Solid dispersions comprising tacrolimus

[0001] The present invention relates to a solid solution containing tacrolimus, with increased bioavailability, in particular a solid solution of tacrolimus in a hydrophilic carrier; a pharmaceutical composition containing a solid solution, and dosage forms containing a solid solution.

BACKGROUND OF THE INVENTION [0002] Tacrolimus, also known as FK-506 or FR-900506, has a chemically tricyclic structure, as follows:

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corresponding to C44H69NO12. Tacrolimus is in the form of white crystals or crystalline powder. It is practically insoluble in water, freely soluble in ethanol and well soluble in methanol and chloroform.

[0003] The production of tacrolimus is described in EP-A-0184162, and tacrolimus analogues are disclosed e.g. in EP-A-0444659 and US 6,387918.

[0004] Tacrolimus is a macrolide compound with useful immunosuppressive, antimicrobial and other pharmacological effects and is useful in the treatment or prophylaxis of rejection reactions in organ and tissue transplantation, graft versus host diseases, autoimmune diseases and infectious diseases. Tacrolimus prolongs host and transplant survival in animal models of liver, kidney, heart, bone marrow and small intestine and pancreas, lungs and trachea, skin, cornea and limb transplants.

[0005] In animals, tacrolimus has been shown to inhibit to some extent humoral resistance and, to a significant extent, cell-mediated responses such as allograft rejection, delayed type hypersensitivity, collagen-induced arthritis, experimental allergic encephalomyelitis and disease. graft versus host.

[0006] Tacrolimus inhibits T cell activation, although the exact mechanism of action is unknown. Experimental evidence suggests that tacrolimus binds to intracellular FKBP-12 protein. The tacrolimus-FKBP-12, calcium, calmodulin and calcineurin complex is then formed and the phosphatase activity of calcineurin is inhibited. This action can prevent the dephosphorylation and translocation of the nuclear factor of activated T cells, a nuclear component that is thought to initiate transcription of the lymphokine forming gene. As a result, T lymphocyte activation is inhibited, i.e., immunosuppression.

[0007] Tacrolimus is significantly metabolised by CYP3A4 in the intestinal wall and liver. Therefore, drugs that affect this isoenzyme may affect the absorption and subsequent removal of systemically absorbed tacrolimus. Inhibition of CYP3A4 may increase tacrolimus levels, while CYP3A4 inducers may increase tacrolimus metabolism and lower tacrolimus levels. Therefore, tacrolimus may be co-administered with one or more CYP3A4 inhibitors to improve its overall bioavailability.

[0008] Typically, tacrolimus is administered orally and is then absorbed from the gastrointestinal tract. It has been observed that absorption is adversely affected by the simultaneous intake of food. Therefore, the rate and extent of tacrolimus absorption was greatest under fasting conditions.

[0009] It is well known that, when administered orally, the absorption and bioavailability of a therapeutically active substance may be influenced by various factors. Such factors include the presence of food in the gastrointestinal tract and, in general, the residence time of the drug substance in the stomach is significantly longer in the presence of food than in the fasted state. When the presence of food in the digestive tract exceeds a certain point on the bioavailability of a drug substance, the drug substance is defined as having a gastrointestinal effect. Food effects are important because absorption and therefore plasma levels become highly variable depending on food intake. Adverse effects on the absorption into the bloodstream may result in the patient being at risk of being insufficiently absorbed for the treatment of the condition to be treated with the drug being administered. On the other hand, very high peak levels sometimes seen under fasting conditions can also cause significant side effects of nephro- or neurotoxic origin as well as side effects of GI (gastrointestinal) and others.

[0010] Absorption of tacrolimus from the gastrointestinal tract after oral administration is rapid with the average time to peak concentration (tmax) about 1-2 hours after administration to healthy subjects or patients with kidney or liver transplantation, but is incomplete and variable. The bioavailability is usually low, at most around 20% after oral administration.

[0011] Commonly observed side effects include vomiting and nausea, but side effects such as tremor, headache, hypertension, renal failure, hyperkalemia, hypomagnesaemia, hyperglycemia, insomnia, diarrhea, constipation, abdominal pain, nephrotoxicity and neurotoxicity are also observed.

[0012] For oral administration, tacrolimus is currently formulated and marketed in the form of soft gelatin capsules containing the equivalent of 0.5, 1 or 5 mg of anhydrous tacrolimus and is commercially available under the trade names Prograf® and Protropic®. The recommended initial daily oral dose for the patient is about 0.1 to 0.2 mg / kg / day. The dose is designed to provide some minimum plasma level of about 5 to about 20 ng / ml. Prograf^® is indicated for the prophylaxis of organ rejection in patients receiving allogeneic liver or kidney transplants.

[0013] D1 discloses a formulation obtained by spraying a solution of tacrolimus, PEG-24 cholesterol ether, monoglycerides and deoxycholic acid in an organic solvent onto embryos in the form of micro-drops, and furthermore relates to improving oral bioavailability.

[0014] D2 discloses sustained-release preparations obtained by dissolving tacrolimus in molten glycerol monostearate or a fatty acid tetraglycerol triester and mixing with HPM or lactose.

[0015] D3 discloses capsules containing a solid dispersion of 20% tacrolimus in HPMC, obtained by the solvent method.

[0016] D4 discloses a method of controlled agglomeration to improve the bioavailability of poorly water-soluble compounds in solid solutions or dispersions.

[0017] There remains a need for new pharmaceutical compositions and / or dosage forms containing tacrolimus that have increased bioavailability. Increased bioavailability may allow a reduction of the dosage units taken by the patient, e.g. to a single daily dose, and may also reduce or exclude the need to take food simultaneously with the dosage form, thus providing greater freedom for patients as to when the drug is being taken. Moreover, it is taken into account that fluctuations in the plasma concentration profile as a function of time can be significantly reduced. In addition, increased bioavailability can also provide a more reproducible (i.e. less variable compared to Prograf®) release profile.

BRIEF SUMMARY OF THE INVENTION [0018] The inventors have now found that the bioavailability of tacrolimus can be significantly increased by dissolving tacrolimus in a hydrophilic or water-miscible carrier in an amount that is effective to use to produce a useful dosage form of the drug. Tacrolimus is known to have very low water solubility, but the present invention provides pharmaceutical compositions and formulations exhibiting very fast in vitro release profiles, i.e. immediate release compositions that are believed to have significantly increased in vivo bioavailability in patients in need thereof.

[0019] Therefore, in a first aspect, the present invention relates to a solid solution containing tacrolimus dissolved in a mixture of polyethylene glycol and poloxamer, wherein tacrolimus is present therein at a concentration between about 0.01% w / w. and up to about 15% w / w to form a solid solution at ambient temperature. It is believed that this solution is capable of releasing at least 50% w / w. tacrolimus content within about 30 minutes when tested in any USP dissolution test using an aqueous dissolution medium.

[0020] In another aspect, the invention relates to a pharmaceutical composition comprising a solid tacrolimus solution and one or more pharmaceutically acceptable excipients, which may be fillers, disintegrants, binders or lubricants. In yet another aspect, the invention relates to dosage forms such as solid oral unit dosage forms containing a solid tacrolimus solution, pharmaceutically acceptable excipients and optionally a pharmaceutically acceptable additive such as flavoring agents, coloring agents, taste masking agents, pH adjusting agents, agents buffering agents, preservatives, stabilizing agents, antioxidants, wetting agents, humidity regulating agents, surfactants, suspending agents, absorption enhancers and release modifying agents. In particular, the present invention relates to a dosage form comprising tacrolimus and release-modifying agents, especially dosage forms exhibiting delayed release, such as solid oral unit dosage forms containing enteric coating. Delaying the release of tacrolimus into the distant part of the duodenum may reduce drug-related gastrointestinal side effects and a relatively high degree of metabolism in the anterior gastrointestinal tract (CYP3A4 mediated metabolism). With the new solid solution according to the invention this is achieved without loss of systemic bioavailability.

DETAILED DESCRIPTION OF THE INVENTION

Definitions [0021] As used herein, the term "active substance" or "pharmaceutically active ingredient" means any ingredient that is intended to provide pharmacological or other direct action in the diagnosis, healing, alleviation, treatment or prevention of the disease, or affect the structure or any function of the human body or other animals. The term includes ingredients that may undergo chemical change during the manufacture of the drug product and are present in the drug product in a modified form intended to provide a particular action or effect.

[0022] In the present context, the term "hydrophilic" describes that something "resembles water", i.e. a hydrophilic molecule or part of a molecule is one that is usually electrically polarized and is capable of forming hydrogen bonds with water molecules, thereby dissolving more easily water than in oil or other "non-polar" solvents.

[0023] In the present context, the term "amphiphilic" refers to a molecule (as a surfactant) having a polar, water-soluble group attached to a water-insoluble hydrocarbon chain. Therefore, one end of the molecule is hydrophilic (polar) and the other is hydrophobic (non-polar).

[0024] As used herein, the term "carrier" means any solvent or carrier fluid in a pharmaceutical product that does not play a pharmacological role. For example, water is a carrier for xylocaine and propylene glycol is a carrier for many antibiotics.

[0025] In the present context, the term "solid dispersion" means a drug or ingredient or active substance dispersed at the level of particles in an inert excipient, carrier, diluent or matrix in solid form, typically a fine particle dispersion. [0026] In the present context, the term "solid solution" refers to a drug or active ingredient or substance dissolved at the molecular level in an inert excipient, carrier, diluent or matrix in the solid state.

[0027] As used herein, the term "analog" means a chemical compound that is structurally similar to another.

[0028] The term "drug" means a compound for use in diagnosing, healing, alleviating, treating or preventing a disease in a human or other animal. [0029] In this context, the term "dosage form" means the form in which the drug is administered to the patient. It can be parenteral, topical, as a tablet, oral (liquid or dissolved powder), as a suppository, inhaled, transdermal, etc.

[0030] As used herein, the term "bioavailability" means the extent to which a drug or other substance becomes available to the target tissue after administration.

[0031] As used herein, the term "bioequivalence" means the scientific basis on which generic and generic names are compared. For example, drugs are bioequivalent if they enter circulation at the same rate when given at similar doses under similar conditions. Parameters often used in bioequivalence studies are tmax, cmax, AUC0-infinity, AUC0-t. Other relevant parameters may be W50, W75 and / or MRT. Therefore, at least one of these parameters can be used to determine if bioequivalence exists. In addition, two compositions are considered bioequivalent in the present context if the value of the parameter used is within 80-125% of the value for Prograf® or a similar commercially available tacrolimus containing product used in the test.

[0032] In the present context, "tmax" means the time to reach maximum plasma concentration (cmax) after administration; AUC0-infinity means the area under the plasma concentration-time curve as a function of time from time 0 to infinity; AUC0-t is the area under the plasma concentration curve versus time from time 0 to time t; W50 is the time when the plasma concentration is 50% or more of Cmax; W75 is the time when the plasma concentration is 75% or more of Cmax; and MRT means the average residence time of tacrolimus (and / or its analogue).

[0033] In the present context, the term "drug" means a compound used to treat a disease, injury or pain. The drug is correctly attributed to "preventive" measures, e.g. in the field of maintaining health, and "therapeutic", e.g. in the field of restoring health.

[0034] As used herein, the term "delayed release" means the release profile of a drug from a pharmaceutical composition or pharmaceutical formulation that, when compared to the immediate release profile, exhibits unchanged Cmax, but simply a time delay from the time of administration to the release of the drug. Therefore, tmax is delayed, at_/2 usually does not change.

[0035] In the present context, the term "erosion" or "eroding" means the gradual degradation of the surface of a material or structure, for example a tablet or tablet coating.

Solid dispersion and / or solid tacrolimus solution [0036] The solid solution of the invention contains tacrolimus dissolved in a mixture of polyethylene glycol and poloxamer at a concentration between about 0.01% w / w. and about 15% w / w, which solution forms a solid solution at ambient temperature (room temperature).

[0037] The active substance is tacrolimus (FK-506 or FR-900506). However, the scope of the present invention includes tacrolimus of any physical form (crystals, amorphous powder, any possible polymorphs, any possible solvates, including hydrate, anhydrate, its complexes, etc.). Any derivative or active metabolite of tacrolimus, its pharmaceutically acceptable salts, solvates, complexes and prodrugs is also included.

[0038] The active substance concentration in the carrier is at most 15% w / w, preferably at most 10% w / w, preferably at most 8% w / w, more preferably at most 6% w / w , even more preferably at most 5% w / w, at most 4% w / w, especially at most 3% w / w, and especially at most 2% w / w; and / or is at least about 0.05% w / w, preferably at least about 0.1% w / w, more preferably at least about 0.5% w / w, especially at least about 0 , 7% w / w, in particular at least about 1% w / w.

[0039] Physically, the combination of active substance and carrier forms a solid solution, i.e. the active substance is dissolved in the carrier at the molecular level. The active substance and the carrier can also form a solid dispersion in which part of the active substance is dissolved at the molecular level. The physical state of the dispersion and / or solution can be determined using a variety of techniques, such as heated table microscopy (HSM), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), optionally in combination with X-ray dispersion (EDX) and powder spectrometry X-ray diffraction. In a preferred embodiment, the active substance is completely dissolved in the carrier to form a solid solution at ambient temperature. [0040] Increasing bioavailability, the area under the curve (AUC), will usually reduce intrinsic and inter-individual variability in drug absorption. This is especially true, especially when low and poor bioavailability is due to poor water solubility. It is believed that the compositions of the invention will show CV (coefficient of variation) in the AUC data having a much lower value than Prograf^® and similar products.

[0041] As mentioned above, one of the basic features of the present invention is that it allows achieving an improvement in bioavailability when the oral administration of the composition of the present invention. Typically, the low oral bioavailability of a drug substance is a barrier to the design of controlled or modified release of drug substance compositions due to the fact that it is almost impossible to achieve effective drug levels over a prolonged period. However, a significant improvement in bioavailability and thus the possibility of designing, for example, delayed release compositions can be achieved using this technology.

[0042] The solid solution of the invention exhibits very rapid immediate release of tacrolimus when the solution containing composition is tested in a USP dissolution test using an aqueous dissolution medium and at least 50% w / w. the active pharmaceutical ingredient is released within about 30 minutes, preferably within 20 minutes, more preferably within 15 minutes; such as at least 75% w / w the active pharmaceutical ingredient is released in about 40 minutes or even better at least 90% w / w the active pharmaceutical ingredient is released in about 60 minutes, preferably within 45 minutes. For example, the test can be performed according to any method and any requirements cited in USP. Thus, the dissolution test can be carried out in an aqueous dissolution medium at a neutral or near neutral pH, for example at pH 6.8 or at acidic pH simulating the pH conditions in the gastrointestinal tract. However, variants with respect to the particular method used and the components contained in the dissolution medium etc. are within the scope of the present invention. A person skilled in the art will know how to perform a suitable dissolution test, e.g. with directions from USP, Ph. Eur. and the like. Suitable in vitro dissolution test conditions include the use of the USP dissolution test (paddle method) and pH 7.5 buffer containing 2.5% SDS and 1 g / ml pancreatin as dissolution medium.

[0043] In other embodiments, the following conditions are met with respect to in vitro dissolution tests:

i) at least about 50% w / w the total amount of tacrolimus is released in about 10 hours, such as e.g. in about 8 hours, in about 6 hours, in about 4 hours, in about 3 hours, in about 2 hours, in about 1 hours, for about 45 min, for about 30 min or for about 15 min, when tested in an in-vitro dissolution test and using a buffer solution pH 7.5 and ii) at least about 50% w / w the total amount of tacrolimus is released within approximately 1.5 hours, such as e.g. within approximately 1 hour, within approximately 0.75 hours, within approximately 0.5 hours, or within approximately 20 minutes when tested in the dissolution test in vitro and using a buffer solution of pH 7.5.

iii) at least about 55% w / w, such as e.g. about 60% w / w or above, about 65% w / w or above, about 70% w / w or above, about 75% w / w or above or about 80% w / w or above, the total amount of tacrolimus is released within about 15 hours, such as e.g. for about 12 hours, for about 10 hours, for 8 hours or for about 6 hours, when tested in an in-vitro dissolution test and using a buffer solution pH 7.5 iv) at least about 55 wt. / w / w, such as e.g. about 60% w / w or above, about 65% w / w or above, about 70% w / w or above, about 75% w / w or above or about 80% w / w or above, the total amount of tacrolimus is released in about 5 hours, such as e.g. in about 4 hours, in about 3 hours, in about 2 hours, in about 1 hour or in about 30 minutes when tested in an in vitro dissolution test and using a buffer solution pH 7.5, and / or

v) at least about 20% w / w, such as e.g. at least about 25% w / w, at least about 30% w / w, at least about 35% w / w. or at least about 40% w / w the total amount of tacrolimus is released within the first 3 hours, such as e.g. within the first 2 hours or within the first hour, when tested in an in vitro dissolution test and using a pH 7.5 buffer dissolution medium.

[0044] In other embodiments, the following conditions are met with respect to in vitro dissolution tests carried out under acidic conditions:

i) at most about 30% w / w, such as e.g. at most about 25% w / w, at most about 20% w / w, at most about 15% w / w. or at most about 14% w / w Tacrolimus is released within 2 hours in an in vitro dissolution test using a dissolution medium with a pH of at most about 5, such as at most about 4.5, at most about 4, at most about 3.5, at most about 3 , at most about 2 or at most about 1.5;

ii) at most about 10% w / w, such as e.g. at most about 7.5% w / w, at most about 5% w / w or at most about 2.5% w / w Tacrolimus is released within 2 hours in an in vitro dissolution test using a dissolution medium with a pH of at most about 5, such as at most about 4.5, at most about 4, at most about 3.5, at most about 3 , at most about or at most about 1.5;

iii) at most about 60% w / w, such as e.g. at most about 50% w / w, at most about 40% w / w or at most about 30% w / w tacrolimus is released within 15 hours, such as e.g. around 12 hours, when tested in an in vitro dissolution test using a dissolution medium with a pH of at most about 4.5, such as e.g. at most about 4.0, at most about 3.5, at most about 3, at most about 2 or at most about 1.5;

iv) at most about 40% w / w, such as e.g. at most about 30% w / w, at most about 25% w / w or at most about 20% w / w Tacrolimus is released within 6 hours when tested in an in vitro dissolution test using a dissolution medium with a pH of at most about 4.5, such as e.g. at most about 4.0, at most about 3.5, at most about 3, at most about 2 or at most about 1.5, and / or

v) at most about 30% w / w, such as e.g. at most about 25% w / w, at most about 20% w / w or at most about 15% w / w Tacrolimus is released within 4 hours when tested in an in vitro dissolution test using a dissolution medium with a pH of at most about 4.5, such as e.g. at most about 4.0, at most about 3.5, at most about 3, at most about 2 or at most about 1.5.

[0045] The carriers for use according to this invention are selected from mixtures of polyethylene glycol and poloxamer.

[0046] The carrier is polyethylene glycol (PEG), especially PEG with an average molecular weight of at least 1500, preferably at least 3000, more preferably at least 4000, especially at least 6000, mixed with the poloxamer in a ratio (by weight) between 1: 3 and 10: 1, preferably between 1: 1 and 5: 1, more preferably between 3: 2 and 4: 1, especially between 2: 1 and 3: 1, especially about 7: 3. A specific example of a useful mixture is a mixture of PEG6000 and poloxamer 188 in a ratio of 7: 3.

[0047] For polyethylene glycols (PEG), the melting point (freezing point or freezing point) increases as the average molecular weight increases. For example, for PEG 400 is in the range of 4-8 ° C, for PEG 600 is in the range of 2025 ° C, for PEG1500 is in the range of 44-48 ° C, for PEG2000 is about 52 ° C, for PEG 4000 is about 59 ° C, for PEG 6000 it is about 65 ° C, and for PEG 8000 it is about 61 ° C.

[0048] Useful poloxamers (also referred to as polyoxypropylene-polyoxyethylene block copolymers) include, for example, poloxamer 188, poloxamer 237, poloxamer 338 or poloxamer 407, or other block copolymers of ethylene oxide and propylene oxide, such as from the Pluronic® and / or Tetronic® series . Suitable block copolymers of the Pluronic® series include polymers with a molecular weight of about 3,000 or more, such as e.g. from about 4,000 to about 20,000 and / or viscosities (Brookfield) from about 200 to about 4,000 cP, such as e.g. from about 250 to about 3,000 cP. Useful examples include Pluronic® F38, P65, P68LF, P75, F77, P84, P85, F87, F88, F98, P103, P104, P105, F108, P123, F123, F127, 10R8, 17R8, 25R5, 25R8 etc. Suitable Tetronic® block copolymers include polymers with a molecular weight of about 8,000 or more, such as from about 9,000 to about 35,000 and / or viscosities (Brookfield) from about 500 to about 45,000 cP, such as e.g. from about 600 to about 40,000. The viscosities given above are determined at 60 ° C for substances, which are pastes at room temperature and at 77 ° C for substances that are solid at room temperature.

[0049] In a preferred embodiment of the present invention, the poloxamer is poloxamer 188, with an average molecular weight of about 8400 and a melting point of about 50-54 ° C. Pharmaceutical compositions [0050] The pharmaceutical composition of the invention comprises the solid solution of the invention and one or more pharmaceutically acceptable excipients, for example one or more excipients useful as fillers, disintegrants, binders and / or lubricants.

[0051] Preferably, the pharmaceutical composition of the invention is in the form of particles, for example in the form of a powder. Preferably, the particulate material obtained is a free-flowing powder, which is therefore easily convertible into e.g. solid dosage forms such as tablets, capsules or sachets. Usually, the particulate material has properties that are useful for making tablets by direct compression without adding large amounts of further additives. A suitable test for testing the flowability of particulate material is the method described in Ph. Eur. to measure the rate of flow of material from a funnel with a nozzle (hole) with a diameter of 10.0 mm.

[0052] The particles may have a weighted geometric mean diameter d_g from about 10 μm to about 2000 μm, preferably from about 20 μm to about 2000 μm, more preferably from about 30 μm to about 2000 pm, more preferably from about 50 μm to about 2000 pm, more preferably from about 60 μm to about 2000 μm, more preferably from about 75 μm to about 2000 pm, more preferably from about 100 μm to about 1500 pm, more preferably from about 100 μm to about 1000 μm, more preferably from about 100 μm to about 700 pm, more preferably from about 50 μm to about 400 pm, more preferably from about 50 μm to about 350 μm, even more preferably from about 50 μm to about 300 μm, especially from about 50 μm to about 250 μm or especially from about 100 μm to about 300 μm. In a preferred embodiment of the invention, the particles have a weighted geometric mean diameter d_g from about 50 μm to about 300 gm.

[0053] Examples of suitable excipients for use in the composition or solid dosage form of the invention include fillers, diluents, disintegrants, binders, lubricants etc. or mixtures thereof. Because the composition or solid dosage form of the invention can be used for a variety of purposes, excipients are typically selected for such different uses. Other pharmaceutically acceptable excipients for suitable use are e.g. acidulants, alkalizing agents, preservatives, antioxidants, buffering agents, chelating agents, coloring agents, complexing agents, emulsifying and / or solubilizing agents, flavoring and flavoring agents, humectants, sweetening agents, wetting agents, etc.

[0054] Examples of suitable fillers, diluents and / or binders include lactose (e.g. spray dried lactose, α-lactose, β-lactose, Tabletose®, various grades Pharmatose®, Microtose® or Fast-Floc®), microcrystalline cellulose (various grades of Avicel®, Elcema®, Vivacel®, Ming Tai® or Solka-Floc^®), hydroxypropyl cellulose, L-hydroxypropyl cellulose (low substituted), hydroxypropyl methyl cellulose (HPMC) (e.g. Methocel E, F and K, Metolose SH from Shin-Etsu, Ltd, such as e.g. 4,000 cP Methocelu E and Metolose 60 SH grades , 4,000 cP Methocel F and Metolose 65 SH grades, 4,000, 15,000 and 100,000 cP Methocel K grades; and 4,000, 15,000, 39,000 and 100,000 Metolose 90 SH grades, methyl cellulose polymers (such like for example. Methocel A, Methocel A4C, Methocel A15C, Methocel A4M), hydroxyethylcellulose, sodium carboxymethylcellulose, carboxymethylene, carboxymethylhydroxyethylcellulose and other cellulose derivatives, sucrose, agarose, sorbitol, starch, maltol, skeleton, dextrose rice starch), calcium phosphate (e.g. basic calcium phosphate, calcium hydrogen phosphate, dicalcium phosphate hydrate), calcium sulfate, calcium carbonate, sodium alginate, collagen, etc.

[0055] Specific examples of diluents are e.g. calcium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, microcrystalline cellulose, powdered cellulose, dextrans, dextrin, dextrose, fructose, kaolin, lactose, mannitol, sorbitol, starch, pregelatinized starch, sucrose, sugar etc.

[0056] Specific examples of disintegrants are, e.g., alginic acid or alginates, microcrystalline cellulose, hydroxypropyl cellulose and other cellulose derivatives, croscarmellose sodium, crospovidone, potassium polacrylate, sodium starch glycolate, starch, pregelatinized starch, carboxymethyl starch.^® and Explo13 tab^®) etc.

[0057] Specific examples of binders are, for example, acacia, alginic acid, agar, calcium carrageenan, sodium carboxymethyl cellulose, microcrystalline cellulose, dextrin, ethyl cellulose, gelatin, liquid glucose, guar gum, hydroxypropyl methylcellulose, methylcellulose, pellet, precursor gelatinized starch etc.

[0058] The composition may also include glidants and lubricants. Examples include stearic acid, magnesium stearate, calcium stearate or other metal stearates, talc, waxes and glycerides, light mineral oil, PEG, glyceryl behenate, colloidal silica, hydrogenated vegetable oils, corn starch, sodium stearyl fumarate, polyethylene glycols, sodium sulphate , sodium acetate etc.

[0059] Other excipients that may be included in the composition or solid dosage form of the invention are e.g. flavoring agents, coloring agents, taste masking agents, pH adjusting agents, buffering agents, preservatives, stabilizing agents, antioxidants, wetting agents, moisture regulating agents, surfactants, suspending agents, absorption enhancers, release modifying agents etc.

[0060] Other additives in the composition or solid dosage form of the invention may be antioxidants such as, e.g. ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorous acid, monothioglycerol, potassium metabisulphite, propyl gallate, sodium formaldehyde sulfate sodium thiosulfate, sulfur dioxide, tocopherol, tocopherol acetate, tocopherol hemisuccinate, TPGS or other tocopherol derivatives etc. The carrier composition may also contain e.g. stabilizing agents. The concentration of antioxidant and / or stabilizing agent in the carrier composition is usually from about 0.9% w / w. up to about 5% w / w

[0061] The pharmaceutical composition or solid dosage form of the invention may also contain one or more surfactants or surfactants. It is believed that such substances play a role in wetting the slightly soluble active substance and therefore contribute to improving the solubility characteristics of the active substance. Suitable excipients for use in the composition or solid dosage form of the invention are surfactants, such as e.g. amphiphilic surfactants, such as those disclosed in WO 00/50007 on behalf of Lipocine, Inc.

[0062] Examples of useful substances as surfactants include

i) polyethoxylated fatty acids, such as e.g. mono or diesters of polyethylene glycol fatty acids or mixtures thereof, such as e.g. or polyethylene glycol diesters with lauric acid, oleic acid, stearic acid, myristic acid, ricinoleic acid, where polyethylene glycol can be selected from PEG 4, PEG 5, PEG 6, PEG 7, PEG 8, PEG 9, PEG 10, PEG 12, PEG 15, PEG 20, PEG 25, PEG 30, PEG 32, PEG 40, PEG 45, PEG 50, PEG 55, PEG 100, PEG 200, PEG 400, PEG 600, PEG 800, PEG 1000, PEG 2000, PEG 3000, PEG 4000, PEG 5000, PEG 6000, PEG 7000, PEG 8000, PEG 9000, PEG 1000, PEG 10,000, PEG 15,000, PEG 20,000, PEG 35,000, ii) fatty acid esters with polyethylene glycol and glycerin, e.g. esters as mentioned above, but in the form of glyceryl esters of individual fatty acids;

iii) esters of glycerol, propylene glycol, ethylene glycol, PEG or sorbitol e.g. with vegetable oils, e.g. with hydrogenated castor oil, almond oil, palm kernel oil, castor oil, apricot seed oil, olive oil, peanut oil, hydrogenated oil from palm kernels and the like, iv) polyglycerized fatty acids, such as e.g. polyglycerol stearate, polyglycerol oleate, polyglycerol ricinoleate, polyglycerol linolate,

v) fatty acid esters of propylene glycol, such as e.g. propylene glycol monolaurate, propylene glycol ricinolate and the like, vi) mono- and diglycerides such as e.g. glyceryl monooleate, glyceryl dioleate, glyceryl mono- and / or dioleate glyceryl, glyceryl caprate etc .;

vii) sterol and sterol derivatives;

viii) polyethylene glycol sorbitan fatty acid esters (PEG sorbitan fatty acid esters) such as PEG esters with different molecular weights given above and different esters from the Tween® series;

ix) polyethylene glycol alkyl ethers, such as, for example, PEG-oleyl ether and PEG-lauryl ether;

x) sugar esters such as, for example, sucrose monopalmitate and sucrose monolaurate;

xi) polyethylene glycol ethers with alkylphenols, such as e.g. from the Triton® X series or

N;

xii) polyoxyethylene-polyoxypropylene block copolymers, such as e.g. from the Pluronic® series, Synperonic® series, Emkalyx®, Lutrol®, Supronic® etc. The general term for these polymers is "poloxamers" and Poloxamer 105 is an appropriate example in this context. , 108, 122, 123, 124, 181, 182, 183, 184, 185, 188, 212,

215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, 403 and 407;

xiii) sorbitan and fatty acid esters, such as from the Span® series or the Ariacel® series, such as, e.g., sorbitan monolaurate, sorbitan monopalmitate, sorbitan monooleate, sorbitan monostearate and the like;

xiv) fatty acid esters with lower alcohols, such as, for example, oleate, isopropyl myristate, isopropyl palmitate, etc .;

xv) ionic surfactants, including cationic, anionic and amphoteric surfactants, such as e.g. fatty acid salts, bile salts, phospholipids, phosphoric acid esters, carboxylates, sulfates and sulfonates, etc.

[0063] When the surfactant or surfactant mixture is present in the composition or solid dosage form of the invention, the concentration of surfactant (surfactants) is usually in the range of about 0.1-80% w / w, such as e.g. from about 0.1 to about 20% w / w, from about 0.1 to about 15% w / w, from about 0.5 to about 10% w / w. or alternatively, from about 0.10 to about 80% w / w, such as e.g. from about 10 to about 70% w / w, from about 20 to about 60% w / w or from about 30 to about 50% w / w.

[0064] In a preferred aspect of the invention, at least one of one or more pharmaceutically acceptable excipients is selected from the group consisting of silicic acid or a derivative or salt thereof, including silicates, silicon dioxide and its polymers; magnesium aluminum silicate and / or magnesium aluminum silicate, bentonite, kaolin, magnesium trisilicate, montmorillonite and / or saponite.

[0065] Such materials are particularly useful as material for the sorption of oil or oily materials in pharmaceuticals, cosmetics and / or food. In a specific embodiment, the material is used as a sorption material for oils or oily materials in pharmaceuticals. A material that has the ability to act as a sorption material for oils or oily materials is also referred to as "oil absorbing material". In addition, in the present context, "absorption" as used means "absorption" as well as "adsorption". It should be understood that when one of these terms is used, it is intended to include the phenomenon of absorption as well as adsorption.

[0066] In particular, the pharmaceutically acceptable excipient may be silicic acid or a derivative or salt thereof, such as, e.g., silicon dioxide or a polymer thereof, as a pharmaceutically acceptable excipient. Depending on the quality of the silicon dioxide used, it can be a lubricant or it can be an oil-absorbing material. The types fulfilling the latter function seem to be the most important.

[0067] In a preferred embodiment, the composition or solid dosage form according to the invention comprises a pharmaceutically acceptable excipient, which is a silicon dioxide product with properties corresponding to Aeroperl^® 300 (available from Degussa, Frankfurt, Germany). As the examples show, Aeroperl® 300 is a very suitable material (including materials with similar or equivalent properties to Aeroperl® 300).

[0068] The use of an oil-absorbent material in the compositions or dosage forms of the invention is very advantageous in the preparation of pharmaceutical, cosmetic, nutritional and / or food compositions when the composition comprises an oil or oily material. One advantage is that you can incorporate a relatively large amount of oil and oily material and you will still be dealing with a material that is solid. Accordingly, solid compositions with a relatively high oil or oily-like material content can be prepared by using the oil-absorbing material according to the invention. In the pharmaceutical field, it is convenient to incorporate a relatively large amount of oil or oil-like material into the solid composition, especially in those situations where the active substance does not have adequate water solubility properties (e.g. has poor solubility in water), stability in the aqueous medium (i.e. degradation in the aqueous medium), oral bioavailability (i.e. has low bioavailability) etc., or in those situations where it is desired to modify the release of the active substance from the composition to achieve controlled, delayed, prolonged and / or pulsed delivery of the active substance. Therefore, in a preferred form, it is used in the preparation of pharmaceutical compositions. [0069] The oil absorbing material for use in processing into solid compositions absorbs typically about 5% w / w. or above, such as e.g. about 10% w / w or above, about 15% w / w or above, about 20% w / w or above, about 25% w / w or above, about 30% w / w or above, about 35% w / w or above, about 40% w / w or above, about 45% w / w or above, about 50 w / w or above, about 55% w / w or above, about 60% w / w or above, about 65% w / w or above, about 70% w / w or above, about 75% w / w or above, about 80% w / w or above, about 85% w / w or above, about 90% w / w or above or about 95% w / w or above oil or oily material and still remains a solid material.

[0070] As can be seen from the examples, the bioavailability obtained after administration of the compositions of the invention significantly improves. Thus, in specific embodiments, the AUC / AUCPrograph® value is at least about 1.5, such as about 1.75 or above, about 1.8 or above, about 1.9 or above, about 2.0 or above, about 2 , 5 or above, about 2.75 or above, about 3.0 or above, about 3.25 or above, about 3.5 or above, about 3.75 or above, about 4.0 or above, about 4, 25 or above, about 4.5 or above, about 4.75 or above or about 5.0 or above, when determining AUC under similar conditions.

[0071] Following oral administration of the pharmaceutical composition of the present invention, the plasma concentration profile as a function of time shows an extended period during which the plasma concentration is maintained in the therapeutic range (i.e. plasma concentration leads to a therapeutic effect) without causing serious adverse effects. side effects. Therefore, a decrease in peak concentration can be observed.

[0072] However, a decrease in peak concentration may not lead to a decrease in therapeutic effect as long as the plasma concentration of tacrolimus is maintained within the therapeutic range.

[0073] The need for simultaneous food intake to ensure sufficient absorption of tacrolimus is believed to be significantly reduced or even completely eliminated when the pharmaceutical composition or dosage form of the present invention is administered.

[0074] Thus, the pharmaceutical compositions of the invention provide significantly higher bioavailability of tacrolimus, which can significantly reduce the number of unit dosage forms administered during the day and reduce or eliminate administration in combination with food intake, which provides a greater degree of freedom for the recipient of the pharmaceutical compositions and, consequently patient acceptance and / or patient compliance can be significantly improved. In addition, the compositions provide a significant reduction in side effects, especially side effects associated with high peak concentration (such as e.g. nephro- and neurotoxicity, diarrhea, constipation, abdominal pain, nausea, etc.) and provide prolonged release of tacrolimus leading to better therapy.

[0075] One of the main challenges in formulating the tacrolimus composition is to avoid an undesirable food effect. In general, tacrolimus is much better absorbed when given orally without food. Therefore, there is considerable variability in bioavailability after administration with or without food. This relationship makes it difficult to give precise guidance on how much to administer, and also requires informing the patient about the mode of administration. The present invention aims to provide compositions in which the adverse food effect is limited. Thus, the present invention provides a composition that does not exhibit a significant adverse food effect when administered to a composition in need of such treatment to a mammal, as evidenced by a value (food AUC / fasting AUC) of at least about 0.85 with a lower confidence level of 90% at least 0.75 .

[0076] In particular, the pharmaceutical composition of the invention has a value (food AUC / fasting AUC) of about 0.9 or above, such as e.g. about 0.95 or above, about 0.97 or above or about 1 or above, yes such as up to about 1.1 or up to about 1.2.

[0077] In addition to tacrolimus, the composition of the invention may also contain an additional therapeutically, prophylactically and / or diagnostically active substance. In particular, combinations of tacrolimus with at least one of the following active substances are of interest: Substances that are indicated for use in organ transplantation, such as steroids, calcineurin inhibitors and / or antiproliferative agents, for example. Specific examples include prednisone, prednisolone, methylprednison, cyclosporin, mycophenolate, azathioprine, sirolimus, everolimus, mycophenolate sodium and FTY720 (developed by the Novartis pharmaceutical company).

Dosage Forms [0078] Useful dosage forms of the invention are solid oral dosage forms containing a solid solution and one or more pharmaceutically acceptable excipients, preferably unit dosage forms.

[0079] The pharmaceutical composition of the invention is in the form of particles and can be used as such. However, in many cases it is more convenient to provide the composition in the form of granules, pellets, microspheres, nanoparticles and the like, or in the form of solid dosage forms including tablets, capsules, sachets and the like. [0080] The solid dosage form of the invention may be a single unit dosage form, or it may be in the form of a multi-depot dosage form containing a plurality of individual units, such as, for example, pellets, beads and / or granules. [0081] Dosage forms may further contain pharmaceutically acceptable additives, such as flavoring agents, coloring agents, taste masking agents, pH adjusting agents, buffering agents, preservatives, stabilizing agents, anti-oxidants, wetting agents, humidity regulating agents, surfactants, suspending agents, absorption enhancers and release modifying agents. [0082] In a preferred embodiment, the dosage form comprises silicic acid or a derivative or salt thereof, including silicates, silicon dioxide and its polymers; and / or magnesium aluminum silicate and / or magnesium aluminum silicate, bentonite, kaolin, magnesium trisilicate, montmorillonite and / or saponite. A particularly useful excipient for use in dosage forms is any product in the form of silicon dioxide with properties corresponding to Aeroperl^® 300 (available from Degussa, Frankfurt, Germany).

[0083] The solid dosage form according to the present invention comprises a particulate pharmaceutical composition as described above. The details and particulars disclosed in this main aspect of the invention relate mutatis mutandis to other aspects of the invention. Therefore, properties with respect to increasing bioavailability, changes in bioavailability parameters, reduction of undesirable food effect as well as release of tacrolimus and / or its analogue etc. the particulate pharmaceutical compositions described herein and / or claimed are analogous to the solid dosage form of the present invention.

[0084] Typically, the concentration of the particulate pharmaceutical composition ranges from about 5 to 100% w / w, such as, e.g., from about 10% to about 90% w / w, from about 15% to about 85% w / w, from about 20% to about 80% w / w, from about 25% to about 80% w / w, from about 30% to about 80% w / w, from about 35% to about 80% w / w, from about 40% to about 75% w / w, from about 45% to about 75% w / w. or from about 50% to about 70% w / w dosage form. In an embodiment of the invention, the concentration of the particulate pharmaceutical composition is 50% w / w. or above dosage form. [0085] The solid dosage form according to the invention is obtained by processing the particulate material according to the invention using techniques well known to the person skilled in the art. Typically, this further includes the addition of one or more of the pharmaceutically acceptable excipients listed herein.

[0086] The composition or solid dosage form of the invention may be designed to release tacrolimus in any suitable manner, provided that bioavailability increases. Thus, the active substance can be released relatively quickly for a faster onset of action, it can be released so that it exhibits zero or first order kinetics, or it can be released in a modified manner to achieve a specific release process. All such routes are considered controlled. Normal formulations are also within the scope of the present invention.

[0087] Recommended dose range for Prograf^® is 0.1 to 0.2 mg / kg / day when given every 12 hours in two divided doses. More importantly, blood levels must be monitored. A typical level for 1-3 months is 7-20 ng / ml, and for 4-12 months the level should be 5-15 ng / ml. These are only indications and may vary depending on the type of transplant and ethnic issues.

[0088] The following data was established for renal transplant patients:

<td>Time after</td><td>Dose</td><td>Caucasian race n = 114 Minimum concentration</td><td>Dose</td><td>Black race n = 56 Minimum concentration</td>

<td>transplant</td><td>(Mg / kg)</td><td>(Ng / ml)</td><td>(Mg / kg)</td><td>(Ng / ml)</td>

<td>Day 7</td><td> 0,18</td><td> 12,0</td><td> 0,23</td><td> 10,9</td>

<td>Month 1</td><td> 0,17</td><td> 12,8</td><td> 0,26</td><td> 12,9</td>

<td>6th month</td><td> 0,14</td><td> 11,8</td><td> 0,24</td><td> 11,5</td>

<td>12 months</td><td> 0,13</td><td> 10,1</td><td> 0,19</td><td> 11,0</td>

[0089] Contemplated recommended doses of the products of the present invention will be from 0.02 mg / kg / day to 0.15 mg / kg / day, with once daily dosing.

Enteric coatings - delayed release [0090] It has been found that the efficacy of oral tacrolimus treatment can be significantly improved by appropriately designing the tacrolimus release profile. On the one hand, relatively high doses of tacrolimus are necessary to avoid transplant rejection, and on the other hand side effects are often too significant, even at therapeutically relevant levels. Therefore, side effects such as acute nausea, vomiting, nephrotoxicity and neurotoxicity are directly related to high peak plasma concentrations. This association has been demonstrated for dogs. In those cases where a lower dose was used to avoid a high peak, the dose-dependent side effects almost ceased to appear at a certain threshold level, and if they occurred, they are much less significant. However, due to dose reduction (without increasing bioavailability), the therapeutically effective level is only maintained for a short time. The present invention solves this problem by providing a pharmaceutical composition or dosage form containing tacrolimus in which the release of tacrolimus is designed to avoid high peak concentrations, while the composition is designed so that overall bioavailability is maintained or increased compared to available commercially available dosage forms containing tacrolimus. In addition, by delaying the release of tacrolimus and at the same time providing a composition in which tacrolimus is at least partially in dissolved form, significant absorption in the distal gastrointestinal tract can be achieved.

[0091] Thus, the dosage form of the invention may further comprise one or more release modifying agents selected from the group consisting of water-miscible polymers, water insoluble polymers, oils and oily materials. [0092] The water insoluble polymer may be ethyl cellulose, cellulose acetate, cellulose nitrate and mixtures thereof. The water-miscible polymer can also be a cellulose derivative selected from the group consisting of hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), methylcellulose, sodium carboxymethylcellulose, hydroxyethylcellulose, poloxamers, polyoxyethylene pyrroleol, polyoxyethylene pyrrole - poly (vinyl acetate) PVP-PVA, polymethacrylic polymers and poly (vinyl alcohol) (PVA), poly (ethylene oxide) (PEO) and mixtures thereof. Examples of particularly useful polymethacrylic polymers are Eudragit^® RS, Eudragit^® RL, Eudragit^® NE and Eudragit^® E.

[0093] The oil or oily material may be hydrophilic and hydrophobic oils or oily materials.

[0094] The hydrophilic oil or oily material may be polyether glycols such as polypropylene glycols; polyoxyethylenes; polyoxypropylenes; poloxamers; polyglycolyzed glycerides such as Gelucire^®, for example Gelucire^® 50/13, Gelucire^® 44/14, Gelucire^® 50/10, Gelucire^® 62/05 and mixtures thereof.

[0095] The hydrophobic oil or oily material may have a melting point of at least about 20 ° C. Useful examples are straight chain saturated hydrocarbons, sorbitan esters, paraffins; fats and oils such as cocoa butter, beef tallow, lard, polyether glycol esters; higher fatty acids such as stearic acid, myristic acid, palmitic acid, higher alcohols such as cetanol, stearyl alcohol, low melting waxes such as glyceryl monostearate, glyceryl monooleate, hydrogenated tallow, myristyl alcohol, substituted stearyl alcohol, and / or unsubstituted monoglycerides, substituted and / or unsubstituted diglycerides, substituted and / or unsubstituted triglycerides, yellow beeswax, white beeswax, carnauba wax, castor wax, Japanese wax, acetylated monoglycerides; NVP polymers, PVP polymers, acrylic polymers and mixtures thereof.

[0096] The oil or oil-like material may also be a sorbitan ester, such as, for example, sorbitan diisostearate, sorbitan dioleate, sorbitan monolaurate, sorbitan monoisostearate, sorbitan monooleate, sorbitan sorbitanate, sorbitan monostearate, sorbitan monostearate, sebitanate, sorbitan sorbitan trioleate, sorbitan tristearate or mixtures thereof.

[0097] The oil or oil-like material may also be a mixture of different oils or oil-like materials, such as, for example, a mixture of hydrophilic and / or hydrophobic materials.

[0098] Other oils or oil-like materials may be solvents or semi-solid excipients, such as e.g. propylene glycol, polyglycolyzed glycerides including Gelucire 44/14, complex fat materials of plant origin including theobromic oil, carnauba wax, vegetable oils such as e.g. . almond oil, coconut oil, corn oil, cottonseed oil, sesame oil, soybean oil, olive oil, castor oil, palm kernel oil, peanut oil, rapeseed oil, grape seed oil etc., hydrogenated vegetable oils such as e.g. hydrogenated peanut oil, hydrogenated palm kernel oil, hydrogenated cottonseed oil, hydrogenated soybean oil, hydrogenated castor oil, hydrogenated coconut oil; natural fat materials of animal origin, including beeswax, lanolin, fatty alcohols including cetyl, stearyl, lauric, myristine, palmitic, stearic fatty alcohol; esters including glycerol stearate, glycol stearate, ethyl oleate, isopropyl myristate; liquid transesterified semi-synthetic glycerides including Miglycol 810/812; fatty acid amides or alcoholamides including stearamidoethanol, coconut oil fatty acid diethanolamide, mono and diglyceride acetic acid esters, mono and di-glyceride citric acid esters, mono and di-glyceride lactic acid esters, mono and diglycerides, polyglycerol and fatty acid esters polyglycerol polyricinolate, fatty acid propylene glycol esters, sorbitan monostearates, sorbitan tristearates, sodium stearoyl lactactates, calcium stearoyl lactylates, diacetyltartaric acid esters with mono and di-glycerides, etc.

[0099] Delayed release of the active substance is desirable to increase the bioavailability of the active substance by providing the ingredient in the gastrointestinal tract, so that release occurs primarily after passage through the stomach. For example, the dosage form of the present invention may be designed to release, after oral administration to a mammal in need thereof, at most about 10% w / w, preferably at most about 7.5% w / w, more preferably at most about 5% w / w, especially at most about 2% w / w the total amount of active substance in the first 3 hours, preferably within 2 hours, more preferably within 1 hour and especially within about 30 minutes after administration.

[0100] In addition, the solid dosage form of the invention may, after oral administration to a mammal in need thereof, release at least about 50% w / w. active substance within 24 hours, preferably within about 20 hours, more preferably within about 18 hours, especially within about 15 hours and especially within about 12 hours. [0101] Delayed release is achieved mainly due to some type of enteric coating. Although the semi-permeable coating will exhibit some type of delayed release, this may not be a sufficiently valuable "delayed" release. In addition, it requires some time to release the content. The coating sought for the present invention is a coating with a pH-dependent effect. This type of coating is very resistant to drug release before reaching a certain pH. With a slight increase in pH, with an increase in pH of about 0.2 to 0.4, the film changes properties and becomes permeable.

[0102] Accordingly, the solid dosage forms according to the invention may exhibit delayed release of the active substance due to an enteric coating using a water-miscible polymer with a pH-dependent water solubility. Examples of pH-sensitive polymers that are relatively insoluble and impermeable at gastric pH, but which are more soluble and permeable at pH in the small intestine and colon include, but are not limited to, polyacrylamides; phthalate derivatives such as acid phthalates of carbohydrates including amylose acetate phthalate, cellulose acetate phthalate, cellulose, cellulose acetate isophthalate, other cellulose ester phthalates phthalate, cellulose ether phthalate, hydroxypropyl cellulose phthalate, hydroxypropylcellulose phthalate, hydroxypropylethylcellulose phthalate, hydroxypropylmethylcellulose phthalate (HMPCP), methylcellulose phthalate, methylcellulose acetate phthalate , polyvinyl acetate phthalate, polyvinyl acetate phthalate, sodium cellulose acetate phthalate, acid starch phthalate; phthalates of other compounds including polyvinyl acetate phthalate (PVAP); other cellulose derivatives including hydroxypropyl methylcellulose acetate succinate (HPMCAS), carboxymethyl cellulose, cellulose acetate trimellitate; alginates; carbomers; polyacrylic acid derivatives such as copolymers of acrylic acid and acrylic ester, poly methacrylic acid and its esters, copolymers of poly (acrylic methacrylic acid), copolymers of methacrylic acid (for example Eudragit^® L and Eudragit^® S); styrene-maleic acid-dibutyl phthalate copolymer, styrene-maleic acid-polyvinyl acetate phthalate copolymer, styrene and maleic acid copolymers; shellac, starch glycolate; polacriclin; vinyl acetate and crotonic acid copolymers and mixtures thereof. Shellac polymers are particularly interesting for pH sensitive polymers; phthalate derivatives, in particular cellulose acetate phthalate, polyvinyl acetate phthalate and hydroxypropyl methyl cellulose phthalate; polyacrylic acid derivatives, especially polymethyl methacrylate mixed with copolymers of acrylic acid and acrylic esters; and copolymers of vinyl acetate and crotonic acid. [0103] The first embodiment of delayed release according to the invention is a "pH-dependent coated dosage form", such as e.g. tablet or capsule. In the case of a tablet, it contains a tablet core containing tacrolimus, e.g. in a solid solution / solid dispersion as a multi-particle product, a controlled release matrix, e.g. of HPMC, a disintegrant, a lubricant and one or more pharmaceutical carriers, the core being coated with a material, preferably a polymer, which is substantially insoluble and impermeable at gastric pH, but which is more soluble and permeable at the pH in the small intestine . Preferably, the coating polymer is substantially insoluble and impermeable at pH &lt;5.0 and soluble in water at pH&gt; 5.0. The tablet core may be coated with an amount of polymer sufficient to ensure that substantially no release of tacrolimus from the dosage form occurs until the dosage form leaves the stomach and remains in the small intestine for about 15 minutes or longer, preferably about 30 minutes or longer, which will provide minimal release of tacrolimus in the duodenum. Mixtures of a pH sensitive polymer with a water insoluble polymer can also be used. The tablets are coated with the polymer in an amount from about 10% to about 80% by weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer comprising about 15% to about 50% of the weight of the tacrolimus-containing tablet core.

[0104] pH-sensitive polymers that are very insoluble and impermeable at gastric pH, but which are more soluble and permeable at the pH of the small intestine and colon, include polyacrylamides, phthalate derivatives such as acid carbohydrate phthalates, amylose acetate phthalate, acetate phthalate celluloses, other cellulose phthalate esters, cellulose ether phthalates, hydroxypropyl cellulose phthalate, hydroxypropyl ethyl cellulose phthalate, hydroxypropyl methyl cellulose phthalate, methylcellulose phthalate, polyvinyl acetate phthalate, polyvinyl acetate phthalate phthalate, sodium cellulose acetate phthalate, acid starch phthalate, styrene-maleic acid copolymer-dibutyl copolymer, styrene-maleic acid-polyvinyl acetate phthalate copolymer, styrene-like copolymer and styrene copolymer acrylic and acrylic esters, poly methacrylic acid and its esters, poly (acrylic methacrylic acid) copolymers, shellac and copolymers of vinyl acetate and crotonic acid.

[0105] Preferred pH sensitive polymers include shellac; phthalate derivatives, in particular cellulose acetate phthalate, polyvinyl acetate phthalate and hydroxypropyl methyl cellulose phthalate; polyacrylic acid derivatives, especially polymethyl methacrylate mixed with copolymers of acrylic acid and acrylic esters; and copolymers of vinyl acetate and crotonic acid.

[0106] The delay time before tacrolimus release after the stomach leaves the dosage form in the form of a "coated pH dependent tablet" can be adjusted by selecting the relative amounts of Eudragit-L^® and Eudragit-S^® in the coating and by choosing the thickness of the coating. Membranes from Eudragit-L^® dissolve at pH above 6.0, membranes from Eudragit-S^®they dissolve at a pH above 7.0, and the mixtures dissolve at an intermediate pH. Because the duodenal pH is about 6.0 and the colon pH is about 7.0, coatings consisting of Eudragit-L mixtures^® and Eudragit-S® provide duodenal protection against tacrolimus. If it is desired to delay the release of tacrolimus until the "coated pH-dependent tablet" containing tacrolimus reaches the colon, Eudragit-S may be used as the coating material^®as described by Dew et al. (Br. J. Clin. Pharmac. 14 (1982) 405-408). To delay the release of tacrolimus by about 15 minutes or above, preferably 30 minutes or above, after the dosage form leaves the stomach, preferred coatings contain from about 9: 1 to about 1: 9 EudragitL® / Eudragit-S®, more preferably from about 9 : 1 to about 1: 4 Eudragit-L® / Eudragit-S®. The coating may comprise about 3% to about 70% by weight of the uncoated tablet core. Preferably, the coating comprises from about 5% to about 50% of the weight of the tablet core.

[0107] The release of the active ingredient from a composition comprising a delayed release coating can also be an enzymatic reaction if, for example, zein or mono / diglyceride mixtures are used as the coating material.

Preparation of compositions and dosage forms according to the invention [0108] The invention also provides a method for producing a solid solution according to the invention, a method comprising the step of dissolving tacrolimus in polyethylene glycol and poloxamer to form a solid solution at ambient temperature.

[0109] The pharmaceutical compositions of the invention can be prepared by any convenient method, such as, e.g., granulation, mixing, spray drying, etc. An example of a useful method is the controlled agglomeration method disclosed in WO 03/004001, i.e. a method that allows controlled particle size increase . This method includes spraying the first composition containing the active ingredient and the molten excipient onto a second solid carrier. Typically, the meltable excipient has a melting point of at least 5 ° C, but preferably its melting point is lower than the melting point of tacrolimus. The melting point of the excipient may range from 10 ° C to 150 ° C.

[0110] The advantage of using the controlled agglomeration method described in WO 03/004001 is that it is possible to use a relatively large amount of molten material on the particulate material without obtaining an undesirable increase in particle size.

[0111] A solid solution can also be obtained, e.g., by using organic solvents or by dissolving the active substance in another suitable medium (e.g., oil or oily material, which is liquid at room temperature or elevated temperature).

[0112] Solid dispersions (solvent method) are prepared by dissolving a mixture of the physical active substance (e.g. drug substance) and excipient or carrier in a conventional organic solvent followed by evaporation of the solvent. Suitable organic solvents include a pharmaceutically acceptable solvent in which the active substance is soluble, such as methanol, ethanol, methylene chloride, chloroform, ethyl acetate, acetone or mixtures thereof.

[0113] Suitable water-soluble carriers include polymers such as polyethylene glycol, poloxamers.

[0114] The solid dispersion is preferably obtained by spray drying, controlled agglomeration, freeze drying or coating on carrier particles or any other solvent removal process. The dried product contains the active substance in the form of a solid dispersion consisting of a molecular dispersion and a solid solution.

[0115] As an alternative to the use of organic solvents, the drug and polymer can be subjected to co-milling or extrusion at elevated temperatures (hot extrusion).

[0116] Pharmaceutical compositions containing tacrolimus in the form of a solid solution can generally be prepared using any suitable known procedure for making pharmaceutical compositions.

[0117] In addition to using the organic solvent based method, solid tacrolimus solutions can be obtained by dissolving tacrolimus in the carrier composition used in the controlled agglomeration method. Stabilizing agents may be added to guarantee the stability of the solid dispersion / solid solution.

Uses [0118] The solid solution according to the invention or the pharmaceutical composition according to the invention can be used to prepare a solid dosage form for oral administration, such as tablets, capsules or sachets, or for the production of granules, pellets, microspheres, nanoparticles.

[0119] Preferably, the solid dispersion or solid solution is used to make a solid immediate-release dosage form or a solid delayed-release dosage form.

[0120] Other uses of the solid dispersion or solid solution of the invention are the use for preparing a topical dosage form.

[0121] Another advantage of the compositions of the invention is the possibility of achieving an effective therapeutic response at a lower dose compared to traditional oral treatment. Therefore, it is believed that the solid dosage form of the invention, administered orally to a mammal in need thereof, at a dose of at most about 85% w / w, such as e.g. at most 80% w / w, at most about 75%, at most about 70% w / w, at most about 65% w / w, at most about 60% w / w, at most about 55% w / w or at most about 50% w / w doses of tacrolumus administered in the form of Prograf^® or a similar tacrolimus-containing product, commercially available, is essentially biologically equivalent to Prograf^® or a similar commercial product containing tacrolimus.

[0122] Each of the dosage forms, compositions, dispersions and solutions of the invention containing tacrolimus may improve the treatment of conditions responsive to tacrolimus treatment.

[0123] Tacrolimus is indicated (or suggested to be used) in the treatment of diseases such as e.g. rejection reactions for organ or tissue transplants such as heart, kidney, liver, bone marrow, skin, cornea, lung, pancreas, small intestine, limb, muscle, nerve, intervertebral disc, trachea, myoblast, cartilage, etc .; graft versus host reactions after bone marrow transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, etc .; infections caused by pathogenic microorganisms (e.g. Aspergillus fumigatus, Fusarium oxysporum, Trichophyton asteroides, etc.); inflammatory or hyperproliferative skin diseases or cutaneous manifestations of the immune-mediated disease (e.g. psoriasis, atopic dermatitis, contact dermatitis, eczema dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, bullous epidermal separation, urticaria, angioedema, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, acne plaques); autoimmune eye diseases (e.g. keratoconjunctivitis, spring conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conic keratitis, corneal epithelial dystrophy, corneal endosperm, ocular pemphigus, Mooren's ulcer, scleritis, Graves' ophthalmopathy, Vogt syndrome -Koyanagi-Harada, dry keratoconjunctivitis (dry eye syndrome), pimple, iritis and ciliary body inflammation, sarcoidosis, ophthalmopathy on the background of endocrine disease, etc.); recurrent obstructive airway disease [asthma (e.g. bronchial asthma, allergic asthma, essential asthma, extrinsic asthma and pulmonary asthma), in particular chronic or persistent asthma (e.g. late asthma and airway hypersensitivity), bronchitis, etc .; inflammation of the mucous membranes or vessels (e.g. stomach ulcer, ischemic or thrombotic blood vessel damage, ischemic bowel disease, inflammation of the small intestine, gangrenous enterocolitis, burn-related intestinal damage, leukotriene-mediated B4 diseases); enteritis / intestinal allergy (e.g. celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis); food allergy diseases with symptoms occurring outside the gastrointestinal tract (e.g. migraine, rhinitis and eczema); kidney disease (e.g., interstitial nephritis, Goodpasture syndrome, hemolytic uremic syndrome and diabetic nephropathy); nervous system diseases (e.g. dermatomyositis, GuillainBarre syndrome, Meniere's disease, polyneuritis, single neuritis, cerebral infarction, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and radiculopathy); ischemic brain disease (e.g. head injury, cerebral haemorrhage (e.g. arachnoid haemorrhage, intracerebral bleeding), cerebral thrombosis, cerebral embolism, cardiac arrest, stroke, transient cerebral ischemia (TIA), hypertensive encephalopathy, cerebral infarction); endocrine diseases (e.g. hyperthyroidism and Basedow's disease); blood diseases (e.g. pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia and anerytroplasia); bone diseases (e.g. osteoporosis); respiratory diseases (e.g. sarcoidosis, pulmonary fibrosis and idiopathic interstitial pneumonia); skin diseases (e.g. dermatomyositis, albinism of the skin, fish scale, photosensitivity and cutaneous T-cell lymphoma); cardiovascular diseases (e.g. arteriosclerosis, atherosclerosis, aortitis syndrome, nodular arteritis and myocarditis); collagen diseases (e.g. scleroderma, Wegener's granulomatosis and Sjogren's syndrome); steatosis, eosinophilic fasciitis; periodontal disease (e.g. damage to gums, periodontium, alveolar bone or tooth cement); nephrotic syndrome (e.g. glomerulonephritis); male pattern baldness, senile alopecia; muscular dystrophy; purulent dermatitis and Sezary syndrome; diseases associated with chromosomal disorders (e.g. Down syndrome); Addison's disease; active oxygen mediated diseases [e.g. organ damage (e.g. ischemic organ circulation disorder (e.g. heart, liver, kidney, gastrointestinal tract, etc.) related to maintenance, transplantation or ischemic diseases (e.g. thrombosis, myocardial infarction, etc.); intestinal diseases (e.g. endotoxic shock, pseudomembranous colitis and drug-induced or radiation colitis); kidney disease (e.g. ischemic acute renal failure, chronic renal failure); lung diseases (e.g. toxic damage due to oxygen or lung drugs (e.g. paracort, bleomycin, etc.), lung cancer and emphysema); eye diseases (e.g. cataracts, iron storage disease (eyeball siderosis), retinitis, pigmentary degeneration, senile plaques, vitreous scarring, corneal burn with alkali); dermatitis (e.g., erythema multiforme, bullous dermatitis associated with linear immunoglobulin A, cement dermatitis); and other diseases (e.g. gingivitis, periodontitis, septicemia, pancreatitis and diseases caused by environmental pollution (e.g. air pollution), aging, carcinogens, cancer metastases and ailment caused by a decrease in atmospheric pressure)]; diseases caused by the release of histamine or leukotriene C4; coronary artery restenosis after angioplasty and prevention of postoperative adhesions; autoimmune diseases and inflammation (e.g. primary mucosal edema, autoimmune atrophic gastritis, premature menopause, male infertility, juvenile diabetes, pemphigus vulgaris, pemphigoid, sympathetic ophthalmic or conjunctivitis, uveitis caused by the lens, idiopathic leukopenia, active chronic hepatitis, idiopathic hepatitis , chronic lupus erythematosus, autoimmune orchitis, arthritis (e.g. deforming arthritis or multi-cartilaginitis); human immunodeficiency virus (HIV) infection, AIDS; allergic conjunctivitis; hypertrophic scars and keloid caused by injury, burns or surgery.

[0124] In addition, tricyclic macrolides such as, for example, tacrolimus, have liver regenerating and / or hepatocyte hyperplasia and hyperplasia stimulating effects. Therefore, the pharmaceutical composition of the invention is useful for increasing the effect of treatment and / or prevention of liver disease [e.g. immunogenic diseases (e.g.

chronic autoimmune liver disease such as autoimmune liver disease, primary biliary cirrhosis or sclerosing cholangitis), partial liver resection, acute liver necrosis (e.g. toxin-induced necrosis, viral hepatitis, shock or anoxia), hepatitis B, non-A and non-hepatitis B, cirrhosis and liver failure (e.g. fulminant hepatitis, late onset hepatitis and exacerbated chronic liver failure (acute liver failure in the course of chronic liver disease)].

[0125] In addition, the composition of the invention is useful for enhancing the effect of preventing and / or treating various diseases due to the useful pharmacological effect of tricyclic macrolides, such as the complementary activity of the chemotherapeutic effect, activity in cytomegalovirus infection, anti-inflammatory activity, inhibition of peptidyl prolyl isomerase activity or rotamases, anti-malarial activity, anti-tumor activity and so on.

Materials and methods

Materials [0126] Tacrolimus (supplied by Eurotrade); serial number RD 03-111

200 mesh lactose monohydrate (from DMV) _®

Granulated Silicon Oxide, Aeroperl 300, (Degussa)

Polyethylene glycol 6000, Pluracol^® E6000 (with BASF)

Poloksamer 188, Pluronic^® F-68 (from BASF)

Glyceryl monostearate, Rylo^® MD50, (from Danisco Cultor), purity Ph. Eur .; series number

4010056276

Avicel PH200 (microcrystalline cellulose) (with FMC)

DCL 11 lactose (from DMV)

Magnesium stearate

Croscarmellose sodium, Ac-Di-Sol^® (from FMC)

Eudragit^® L30D.55 (from Degussa); Lot No. 1220314079

Triethyl citrate (from Merck); serial number RD03-122

Anti-foaming emulsion (with Dodge)

Mikrotalk [0127] Tablets, capsules and granules can be enteric coated with various types of polymers such as hydroxypropyl methylcellulose acetate succinate (Aqoat), cellulose acetate phthalate CAP, HPMCP hydroxypropyl methylcellulose phthalates or copoly-

<td colspan="2">methacrylic acid units such as Eudragit L30D, Eudragit 100 / S, Eudragit 100 / L. [0128] Comparison of known tacrolimus preparations in vivo studies:</td>

<td colspan="2">Prograf^© hard gelatin capsules, manufactured by Fujisawa Ireland Ltd.</td>

<td>Ingredients</td><td>mg</td>

<td>Tacrolimus, anhydrous</td><td> 1,0</td>

<td>Gelatin</td><td> 6,9</td>

<td>Hypromellose</td><td> 1,0</td>

<td>Lactose monohydrate</td><td> 24,7</td>

<td>Magnesium stearate</td><td> 0,3</td>

<td>Shellac</td><td>as much as needed</td>

<td>Soy Lecithin</td><td>as much as needed</td>

<td>Red iron oxide (E172)</td><td>as much as needed</td>

<td>Titanium dioxide (E171)</td><td>as much as needed</td>

<td>Dimethicon (E900)</td><td>as much as needed</td>

methods

Determination of mass variability [0129] The tablets prepared according to the examples were subjected to a mass variability test, carried out in accordance with Ph. Eur.

Determination of average tablet hardness [0130] The tablets prepared according to the examples were tested for tablet hardness using a Schleuniger Model 6D apparatus, carried out in accordance with the general instructions of the apparatus.

Determination of disintegration time [0131] Tablet disintegration time, i.e. the time to break down into particles or agglomerates, was determined according to Ph. Eur.

Determination of the weighted geometric mean diameter d_g [0132] The geometric weighted average diameter was determined using a laser diffraction method by dispersing the obtained particulate material (or starting material) in air. The measurement was made at a dispersive pressure of 1 bar in a Sympatec Helos device that records the distribution of the equivalent spherical diameter. This distribution adapts to the normal volume-size logarithm distribution.

[0133] In the present specification, "geometric weighted average diameter" means the average diameter from the log of normal volume-size distribution.

In Vitro Dissolution Studies [0134] The following methods were used to test the compositions and dosage forms of the invention.

Test 1:

[0135] In vitro dissolution testing according to USP Method A, delayed release articles (paddle method according to USP; rotation speed: 50 rpm; 37 ° C; after 2 hours in acidic medium, the medium was changed to pH 6 phosphate buffer 8).

Test 2:

[0136] In vitro dissolution test in an aqueous dissolution medium with a pH set to 4.5 (900 ml water with 0.005% HPC (hydroxypropyl cellulose), pH set to 4.5; paddle method according to USP; rotation speed: 50 rpm for a minute).

In vivo studies in beagle dogs [0137] In vivo studies to determine the bioavailability of the compositions of the present invention relative to the bioavailability of a commercially available tacrolimus product, i.e. Prograf®, were performed in beagle dogs.

[0138] The experiments were carried out in Denmark on male beagle dogs with a body weight

12 - 18 kg (initial body weight). The studies were conducted using an open-ended, non-randomized, cross-over method. The dogs received Primperan 5 mg / ml (anti-emetic) for injection before administration and tacrolimus, orally, at a dose of 0.5 to 4 mg.

[0139] The dogs were fasted for 10 hours prior to drug administration (had free access to water) and were fed 5 hours after drug administration (had free access to water). Each dog received a specific dose of tacrolimus regardless of the dog's body weight.

[0140] Blood samples were taken from the external jugular vein at the following time points: Prior to drug administration, 1, 1.5, 2, 3, 4, 6, 8, 12 and 24 hours after drug administration.

4 ml of blood was collected, mixed with EDTA, and then the samples were frozen (-80 ° C). Blood samples were analyzed using the currently used LC / MS extraction and the results were given in ng / ml.

[0141] The obtained tacrolimus concentration profiles in whole blood were processed using WinNonlin® software (Pharsight, California; USA) for pharmacokinetic studies to calculate pharmacokinetic parameters. All data was corrected for the dose administered.

[0142] The following examples are provided to illustrate the invention and not to limit the scope of the invention.

[0143] The pharmaceutical compositions and dosage forms of the invention are illustrated in Examples 1-4, also providing the results of in vitro dissolution tests. The results of in vivo comparative studies on beagle dogs (blood plasma concentration) are given in Examples 5-6.

EXAMPLE 1 [0144] An immediate release tablet with increased bioavailability

The composition of the tablet:

<td></td><td> %</td><td>mg</td>

<td>tacrolimus</td><td> 0,50</td><td> 1,00</td>

<td>200 mesh lactose</td><td> 49,75</td><td> 100,00</td>

<td>PEG 6000</td><td> 34,48</td><td> 69,30</td>

<td>Poloxamer 188</td><td> 14,78</td><td> 29,70</td>

<td>Magnesium stearate</td><td> 0,50</td><td> 1,01</td>

<td>Together</td><td> 100,00</td><td> 201,01</td>

[0145] Tacrolimus was dissolved in polyethylene glycol 6000 and poloxamer 188 (w / w ratio 70:30) at 70 ° C. The solution was sprayed onto 250 g lactose in a Strea-1 fluidized bed. The granulated product was sieved through a 0.7 mm sieve and mixed with magnesium stearate for 0.5 minutes in a Turbula mixer. The mixture was compressed into 8 mm tablets with 1 mg of active substance (200 mg tablet) in the shape of a shell.

[0146] Average disintegration time: 20 minutes. Hardness: 45 N.

EXAMPLE 2 [0147] An immediate release tablet based on PEG 6000 / Poloxamer 188

The composition of the tablet:

<td>Substance</td><td> %</td><td>mg</td>

<td>tacrolimus</td><td> 1,98</td><td> 2,00</td>

<td>Lactose monohydrate, lactose 200 mesh</td><td> 40,50</td><td> 40,91</td>

<td>PEG 6000</td><td> 33,26</td><td> 33,60</td>

<td>Poloxamer 188, Lutrol 68</td><td> 14,40</td><td> 14,40</td>

<td>Magnesium stearate</td><td> 0,50</td><td> 0,51</td>

<td>Talc</td><td> 4,50</td><td> 4,55</td>

<td>Substance</td><td> %</td><td>mg</td>

<td>Croscarmellose sodium, Ac-Di-Sol</td><td> 5,00</td><td> 5,05</td>

<td></td><td> 100,00</td><td> 101,01</td>

[0148] Tacrolimus was dissolved in PEG 6000 at a temperature above 80 ° C. Poloxamer 188 was added and the solution heated to above 80 ° C. Using a Phast FS1.7 feeding unit, the solution was sprayed on 200 g lactose monohydrate in a Phast FB100 fluidized bed. The obtained granulate was passed through Comill, No. 1397 sieve, 4500 rpm and mixed with croscarmellose sodium for 3 minutes in a Turbula mixer.

[0149] Magnesium stearate and talc were sieved through a No. 300 sieve and mixed in a Turbula mixer for 3 min. The granules were mixed with magnesium stearate: talc (1: 9) for 0.5 minutes in a Turbula mixer.

[0150] The resulting mixture was compressed into 6 mm tablets with 2 mg active ingredient (100 mg tablet) in the shape of a shell.

[0151] Average disintegration time: 7 minutes. Hardness: 65 N [0152] The tablets were subjected to an in vitro dissolution test in dissolution medium: 900 ml, 0.005% HPC (hydroxypropyl cellulose) aqueous medium adjusted to pH = 4.5, USP paddle method; rotational speed: 50 rpm; and the following dissolution profile was found:

<td>Time (minutes)</td><td>% release</td><td>Relative standard deviation (Rsd)%</td>

<td> 0</td><td> 0</td><td> 0</td>

<td> 5</td><td> 27,2</td><td> 15,1</td>

<td> 10</td><td> 49,1</td><td> 10,9</td>

<td> 20</td><td> 80,7</td><td> 8,0</td>

<td> 35</td><td> 98,9</td><td> 5,4</td>

<td> 42</td><td> 102,7</td><td> 3,6</td>

<td> 52</td><td> 104,9</td><td> 2,0</td>

EXAMPLE 3

Immediate Release Tablets of Example 2 with enteric coating [0153] The enteric coating is based on the Eudragit L30D-55 acrylic polymer. Eudr20 agit L30D is supplied as an aqueous latex suspension forming an insoluble film upon evaporation of water during coating. The polymer is insoluble at pH values ​​below 5.0 and easily soluble at pH values ​​above 6.0. Tablets prepared as described in Example 2 were coated with the following film-forming composition:

<td>Substance</td><td>% w / w</td>

<td>Eudragit L30D-55</td><td> 40</td>

<td>Water</td><td> 52</td>

<td>Triethyl citrate</td><td> 1,8</td>

<td>Anti-foaming emulsion</td><td> 0,2</td>

<td>Talc (micro)</td><td> 6</td>

<td>Together</td><td> 100</td>

The amount of coating polymer (Eudragit) applied is based on the calculation of mg poly₂ film forming meru per cm of tablet surface. The thickness of the enteric coating was

80 um. The thickness of the coating applied was confirmed by measuring the increase in tablet height with a digital micrometer. The film coating process was carried out in a fluid bed Phast FB100 equipped with a Wurster type insert at air temperature₃ at an inlet of 50 ° C, an inlet air flow rate of 100 m per hour, a product temperature of 38 ° C and a feed rate of 15 g / min.

[0154] The coated tablets were cured in an oven at 30 ° C for 48 hours. Alternatively, coated tablets can be more effectively cured at 40 ° C for 24 hours.

[0155] Enteric-coated tablets were subjected to in vitro dissolution tests using two different dissolution / test centers.

[0156] Using the dissolution medium / test: 900 ml aqueous medium with 0.005% HPC (hydroxypropyl cellulose) adjusted to pH = 4.5 (USP paddle method; rotation speed: 50 rpm), the following dissolution profile was obtained:

<td>Time (hours)</td><td>% release</td><td>Relative standard deviation (Rsd)%</td>

<td> 0</td><td> 0</td><td> 0</td>

<td> 4</td><td> 0,8</td><td> 32,3</td>

<td> 8</td><td> 0,4</td><td> 61,1</td>

<td> 15</td><td> 11,0</td><td> 17,3</td>

<td> 17</td><td> 13,2</td><td> 12,1</td>

[0157] Using the dissolution medium / test: USP, Method A, delayed release products (USP paddle method; rotation speed: 50 rpm), the following dissolution profile was obtained:

<td>Time (minutes)</td><td>% release</td><td>Relative standard deviation (Rsd)%</td>

<td> 0</td><td> 0</td><td>NB</td>

<td> 120</td><td> 0</td><td>NB</td>

<td> 155</td><td> 84,8</td><td> 12,8</td>

<td> 165</td><td> 102,9</td><td>NB</td>

<td> 175</td><td> 101,0</td><td> 3,5</td>

EXAMPLE 4 [0158] The following tacrolimus preparation was prepared as described in Example 2:

<td>Substance</td><td> %</td><td>mg</td>

<td>tacrolimus</td><td> 2,09</td><td> 2,10</td>

<td>Lactose monohydrate, 200 mesh</td><td> 42,75</td><td> 42,95</td>

<td>PEG 6000</td><td> 35,11</td><td> 35,28</td>

<td>Poloxamer 188, Lutrol 68</td><td> 15,05</td><td> 15,12</td>

<td>Magnesium stearate</td><td> 0,50</td><td> 0,50</td>

<td>Talc</td><td> 4,50</td><td> 4,52</td>

<td></td><td> 100,00</td><td> 100,48</td>

The mixture was compressed into 6 mm tablets with 2.1 mg active ingredient (100 mg shell-shaped tablet). Average tablet hardness: 41 N.

EXAMPLE 5

In vivo test for immediate release preparations in dogs [0159] The following tacrolimus preparation was prepared as described in Example 2:

<td>Substance</td><td> %</td><td>mg</td>

<td>tacrolimus</td><td> 0,76</td><td> 0,5</td>

<td>200 mesh lactose</td><td> 49,14</td><td> 32,43</td>

<td>PEG 6000</td><td> 34,73</td><td> 22,92</td>

<td>Poloxamer 188</td><td> 14,88</td><td> 9,82</td>

<td>Magnesium stearate</td><td> 0,50</td><td> 0,33</td>

<td>Together</td><td> 100,00</td><td> 66,00</td>

66 mg of granules were weighed into hard gelatin capsules.

[0160] An in vivo study of this preparation, 0.5 mg on a beagle dog, was performed as described above in the Methods, for Prograf®, 4 x 1 mg (lot # 1 C56050), obtaining the following results:

Blood concentration (ng / ml) in dog No. F1182 after administration of the preparation:

<td rowspan="2">Time (H)</td><td colspan="2">Preparation</td>

<td>Prograf (4 mg)</td><td>According to the invention, the dose adjusted to 4 mg</td>

<td></td><td></td><td></td>

<td> 0</td><td> 0</td><td> 0,0</td>

<td> 0,5</td><td> 0,5</td><td> 10,5</td>

<td> 1,0</td><td> 5,5</td><td> 44,1</td>

<td> 1,5</td><td> 4,1</td><td> 34,3</td>

<td> 2,0</td><td> 4,0</td><td> 21,0</td>

<td> 3,0</td><td> 4,6</td><td> 10,5</td>

<td> 4,0</td><td> 4,0</td><td> 9,1</td>

<td> 6,0</td><td> 2,7</td><td> 4,9</td>

<td> 8,0</td><td> 2,0</td><td> 4,2</td>

<td> 12,0</td><td> 2,0</td><td> 3,5</td>

<td> 24,0</td><td> 0,7</td><td> 2,1</td>

Relative bioavailability based on AUC (according to the invention / Prograf): 293%.

EXAMPLE 6

In vivo test for immediate release preparations in dogs [0161] The following tacrolimus formulation according to the invention was prepared as described in Example 2:

<td>Substance</td><td> %</td><td>mg</td>

<td>tacrolimus</td><td> 1,86</td><td> 0,50</td>

<td>Monohy lactose 200 mesh drate</td><td> 43,56</td><td> 11,72</td>

<td>PEG 6000</td><td> 31,21</td><td> 8,40</td>

<td>Poloxamer 188, Lutrol 68</td><td> 13,37</td><td> 3,60</td>

<td>Magnesium stearate</td><td> 0,50</td><td> 0,13</td>

<td>Talc</td><td> 4,50</td><td> 1,21</td>

<td>Croscarmellose sodium, Ac-Di-Sol</td><td> 5,00</td><td> 1,35</td>

<td></td><td> 100,00</td><td> 26,92</td>

[0162] It was compressed into 4 mm tablets with 0.5 mg active ingredient (27 mg tablet in the shape of a shell).

[0163] An in vivo study of this preparation, 0.5 mg on a beagle dog, was performed as described above in the Methods, in relation to Prograf®, 0.5 mg capsules (lot no .: OC512OD), obtaining the following results: blood (ng / ml) in dog No. 1 after administration of the preparation:

<td rowspan="2">Time (H)</td><td colspan="2">Preparation</td>

<td>Prograf (0.5 mg)</td><td>Preparation B (0.5 mg)</td>

<td> 0</td><td> 0</td><td> 0</td>

<td> 0,5</td><td> 0,95</td><td> 0,04</td>

<td> 1,0</td><td> 0,84</td><td> 1,56</td>

<td> 1,5</td><td> 0,55</td><td> 4,68</td>

<td> 2,0</td><td> 0,40</td><td> 9,11</td>

<td> 3,0</td><td> 0,26</td><td> 2,82</td>

<td> 4,0</td><td> 0,18</td><td> 2,46</td>

<td> 6,0</td><td> 0,18</td><td> 1,10</td>

<td> 8,0</td><td> 0,14</td><td> 1,25</td>

<td> 12,0</td><td> 0,11</td><td> 0,74</td>

<td> 24,0</td><td> 0,06</td><td> 0,40</td>

<td> 25,0</td><td> 0,06</td><td> 0,44</td>

Relative bioavailability based on AUC (according to the invention versus Prograf): 742%.