CZ233799A3 - Peroral pharmaceutical preparations of cyclosporin - Google Patents

Abstract

Oral Cyclosporinocide with Improved Biological the availability of cyclosporine containing cyclosporin is at least one C 1-4 alkanol solvent, at least one nonionic solvent a surfactant selected from the group consisting of polyoxyethylene alcohols of monoester fatty acids ethoxylated C ^ poly polyols; and at least one polyglycol molecular weights in the range of 800 to 10,000 IU.

Description

Technical field

The present invention relates to oral cyclosporin pharmaceutical preparations.

BACKGROUND OF THE INVENTION

Despite efforts to prevent implant rejection upon donor tissue contact with the host, in most transplants where the donor organ is introduced into the host, immunosuppressive therapy plays a critical role in maintaining the donor organ viability in the host. Many immunosuppressive agents such as azathioprine, methotrexate, cyclophosphamide, FK-506, rapamycin and corticosteroids have been used in transplants. Substances of increasing importance in immunosuppressive therapy due to the preferential effect on T-cell mediated reactions are cyclosporins.

Cyclosporins belong to the group of cyclic polypeptides containing eleven amino acids and are metabolic products of the fungus Tolypocladium Inflatum Gams. It has been observed that cyclosporins reversibly inhibit immunocompetent lymphocytes, particularly T cells in the Gq or Gj phase of the cell cycle. It has also been observed that cyclosporins reversibly inhibit the production and release of lymphokines. Although a number of cyclosporins are known, Cyclosporin A has the most widespread use.

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Cyclosporin A has been reported to prolong the persistence of allogeneic transplants of skin, heart, kidney, pancreas, bone marrow, small intestine and lung. In allogeneic transplants, cyclosporin A suppresses humoral immunity and largely cellular immune response, such as allograft rejection, prolonged hypersensitivity, experimental allergic encephalomyelitis, Freund's ajuvant arthritis, and implant versus disease. host. Although significant success has been achieved with Cyclosporin A, drug administration after transplantation must continue because the benefits of cyclosporine therapy are reversible and implant discontinuation upon discontinuation of treatment.

Although ciclosporin formulations have been developed for both oral and intravenous administration, oral administration is preferred for both ease of administration and better drug acceptance. In addition, intravenous administration of ciclosporin may lead to anaphylactic reactions, a side effect that is not likely to occur with oral administration. Currently, oral cyclosporin preparations are available in soft capsule and solution form and are sold under the trade names SANDIMMUNE and NEORAL ™.

In oral therapy with ciclosporin products, the medical staff and the manufacturer must be aware of many problems. The bioavailability of ciclosporin in oral preparations is limited because ciclosporin is insoluble in water and tends to crystallize in an aqueous environment. The concentration of cyclosporin in oral preparations is limited due to the hydrophobic nature of the cyclosporin molecule. Another problem is stability when adjusting dosage forms and storing oral preparations. Soft gelatin capsules of cyclosporin preparations must be airtight, which is both space-intensive and expensive. Cyclosporin formulations are unstable at low temperatures as ciclosporin crystallizes.

Therefore, it is still desirable to provide cyclosporin formulations that address at least some of the above problems. An ideal oral formulation should provide high bioavailability

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· Cyclosporin, a high concentration of cyclosporin and should allow the formation of liquid drug forums and hard capsules.

Relevant literature:

Physicides Desk Reference (1994), pp. 2071-2074 describes oral cyclosporin preparations currently marketed under the trade name SANDIMMUNE.

Oral cyclosporin preparations are also disclosed in NEORAL, (1995) (Sandoz Pharmaceuticals Corporation, East Hanover, New Jersey, 07936).

U.S. Pat. patents dealing with cyclosporins and derivatives thereof are 4,220,641; 4,639,434; 4,289,851 and 4,384,996. No. 5,047,396 discloses an intravenous preparation of cyclosporin. U.S. Pat. 4,388,307; Nos. 4,970,076 and 4,990,337 disclose oral cyclosporin preparations.

The preparation of oral hard capsules is described in U.S. Pat. 4,822,618; 4,576,284; 5,120,710 and 4,894,235.

SUMMARY OF THE INVENTION

The invention provides oral cyclosporin pharmaceutical compositions and methods for their use in immunosuppressive therapy. In the present compositions, the cyclosporin is present in an orally acceptable carrier comprising at least one C 2-3 alkanol solvent and at least one nonionic surfactant. The present pharmaceutical compositions may further comprise one or more additional solvents which are esters of fatty acids and diols, preferably polyglycols. Cyclosporin preparations can be prepared in the form of hard capsules. The formulations are essentially anhydrous, which reduces the tendency of cyclosporin to clot and increases its bioavailability.

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Brief description of the pictures

Giant. 1 depicts the maximum cyclosporin concentration (C _max ) achieved in rats when administering several oral formulations of the present invention, C _max is the relative value relative to C _max for SANDIMMUNE ORAL (SO).

Giant. 2 shows the time taken for C _max (T _max ) for the individual formulations shown in FIG. 1, T _max is the relative value relative to T _max for SANDIMMUNE ORAL (SO).

Giant. 3 shows the relative areas under the concentration-time curve (AUC) for the individual formulations shown in FIG. 1, the AUC is the relative value relative to the AUC for SANDIMMUNE ORAL (SO).

Giant. 4 shows the maximum cyclosporin concentration (C _max ) achieved in humans by administering several oral formulations of the present invention, as well as C _max for SANDIMMUNE ORAL (SO) in solution (Sand).

Giant. 5 shows the time taken for C _max (T _max ) for the individual formulations shown in FIG. 4.

Giant. 6 shows the relative areas under the concentration-time curve (AUC) for the individual formulations shown in FIG. 4.

Description of a specific embodiment of the invention

The present invention provides oral cyclosporin pharmaceutical compositions with improved bioavailability that can be prepared

- 5 in the form of capsules, in particular hard capsules. In the present composition, the cyclosporin is present in an orally acceptable carrier comprising at least one C 2-3 alkanol solvent and at least one nonionic surfactant. The present pharmaceutical compositions may further comprise one or more additional solvents which are esters of fatty acids and diols, preferably one or more polyglycols. All components of the present composition are pharmaceutically acceptable. In addition to the high bioavailability, different batches of the present compositions guarantee reproducible absorption. The present compositions are used in immunosuppressive therapy.

There are a large number of cyclosporins with immunosuppressive effects that can be administered in oral preparations. These are cyclosporins of the types: Cyclosporin A, Cyclosporin B, Cyclosporin C, Cyclosporin D and Cyclosporin G, as well as their synthetic analogues. See Merck Index (1989) 2759. The present oral formulations are particularly preferred for the delivery of Cyclosporins A. When administered, the concentration of Cyclosporins is in the range of 50-150mg / ml, usually 100-150mg / ml, based on the volume of the carrier components of the formulation.

The carrier of the formulation comprises the components: an alkanol solvent consisting of up to three different C2-3 alkanols bearing 1-2 hydroxyl groups so that no more than 1 hydroxyl group falls on 1.5 carbon atoms. Suitable alkanols are ethanol and propylene glycol, preferably ethanol. The total amount of alkanol solvent is at least 1% by volume, usually 3% by volume, up to 95% by volume, usually in the range of 5 to 75% by volume, more often 5 to 60% by volume, even more often 10 to 60% by volume due to the volume of the preparation. When the alkanol solvent is ethanol, it is present in an amount of 5 to 25 vol%, usually 5 to 20 vol%, and more often 10 to 25 vol%, based on the volume of the formulation. When the alkanol solvent is propylene glycol, it is present in an amount of 5 to 90% by volume, usually 5 to 85% by volume, and more often 10 to 50% by volume, based on the volume of the formulation.

The orally acceptable carrier of the pharmaceutical composition also comprises at least one nonionic surfactant, more often not.

- more than two nonionic polyoxyalkylene surfactants. The polyoxyalkylene surfactants have a hydrophilic-lipophilic balance (HLB) in the range of 5 to 20, usually 8 to 16. The nonionic polyoxyalkylene surfactants in the present composition are preferably polyoxyethylene compounds. Suitable polyesters of the oxyethylene compound are ethoxylated alcohols, i.e. polyoxyethylene alcohols or fatty polyoxyethylene alcohols, wherein the alcohol moiety typically contains 10 to 18, more typically 10 to 14 carbon atoms, ether and ester substitution derivatives thereof; and polyoxyethylene derivatives of partial esters of fatty acids, usually monoesters and C4-6 polyols, usually C8 polyols, wherein the polyol may be polyol anhydrides such as sorbitan. The fatty acid moiety typically contains 10 to 18 carbon atoms. The number of ethylene oxide groups ranges from 2 to 30, usually from 2 to 25. Preferred surfactants are polyoxyethylene (4) lauryl ether (BRIJ 30) and polyoxyethylene (20) mono sorbitan monooleate (TWEEN 80). The total amount of nonionic surfactants in the present composition is 5 to 80 vol%, usually 5 to 70 vol%, more often 5 to 65 vol%, preferably 5 to 60 vol%, more preferably 20 to 75 vol%, and often 50 to 80% by volume of the formulation. When TWEEN 80 is included in the formulation, it is usually 5 to 80% by volume, more often 5 to 70% by volume, preferably 5 to 60% by volume, and more preferably 10 to 50% by volume of the formulation. If BRIJ 30 is included in the formulation, it is usually 10 to 45% by volume, more often 15 to 40% by volume.

Optionally, the present compositions further comprise one or more additional solvents, usually no more than three different solvents, more typically no more than two different solvents. Suitable co-solvents include fatty acid esters and diols, wherein the co-solvent comprises 100% fatty acid ester or 100% diol or a combination thereof. In formulations containing only a fatty acid ester or only a diol, it may be a combination of different fatty acid esters or a combination of different diols. The total amount of co-solvents in the formulation is in the range of 20 to 80% by volume, usually 25 to 75% by volume. If the co-solvents are present in the formulation, the ratio of co-solvents to solvents is usually in the range 1: 1 to 15: 1. more often 1: 1 to 13: 1.

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The fatty acid esters that may serve as additional solvents typically contain a C 12-18 carbon chain, more often ¢ 443, are usually mono-esters of fatty acids and lower alkanols. Suitable fatty acid esters usually contain an even number of carbons in the chain, the chain being saturated or unsaturated, but usually has no more than two unsaturated positions. The fatty acids are usually of plant or animal origin, for example palmitic, stearic, palmitoleic, linoleic, linolenic and the like, especially myristic and oleic acids. The alcohol part of the fatty acid ester is usually a lower alkanol C2-4, usually C2-3, branched or unbranched. Suitable fatty esters are isopropyl myristate and ethyl oleate. When isopropyl myristate is present, it is present in an amount of 15 to 75% by volume, ethyl oleate is present in an amount of 15 to 75% by volume, based on the volume of the formulation. The volume fraction of the fatty acid ester to the surfactant in the formulation is in the range of 1: 1 to 8: 1, usually not exceeding 5: 1. The fatty acid ester is preferably anhydrous.

In addition to the additional fatty acid esters, the present compositions further optionally comprise diols. The diols are preferably anhydrous. Diols as additional solvents are usually in liquid phase at physiological temperatures, such as diols, often C 18-20 diols, diols may be polyoxyalkylenediols containing C 2-3 alkylene, but diols containing substantially more carbon atoms may also be useful. The size of the diol molecules suitable as co-solvents is usually in the range of 200 to 10,000 hU, often 300 to 10,000 hU, more often 400 to 10,000 hU, preferably 800 to 10,000 hU. Particularly preferred diols are the ethyl englycols, especially polyethylene glycol 200 (PEG200), polyethylene glycol 300 (PEG300), polyethylene glycol 400 (PEG400), polyethylene glycol 600 (PEG 100X polyethylene glycol 1000 (PEG 1000f polyethylene glycol 3400 (PEG340Q), polyethylene glycol 8000 (PEG 100)) and the like. Diols, especially higher molecular weight polyethylene glycols (e.g. polyethylene glycols having a molecular weight of 1000 or more), act as water absorbents in the composition, and are present in the composition, forming 5 to 60% w / v diols, more often 5 to 55% w / v. compositions, thereby substantially reducing the possibility of precipitation of cyclosporin from the composition caused by free water molecules.

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The co-solvents further impart suitable physical properties to the formulation such as viscosity, stability and the like. Optionally, the composition further comprises other agents affecting its physical properties such as thickeners, suspending agents, solidifying agents, and the like, including agents such as acacia resin, carboxymethylcellulose, hydroxypropyl cellulose, lecithin, methylcellulose, high molecular weight polyethylene glycols (e.g. 1000 to 10,000 IU, often 1000 to 8,000 IU, more often 1000 to 6000 IU), povidone, sodium alginate, tragacanth and the like. The present compositions also contain minor ingredients providing various functions such as enzyme inhibitors, preservatives, antioxidants, antimicrobial agents, stabilizers, flavorings, etc. The total amount of thickeners and other additives usually does not exceed 55% by weight, often 45% by weight, more often 25% by weight. . preparation. The formulations further comprise various excipients, as is conventional in the art. All of the ingredients of the present composition are preferably anhydrous.

The substantially anhydrous formulations are those containing less than 1.0 vol% water, more preferably less than 0.1 vol% water.

The present formulations are stable over a wide range of temperatures, the stability of the formulation being understood to mean its physical integrity, i.e., there is no crystallization of the cyclosporin active ingredient from the formulation. A stable temperature range includes low storage temperatures, typically 0-15 ° C, more typically 2-8 ° C.

The present compositions may be administered in liquid form as well as in capsules, such as hard and soft capsules. Methods for preparing hard capsules containing a liquid formulation are known in the art, e.g. Nos. 4,822,618 and 4,576,284, incorporated herein by reference. Generally, hard capsules suitable for the present purposes consist of two parts: a capsule and a cap. The cap can be tightly closed by the cap and the whole creates a space of precisely defined volume. The capsule and cap are made of a hydrophilic polymer such as starch or gelatin. In preparing hard capsules, the liquid preparation is poured into the capsule and the cap is closed with a cap. The closure must be tight to prevent leakage

The capsule formulation, see E.P., &lt; tb &gt; ______________________________________ &lt; tb &gt; 116,744, incorporated herein by reference. The capsules must also remain stable in the acidic environment of the stomach and must therefore be coated with an enteric coating to avoid degradation. Various enteric coatings are known in the art, see e.g. No. 5,206,219, incorporated herein by reference.

The present compositions can be administered to patients in conjunction with other immunosuppressive agents that can be administered with the cyclosporin compositions of the present invention. These include, for example, rapamycin, FK-506, mycolenolic acid, analogs and derivatives thereof, azathioprine, methotrexate, cyclophosphamide, corticosteroids, and other immunosuppressants and analogues thereof.

The present compositions have utility in immunosuppressive therapy. Immunosuppressive therapy is indicated for a wide range of diseases such as idiopathic nephrotic syndrome, insulin-dependent type I diabetes, Behcet's syndrome, active Crohn's disease, aplastic anemia, advanced corticosteroid-dependent asthma, psoriasis, rheumatoid arthritis, and other diseases where the immune system it can play a pathogenic role. Of great interest is the use of the present compositions in transplants of both allogeneic and xenogeneic organs, tissues or cells where immunosuppression is desirable in order to maintain the viability of the transplanted organ, tissue and cells after trasplantation, i.e. immunosuppression prevents implant rejection or implant vs. disease disease. a host, e.g., after a bone marrow transplant.

Using the present compositions for immunosuppressive therapy, an effective amount of cyclosporin is orally delivered to the patient to achieve the desired level of immunosuppression, according to the particular condition being treated. In transplants, the initial dose of ciclosporin is usually given before or after surgery. Following transplantation of the donor organ to the host, cyclosporin is administered repeatedly, i.e., chronically, to the patient to maintain immunosuppression. The initial dose is administered 4 to 12 hours prior to transplantation in an amount of 5 to 18 mg / kg body weight, usually 5 to 15 mg / kg. After surgery, usually the same daily doses are administered for a further 1 to 3 weeks, usually 1 to 2 weeks. Later, doses are reduced to 3 to 10 mg / kg daily, usually 3 to 10 mg / kg / day, usually 3 to 10 mg / kg daily, usually 3 to 10 mg / kg.

- 10φφ φφ up to 6 mg / kg. The rate of dose reduction to maintenance levels is 3 to 8% per week, usually 5% per week. Dosage is determined by blood level to maintain a concentration of 100 to 350 ng / ml by HPLC, RIA, ELIS A or TDx assays. The present compositions may be co-administered with other agents where such co-treatment is recommended and known in the art. For example, the present compositions may be administered together with adrenocorticosteroids, azathioprine, and the like.

Administration of the present compositions in transplantation results in prolonged viability of the transplanted organ in the recipient, by suppressing the host's immune response to the donor organ. By prolonging viability, it is meant that the donor organ remains functional in the recipient's body for a longer period of time than would be the case without immunosuppressive therapy. Extending viability means maintaining viability indefinitely. A donor organ can be considered viable as long as it remains functional in the host's physical environment.

The present compositions can also be prepared as aqueous colloidal dispersions of cyclosporin nanoparticles that retain good bioavailability. In this case, the nanoparticles are substantially spherical, the cyclosporin is contained in them in an amorphous form, and their average size is usually less than 1000 nm, greater than 50 nm, usually in the range of 200 to 800 nm, often 200 to 600 n. 50 wt. The particle size must be within the specified size range. Larger particles are present as nanoparticle aggregates with an average diameter of less than 50 µm, more often less than 25 µm. Aggregates generally do not comprise more than 40 wt. of the total amount of cyclosporin.

The formulation must contain sufficient amorphous cyclosporin particles for the desired therapeutic effect. Since the preparation is only formed in an aqueous environment prior to administration, or directly in the digestive juices, the concentration cannot be determined because no stomach dilution is known. Formulations ⁷ diluted prior to oral administration typically contain from 0.01 to 2.5% by weight. cyclosporin, more often

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0.01 to 0.5 wt. The stirring temperature is in the range of 10 ° C to 20 ° C, usually 20 ° C to 40 ° C. Stirring is usually performed for a sufficient time to form a solution.

The colloidal amorphous nanoparticle suspension is sufficiently stable to allow some delay before administration, usually within 6 h, more often 3 h.

Further details regarding the preparation and use of nanoparticulate cyclosporin formulations can be found in PCT / US97 / 04627.

The following examples are for illustrative purposes only and are not to be construed as limiting the invention.

DETAILED DESCRIPTION OF THE INVENTION

Several cyclosporin formulations of the present invention have been prepared. The bioavailability of ciclosporin in these formulations was studied in both rats and humans.

Oral cyclosporin preparations

The following Cyclosporin A preparations were prepared. In all formulations, a specified amount of Cyclosporin A, a surfactant, and ethanol or propylene glycol were placed in a 1 L volumetric flask. The final volume was supplemented to 1 L with an appropriate volume of fatty acid ester and / or diol.

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Preparation Ingredients 19 Dec CsA EtOH Tween 80 IM 100 mg 0.1 ml 300 mg to 1 ml (10% w / v) (10%) (0.278 ml) <(0.622 ml) (531 mg) 20 May CsA EtOH Brij 30 IM 100 mg 0.05 ml 350 mg to 1 ml (10% w / v) (5%) (0.368 ml) <(0.582 ml) (496 mg) 21 CsA PG Brij 30 IM 100 mg 0.05 ml 350 mg to 1 ml (10% w / v) (5%) (0.368 ml) <(0.582 ml) (496 mg) 22nd CsA EtOH Tween 80 EO 100 mg 0.1 ml 300 mg to 1 ml (10% w / v) (10%) (0.278 ml) <(0.622 ml) (541 mg) 23 CsA EtOH Brij 30 EO 100 mg 0.05 ml 350 mg to 1 ml (10% w / v) (5%) (0.368 ml) <(0.582 ml) (506 mg) 24 CsA PG Brij 30 EO 100 mg 0.05 ml 350 mg to 1 ml (10% w / v) (5%) (0.368 ml) <(0.582 ml) (506 mg)

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Preparation Ingredients 33 CsA EtOH Brij 30 100 mg 0.1 ml 350 mg (10% w / v) (10%) (0.368 ml) IM to 1 ml <(0.742 ml) (633 mg) 34 CsA EtOH Brij 30 EO 100 mg 0.1 ml 150 mg to 1 ml (10% w / v) (10%) (0.158 ml) <(0.742 ml) (646 mg) 35 CsA EtOH Tween 80 PG 100 mg 0,105 ml 500 mg to 1 ml (10% w / v) (10.5%) (0.463 ml) <(0.437 ml) (453 mg) 36 CsA EtOH Tween 80 PG EO 100 mg 0.1 ml 300 mg 100 mg to 1 ml (10% w / v) (10%) (0.278 ml) (0.097 ml) <(0.525 ml) (465 mg) 37 CsA EtOH Tween 80 PEG 400 EO 100 mg 0.1 ml 300 mg 100 mg to 1 ml (10% w / v) (10%) (0.278 ml) (0.088 ml) <(0.534 ml) (464 mg)

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Preparation Ingredients 38 CsA 100 mg (10% w / v) EtOH 0.1 ml (10%) Brij 30 300 mg (0.316 ml) PG 100 mg (0.097 ml) EO to 1 ml <(0.487 ml) (424 mg) 39 CsA 100 mg (10% w / v) EtOH 0.1 ml (10%) Brij 30 300 mg (0.316 ml) PG 200 mg (0.193 ml) EO to 1 ml <(0.391 ml) (340 mg) 40 CsA 100 mg (10% w / v) PG 300 mg (0.290 ml) Brij 30 300 mg (0.316 mi) EO to 1 ml <(0.394 ml) (343 mg) 41 CsA 100 mg (10% w / v) EtOH 0.05 ml (5%) Brij 30 150 mg (0.158 ml) Tween 80 100 mg (0.093 ml) EO to 1 ml <(0.649 ml) (565 mg) 42 CsA 100 mg (10% w / v) PG 0.05 ml (5 wt.%) Brij 30 150 mg (0.158 ml) Tween 80 100 mg (0.093 ml) EO to 1 ml <(0.649 ml) (565 mg)

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Preparation Ingredients 44- fluid CsA EtOH Tween 80 peg ₄₀₀ 100 mg 0.125 ml 400 mg to 100 ml (10% w / v) (12.5%) (0.371 ml) <(0.529ml) (601mg) 44-hard CsA 100 mg (10% w / v) capsule EtOH Tween 80 PEG400 PEG1000 0.125 ml 400 mg 0.25 ml 0.25 ml (12.5%) (0.371 ml) 45 CsA EtOH Tween 80 100 mg 0,10 ml 300 mg (10% w / v) (10%) (0.278 ml) PG about 250 mg (0.243 ml) PEG400 about 250 mg (0.220 ml) 46 CsA EtOH Tween 80 PG 100 mg 0.10 ml 100 mg to 1.0 ml (10% w / v) (10%) (0.093 ml) (0.807 ml) 48 CsA EtOH Tween 80 100 mg 0,10 ml 200 mg (10% w / v) (10%) (0.278 ml) PG about 250 mg (0.243 ml) peg ₄₀₀ about 250 mg (0.220 ml)

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Preparation Ingredients 49 CsA EtOH Tween 80 PG 100 mg 0,10 ml 600 mg to 1.0 ml (10% w / v) (10%) (0.558 ml) (0.342 ml) 50 CsA EtOH Tween 80 PG 100 mg 0,10 ml 300 mg to 1.0 ml (10% w / v) (10%) (0.278 ml) (0.622 ml) 51 CsA EtOH Tween 80 PG 100 mg 0,10 ml 200 mg to 1.0 ml (10% w / v) (10%) (0.186 ml) (0.714 ml) 52 CsA EtOH Tween 80 PG 100 mg 0.05 ml 400 mg to 1.0 ml (10% w / v) (5%) (0.371 ml) (0.579 ml) 100 ALIGN! CsA EtOH Tween 80 peg ₃₀₀ PEG 100 125 mg 0.125 ml 200 mg about 0.44 mg about 0.11 mg (12.5% w / v) (12.5%) (0.186 ml) 101 CsA EtOH Tween 80, PEG _300, PEG ₈ ooo 125 mg 0.125 ml 100 mg about 0.52 mg about 0.13 mg (12.5% w / v) (12.5%) (0.093 ml)

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Preparation Ingredients 102 CsA 125 mg (12.5% w / v) EtOH 0.125 ml (12.5%) Tween 80 100 mg (0.093 ml) peg ₃₀₀ about 0.585 mg ΡΕθδοοο about 0.065 mg 103 CsA 125 mg (12.5% w / v) EtOH 0.125 ml (12.5%) Tween 80 400 mg (0.372 ml) PEG ₃₀₀ about 0.28 mg PEG ₃ 4oo about 0.07 mg 104 CsA 125 mg (12.5% w / v) EtOH 0.15 ml (15%) Tween 80 100 mg (0.093 ml) peg ₃₀₀ about 0.496 mg PEGgooo about 0.124 mg 105 CsA 125 mg (12.5% w / v) EtOH 0.15 ml (15%) Tween 80 400 mg (0.372 ml) PEG ₃₀ o about 0.256 mg PEG _34O o about 0.064 mg 106 CsA 125 mg (12.5% w / v) EtOH 0.125 ml (12.5%) Tween 80 400 mg (0.372 ml) peg ₃₀₀ about 0.329 mg PEG ₈ ooo about 0.021 mg

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Preparation Ingredients 107 CsA EtOH Tween 80 112 mg 0.158 ml 800 mg (11.2% w / v) (15.8%) (0.744 ml) 108 CsA 112 mg (11.2% w / v) EtOH 0.158 ml (15.8%) Tween 80 Peg ₃₀₀ PEGgooo 700 mg about 0.105 mg about 0.045 mg (0.651 ml) 109 CsA 112 mg (11.2% w / v) EtOH 0.158 mi (15.8%) Tween 80 Peg ₃₀₀ PEGgooo 700 mg about 0.075 mg about 0.075 mg (0.651 ml) 110 CsA 112 mg (11.2% w / v) EtOH 0.158 ml (15.8%) Tween 80 Peg ₃₀₀ PEGgooo 600 mg about 0.20 mg about 0.05 mg (0.558 ml) 111 CsA 112 mg (11.2% w / v) EtOH 0.158 ml (15.8%) Tween 80, PEG _300, PEG ₈ ooo 600 mg about 0.15 mg about 0.10 mg (0.558 ml)

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Preparation Ingredients 112 CsA EtOH Tween 80 peg ₃₀₀ PEG 500 112 mg 0.158 ml 500 mg about 0.315 mg about 0.035 mg (11.2% w / v) (15.8%) (0.465 ml) 113 CsA EtOH Tween 80 peg ₃₀₀ PEG ₃₄ 112 mg 0.158 ml 500 mg about 0.245 mg about 0.105 mg (11.2% w / v) (15.8%) (0.465 ml) 114 CsA EtOH Tween 80 PEG _{3 and} PEG ₈ ooo 125 mg 0.158 ml 300 mg about 0.385 mg about 0.165 mg (12.5% w / v) (15.8%) (0.279 ml) 115 CsA EtOH Tween 80 peg ₃₀₀ PEG 100 125 mg 0.158 ml 300 mg about 0.33 mg about 0.22 mg (12.5% w / v) (15.8%) (0.279 ml) 116 CsA EtOH Tween 80 peg ₃₀₀ PEG 500 125 mg 0.158 ml 400 mg about 0.225 mg about 0.225 mg (12.5% w / v) (15.8%) (0.372 ml)

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PG = propylene glycol; EtOH = ethanol; Brij 30 = polyoxyethylene (4) lauryl ether; Tween BO polyoxyethylene (20) monosorbitan monooleate; IM = isopropyl myristate; EO = ethyl oleate

2. Bioavailability studies for formulations 19-24 and 33-42

The bioavailability of ciclosporin in formulations 19-24 and 33-42 was studied. The following pharmacokinetic parameters were determined as a measure of bioavailability: (a) maximum cyclosporin blood concentration (C _max ); (b) the time taken to reach C _max (T _max );

(c) the relative area under the concentration-time curve (AUC). Except for Formulations 19-24 and 33-42, the same determinations were made for cyclosporin in SANDIMMUNE Oral Solution (SO) for comparative purposes. Experimental animals were prepared for testing the above preparations. Sprague Dewley rats not yet in contact with CsA weighing 250-350 g were fed a pelletized standard diet (Agway 3000, Granviile Mili, Greensboro, NC) and water ad libitum. One day before the experiment, a rubber cannula was inserted into the right jugular and right femoral vein under light ether anesthesia. After overnight interruption of food supply, CsA was administered via a dosing device.

After administration, 200 z / l jugular vein blood samples were collected into 0.5 ml polypropylene microcentrifuge tubes containing 0.3 mg lyophilized EDTA / Na and heated for 10 s. Blood samples from animals treated with the present compositions were collected at 0.5, 1, 2, 4, 8, 12, 24, 36, 48 and 72 hours after administration.

CsA, including some metabolites, was determined in the blood by a fluorescent polarization immunoassay (FPI) (TDx, Abbot Lab). Briefly, 150 L of blood sample was quantitatively transferred to 1.5 ml microcentrifuge tubes. Cells were lysed and dissolved with 50 L of surfactant-containing solubilizing agent. The proteins were then precipitated with 300 µl of acetonitrile. After centrifugation, the FPI test on the TDx Autoanalyzer was performed on the supernatant according to the procedure described in FIG

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99 Abbott Diagnostics recommendations. Since the TDx assay was originally developed for human blood, some procedures have been modified. A number of standard solutions of known CsA concentration were prepared by adding a known amount of CsA to EDTA-reacted rat blood. Where it was expected that the CsA concentration would exceed 1.0 Mg / ml, the samples 10x were diluted with 0.1 mol.H phosphate buffer pH 7.0. For diluted samples, i.e. 10% rat blood and 90% phosphate buffer, another calibration curve was plotted using a series of standard solutions containing a known amount of CsA.

Pharmacokinetic parameters were obtained from non-compartmental analyzes. The maximum concentration (C _max ) and the time to reach it (T _max ) were determined by assessing the concentration-time profile of concentration for each rat. The area under the concentration-time curve (AUC) from time 0 to the end of the measurement (AUC () - t) was calculated by the linear trapezoidal projection method. The residual area under the end of the curve (AUCq.qq) was determined as the ratio of the last observed concentration (C) to the first order rate constant describing the terminal elimination phase of the concentration profile (λ _ζ ). The rate constant λ _ζ was determined from the log-linear regression of the concentration-time data (ie the last 3 to 5 data according to the analyzed profile). The total area (AUC ^ QQ) was calculated as the sum of AUC t and AUC _t · _oo

The results for each formulation were compared to those obtained for SO and are shown in Figures 1-3. The results show that most preparations lead to higher bioavailability of cyclosporins over the comparative SANDIMMUNE Oral Solution (SO), as indicated by higher AUC values for the present preparations.

3. Bioavailability studies for formulations 35, 43-46 and 48-52 in humans

Testing was performed on a group of 48 healthy men aged 19 to 55 years with a weight differing by a maximum of 20% from the ideal weight. Single-dose, non-food, random, double-dose studies were conducted

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9 9 9 9 9 ft · · * · <ftft * ft · ft * 4 «·· blind and triple crossed. 48 men were randomly divided into 6 groups of

8. Each group received one 300 mg dose of ciclosporin from the above, or SANDIMMUNE Oral Solution (SO), on three different occasions, after a seven-day break.

Test subjects were asked not to eat 10 h before and 4 h after administration. They could drink water to taste except 1 hour before and 2 hours after administration. 15 ml blood samples were taken prior to administration. The following doses were prepared for administration: 3 ml aliquots of the preparation (300 mg) were mixed with 200 ml diluent and administered orally. 10 ml blood samples were taken at time t = 0, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 16, 20 and 24 h. ml blood sample.

Cyclosporin A concentration in total blood was determined by TDx (Abbott Diagnostics, N. Chicago, IL) according to the manufacturer's instructions.

Non-compartmental pharmacokinetic analysis was performed using standard procedures. The maximum concentration (C _max ) and the time to reach it (T _max ) were determined from the concentration time course data. The area under the concentration-time curve (AUC) was calculated by the linear trapezoidal projection to the last concentration point at the sensitivity limit (25 ng / ml) and extrapolated to infinity. The observed C _max , T _max and AUC values were averaged. Mean values for each formulation are shown in Figure 4-6. The results show that for each test preparation C _max was achieved twice as fast as for the comparative SANDIMMUNE (SO). The observed AUC of the test products was at least 2000 ng h / ml higher than that of SANDIMMUNE (SO) under the same conditions. It follows that formulations 35, 43-46 and 48-52 provide better bioavailability than SANDIMMUNE Oral Solution (SO).

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Physical characteristics of liquid formulations 35 and 44 at low temperatures.

The liquid preparations of cyclosporin 35 and 44 and the comparative NEORAL Oral Solution were stored at different temperatures and their physical properties determined. In particular, the liquid formulations of Cyclosporin A 35 and 44 and NEORAL Oral Solution were stored overnight at 10 ° C and 2 to 8 ° C. formulation 44 and NEORAL solidified, formulation 35 not, only viscosity increased. Chilled formulation 35 showed no signs of cyclosporine precipitation. After removal of the formulations into the ambient temperature environment, formulation 44 thawed earlier than the comparative NEORAL Oral Solution.

During malting at 2 to 8 ° C, NEORAL Oral Solution solidified into a paste, preparations 35 and 44 remained in the form of viscous liquid preparations and showed no signs of cyclosporin A precipitation.

From these results and considerations, it is apparent that the novel cyclosporin formulations provide high bioavailability. Despite the high concentration of cyclosporin, they are stable when stored at low temperatures in freezers. The present compositions may be administered in the form of capsules, including hard capsules, for ease of storage and use.

All publications and patent applications cited in this specification are incorporated by reference.

Although the present invention has been illustrated in more detail by way of example for clarity and understanding, it will be apparent to those skilled in the art that certain changes or modifications may be made within the scope of the present disclosure without departing from the spirit or scope of the claims.

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-24Industrial applicability

The invention provides pharmaceutically useful oral cyclosporin formulations and methods for their use in immunosuppressive therapy. Due to suitable ingredients, the present compositions are characterized by high stability and bioavailability of cyclosporine.

Claims (15)

PATENT CLAIMS An oral cyclosporin preparation comprising: cyclosporin; at least one C 2-3 alkanol solvent at least one nonionic surfactant selected from the group consisting of polyoxyethylene alcohols and monoesters of fatty acids with ethoxylated C 4.5 polyols; and at least one polyglycol having a molecular weight in the range of 800 to 10,000 h. The composition of claim 1, wherein the polyglycol comprises polyethylene glycol. A formulation according to claim 2, wherein the alkanol solvent comprises 5 to 75% v / v. preparation. The composition of claim 2, wherein the alkanol solvent is 10-25% v / v. preparation. The formulation of claim 4, wherein the alkanol solvent is absolute ethanol. - 266. The composition of claim 2, wherein said at least one nonionic polyoxyalkylene surfactant is from 5 to 80% w / v. preparation. The composition of claim 2, wherein said at least one nonionic polyoxyalkylene surfactant is from 10 to 50% w / v. preparation. 8. The composition of claim 2, wherein said at least one nonionic polyoxyalkylene surfactant is from about 20% to about 75% v / v. preparation. The composition of claim 2, wherein said at least one nonionic polyoxyalkylene surfactant is selected from the group consisting of polyoxyethylene (20) mono sorbitan monooleate and polyoxyethylene (4) lauryl ether. The composition of claim 2, wherein said at least one polyethylene glycol is a polyethylene glycol having a molecular weight in the range of 1000 to 8000 i. . 11. The composition of claim 2 wherein said at least one polyethylene glycol is a mixture of two different polyethylene glycols of different molecular weights. Composition according to claim 2, characterized in that said at least one polyethylene glycol constitutes 20 to 80% w / v. preparation. A formulation according to claim 1 wherein the concentration of cyclosporin is 100 mg / mL, absolute ethanol is 12.5% v / v, and polyoxyethylene (20) mono sorbitan monooleate 400 mg / mL. • · ·· -π 14. A formulation according to claim 1, characterized in that it is adjusted in hard capsules. 15. An oral cyclosporin preparation comprising cyclosporin and a pharmaceutically acceptable carrier, which is substantially anhydrous. Use of an oral cyclosporin preparation according to any one of claims 1 to 15.