Pharmaceutical cyclosporin composition

Pharmaceutical compositions containing cyclosporin

Full description

Background of the invention

The present invention relates to modern galenic formulations, in particular modern galenic formulations comprising the active ingredient and one or more elements selected from an unbroken cyclic poly-N-methylated polymer of the cyclosporin group. See, for example, British Published Patents Nos. 770-2222 and 2228198 and their counterparts throughout the world . As discussed in the British patent literature, cyclosporins present significant specific difficulties for administration in general and the galenic formulation in particular, including issues of drug bioavailability and variation in patient response to dose.

To address these difficulties, in British Patent No. 770 2222, Galenic formulations are provided that include cyclosporin as the active ingredient and which take the form of, among other things, a microemulsion or more, a first of a microemulsion. These preparations contain: 1) a hydrophilic phase 2) a lipophilic phase 3) a surfactant. The components of the hydrophilic phase mentioned represent in particular well-known products commercially available under the trade names Transcutol and glycofurol, and also 1,2 propylene glycol. The preferred components of the lipophilic phase are medium chain fatty acid triglycerides as they are known and commercially available under the brand names Miglyol, Captex, Myritol, Capmul, Captex, Neobee, Mazol and the most preferred is Miglyol 812. Particularly suitable surfactant components include reaction products of natural or hydrogenated vegetable oils and ethylene glycol such as those commonly available commercially under the trade names Cremophor and Nikkol. Cremophor RH40 and 40-Nikkol HCO products are especially preferred.

British Patent No. 198 2228 proposes an alternative method to address the difficulties of cyclosporin administration. In particular, it reveals oil-based compositions in which the oil component includes a combination of tri-glyceride and (I) esters of parts of glycerol (partial esters), or (1,2-propylene glycol (ii) partial or complete, or (sorbitol (iii) esters, partial or complete). The discovered preparations include Also a surfactant component, eg Cremopher RH40, but preferably free of any

Fat-loving ingredients such as ethanol. Prescription and representative preparations are free of such ingredients.

General description of the invention

According to this invention, it has now been found that it is possible to obtain stable compositions of galene cyclosporins with interesting bioavailability properties and low rates of variation in bioavailability parameters per individual or between individuals. Since these compositions are new, the present invention presents in its broadest sense:

A pharmaceutical composition containing cyclosporin as the active ingredient in a carrier medium containing:

1) 1,2-propylene glycol.

2) a mixed C12-C20 fatty acid mono-, di-, tri-glyceride

3) Surfactants

Which forms the first concentrate for microemulsion preconcentrate.

The phrase “pharmaceutical preparation” and “phamaceutical composition” is used here and in the attached protections to mean that it defines preparations in which the individual components are themselves pharmaceutically acceptable, for example, when intended for oral administration, they are suitable or acceptable for oral administration.

Cyclosporins to which this invention applies are any of those that have a pharmaceutical use, for example, as immunosuppressive agents, anti-parasitic agents, and agents for reversal of multi-drug resistance, as known and described in Previous areas, especially Cyclosporins A (also known in this study as Cyclosporin G, (Ciclosporin,

[O-(2-hydroxyethyl)-(D)Ser]8-Ciclosporin, and [Val]2-Ciclosporin-1[3,-deshydroxy-3,-keto-MeBmt].

Component (2) in the inventor's formula consists of a mixture of mono-, di-, and tri-glycerides containing fatty acids C12-20, especially C16-18. The fatty acid component of the aforementioned mixture may include saturated or unsaturated fatty acids. In general, it is preferable for it to consist mostly of unsaturated fatty acids, especially C18 unsaturated fatty acids, for example linoleic, linolenic, oleic, and it is most appropriate for component (2) to include at least 60%, preferably 75%, and more preferably 85%. Or more by weight, C18 unsaturated fatty acid with mono-, di-, and triglycerides, such as linoleic and linolenic. It is more appropriate to include less than 20%, for example, about 15% or 10%, by weight, or less, saturated fatty acids with mono-, di-, and triglycerides, such as palmitic and stearic.

It is preferable that the components (2) in the preparations of this invention contain a majority of mono-, di-, and tri-glycerides, for example, at least 50%, and preferably at least 70%, for example, 75%, 80%, 85%, by weight, or more of mono-, di-, and triglycerides. Glyceride is estimated based on the total weight of the ingredient (2).

Components (2) of the patented preparations should preferably include from about 25 to about 50%, and preferably from about 30 to about 40%, for example 35 to 40% monoglycerides, estimated on the basis of the total weight of the component (2).

Components (2) of the preparations of the invention should preferably include from about 30 to about 60% and preferably from about 40 to 55%, for example: about 48 to 50% di-glycerides estimated on the basis of the total weight of the component (2).

Components (2) of the preparations of the invention will be suitable to contain at least 5%, for example from about 7.5 to 15%, and for example from 9 to 12% by weight, of triglycerides.

Components (2) in the preparations of the invention may be prepared by mixing mono, di or tri-glycerides individually in an appropriate ratio. In general, it is common for the products of transesterification to include vegetable oils, for example almond oil, peanut oil, olive oil, peach oil, palm oil, and preferably palm oil.

Corn, sunflower oil, or safflower oil, and corn oil with glycerol is more preferred.

These transesterification products can be obtained by heating the selected glycerol vegetable oil at a high temperature in the presence of the appropriate catalyst under an inert atmosphere of continuous stirring, for example, in a stainless iron reactor to achieve transesterification or glycerol analysis. In addition to their mono-, di- and tri-glyceride components, these conversion esterification products will also include small amounts of free glycerol. It is preferable that the amount of free glycerol present in the components (2) for use in the patented preparations be less than 10%, and More than 5% is preferred, and the most preferable is 1 or 2% by weight, depending on the total weight of glycerol with mono-, di- and tri-glyceride.

It is preferable to remove some of the glycerol first, for example by distillation (to give a portion completely free of glycerol) when gelatin capsules are to be manufactured.

Components (2) Particularly suitable for use in the preparations of the invention will contain components. The following are the recommended quantities by weight based on the total weight of the ingredient (2):

Mono-glycerides: 25 or 30 to 50%, especially 30 to 40%. di-glycerides: 30 or 40 to 60%, especially 40 to 55%, for example 45 to 55%. mono plus di-glycerides: > 75%, especially > 80%, for example, is about 85%. Tri-glycerides: at least 5%. free glycerol: < 5%, preferably < 2% or < 1%.

Components (2) particularly suitable for use in the preparations of the invention are the products of the transesterification of corn oil and glycerol, for example, as is commercially available under the trade name Maisine. These compounds consist mostly of linoleic and oleic acid mono-, di-and tri-glyceride, with small amounts of palmitic and stearic acid mono-, di-and tri-glyceride (corn oil itself consists of about 56% by weight linoleic acid, 30 oleic acid%, about 10% palmitic acid, about 3% Saint-Priest).

Available at Maisine Company. Physical properties of Maisine

Etablissements Gattefosse, of 36, Chemin de Genas, POBox 603, 69804 Saint-Priest, cedex (France)}

These are: Approximate composition:

Free glycerol: 10% at most (typical 3.9 - 4.9% or in combinations

Completely free of glycerol, about 0.2%.

Mono-glycerides: about 35% (typical is 30-40%, or in groups ((completely devoid of glycerol)), for example, 32-36%, about 36%). di-glycerides: about 50% (or in groups & completely devoid of glycerol & about 46-48%).

Tri-glycerides: about 10% (or in groups completely free of glycerol, about 12-15%). Free oleic acid content: about 1%.

Other physical properties of Maisine are: acid value is at most about 2, iodine no is about 85-105, saponification number is about 150-175,

(768 .Fiedler & Lexikon der Hilfsstoffe”, 3rd revised and expanded edition (1989) vol. 2, p)

The typical fatty acid content of Maisine is: palmitic acid: about 11%, stearic acid: about 2.5%, oleic acid: about 29%, linoleic acid: about 56%, other acids: about 1.5%.

It is especially preferable that ingredient (2), for example, corn oil transesterified with glycerol, be clear. This is after keeping a sample of it in the refrigerator between 2 and 8 °C for 24 hours. The sample will be clear at room temperature one hour after removing it from the refrigerator.

It is preferable that ingredients (2) contain only a small amount of saturated fatty acids. Components (2) that meet these requirements can be obtained, for example, from commercially available products, for example, obtained by methods such as the separation processes known in the patent, for example, freezing processes combined with separation processes such as

٠٠

To centrifuge, to remove the contents of the saturated fatty acid and stabilize the content of the unsaturated fatty acid component. The typical total content of a saturated fatty acid component will be < 15%, for example < 10% or 5% by weight based on the total weight of the component (2). A decrease in the content of the saturated fatty acid component in the mono glyceride portion of components (2) can be observed after the separation process.

Therefore, it is preferable for the ingredients (2) to contain smaller amounts of saturated fatty acids (such as palmitic acid, stearic acid) and relatively larger amounts of unsaturated fatty acids (such as oleic and linoleic acids) from the starting material.

Typically the components (2) embodied in this invention preferably contain:

32 - 36% mono-glycerides 45 - 55% di-glycerides and

12 - 20% tri-glycerides

By weight based on the total weight of the ingredients (2).

In addition, distinctive components are preferred, which include the following: stearic acid content, as determined as methyl ester by chromatographic analysis.

chromatography:

<img file="SA182B1_D0001.tif" />

Referring here to the components (2), which agree with the features drawn above as ((

Refined glycerol-transesterified corn oils. Ingredients (2) Freshly prepared according to the preferred contents, they look delicious and remain delicious when stored at a temperature of 20°C - 25°C for more than 20 days.

I have proposed “refined corn oils transesterified with glycerol” for the preparation of preparations of this invention. It can also be used to dissolve other active substances and has the property of remaining unchanged, for example remaining clear for a long time.

It is preferable for the components (3) in the patented preparations to have an HLB of at least 10%.

Suitable examples of components (3) in the patented preparations are:

3.1: Products of the reaction of natural or hydrogenated castor oil and ethylene oxide. These products can be obtained by known methods, for example by reacting natural or hydrogenated castor oil with ethylene oxide, for example, at a molar ratio of about 1:35 to about 1:06, with the optional elimination of the polyethylene glyccol component from the product, for example according to the methods discovered in and 1,182,388 Auslegeschriften'

1,518,819 German. The most suitable raw materials are available under the trade name Cremophor. The most suitable products are Cremophor RH40 with a saponification number of about 50-60, acid number &lt; 1, iodine number &lt; 1, Water content (Fisher) &lt; 2%, an nD60 is about 1.453, and an HLP is about 14-16, Cremophor RH40 has a saponification number of about 40-50, an acid number &lt; 1, iodine number &lt; 1, water content (Fischer) 4.5-5.5%, nD60 is about 1.453-1.457, HLP is about 15-17, Cremophor EL has a molecular weight (measured by steam osmometry) of about 1630, and a saponification number of about 65 - 70 and the acid number is about 2, the iodine number is about 28 - 32, and the nD60 is about 1.471.

,(1989) revised and expanded edition rd3 ,”cf Fiedler, &amp; Lexikon der Hilfstoffe)

(326.Vol. 1, p

Also, there are different materials suitable for use in this group available under the trade name Nikkol (eg 40-Nikkol HCO and 60-Emulgin), HCO (eg Mapeg), Emulgin RO40 (eg Incrocas), Mapeg CO-40h (eg 40 CFFiedler) (Incrocas). ). The aforementioned product 60-Nikkol HCO is the product of the reaction of ethylene hydrogenated castor oil and oxide and has the following properties: acid value about 0.3, saponification number about 47.4, hydroxy value about 42.5, PH (5%) about 4.6, color APHA about 40, melting point .mp about 36 C, freezing point about 32.4 C, moisture content H2O (%, KF) = 0.03.

3.2. Polyoxyethylene-sorbitan-fatty acid esters, for example, mono-and tri-lauryl, palmityl stearyl and oleyl esters, for example of the well-known type that is commercially available under the trade name CFFiedler, Loc. Cit. P.1300-1304) Tween) includes Tween products

20 (polyoxyethylene (20) sorbitanmonolaurate),

21 (polyoxyethylene (4) sorbitanmonolaurate),

40 [polyoxyethylene (20) sorbitanmonopalmitate], 60 [polyoxyethylene (20) sorbitanmonostearate],

65 [polyoxyethylene (20) sorbitantristearate],

80 [polyoxyethylene (20) sorbitanmonooleate),

81 [polyoxyethylene (5) sorbitanmonooleate],

85 [polyoxyethylene (20) sorbitantrioleate].

The preferred products from the above group for use in patent preparations are Tween 40 and Tween 80

3.3 Polyethylene esters and fatty acids: polyoxyethylene fatty acid esters, for example, polyoxyethylene stearic acid esters, the type known and commercially available. Under the trade name cfFiedler, Loc.cit.2, p.834-835 (Myrj), the preferred product particularly from this group for use in patents is the product Myrj 52, which has a D25 of about 1.1, an MP of about 40-44 C, and an HLB value of about 16.9, the acid value is about 0-1, and the saponification number is about 25-35.

3.4 Conjugated polymers and a group of conjugated polymers of polyoxyethylene (polyoxypropylene), for example, of the well-known type that is commercially available under the trade names Pluronic, Emkalyx, and cfFeedler, loc.cit, 2 p.959 (poloxamer). A particular favorite product from this group for use in patent preparations is

Pluronic F68 product, has an MP of about 52 M, and a molecular weight of about 6800-8975. Another preferred product from this group for use in patents is Poloxamer 188.

3.5 Dioctylsulfosuccinate or di-[2-ethylhexyl]-succinate (107-108 .1, ,P.cf Fiedler, loc. Cit.)

3.6 Phosphorlipids, especially CF Fiedler, Loe. Cit., 2, p. 943 - 944 (Lecithins) Lecithins suitable for use in the preparations of the invention include in particular soybean Lecithins.

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

propylene glycol dilaurate, propylene glycol hydroxystearate, propylene glycol isostearate, propylene glycol laurate, propylene glycol ricinoleate, propylene glycol stearate, and so on (809-808 .cfFiedler ,loc,cit,2,p).

3.8 Sodium lauryl sulfate.

For this invention, it is preferable to use components as mentioned under (3.1) above. It is preferable for components (1), (2), and (3) to be present in the patented preparations in proportion to each other, such that the preparation is a micro-emulsion as the first concentration, meaning that it has the properties of a system: a micro-emulsion as the first concentration, as described in the bulletin. British Patent No. 2,222,770 on pages 11 to 12, is included here by reference for the purpose of identifying this system. Therefore, it is preferable for the patented preparations to have a microemulsion as the first concentration, especially the type that gives an O/W (oil-in-water) microemulsion. This invention is also understood to include preparations comprising components (1), (2) and (3) with (4) water, which are microemulsions.

Also, as stated in British Patent Publication No. 2222770, the hydrophilic phase of microemulsion systems is the first concentration, i.e. component (1)

In preparations of this invention one or more additional substances may be included as hydrophilic phase components, for example low-length alkanols (such as C1-5), especially ethanol. These components will be present

Generally as a partial replacement for component (1). Although the use of ethanol in the preparations of this invention is not necessary, it has been found to have a special advantage when the preparations are made in the form of gelatin capsules, for example, to improve storage properties, especially to reduce the risk of cyclosporin precipitation after the encapsulation steps. Thus, the shelf life during storage can be extended by using low-length alkanol as an additive to the hydrophilic phase.

It is convenient to have a component as a hydrophilic phase, i.e., component (1), 1,2-propylene glycol, or component (1) combined with any other hydrophilic phase component, such as ethanol, in the preparations in an amount from 1 or 2.5 to 50 % , preferably from 5 to 40, preferably more than 10 to 35%, for example higher than 15%, for example from about 20 to 30%, of the weight based on the total weight of the component (or components) of the hydrophilic phase with components (2) And (3).

When a hydrophilic phase is used as a cofactor, it is appropriate for the cofactor to be present in an amount of about 20%, preferably 10 or 15%, for example 5 to 10 or 15% by weight based on the total weight of the preparation. Thus, it is appropriate to have the auxiliary component present in an amount of 25 to 75% by weight, based on the total weight of the components of the hydrophilic phase (for example, 1,2-propylene glycol plus ethanol). It is more preferable for it to be present in an amount less than 50%, for example, from 25 to 50% over For example, about 30, 40 or 50%. It will be appropriate for the presence of ingredient (2) in the preparations of the invention in an amount from 5 to 65%, preferably from 15 to 45%, preferably more than 20 to 40%, for example from about 25 to about 35%, based on the total weight of the ingredient (or Components of the hydrophilic phase, in addition to components (2) and (3).

It will be appropriate to have component (3) in the preparations of the invention 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 component (or components) of the hydrophilic phase in addition to the components (2). And (3).

It is appropriate for the inventions to contain from about 1 or 2 to 30%,

Preferably from 5 to 20 or 25%, preferably more than 7.5 to 15%, for example about 10% by weight of cyclosporin based on the total weight of the preparation. Brief explanation of the drawings:

The attached Figure 1 represents a three-dimensional drawing of the relative concentration of the hydrophilic phase component, 1,2-propylene glycol, component (2), for example ((corn oil refined by transesterification with glycerol)) and component (3), for example, Gremophor RH40, in preparations according to this invention. It contains 10% cyclosporin (eg Ciclosporin) by weight. The relative concentrations of the carrier components increase in the direction indicated by the arrows from 0 to 100%.

In preparations according to this invention, the relative sizes of the components of the hydrophilic phase, component (2) and component (3), are proportional to their presence in the shaded area. Thus, the defined preparations form a microemulsion as an initial concentration with high stability, capable, when added to water, of producing a microemulsion with particles of average size &lt; 1,500 Angstroms and fixed for more than 24 hours. In comparison preparations in zones B, A and C give aqueous systems susceptible to (A) coloration (B), discoloration (C) phase separation) and turbidity respectively. Preparations according to this invention contain components of the hydrophilic phase, component (2) and component (3), the amounts of which are proportional to what is defined by line X in Figure 1. Detailed description:

Once the 1,2-propylene component is partially replaced by ethanol as described here before, the region However, this upward movement is only a small percentage and does not change the aforementioned chart radically.

The preparations of the invention show good stability properties, for example, as achieved in standard stability experiments, for example, they have stability during storage ranging up to three years or

more.

Preparations according to the invention may include additives or other active substances

(For example, butyle hydroxyl, butyl hydroxy anisole (BHA), antioxidants ascorbyl palmitate (toluene (BHA), and tocopherols such as α-tocopherol (Vitamin E)) and/or preservatives, for example in an amount of 0.05 to 1% by weight based on the total weight of the preparations Or sweeteners or flavorings, for example in an amount of about 2.5 or 5% by weight, based on the total weight of the preparation.

It has been found that the preparations according to this invention have distinctive properties, especially when given orally, for example with regard to stability rates and a high level of bioavailability, as demonstrated by standard bioavailability experiments, for example in healthy patients using a monoclonal kit to determine cyclosporin levels, for example, as described in the following examples mentioned here. . In particular, preparations according to this invention provide an improved oral administration of cyclosporin (eg, Ciclosporin) because they do not exhibit the significant interaction with food, which we have observed with commercially available ciclosporin formulations, especially their interactions with high-fat foods. Also, the variability of pharmacokinetic parameters is significantly less with preparations according to this invention than with commercial oral forms of ciclosporin, whether per person or between individuals, especially the difference between pharmacokinetic parameters with and without food administration, or even between daytime absorption and absorption. During the night, all these differences can be eliminated according to this invention. Accordingly, the new formulation according to the present invention has standardized pharmacokinetics and blood levels that make it more predictable and surprising, and the new pharmaceutical form can reduce the problems of abuse and irregular absorption of ciclosporin. In addition, the preparation of this invention can provide improved bioavailability in patients suffering from malabsorption. For example, liver transplant patients or pediatric patients. In particular, it was found that these preparations are compatible with tensile raw materials, for example, bile salts present in the digestive tract. This is because it spreads completely in the comprehensive aquatic systems of these natural raw materials, and thus it is able to give a precise emulsion in place that does not change and is not characterized by cyclosporin precipitation or disturbance.

Another for special precise configuration. When administered orally, the function of these systems remains independent of and/or is not impaired by the relative presence or absence of bile salts at any given time or in any individual taking them.

The preparations of the invention may be well tolerated, for example as indicated in clinical trials over a period of 4 weeks. It is preferable for the preparations of this invention to be in the form of a dosage unit, for example by depositing them inside a capsule to be given orally, for example the shell of a gelatin or hard capsule, but if it is desired in the form of a syrup solution where the preparations are in the form of a dosage unit, it will be appropriate for each dosage unit to contain Between 10 and 200 mg of cyclosporin, and it is more preferable to take between 10 and 150 mg, for example 15, 20, 25, 50 or 100 mg of cyclosporin. These dosage units are suitable for administration x3, x2, x1 to x5 times daily (for example, depending on the specific purpose of treatment, stage of treatment, etc.).

Alternatively, preparations according to this invention suitable for oral administration may contain (4) water or any aqueous system to give micro-saponification systems suitable for syrup.

In addition to the above, this invention also gives a method for preparing a pharmaceutical preparation as known here before. This method includes completely mixing component (1), component (2), and component (3) as known here before, and when it is required to formulate the preparation in the form A dosage unit, for example depositing the said preparation in gelatin, for example gelatin or hard capsules.

In a more specific area, this invention provides a method for producing a pharmaceutical preparation as previously known herein in the form of ((microemulsion with initial concentration)) or microemulsion. This method includes bringing component (1), component (2) and component (3) optionally with Other special components or additives with a hydrophilic phase co-component, ethanol, to a complete mixture in proportion to the components (1), (2), (3), so that a microemulsion of initial concentration is obtained, and when required, the composition of the preparation in the form of a dosage unit or Mixing the aforementioned preparation with sufficient water or sufficient aqueous solvent medium so that an emulsion is obtained precise.

The following examples are illustrative of preparations according to the invention in the form of a dose unit suitable for use, for example, to prevent rejection of transplanted organs or to treat autoimmune disease by administering 1 to 5 dose units/day. Examples are described with specific reference to cyclosporin. In general, equivalent preparations can be obtained using any cyclosporine, specifically Ciclosporin-8[O-(2-hydroxyethyl)-(D)- Ser] (hereafter referred to as compound Z). Example 1:

Preparation &amp; Refined corn oil esterified by transesterification with glycerol &amp; Cooling the corn oil esterified by transesterification with glycerol and completely free of glycerol (as it was necessary after heating to give a clear mixture) slowly to a temperature of +20°C and kept at this temperature for one night. In the first centrifuge step, at a speed of 12,000 G and a flow rate of 103 kg/h in a continuous flow centrifuge, a liquid phase (62 kg/h), a precipitate phase (41 kg/h), and a precipitate phase (35 8 kg/h).

The liquid phase is corn oil refined by transesterification with glycerol. Alternatively, an improved product can be obtained by centrifuging in three steps, for example at +20°C, +10°C and +5°C.

The method is characterized by reducing the percentage of the mono-glyceride component in refined corn oil by transesterification with glycerol compared to the starting material (for example, 35.6% compared to 38.3%).

The optimal analytical comparison between the precipitate and the clear solution is as follows:

<img file="SA182B1_D0002.tif" />

The typical content of ingredients in the refined product can be obtained from these preparations as listed in the following table:

Component composition (w%/w)

<img file="SA182B1_D0003.tif" />

Example 2:

Preparation of oral unit dosage forms

<img file="SA182B1_D0004.tif" />

Cyclosporin is dissolved in (1) and mixed at room temperature and added (2)

(3) To the solution obtained by mixing again, the resulting mixture is deposited in

Gelatin capsules, size 1, for example, riveted. Using Qulati-Seal technology.

Formulations of 50 and 100 mg Ciclosporin are similarly prepared using the following recommended ingredients in recommended amounts:

In this example, refined oil = corn oil refined by transesterification with glycerol as described in Example 1, or Maisine, for example, Maisine is completely free of gylcerol. Formulations containing 100 mg cyclosporin, eg Ciclosporin.

<img file="SA182B1_D0005.tif" />

A preparation containing 50 mg ciclosporin

<img file="SA182B1_D0006.tif" />

Equivalent preparations can be made, as previously indicated, containing Compound Z instead of ciclosporin. Therefore, preparation W can be made containing 50 mg of compound Z instead of ciclosporin. Example 3:

Bioavailability in dogs

To compare the biopharmaceutical properties of preparations according to this

The invention is with the marketed Ciclosporin soft gelatin capsules. The preparations were compared after oral administration to 21 male Beagle dogs in a crossover design. The pharmacokinetic profile of Ciclosporin within 24 hours in the blood. Areas under the blood concentration versus time curves (AUC) were determined and Cmax and Tmx (maximum concentration with maximum time) were determined. Body: Dose of 100 mg Ciclosporin/dog

<img file="SA182B1_D0007.tif" />

<img file="SA182B1_D0008.tif" />

Total 1075 mg/dose

giving medicine:

01 male beagle dogs weighing approximately 12 kg completed the experiment successfully. Food was withdrawn twenty hours before drug administration, but the animals were allowed to drink water freely until the start of the experiment. The dose was given to animals with gastric fistula, early in the morning (approximately eight in the morning) and followed by 20 ml of 0.9% NaCl solution. After three hours, the animals were again allowed to eat food and water freely. Between every two experiments in one animal, a period of one week is allowed for purification.

Taking blood samples:

2 blood samples (1 and 5 ml for blank sample) were taken from the cephalic vein (foream) using sterile needles (diameter about 1.2 mm) and collected in 5 ml plastic tubes containing EDTA at 15 minutes, 30 minutes, 1. 1, 2, 3, 4, 6, 8, 12, and 24 hours after oral administration. Blood samples were stored at approximately -18 degrees Celsius until the drug was measured. The drug samples were analyzed by radioimmunoassay (RIA) for ciclosporin Drug concentrations in dogs are in the attached figure (2). Curves of areas under blood drug concentration against time (AUC) were calculated using the trapezoidal rule. An analysis of variance (27) was conducted and the average Tmax, Cmax, and AUC were compared statistically using Tukey’s test. The results he obtained are shown in the following table:

<img file="SA182B1_D0009.tif" />

The body weight and behavior of the animals were maintained during the study. No weight loss was observed

the body.

Conclusion:

The preparation according to this invention (Preparation 1) has a higher estimated bioavailability (a factor of 1.5) than commercial Ciclosporin soft gelatin capsules.

Figure (2) shows the average of all ciclosporin concentrations as determined by a monoclonal RIA after a single oral dose of Preparation Example 4: Bioavailability in humans:

The bioavailability of ciclosporin as determined after administration of commercial soft gelatin ciclosporin capsules, the preparation according to this invention, was compared.

Dosage form: 100 mg ciclosporin per capsule

<img file="SA182B1_D0010.tif" />

Preparation No. 8 (according to Example 2, containing refined corn oil transesterified with glycerol) in a soft gelatin capsule. Method:

Forty-eight healthy males completed the study. Each participant received four of the eight doses (two doses of preparation 8, and the same two doses of preparation x).

The participants were randomly divided into two subgroups consisting of twenty-four people, each according to a parallel design. People in group received (1)

Doses of 200 mg and 600 mg ciclosporin, and people in group (2) received

400 1 mg and 800 mg.

The trial was conducted within each group based on a four-way balanced cross-over design with a two-week purification period between each study. Samples for ciclosporin were taken in the blood one minute (s) before taking the drug and then 15 s, 30 s, 45 s, 1 h, 1.5 h, 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 6h, 8h, 20h, 12h, 16h, 20h, 24h, 28h, 32h 36 hours, 40 hours and 48 hours after taking the medicine.

Different concentrations of ciclosporin in blood were estimated for each blood sample by the RIA method. The limit of quantity was 12.5 ng/ml.

<img file="SA182B1_D0011.tif" />

The relative bioavailability of preparation 8 compared to preparation

<img file="SA182B1_D0012.tif" />

Conclusion:

The preparation according to this invention (Preparation 8) has a significantly higher bioavailability in humans than the commercial preparation (Preparation X) by at least 1.7 times.

Attached Figure 3 gives a plot of the average AUC (0-48h) values for Preparation .

The degree of absorption of the preparation 8 (in terms of AUC(0-48h) values) appears to be unsupported

depends on the dose, while the degree of absorption of preparation X decreases with increasing dose (see Figure 3).