individual components or agents are themselves pharmaceutically acceptable, for example, suitable or acceptable for oral use where oral administration is contemplated ,

Cyclosporins to which the present invention relates are all those having pharmaceutical utility, for example, as immunosuppressive agents, anti-parasitic agents, and multi-drug resistance-eliminating agents as known and described in the literature, in particular cyclosporin A (also known and hereafter referred to as cyclosporin), cyclosporin G, [O- (2-hydroxyethyl) - (D) ser]^8th-Ciclosporin, and [3'-Deshydroxy-3'-keto-MeBmt] '- [Val]²Ciclosporin.

The components (2) in the compositions according to the invention preferably comprise mixtures of C 12-2o fatty acid mono-, di- and tri-glycerides, in particular C 16-16 fatty acid mono-, di- and tri-glycerides. The fatty acid component of said mixed mono-, di- and tri-glycerides may contain both saturated and unsaturated fatty acid residues. Preferably, however, they will consist predominantly of unsaturated fatty acid residues, in particular of Ci 8 unsaturated fatty acid residues, such as linoleic, linoleic and oleic acid residues. Suitably, component (2) will be at least 60%, preferably at least 75%, more preferably 85% or more, by weight, of a cis unsaturated fatty acid, for example linolenic, linoleic and oleic acid mono-, di- and tri -Glycerides included. Suitably, they are less than 20%, for example about 15% or 10%, based on the

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Weight or less of saturated fatty acid, eg palmitic and stearic acid mono-, di- and tri-glycerides.

The components (2) of the compositions according to the invention will preferably be predominantly composed of mono- and di-glycerides. For example, they will contain at least 50%, especially at least 70%, eg 75%, 80% or 85% or more of mono- and di-glycerides, based on the total weight of component (2).

The components (2) of the compositions of this invention will suitably contain from about 25 to about 50%, preferably from about 30 to about 40%, eg 35 to 40%, of mono-glycerides, based on the total weight of component (2).

The components (2) of the compositions of this invention will suitably contain from about 30 to about 60%, preferably from about 40 to 55%, for example about 48 to 50% of di-glycerides, based on the total weight of component (2).

The components (2) of the compositions of this invention will suitably contain from at least 5%, eg from about 7.5 to about 15%, for example 9 to 12%, of tri-glycerides by weight.

The components (2) of the compositions according to the invention can be prepared by mixing together individual mono-, di- or tri-glycerides in a suitable relative ratio. In practice, however, they conveniently contain transesterification products of vegetable oils, for example almond oil, peanut oil, olive oil, peach oil, palm oil or, preferably, corn oil, sunflower oil or safflower oil, but especially corn oil, with glycerol.

Such transesterification products are generally obtained by heating the selected vegetable oil with glycerol in the presence of a suitable catalyst under inert gas atmosphere and constant stirring in, for example, a stainless steel reactor to achieve transesterification or glycerolysis. In addition to their mono-, di- and tri-glyceride components, such transesterification products will generally also contain minor amounts of free glycerin. The amount of free glycerol in components (2) intended for use in the compositions according to the invention is preferably less than 10%, more preferably less than 5%, in particular approx. 1 or 2%, based on the total weight of the amount of free glycerol and mono-, di- and tri-glycerides.

When it is intended to prepare soft gelatin capsules, it is preferable to first remove another portion of the glycerine, for example by distillation, to obtain a substantially glycerol-free batch.

Particularly suitable components (2) for use in compositions according to the invention will accordingly comprise the following components in the stated amounts by weight, based on the total weight of component (2):

Mono-glycerides: 25 or 30 to 50%, preferably 30 to 40%.

Di-glycerides: 30 or 40 to 60%, in particular 40 to 55%, for example 45 to 55%.

Mono- and di-glycerides:&gt; 75%, preferably&gt; 80%, eg approx. 85%.

Tri-glycerides: at least 5%.

Free glycerin: &lt;5%, preferably &lt;2% or &lt;1%.

Particularly suitable components (2) for use in the compositions according to the invention are the transesterification products of corn oil and glycerol, as are commercially available, for example, under the trade name Maisine. These products are preferably composed of linoleic and oleic acid mono-, di- and tri-glycerides together with minor amounts of palmitic and stearic acid mono-, di- and tri-glycerides (corn oil itself is composed of ca. 56 % By weight of linoleic acid, 30% by weight of oleic acid, approx. 10 Weight% palmitin and approx. 3% stearic acid components). Physical characteristics for maize [available from Etablissements Gattefosse, in 36, Chemin de Genas, POBox 603, 69804 Saint-Priest, Cedex (France)] are:

approximate composition of free glycerin

Mono-glycerides

Di-glycerides tri-glycerides free oleic acid

Additional physical saponification number = approx.

- Max. 10% (typically 3.9-4.9% or, in essentially glycerol-free batches, about 0.2%)

about 35% (typically 30-40% or, in essentially glycerine-free batches, for example 32-36%, eg about 36%)

- about 50% (or, in "substantially glycerol-free batches about 46-48%)

- about 10% (or, in "essentially glycerin-free batches, about 12-15%)

- about 1%

Characteristics for Maisine are: acid number = max. about 2, iodine number = about 85-105, 150-175 (Fiedler Lexicon of excipients, 3rd revised and expanded edition (1989) Volume 2, page 768). The fatty acid content in Maisine is typically: palmitic acid - about 11%;

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Stearic acid - about 2.5%; Oleic acid - about 29%; Linoleic acid - about 56%; others - about 1.5%.

It is particularly preferred that component (2), for example, a corn oil interesterified with glycerine, is a clear liquid. For example, a sample thereof remains clear at room temperature for one hour after the sample has been taken out of the refrigerator, where it has been stored, for example, between + 2 and + 8 · C for 24 hours.

Preferably, the components (2) have a low content of saturated fatty acids. Components (2) which satisfy these requirements can be obtained, for example, from commercially available products. For example, they can be prepared by separation techniques known per se, for example freezing processes, which are coupled with separation techniques, eg Centrifugation can be obtained, wherein the saturated fatty acid components are removed, or the unsaturated fatty acid components are enriched. Typically, the total content of saturated fatty acid components will be less than 15%, for example less than 10% or less than 5%, based on the total weight of component (2). A reduction in the content of the saturated fatty acid component in the mono-glyceride fraction of component (2) can be observed after carrying out the separation techniques.

The components (2) thus preferably contain lower amounts of saturated fatty acids (eg, palmitic and stearic acids) and relatively larger amounts of unsaturated fatty acids (eg, oleic and linoleic acids) than the starting material.

Typical preferred components (2) according to a preferred embodiment of the invention may include:

32-36% mono-glycerides,

45-55% di-glycerides and

12-20% tri-glycerides based on the total weight of component (2).

Other preferred features are as follows:

<td colspan="2">Fatty acid content as determined as methyl ester by chromotography</td>

<td>Linoleic acid methylester</td><td>53-63%</td>

<td>Oleic acid methylester</td><td>24-34%</td>

<td>Linolenic acid methylester</td><td>0- 3%</td>

<td>Arachninsäure-methylester</td><td>0- 3%</td>

<td>Palmitic acid methylester</td><td>6-12%</td>

<td>Stearic acid methylester</td><td>1- 3%</td>

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

<td>Hydroxyl value</td><td>140-210</td>

<td>iodine</td><td>110-120</td>

<td>peroxide</td><td>&lt;4.0</td>

<td>Free glycerin</td><td>&lt;1.0</td>

<td>acid number</td><td>about 2</td>

<td>Saponification number approx.</td><td>150-185</td>

The components (2) which agree with the numerical values ​​given above are hereafter referred to as refined glycerol transesterified corn oils. Freshly prepared components (2) according to the preferred embodiments have a clear appearance and remain clear at a storage temperature of +20 to + 25'C for more than 20 days.

The refined glycerol transesterified corn oils have been specifically suggested for the preparation of the compositions of this invention. However, they may also have additional use for solubilizing other actives, and they have the advantage of remaining stable for a long time, such as staying clear. They represent a further aspect of the present invention. Accordingly, in a further aspect, the invention provides a transesterification product of corn oil and glycerine consisting predominantly of linoleic and oleic acid mono-, di- and tri-glycerides which have been treated so that the content of unsaturated fatty acid components of the mono-, Di- and tri-glycerides has been increased so that the content of linoleic acid and oleic acid mono-, di- and tri-glycerides total 85% or more of the total composition.

The components (3) in the compositions according to the invention preferably have an HLB value of at least 10.

Examples of suitable components (3) in the agents according to the invention are:

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3.1 Reaction products of natural or hydrogenated castor oil and ethylene oxide. Such products are obtained in a known manner, for example by reaction of a natural or hydrogenated castor oil with ethylene oxide, for example in a molar ratio of about 1:35 to about 1:60, where desired the polyethylene glycol component can be removed from the product. Such production processes are described, for example, in DE-A 1 182 388 and 1 518 819. Particularly suitable are various surfactants, such as ethoxylates of fatty alcohols or hydrogenated castor oil or Nonylphenol, which are available under the trade name Cremophor. Very suitable are the following products:

Cremophor RH40 with a saponification number of about 50-60, an acid number &lt;1, ​​an iodine value &lt;1, a water content (Fischer) &lt;2%, an n_D⁶⁰ of about 1.453 - 1.457 and to HLB value of about 14-16: Cremophor RH60 having a saponification number of about 40-50, an acid number &lt;1, ​​an iodine value &lt;1, a water content (Fischer) of 4.5- 5.5%, a no²⁵ from about 1.453 - 1.457 and to HLB value of about 15-17; and Cremophor EL having a molecular weight (vapor osmometry) of about 1630, a saponification number of about 65-70, an acid number of about 2, an iodine number of about 28-32, and an n_D²⁵ from about 1,471. (see Fiedler Lexicon der Hilfsstoffe, 3rd revised and expanded edition (1989), Volume 1, p. 326).

So suitable for use in this class are the various surfactants available under the trade name Nikkol (eg Nikkol HCO-40 and HCO-60) (cf. Fiedler). Nikkol HCO-60 is a reaction product between hydrogenated castor oil and ethylene oxide which has the following characteristics: acid value of 0.3; Saponification number of 47.4; Hydroxy value of 42.5; pH (5%) of 4.6; Color APHA = 40; Melting point of 36.0 * C; Freezing point of 32.4 * C; H2O content (%, KF) = 0.03;

3.2 Polyoxyethylene sorbitan fatty acid esters, for example mono- and tri-lauric, palmitic, stearic and oleic acid esters, for example of the known type available under the trade name Tween (Fiedler, loc. Cit., 2, p. 13001304) including the products

Tween 20 [polyoxyethylene (20) sorbitan monolaurate],

Tween 21 [polyoxyethylene (4) sorbitan monolaurate],

Tween 40 [polyoxyethylene (20) sorbitan monopalmitate],

Tween 60 [polyoxyethylene (20) sorbitan monostearate],

Tween 65 [polyoxyethylene (20) sorbitan tristearate],

Tween 80 [polyoxyethylene (20) sorbitan monooleate],

Tween 81 [polyoxyethylene (5) sorbitan monooleate] and Tween 85 [polyoxyethylene (20) sorbitan monooleate].

Particularly preferred products of this class for use in the compositions according to the invention are the above products Tween 40 and Tween 80.

3.3 Polyoxyethylene fatty acid esters, for example polyoxyethylene stearic acid esters of the known type, which are obtainable under the trade name Myrj (Fiedler, loc. Cit., 2, pp. 834-835). A particularly preferred product of this class for use in the compositions of this invention is the product Myrj 52, which has a density of 025 = about 1.1, a melting point of = about 40-44'C, an HLB value = about 16.9 , an acid number of = about 0-1 and a saponification number of = about 25-35 has.

3.4 polyoxyethylene-polyoxypropylene copolymers and block co-polymers, for example of the known type commercially available under the trade names Pluronic, Emkalyx and Poloxamer (see Fiedler, loc. cit., 2, p. 959). A particularly preferred product of this class for use in the compositions according to the invention is the product Pluronic F68, which has a melting point of approx. 52 C and a molecular weight of approx. 6800-8975 has. Another preferred product of this class for use in the compositions of this invention is the product poloxamer 188.

3.5 dioctylsulfosuccinate or di- (2-ethylhexyl) -succinate (see Fiedler, loc cit, pp. 107-108).

3.6 Phospholipids, especially lecithins (cf., Fiedler, loc. Cit., 2, pp. 943-944). Suitable lecithins for use in the compositions of the invention include in particular soybean lecithins.

3.7 propylene glycol mono- and di-fatty acid esters such as propylene glycol dicaprylic acid ester (also known and commercially available under the trade name Miglyol 840),

Propylene glycol dilaurinsäureester,

Propylene glycol hydroxystearinsäureester,

Propylene glycol isostearic acid ester,

Propylene glycol Iaurinsäureester,

Propylene glycol rizinusölfettsäureester,

Propylene glycol stearic acid ester, etc. (see Fiedler, loc.cit., 2, pp. 808-809).

3.8 sodium lauryl sulfate.

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For use in the context of the present invention, the components as described in item (3.1) are most preferred.

Components (1), (2) and (3) are preferably present in the compositions of the invention in such relative proportions that the agent is a microemulsion preconcentrate, ie having the characteristics of a microemulsion preconcentrate which is described in GB -A 2 222 770 on pages 11 to 12 are described. The definition of this reference is hereby considered part of this description for the purposes of defining such a system. The compositions of the invention are thus preferably microemulsion preconcentrates, particularly of the type providing o / w (oil in water) microemulsions. According to the invention, it is understood that agents which are microemulsions containing components (1), (2) and (3) together with (4) water also form part of the present invention.

As also indicated in GB-A 2 222 770, the hydrophilic phase of a microemulsion preconcentrate, ie Component (1) in the inventive compositions, also include one or more additional components as components of the hydrophilic phase, for example, lower (eg Ci-s) alkanoic acid, especially ethanol. Such components will generally be present as a partial replacement of component (1). Although the use of ethanol in the compositions of this invention is not critical to the invention, it has been found that it may be of particular advantage to process the compositions in encapsulated form in soft gelatin, for example, to improve storage characteristics, particularly to reduce risk in that the cyclosporin crystallizes out after the encapsulation process. Thus, the storage stability can be extended by using a lower alcohol as an additional component of the hydrophilic phase,

Suitably, the hydrophilic phase, ie component (1), 1,2-propylene glycol, or component (1) and further co-components of the hydrophilic phase, eg ethanol, in the inventive compositions in an amount of 1.0 or 2.5 up to 50%, preferably from 5 to 40%, in particular from 10 to 35%, eg more than 15%, for example from about 20 to about 30,% based on the total weight of the hydrophilic phase components together with components (2) and (3) , to be available.

If a co-component is used in the hydrophilic phase, then this co-component, for example ethanol, is suitably in an amount up to about 20%, preferably up to about 10 or 15%, eg from about 5 to 10 or 15%, based on the total weight of the agent. Thus, this co-component is suitably in an amount of about 25 to 75%, based on the total weight of the hydrophilic phase component (eg 1,2-propylene glycol and ethanol). Preferably, it is in an amount of less than 50%, for example from 25 to 50%, for example about 30, 40 or 50% available.

Suitably component (2) in the compositions according to the invention is in an amount of from 5 to 65%, preferably from 15 to 45%, in particular from 20 to 40%, for example from about 25 to about 35%, based on the total weight of the components hydrophilic phase and components (2) and (3).

Suitably, the component (3) in the inventive compositions in an amount of 25 to 75%, preferably from 30 to 60%, for example from about 55 or 60%, based on the total weight of the components of the hydrophilic phase and the components (2) and (3) exist.

Suitably the agents of the invention will contain from about 1 or 2 to 30%, preferably from 5 to 20 or 25%, especially from 7.5 to 15%, eg about 10% cyclosporin, based on the total weight of the agent.

The attached drawing I shows a three-component diagram of the relative concentrations of the hydrophilic phase, ie 1,2-propylene glycol, component (2), eg refined glycerol transesterified corn oil and component (3), eg Cremophor RH40, in mixtures according to the invention 10% cyclosporin (eg ciclosporin), based on the total weight. The relative concentrations of the carrier components increase in the direction of arrows 0 to 100%.

For agents according to the present invention, the relative proportions of the hydrophilic phase component (s), component (2) and component (3) are conveniently within the hatched area X. So defined agents are microemulsion preconcentrates of high stability, which upon addition of water are capable of forming microemulsions with an average particle size of &lt;150 nm and which are stable for periods longer than 24 hours. In contrast, agents in the regions Α, B and C in aqueous systems (A) are subject to discoloration, (B) to phase separation and (C) to cloudiness. Compositions according to the invention which contain the hydrophilic phase components and the components (2) and (3) in the relative proportions as defined by the line X of the drawing I are particularly preferred.

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In the event that the I, 2-propylene glycol component is partially replaced by ethanol as described above, the region X in the diagram of the drawing I is slightly pushed upwards, ie towards higher concentrations of component (3). However, this shift is only a few percent shift and does not significantly change the graph.

The compositions of the invention showed good stability characteristics, as shown for example by standard stability tests. For example, they have a shelf life of up to 3 years or even longer.

The compositions according to the invention may also contain further additives or additives, for example antioxidants such as Ascorbyl palmitate, butyl-hydroxy-anisole (BHA), butyl-hydroxy-toluene (BHT) and tocopherols, eg α-tocopherol (vitamin E) and / or preservatives, for example in an amount of about 0.05 to 1%, based on the total weight of the composition, or sweeteners or perfumes, for example in an amount of up to about 2.5 or 5% , based on the total weight of the agent.

It has been found that the agents of the present invention exhibit particularly advantageous properties when administered orally, for example in terms of both uniformity and high level of bioavailability achieved as indicated in standard bioavailability tests, for example in healthy patients using a specific monoclonal test system to determine cyclosporin levels, such as in the following examples. In particular, the compositions of the invention provide an improved oral dosage form for cyclosporins (eg Ciclosporin), because this form demonstrates the lack of any significant dietary interaction observed in the commercially available form of cyclosporine, particularly in the context of fatty, rich foods. In addition, the inter-individual and intra-individual variations in the pharmacokinetic parameters in the compositions of the invention may be substantially lower than in the commercial oral form of cyclosporin. In particular, the difference between the pharmacokinetic parameters at ingestion and without food intake or even between the absorption by day and the absorption at night can be eliminated by administering an agent according to the invention. Thus, the novel novel agents, the pharmacokinetic parameters, for example the absorption and blood concentration are surprisingly more predictable. This new galenic form can eliminate the administration problems of erratic absorption of cyclosporin. In addition, the agent of the invention may show improved bioavailability in patients having disorders of absorption, eg Liver transplant patients or pediatric patients. In particular, it has been found that such agents are compatible with surfactant materials, for example bile salts, which are present in the gastrointestinal tract. That is, they are completely dispersible in aqueous systems containing such natural surfactants and are thus capable of providing in situ microemulsion systems which are stable and show no precipitation of the cyclosporin or other disturbances of the fine distribution structure. The efficacy of such systems when administered orally remains independent of and / or unaffected by the presence or absence of bile salts at a particular time or in a particular individual.

The compositions according to the invention are well tolerated, as indicated, for example, in clinical trials over 4 weeks.

The compositions according to the invention are preferably prepared in a single-dose form, for example by filling them in orally administrable capsules, for example soft or hard gelatin capsules. If desired, they can also be produced in the form of a drink solution. If the agents are in unit dosage form, each single dose is suitably between 10 and 200 mg cyclosporin, preferably between 10 and 150 mg, eg 15, 20, 25, 50 or 100 mg of cyclosporin. Such finite dosage forms are suitable for 1, 2 or 3 times to 5 times daily administration (for example depending on the specific purpose of the therapy, the particular therapy section, etc.).

Optionally, the compositions of the present invention which are suitable for oral administration may contain as component (4) water or any other aqueous system, thereby providing a microemulsion system suitable for drinking.

Further, the present invention provides a process for producing a pharmaceutical agent according to the foregoing definitions, which comprises intimately mixing the above-defined component (1), component (2) and component (3) and, if necessary, the obtained mixture in single dosage form, for example by filling the agent in gelatin capsules, for example in hard or soft gelatin capsules.

In a particular embodiment, the present invention provides a process for the preparation of a pharmaceutical agent as defined above in the form of a microemulsion preconcentrate or

AT 403 436 Β of a microemulsion ready, that consists in that a component (1), a component (2) and a component (3), if desired, together with other components or additives, in particular with a co-component of the hydrophilic phase, for example, ethanol, in such ratio of components (1), (2) and (3) intimately mixed, that a microemulsion preconcentrate is obtained and, if desired, the resulting agent is filled in single dosage form or the resulting agent is treated with an appropriate amount of water or a suitable aqueous solvent medium, so that a microemulsion is obtained.

The following examples illustrate the compositions of this invention in unit dosage form suitable for use, for example, to prevent rejection of transplants or to treat autoimmune diseases when administered from 1 to 5 individual doses per day. The examples describe in particular the use of cyclosporin. However, similar mixtures can be obtained by substituting any other cyclosporin, especially [0- (2-hydroxyethyl) - (D) -Ser].^8th-Ciclosporin (hereinafter called compound Z) uses.

EXAMPLE 1:

Production of refined glycerol transesterified corn oil.

Substantially glycerol-free, glycerol-transesterified corn oil (if necessary, after heating to obtain a clear mixture) is slowly cooled to a temperature of + 20 ° C. and stored at this temperature for one night. In a first centrifugation at an acceleration of 12,000 G and a throughput of 103 kg / h in a continuous centrifuge, one obtains a liquid phase (62 kg / h) and a sediment-containing phase (41 kg / h). The liquid phase is slowly cooled to + 8 * C and stored at this temperature for one night. In a second centrifugation step with an acceleration of 12,000 G and a throughput of 112 kg / h, a liquid phase (76.2 kg / h) and a sediment-containing phase (35.8 kg / h) are obtained. The liquid phase is the refined glycerol-transesterified corn oil. " Optionally, an improved product can be obtained by performing the centrifugation in three steps, for example at + 20'C, + 10'C and + 5'C.

The process is characterized by a slight percentage reduction in the mono-glyceride component in the refined glycerol transesterified corn oil compared to the starting material (eg 35.6% compared to 38.3%).

A typical analytical comparison between the sediment and the clear solution is as follows:

<td>connection</td><td>Sediment (%)</td><td>Clear solution (%)</td>

<td>1. Mono-palmitic acid ester</td><td>19.1</td><td>3.4</td>

<td>2. mono-linoleic acid ester + mono-oleic acid ester</td><td>23.4</td><td>27.0</td>

<td>3. mono-stearic acid ester</td><td>5.7</td><td>&lt;2</td>

<td>4. di-linoleic acid ester + di-oleic acid ester</td><td>35.4</td><td>44.7</td>

<td>5. other di-glycerides</td><td>7.7</td><td>10.4</td>

<td>6. Tri-glycerides</td><td>8.7</td><td>12.5</td>

The following table gives typical amounts of the components in the refined product obtained according to this method:

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<td colspan="2">COMPOSITION OF COMPONENTS (% by weight)</td>

<td>components</td><td>refined glycerol transesterified corn oil</td>

<td>Glycerides: mono</td><td>33.3</td>

<td>di</td><td>52.1</td>

<td>Tri</td><td>14.6</td>

<td>fatty acids:</td><td></td>

<td>Palmitic acid (C16)</td><td>7.8</td>

<td>Stearic acid (C18)</td><td>1.7</td>

<td>Oleic acid (CI 8: 1)</td><td>31.6</td>

<td>Linoleic acid (C18: 2)</td><td>57.7</td>

<td>glycerine</td><td>&lt;1%</td>

EXAMPLE 2

Preparation of oral dosage forms

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

<td>Cyclosporin, eg cyclosporin</td><td>100</td>

<td>1) 1,2-propylene glycol</td><td>200</td>

<td>2) refined oil</td><td>320</td>

<td>3) Cremophor RH40</td><td>380</td>

<td>Total</td><td>1,000</td>

In this example, refined oil = refined glycerin transesterified corn oil is as described in example 1 or for maizeine, eg essentially glycerine-free maize.

The cyclosporin is dissolved in (1) with stirring at room temperature, and (2) and (3) are added to the resulting solution with further stirring. The resulting mixture is filled into hard gelatin capsules of size 1 and sealed, eg using the Quali-Seal technique.

Agents containing 50 and 100 mg of cyclosporin are prepared in an analogous manner by using the following indicated ingredients in the indicated amounts.

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MEDIUM containing 100 mg of cyclosporin, eg ciclosporin

<td>ί COMPOSITION I</td><td>-1-r 1 2 I 1</td><td>3</td><td>τ- 1 4 J</td><td>-r 5 1 L</td><td>1 6 1</td>

<td>1 (COMPONENT I</td><td>i AMOUNT</td><td></td><td colspan="2">(Mg / capsule)</td><td></td>

<td>1 (1) 1,2-propylene glycol</td><td>ί 200 ί - yy</td><td>270</td><td>[180 j -</td><td>180 | j</td><td>90 i</td>

<td>| 2) refined oil 1</td><td>ί 350 ί 1 1</td><td>180</td><td>(180 1</td><td>360 | 1</td><td>360 |</td>

<td>I | 3) Cremophor RH40 1</td><td>1 350 1 - -1_L</td><td>450</td><td>1 | 540 J</td><td>360 | L</td><td>450 ί 1</td>

<td>(COMPOSITION 1</td><td>-1-r 1 7 i I!</td><td>θ! _Ι_</td><td>- 9</td><td>-1 1ο 1 ι</td>

<td>1 (COMPONENT 1</td><td>( AMOUNT</td><td colspan="3">(mg / capsule) |</td>

<td>1 | 1) 1,2-propylene glycol f</td><td>1 ι 1 150 1 1</td><td>100 ί</td><td>200 -</td><td>200 ί</td>

<td>1 | la) ethanol 1</td><td>ί where ί Ι 1</td><td>100 ί</td><td>100</td><td>100 ί j</td>

<td>1 .· (2) refined 01 ι</td><td>1 ! I 345 ( ί</td><td>320 ί</td><td>320</td><td>290 ί</td>

<td>1 | 3) Cremophor RH40 ι</td><td>ί 405 ( _1___ "-1.</td><td>380 ί ι</td><td>380</td><td>360 ί ι</td>

MEDIUM containing 50 mg cyclosporin

<td>COMPOSITION</td><td>A</td><td>B</td><td>C</td><td>D</td><td>e</td><td>F</td>

<td>COMPONENT</td><td colspan="6">QTY (mg / capsule)</td>

<td>1) 1,2-propylene glycol</td><td>100</td><td>135</td><td>45</td><td>90</td><td>100</td><td>50</td>

<td>1a) ethanol</td><td></td><td></td><td></td><td></td><td></td><td>50</td>

<td>2) refined oil</td><td>160</td><td>90</td><td>180</td><td>180</td><td>67</td><td>160</td>

<td>3) Cremophor RH40</td><td>190</td><td>225</td><td>225</td><td>180</td><td>167</td><td>190</td>

As already indicated above, similar agents can be prepared which contain the compound Z instead of cyclosporin. Thus, the composition D can be prepared which contains 50 mg of compound Z instead of cyclosporin.

EXAMPLE 3: Bioavailability in Dogs

The biopharmaceutical properties of the compositions according to the invention were compared with those in

Commercially available soft gelatin capsule of ciclosporin. The forms were compared after oral administration to 12 male beagle dogs in a cross-over arrangement. The pharmacokinetic profile of ciclosporin was determined in whole blood over 24 hours. The area under the blood concentration curve plotted against time, (AUC), C_Max and T._Max were set.

AT 403 436 Β

Forms: dose 100 mg ciclosporin per dog

<td colspan="2">Medium X (commercial form, soft gelatin capsule)</td>

<td>cyclosporine Labrafil ethanol corn oil Total</td><td>100 mg 300 mg 100 mg 416 mg 926 mg / dose</td>

Agent I according to the present invention

<td>cyclosporine</td><td>100 mg</td>

<td>1) 1,2-propylene glycol</td><td>75 mg</td>

<td>1a) ethanol</td><td>150 mg</td>

<td>2) refined glycerol transesterified corn oil</td><td>345 mg</td>

<td>3) Cremophor RH40</td><td>405 mg</td>

<td>Total</td><td>1075 mg / dose</td>

Administration of the drug Male beagle dogs weighing approximately 12 kg were successful in completing the trial. Twenty hours before the administration of the drug, the animals are deprived of food but continue to have free access to water until the beginning of the experiment. The dosage forms are administered to the animals early in the morning (approximately 8.00 h) by a probe rinsed with 20 ml of 0.9% saline. Three hours after administration, the animals were again given free access to water and food. Between two administrations on the same animal a flushing time of one week was necessary.

Blood sample patterns:

Blood samples of 2 ml (or 5 ml for the blank) were removed from the cephalic vein (forearm) with a sterile needle (diameter ca. 1.2 mm) and collected in 5 ml plastic tubes containing EDTA. These blood samples were 15 min. before and then taken 30 min., 1, 1,5, 2, 3, 4, 6, 8, 12 and 24 hours after the oral administration of the active ingredient. The blood samples were taken at approx. -18'C stored until the determination of the active substance. Blood samples were analyzed by ciclosporin-specific radioimmunoassay (RIA). The mean blood levels of ciclosporin in dogs are shown graphically in attached Figure II. The area under the blood concentration versus time (AUC) curves are calculated using the trapezoidal rule.

An analysis of the deviation (CV) is performed and the mean AUCs, Cmax and Tmax are statistically compared by the Tukey test. The results obtained are shown in the following table.

<td rowspan="2">medium</td><td colspan="2">AUC 0-24 h</td><td colspan="2">Umax</td><td colspan="2">Tmax</td>

<td>Mean value [ng.h / ml]</td><td>CV [%]</td><td>Mean value [ng / ml]</td><td>CV [%]</td><td>Mean value [h]</td><td>CV [%]</td>

<td>X</td><td>6695</td><td>27</td><td>1053</td><td>25</td><td>1.3</td><td>20</td>

<td>I</td><td>10064</td><td>24</td><td>1539</td><td>18</td><td>1.6</td><td>29</td>

The behavior and body weight of the animals was controlled during the examination. There was no loss of body weight.

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Conclusion:

The agent according to the invention (agent I) has a significantly higher bioavailability (factor 1.5) than the commercially available soft gelatin capsule of cyclosporin.

Drawing II shows the average concentrations of cyclosporin in whole blood as found by a specific monoclonal RIA after a single oral administration of Agent X and Agent I, each at a dose of 100 mg. Concentration in blood (in ng / ml) is applied vertically, time horizontal.

EXAMPLE 4: Bioavailable in man

The bioavailability of cyclosporin is compared as determinable after administration of the commercial soft gelatin capsule of cyclosporin and an agent of the present invention.

Form administered: 100 mg ciclosporin per capsule

<td colspan="2">Medium X (hand-shaped, soft geiatine capsule)</td>

<td>cyclosporine</td><td>100 mg</td>

<td>Labrafil</td><td>300 mg</td>

<td>ethanol</td><td>100 mg</td>

<td>corn oil</td><td>426 mg</td>

<td>Total</td><td>926 mg / capsule</td>

Means No. 8 (according to Example 2 containing refined glycerol transesterified corn oil) in a soft gelatin capsule.

Method healthy male subjects completed the investigation. Each of the participants received 4 of the 8 administrations (2 agent 8 doses and the same 2 x agent doses).

Participants were randomly assigned to two subgroups, each consisting of 24 subjects, each corresponding to a parallel study. Group I subjects received doses of 200 mg and 600 mg ciclosporin, and subjects in group II received 400 mg and 800 mg, respectively.

Within each of the two groups, the trial was conducted on the basis of a balanced 4-way cross-over study with a 2 week washout period between each treatment.

Blood samples for ciclosporin in whole blood were taken 1 minute before drug intake and then 15 min, 30 min, 45 min, h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 6 h, 8 h, 10 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h and taken 48 h after the active ingredient intake ,

The individual concentrations of ciclosporin in whole blood were determined in each blood sample by a specific RIA method.

The limit of quantification was 12.5 ng / ml.

Blood levels and associated AUC₍₀Ciclosporin-48 hf values ​​were significantly higher at all dose levels after administration of agent 8 than after administration of agent X. The highest concentrations (Cmax) of the 200 mg, 400 mg, and 600 mg dose levels appeared a little earlier after administration of the agent 8 (see following table).

AT 403 436 Β

table

<td colspan="4">Bioavailability of cyclosporin in humans</td>

<td colspan="4">Mean value (± SD) of the AUC₍o-48 h&gt;. C_Max and T._Max after a single oral administration of various doses of the agent X and the agent 8</td>

<td>shape</td><td>AUC ") - 48 h) [ng.h / ml]</td><td>C_Max [Ng / ml]</td><td>T_Max [H]</td>

<td>200 mg mean X</td><td>2028 ± 608</td><td>558 ± 228</td><td>2.1 ± 0.7</td>

<td>200 mg mean 8</td><td>3468 ± 1000</td><td>1025 ± 218</td><td>1.5 ± 0.4</td>

<td>400 mg mean X</td><td>3326 ± 1115</td><td>785 ± 252</td><td>2.1 ± 0.9</td>

<td>400 mg mean 8</td><td>6944 ± 1468</td><td>1557 ± 286</td><td>1.4 ± 0.4</td>

<td>600 mg mean X</td><td>4501 ± 1217</td><td>917 ± 236</td><td>2.3 ± 1.0</td>

<td>600 mg mean 8</td><td>9689 ± 2282</td><td>1812 ± 400</td><td>1.7 ± 0.6</td>

<td>800 mg mean X</td><td>5209 ± 1554</td><td>1045 ± 264</td><td>2.4 ± 1.0</td>

<td>800 mg mean 8</td><td>12162 ± 3059</td><td>2143 ± 576</td><td>2.1 ± 0.8</td>

Due to the mean ratios of the AUC ₍₀-4β hf values, the relative bioavailability of Agent 8 versus Agent X was rated between 177 and 233%, depending on the dose administered (see the table below).

table

<td colspan="3">Relative bioavailability of agent 8 versus agent X.</td>

<td>Dose of [mg]</td><td>Mean Ratio of AUC j-48 h) Mean 8 vs. Mean X</td><td>Conversion factor: mean X vs. Mean 8</td>

<td>200</td><td>1.70</td><td>0.59</td>

<td>400</td><td>2.09</td><td>0.48</td>

<td>600</td><td>2.15</td><td>0.47</td>

<td>800</td><td>2.33</td><td>0.43</td>

Conclusion:

The agent according to the invention (agent 8) has a significantly higher bioavailability in humans, which is at least the factor 1.7 when compared with the commercial form (agent X).

The attached Fig. 3 shows a graphical representation of the average AUC "hj values ​​of the means X (empty triangles) with respect to those of the means 8 (filled circles). The AUC values ​​(ng.h / ml) of ciclosporin are vertical, the doses applied horizontally as obtained from Example 4.

The extent of absorption of agent 8 (in terms of AUC₍₀-48hr values) appeared to be dose-independent while that of drug X decreased with increasing dose (see Figure 3).