Once daily oral dosage form comprising tacrolimus

The present invention relates to a sustained release oral dosage form comprising tacrolimus active ingredient for use in once-daily immunosuppressive therapy in a patient in need thereof, which dosage form releases the active substance for a very prolonged period of time and which ensures high bioavailability in vivo and improved pharmacokinetic profile compared to conventional dosage forms.

[0002] Tacrolimus, also known as FK-506 or FR-900506, is an active ingredient in Prograf®, Protopic® and Advagraf® approved by the European Agency for the Evaluation of Medicinal Products (EMEA) on April 23, 2007. During the development of Advagraf The product was known as MR4. Detailed information on Advagraf is provided in the EPAR (European Public Assessment Reports) for authorized medicinal products for human use, including the scientific discussion paper published by the EMEA on approval and product information (etiquette, 25/01/2008 Advagraf -HC-712- T-03). Tacrolimus (Prograf®) was approved by the FDA in April 1994 under number NDA 050708 to prevent organ rejection in patients receiving allogeneic liver transplants. It is also approved in the European Union, Japan, Canada and Switzerland and in many other countries under the same brand. It is approved for prophylaxis of organ rejection in patients receiving allogeneic liver, kidney or heart transplants. It is estimated that 72% of all kidney transplant recipients and 89% of all liver transplant recipients receive tacrolimus.

[0003] Tacrolimus, administered as Prograf capsules, exhibits a large inter- and intra-individual variability in its absorption and metabolism. Because of this variation, the standard dosage is not an accurate predictor of concentration. In clinical use, dose adjustments for tacrolimus are often required based on monitoring of tacrolimus blood levels. Tacrolimus occurs in the form of white crystals or crystalline powder. It is practically insoluble in water, easily soluble in ethanol and soluble in methanol and chloroform.

[0004] The production of tacrolimus is described in EP-A-0184162, and tacrolimus analogs are disclosed e.g. in EP-A-0444659 and US 6387918.

[0005] Tacrolimus is a macrolide compound with useful immunosuppressive activity, antimicrobial activity and other pharmacological activities and is valuable in the treatment or prophylaxis of rejection reactions in organs or tissue transplants, graft-versus-host diseases, autoimmune diseases and infectious diseases.

[0006] Tacrolimus inhibits the activation of T lymphocytes, although the exact mechanism of action is unknown. Experimental evidence suggests that tacrolimus binds to the intracellular FKBP-12 protein. Next, the tacrolimus-FKBP-12 complex, calcium, calmodulin and calcineurin are formed and the phosphatase activity of calcineurin is inhibited. This effect may prevent dephosphorylation and translocation of the nuclear factor of activated T cells, a nuclear component that is believed to initiate gene transcription for lymphokine formation. As a result, the activation of T lymphocytes occurs, i.e. immunosuppression.

[0007] Tacrolimus is extensively metabolised by the CYP3A4 isoenzyme in the intestinal and liver walls. CYP3A4 is present or expressed in all parts of the gastrointestinal tract, including the colon. It has been observed that the absorption is negatively affected by simultaneous food intake. Thus, the rate and extent of tacrolimus absorption was greatest in fasting conditions.

[0008] Tacrolimus is known to cause significant undesirable effects, of nephro- or neurotoxic origin, as well as side effects of Gl and others.

[0009] Absorption of tacrolimus from the gastrointestinal tract after oral administration is rapid with mean time to reach peak concentration (tmax) approximately 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 and is at most about 20% after oral administration.

Frequently 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.

[0011] For oral administration, tacrolimus is currently formulated and marketed in the form of soft gelatin capsules containing 0.5, 1 or 5 mg of anhydrous tacrolimus equivalent and is commercially available under the tradenames Prograf® and Protropic®. The recommended initial oral dosage for the patient is about 0.1 to 0.2 mg / kg / day. The dose is intended to provide a certain 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. Detailed information on clinical pharmacology, pharmacokinetics and clinical trials are described on the label approved by the FDA on April 27, 2006 for Prograf, NDA No. 50708.

[0012] There remains a need for new pharmaceutical compositions and / or dosage forms containing tacrolimus exhibiting improved bioavailability and improved pharmacokinetic properties. Increased bioavailability in combination with a sustained release formulation may allow reduction of unit doses taken by the patient, e.g. up to a single dose per day, without the risk of no clinical effect due to low doses in the last part of the dosing interval. In addition, fluctuations in plasma concentration versus time profile can be significantly reduced. In addition, increased bioavailability may lead to more reproducible (i.e., less variable compared to Prograf^®) release profile.

[0013] Sustained release tacrolimus formulations are described in WO99 / ​​49863 (Fujisawa Pharmaceutical Co.), inter alia, conferred as US patents No. 6,440,458, US 6,576,259 and US 6,884,433 on a formulation in which the dissolution time is 63 , 2% (T63.2%) of tacrolimus is between 0.7 and 15 hours. However, the formulation in which 63.2% is released over 42 minutes appears to be only slightly different from the conventional immediate release formulation of tacrolimus, having 68.4% released within 30 minutes. It is clearly stated that when the formulation has a T63.6 value of more than 15 hours, the release of the active substance will be so delayed that the active substance will be eliminated from the body before an effective concentration in the blood is reached. The most preferred embodiments are sustained-release formulations of T63.6 of 2-5 hours. The formulations prepared according to the examples of the application have a T63.6% value of 1.9, the fastest-release formulation, up to 8.2 hours for the slowest-release formulation. In addition, tacrolimus is found to be perfectly absorbed, and its absorption capacity is suppressed with sustained-release formulations. From the examples, improved bioavailability can be achieved with all of the formulations tested. The T63.6% values ​​disclosed for these formulations were 3.0, 3.3, 2.0 and 2.5, respectively. 2 hours for the slowest-release formulation. In addition, tacrolimus is found to be perfectly absorbed, and its absorption capacity is suppressed with sustained-release formulations. From the examples, improved bioavailability can be achieved with all of the formulations tested. The T63.6% values ​​disclosed for these formulations were 3.0, 3.3, 2.0 and 2.5, respectively. 2 hours for the slowest-release formulation. In addition, tacrolimus is found to be perfectly absorbed, and its absorption capacity is suppressed with sustained-release formulations. From the examples, improved bioavailability can be achieved with all of the formulations tested. The T63.6% values ​​disclosed for these formulations were 3.0, 3.3, 2.0 and 2.5, respectively.

[0014] The inventors of the present application are found in the patent application WO 2005/020993. They also tested various preparations of tacrolimus in dogs and minipast Beagles, showing however that both the rapid release tablet (example 18) and the slow tablet release (example 19) may lead to improved bioavailability compared to Prograf®. This indicates that improved bioavailability may be associated with tacrolimus in the dissolved state in the dosage form, which also appears in WO 2005/020994 by the same inventors regarding tacrolimus-containing solid dispersions. Rapid release Prograf® powder contains tacrolimus in a physical mixture of HPMC, lactose,

[0015] The sustained release oral dosage form comprises tacrolimus once daily (Advagraf®) M. Roy First et al., Yonsei Medical Journal, vol. 45, no. 6, pp. 1127-1131.

[0016] One serious problem with modified or extended dosage forms is the difficulty in obtaining sufficient absorption in the lower gastrointestinal tract when oral dosage forms entering the large intestine can be easily excreted before substantial release. The release is generally smaller due to the lack of fluids and the physical interaction of the dosage forms with increasingly constant contents of the large intestine. In addition, the absorption surface is several times smaller than the absorbent surface of the small intestine, and this factor increases the time in which the released active substance is degraded and trapped in the solids present in the colon.

[0017] It is generally accepted that too much excretion can seriously affect bioavailability, even for substances that require good penetration into the large intestine. In the case of substances that are substrates for CYP3A4, the benefits of a lower concentration of metabolisation enzymes in the lower glory can be expected from the point of view of bioavailability. On the other hand, the relative higher concentration of the transporter system, Glycoprotein P in the lower GI, including the large intestine, usually counteracts the action of low CYP3A4 enzymes because the molecules that enter enterite are transported back to the large intestine by the transporter. Tacrolimus is a known substrate for these mechanisms, both CYP3A4 metabolism and the P-glycoprotein transport system. According to increased bioavailability can not be correlated with prolonged release in a straight linear fashion. The release can be carefully adjusted to eliminate several counteracting factors. These factors include lower intestinal absorption area, lower fluid content, higher solids content, bacterial degradation, higher P-glycoprotein transporter effect, lower mobility, differences in mucosal barriers and / or mucosal composition and differences In pH along the large intestine compared to from the small intestine. Accordingly, the control and release time of the sustained release dosage form in vivo to achieve predictable release under the various physical conditions along the GI pathway is a challenge, especially considering that immunosuppressive treatment in patients requiring transplantation requires blood concentration within very narrow limits to balance effectiveness (no rejection) and side effects (infection, nephrotoxicity, metabolic and cardiovascular diseases, etc.). Providing an improved daily treatment formulation in which the release is extended to an accurate level where the resulting pharmacokinetic parameters are fully optimized without compromising safety, i.e. if the variant and interindividual variation is high for important pharmacokinetic parameters, if the correlation between the minimum Concentration and bioavailability is not present, which is an important factor in the treatment of narrow therapeutic drugs, such as tacrolimus, to balance effectiveness (no rejection) and side effects (infection, nephrotoxicity, metabolic and cardiovascular diseases, etc.). Providing an improved daily treatment formulation in which the release is extended to an accurate level where the resulting pharmacokinetic parameters are fully optimized without compromising safety, i.e. if the variant and interindividual variation is high for important pharmacokinetic parameters, if the correlation between the minimum Concentration and bioavailability is not present, which is an important factor in the treatment of narrow therapeutic drugs, such as tacrolimus, to balance effectiveness (no rejection) and side effects (infection, nephrotoxicity, metabolic and cardiovascular diseases, etc.). Providing an improved daily treatment formulation in which the release is extended to an accurate level where the resulting pharmacokinetic parameters are fully optimized without compromising safety, i.e. if the variant and interindividual variation is high for important pharmacokinetic parameters, if the correlation between the minimum Concentration and bioavailability is not present, which is an important factor in the treatment of narrow therapeutic drugs, such as tacrolimus,

Wherein the failure of treatment is closely related to organ rejection, and dose adjustments must take place under safe conditions. Further factors that reduce the risk of clinical success in the preparation once a day in organ transplantation include a high incidence of gastrointestinal complications affecting digestive system parameters, including administration times, pH, bacterial composition and other GI functions. These complications include nausea, vomiting and very often diarrhea. The present invention provides a sustained release oral dosage form comprising from 0.1 mg to 15 mg tacrolimus in a hydrophilic or water miscible carrier for a single daily immunosuppressive therapy in need of a patient, polyvinyl alcohol (PVA), polymethacrylic polymers, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), methylcellulose, sodium carboxymethylcellulose, hydroxyethylcellulose, pectins, cyclodextrins, galactomannans, alginates, carrageenans, xanthan gums and mixtures thereof. In another embodiment, the present invention provides a sustained release oral dosage form comprising from 0.1 mg to 15 mg tacrolimus in a hydrophilic or water miscible carrier, for a one-day immunosuppressive therapy of a patient in need thereof, which dosage form releases the active substance With a significant zero-release profile in the extended period of time determined by the release from an 8-hour time point to a 15-hour time-point, galactomannans, alginates, carrageenans, xanthan gums and mixtures thereof. In another embodiment, the present invention provides a sustained release dosage form of the invention for use in immunosuppressive treatment of a patient in need thereof in a once daily schedule. Preferred embodiments are set forth in the dependent claims.

[0018] Accordingly, the present inventors surprisingly found that a dosage form that releases tacrolimus in a very long and controlled period is able to administer tacrolimus in vivo in such a way that tacrolimus is sufficiently well absorbed at the same time to understand That tacrolimus is not lost in the lower gastrointestinal tract, the release is slow enough to allow a very low rate of absorption, with the maximum concentration being controlled

At a lower value, the minimum concentration increases the efficacy of the treatment in the full dosage range of Z 24 hours. It is very important that the minimum concentration achieved 24 hours after administration of the sustained release preparation of the present invention is very predictable and can be used as a marker of absolute bioavailability, because a high correlation between the minimum concentration and the observed reality of bioavailability is obtained in the last 24 hours before measurement minimum concentration. The minimum concentration can therefore be safely used as a dosing and adjustment tool during treatment.

[0019] Conventional in vitro dissolution methods are believed to correlate or at least reflect the actual in vivo release profile of a human. Accordingly, the in vitro difference in the release rate between the two formulations tested under the same conditions should reflect the difference in the in vivo release rate. However, exceptions may apply if, for example, one formulation has a pH-dependent release and the other does not, and the actual pH values ​​for the tests are not selected to detect such a difference. A clear example is the study of an enterally coated formulation at high temperature, which provides immediate in vitro release and delayed release in vivo. In addition, comparing two extended formulations with different release mechanisms, for example, an osmotically releasing release mechanism compared to an erosive dependent release mechanism, the same in vitro release profiles for both products can theoretically exhibit different in vitro profiles, while decreasing or Increasing the rate of dissolution will reflect in vivo for each product. Accordingly, while the evidence is not contradictory, and provided that the methods are performed according to the pharmacopoeia recipe, conventional dissolution methods are useful tools for differentiating between formulations and corresponding in vivo properties. In accordance with the present invention, the present invention provides in a first aspect an oral sustained release dosage form comprising tacrolimus as an active substance in immunosuppressant treatment once daily for a patient in need thereof, which dosage form releases the active substance over a very long period of time. In a further aspect, the release is characterized by a significant zero zero for most releases.

[0020] Typical in vitro dissolution methods include the methods described in the US Pharmacopeia (USP) as the official standardization body for all prescription and non-prescription drugs in the United States and similar pharmacopoeias for Europe and Japan. Preferred methods include USP dissolution I (basket) method II and method (stirrer) at 50 rpm, use of HPC to prevent adherence of the drug to the device and pH for stability reasons of 4.5. Since tacrolimus is not protonated, pH does not affect the solubility of the drug, however, pH modification may be important if the pH is pH-sensitive, inactive excipient at pH 4.5, pH is not usually used in the formulation. present in the Gl tube. Correspondingly, it can be important to describe the extent of release elongation using alternative dissolution methods. In addition, the extended formulation can be further characterized by other dissolution methods, including methods with different speeds of rotation, different pH values, the use of dissolution media simulating Gl conditions (e.g., Simulation of fasting and administration media, FaSSIP and FeSSIP medicines), Application of additives to the dissolution medium , such as SLS to increase wettability or solubility of tacrolimus, thereby reducing the total measured dissolution time (increasing dissolution rate).

[0021] The inventors have found that the bioavailability of tacrolimus is significantly increased and pharmacokinetic parameters are significantly improved when tacrolimus is administered to a mammal in a sustained release composition in which the release and release time of the active ingredient, i. E. Release profile in in vitro and in vivo, is extended by more than 15 hours as measured by conventional dissolution methods used in tacrolimus dosage forms and measured in vivo by pharmacokinetic parameters of clinical significance and relevant for proving the prolongation of in vivo release. These pharmacokinetic parameters include: a significant prolonged time to reach maximum concentration; Low maximum concentrations; High minimum concentrations, longer average stay,

[0022] This specification also describes the use of sustained release compositions for a more safe immunosuppressive treatment due to an improved pharmacokinetic profile obtained in healthy subjects and patients, and has been demonstrated in several single dose and steady pharmacokinetic studies compared to a conventional commercial dosage form. Safe immunosuppressive treatment also applies to the specific conversion dose regimen of treatment twice daily with Prograf®, in which the conversion is to be carried out at a dose of 1: 0.66-0.80 (depending on the nearest available tablet volume). Such a dosing regimen, giving a comparable mean blood concentration in the dose interval measured before and after the conversion,

Definitions [0023] As used herein, the term "active ingredient" or "active pharmaceutical ingredient" means any ingredient that is intended to provide pharmacological or other direct effect in the diagnosis, treatment, alleviation, treatment or prevention of disease or affecting the structure or any function human body or other animals. The term includes those components that may chemically change in the manufacture of a medicinal product and are present in a modified drug pharmaceutical product to determine a specific activity or effect.

In the present context, the term "hydrophilic" describes that something "likes water", i.e. a hydrophilic molecule or part of the molecule is that it is typically electrically polarized and capable of forming hydrogen bonds with water molecules, which enables easier dissolution of Water. than in oil or other "non-polar" solvents.

In the present context, the term "amphiphile" describes a molecule (as a surfactant) having a polar water-soluble group attached to a water-insoluble hydrocarbon chain. Thus, one end of the molecule is hydrophilic (polar) and the other end hydrophobic (non-polar).

[0026] In the present context, the term "hydrophobic" means a compound that tends to be electrically neutral and nonpolar, and thus prefers other neutral and nonpolar solvents or molecular environments.

[0027] As used herein, the term "vehicle" means any solvent or vehicle fluid in a pharmaceutical product that has no pharmacological role. For example, water is a carrier for xylocaine and propylene glycol as a carrier for many antibiotics.

[0028] In the present context, the term "solid dispersion" means a drug or active ingredient or substance dispersed at the level of solid particles in an inert carrier, carrier, diluent or matrix in solid state, i.e. Typically in a fine dispersion of particulate matter. .

[0029] In the present context, the term "solid solution" means a drug or active substance or substance dissolved at the molecular level in an inert carrier, carrier, diluent or matrix in the solid state.

[0030] The term "drug" means a compound intended for use in the diagnosis, treatment, alleviation, treatment or prophylaxis of diseases in a human or other animal.

[0031] In this context, the term "dosage form" means the form in which the drug is delivered to the patient. This could be parenteral, topical, tablet, oral (liquid or dissolved in powder), suppository, inhalation, transdermal, etc.

[0032] As used herein, the term "bioavailability" means the degree to which a drug or other substance becomes available to the target tissue after administration. As used herein, the term "bioequivalence" means the scientific basis for comparing generic and branded drugs. For example, drugs are bioequivalent if they are put into circulation at the same rate when given at similar doses under similar conditions. Parameters often used in bioequivalence studies are tMaks, CMaks, AUC0-infinity, AUC0-t. Other relevant parameters may be W50, W75 and / or MRT. Accordingly, at least one of these parameters can be used to determine biodiversity. Furthermore, in the present context, the two compositions are considered to be bioequivalent,

[0033] In the present context, "tMax" means the time to reach the maximum plasma concentration (okMax) after administration; AUC0-infinityThis area under the plasma concentration versus time curve from time 0 to infinity; AUC0-t Indicates the area under the plasma concentration versus time curve from time 0 to time t; W50 Indicates the time at which the plasma concentration is 50% or more of CMaks; W75 is the time at which the plasma concentration is 75% or more of CMaks; And MRT means the average residence time of tacrolimus. Swing means (CMaks-CMin) / CMin I hesitation (CMaks-CMin) / Average. The fluctuation is suitable for comparing dosage forms that provide different bioavailabilities.

[0034] In this context, the term "medicine" means a compound used to treat a disease, injury or pain. Medicine is fairly divided into "prophylaxis", ie the art of health and "therapeutic", ie the art of restoring health.

[0035] In the present context, the terms "controlled release" and "modified release" are intended to be equivalents comprising any type of tacrolimus release from a composition of the invention that is suitable for obtaining a specific therapeutic or prophylactic response after being administered to the subject. The person skilled in the art knows how controlled release / modified release differs from the release of ordinary tablets or capsules. The terms "release in a controlled manner" or "release in a modified manner" have the same meaning as given above. The terms include slow release (which results in lower CMaks and later tMax, but 1½ is unchanged), extended release (which results in lower CMaks, later tMaks, but apparently you₂ It's longer); Delayed release (resulting in unchanged CMaks, but delay time and, respectively, tMaks is delayed, and% is unchanged) and pulsed release, vesicle release, sustained release, sustained release, time optimized release, rapid release (for better results)

Initial operation) etc. Included in the terms are also e.g. Using specific conditions in the body, e.g. Various enzymes or changing pH to control the release of the drug substance. The term sustained release release is chosen because it is believed that the term most correctly includes the in vivo release of the product.

[0036] In this context, the term "erosion" or "eroding" means the gradual distribution of the surface of a material or structure, for example a tablet or a tablet coating. As used herein, the term generally means the dissolution of the polymer responsible for the elongation of release is faster than the dissolution of the active ingredient, whereby the polymer dissolves more quickly than the active substance. In other words, the release is mainly controlled by erosion, not by dissolving the active ingredient in the polymer matrix system.

[0037] This specification discloses pharmaceutical products to improve the treatment of conditions that respond to tacrolimus treatment, especially in cases where an immunosuppressive effect is desired.

[0038] In the context of the present invention, tacrolimus is in any physical form (crystals, amorphous powder, any possible polymorphs, optional solvates, including hydrate, anhydrate, complexes, etc.). Pharmaceutically acceptable salts and solvates are also included. However, it is believed that smaller particle sizes on the micro and nano scale and a favorable molecular solution will contribute to the predictable and constant release of tacrolimus in vivo.

[0039] It is generally accepted that the basket dissolving device may be more suitable for capsules, and the paddle dissolution apparatus is more suitable for disintegrating tablets. However, the most suitable dissolution device can be easily recognized by examining whether the one or the other device achieves the highest compatibility.

[0040] In a preferred embodiment, the in vivo release takes place at a constant characteristic value, whereby a significant release profile of zero can be obtained over a longer period of time. Because sufficient release is required at a time when the dosage form could reach the large intestine, such an appropriate period at which zero release is desired can be determined by release from 8 hours to 15 hours when tested according to the USP II dissolution test (shoulder blade) ) Or USP I dissolution (sediment) test in medium at pH 4.5 and containing 0.005% hydroxypropylcellulose and 50 rpm. Because the solid dosage form can leave the stomach soon after ingestion or can be stopped there for a few hours before reaching the Gl tube, it is also desirable for the more initial release to be well controlled,

[0041] In another embodiment of the invention, the addition of a surfactant to the release medium provides a release rate, wherein the release of at most 80% of the active compounds is prolonged for at least 7 hours, e.g. at least 8 hours, e.g. at least 9 hours , e.g. at least 10 hours, e.g. at least 11 hours, e.g. at least 12 hours, e.g. at least 13 hours after an in vitro test. According to the USP II dissolution test (vane) or the USP I dissolution test (cage) in medium at pH 4.5 and containing 0.005%

[0042] As mentioned before, if the release is too stretched, the dosage form can be separated before complete discharge or release is carried out too much; far to ensure sufficient distribution. Accordingly, the dosage form content should be released with the indications indicated herein, but can not be extended beyond a maximum of 24 hours, e.g. within 23 hours at most, e.g. within a maximum of 22 hours, e.g. at a maximum of 21 hours, such as at most 21 hours, such as a maximum of 21 hours, such as a maximum of 18 hours, such as a maximum of 17 hours, such as a maximum of 16 hours, calculated on 80% of the content and Addition 0,

Alternatively or additionally, the dosage form fulfills the condition in which 63.5% of the active ingredient release is prolonged within a maximum of 20 hours, such as at most 18 hours. Shorter dissolution times may also be preferred as an upper limit, such as at most 16 hours, such as at the most 15.5 hours, also in In vitro. According to the USP II dissolution test (shoulder) or the USP I dissolution test (basket) in the environment at pH 4.5 and containing 0.005% hydroxypropylcellulose and a turnover of 50 rpm.

[0044] Sustained release oral dosage, wherein release is initiated within 120 minutes, e.g. within 90 minutes, such as within 60 minutes after deposition of a dosage form in a dissolution apparatus during an in vitro test. According to the USP II dissolution test (paddle) or dissolution test (basket) USP I in an environment of pH 4.5 and containing 0.005% hydroxypropylcellulose, and a rotation of 50 rpm indicates a composition that will provide a predictable release profile. Until an early release, such as within the first two hours are not fast. If release does not occur soon after administration, the patient is exposed to low concentrations. The patient is generally titrated according to the blood values ​​observed immediately before the daily intake of the minimum concentration observed during the day. The release delay will be ensured by a later unknown minimum concentration.

[0045] The following release properties are considered to be within the scope of the invention:

A) sustained release oral dosage form that releases at most about 20% wt / wt. Active substance within 1 hour or within 2 hours, or within 3 hours, or within 4 hours or within 5 hours, In vitro. According to the USP II dissolution test (shoulder) or the USP I dissolution test (basket) in the environment at pH 4.5 and containing 0.005% hydroxypropylcellulose and a turnover of 50 rpm.

B) a sustained release oral dosage form that releases 40% w / w of Active substance in 10 to 14 hours, e.g. in about 11 to 13 hours, In vitro According to the USP II dissolution test (spatula) or USP I dissolution test (basket) in an environment of pH 4.5 and containing 0.005% hydroxypropylcellulose and 50 revolutions per minute.

C) sustained release oral dosage form that releases 20% w / w of The total amount of active substance released over a period of 6 to 10 hours, e.g. in about 7 to 9 hours, in vitro. According to the USP II dissolution test (paddle) or the USP I dissolution test (basket) in an environment of pH 4.5 and containing 0.005% hydroxypropyl cellulose and 50 revolutions per minute.

EP 2 167 033 B1

D) Sustained release oral dosage form that releases 50% w / w of Active substance in 13 to 17 hours, e.g. in about 14 to 16 hours, in vitro According to the USP II dissolution test (shoulder) or the USP I dissolution test (basket) in a pH 4.5 environment and containing 0.005% hydroxypropylcellulose and 50 revolutions per minute.

E) sustained release oral dosage form in which the release profile is substantially linear over a period of from 4 to 8 hours, defined as a gradient or slope in the range of 25% slope or slope measured at 6 o'clock, e.g. in the range of 15%, preferred W 10%.

F) a sustained release oral dosage form in which the release profile is substantially linear over a period of from 6 to 10 hours, defined as a gradient or slope in the range of 25% gradient or slope measured at 8 o'clock, e.g. in the range of 15%, preferably in 10%.

G) a sustained release oral dosage form in which the release profile is substantially linear over a period of from 8 to 12 hours, defined as a gradient or slope in the range of 25% inclination or slope measured at 10 o'clock, e.g. in the range of 15%, preferred W 10%.

H) Oral sustained release oral dosage form in which the release profile is substantially linear over the release period from the point where 20% is released to the time point at which 50% is released as a gradient or slope at 80% of the W time point within 25% of the slope or slope measured at a 20% time point.

I) A sustained release dosage form according to any one of the preceding claims, wherein the release elongation mechanism is not through the penetration control coating.

[0046] It is contemplated that the release profile profile defined above significantly increases the bioavailability of tacrolimus in mammals because all or most of the active ingredient is actually released in the gastrointestinal tract, such that substantially CYP3A4 metabolism is avoided or at least significantly reduced. Furthermore, it is believed that this effect is correlated or at least reflected in the in vitro dissolution profile in the dosage forms of the invention, which profile is easily noticed when administering the dosage forms by the conventional in vitro dissolution method.

[0047] The desired release profile of the dosage form can be achieved by combining one or more of the following possibilities.

I) coating the composition with an enteric coating; And / or

Ii) use of a pharmaceutical composition comprising a solid dispersion or a solid solution of the active ingredient, i.e. tacrolimus, in a hydrophilic or water miscible carrier and one or more modifying releasing agents.

[0048] The enteric-coated preparation may, however, have the disadvantage of delaying release without increasing the release, and therefore should be used in conjunction with the expanding technology.

[0049] In yet another embodiment of the invention there is provided a tacrolimus extended release pharmaceutical composition comprising the active ingredient dissolved or dispersed in a hydrophilic or water miscible vehicle as described above, preferably a vehicle selected from polyethylene glycols, polyoxyethylene oxides, poloxamers, polyoxyethylene stearates , poly-epsilon caprolactone, polyglycolized glycerides such as Gelucire^®, And mixtures thereof, more preferably polyethylene glycol optionally in admixture with a poloxamer. A specific example of a useful mixture is a mixture of 70% w / w. Polyethylene glycol 6000 (PEG6000) and 30% w / w. Poloxamer 188.

[0050] The present specification also discloses a particulate pharmaceutical composition comprising tacrolimus together with one or more pharmaceutically acceptable excipients, wherein the composition after oral administration to a mammal in need thereof exhibits an AUC / AUCPrograf® Value of at least about 1.3 , AUC values ​​are determined under similar conditions.

[0051] As the examples show, the availability of bioavailability after administration of the composition disclosed herein is much better. Thus, in particular embodiments, the AUC / AUCPrograf® value is at least about 1.25, e.g. about 1.5 or more, about 1.8 or more, about 1.9 or more, about 2.0 or more, and the AUC values ​​are determined under similar conditions.

[0052] Following oral administration of the pharmaceutical composition as disclosed herein, it is contemplated that the plasma concentration depending on the time profile shows a prolonged period during which the plasma concentration is maintained in the therapeutic window (i.e., the plasma concentration leads to a therapeutic effect) without leading to serious undesirable side effects. There is also a reduction in peak concentration. Accordingly, the present specification discloses a particulate pharmaceutical composition comprising tacrolimus together with one or more pharmaceutically acceptable excipients, wherein the composition after oral administration to a mammal in need thereof releases tacrolimus in a controlled manner and exhibits a Cmaxthat of at most 80% Cmax. for Prograf® tablets, e.g., at most 75%, at most about 70%,

[0053] In the present context, controlled release and extended release terms are intended to be equivalent to include any type of tacrolimus release from a composition of the invention that is suitable for achieving a particular therapeutic or prophylactic response when administered to a patient. The person skilled in the art knows how the controlled release / extended release release is different from the release of ordinary tablets or capsules. The terms "release in a controlled manner" or "release in an extended manner" have the same meaning as stated above.

[0054] The terms controlled release / extended release include slow release (resulting in lower Cmax and later tmax, but t% remains unchanged), extended release (resulting in lower Cmax, later tmax, but apparently t% and longer); Delayed release (resulting in an unchanged Cmax, but a delay, therefore tmax is delayed, at% remains unchanged), as well as pulsed release, vesicle release, sustained release, sustained release, optimized for safer release, rapid release (to get a better start of action ) etc. Included in the terms is also eg. The use of specific conditions in the body, e.g. Various enzymes or pH changes to control the release of the drug substance.

More specifically, after oral administration to a mammal, including a human, a pharmaceutical composition containing a dose of 5 mg of tacrolimus, tacrolimus is released in a controlled manner and shows a Cmax which is at most 30 ng / ml Such as, e.g., At most 25 ng / ml or at most about 20 ng / ml. [0056] However, a reduction in peak concentration can not lead to a reduction in the therapeutic effect as long as the plasma concentration of tacrolimus is maintained in the therapeutic window. Accordingly, the present invention also discloses a pharmaceutical composition wherein W 50 is at least about 2 hours, e.g., at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about About 7 hours, at least about 8 hours, at least

About 9 hours, about 10 hours or more, about 11 hours or more, about 12 hours or more, about 13 hours or about 14 hours or more.

[0057] Furthermore, or further, the composition according to the invention has a Cdiff = [Cmax-Ct (t = 12 hours)] lower than Prograf® under the same conditions. If the Cdiff for Prograf® is set to 100, then the Cdiff of the compositions of the invention is typically 90 or less, e.g. about 85 or less, about 80 or less, about 75 or less, about 70 or less, about 65 or less, about 60 or less, about 55 or less, about 50 or less, about 45 or less, or about 40 or less.

[0058] More specifically, after oral administration to a mammal, including a human, of a pharmaceutical composition of the invention containing 5 mg of tacrolimus, tacrolimus is released in a controlled manner and exhibits a Cdiff of about 20 ng / ml or less, such as, e.g., about 15 ng / ml. or less, about 13 ng / ml or less or about 10 ng / ml or less.

[0059] The pharmaceutical composition described herein releases tacrolimus in a controlled manner to extend the therapeutic effect of tacrolimus. In one aspect, the release may depend on pH, i.e. the release generally occurs after passage of the stomach. Such pH-dependent release is mainly provided by an enteric coating as described herein. The release may also be independent of pH, e.g. by providing the composition with a controlled release coating such as e.g. a cellulosic coating, e.g. Ethylcellulose or by providing a composition in the form of a matrix composition such as a hydrophilic cellulose polymer matrix, e.g. based on HPMC. You can of course use a combination.

[0060] In general, a change in the bioavailability and / or alteration of other bioavailability parameters is usually determined in in vivo studies in a suitable animal model examining the compositions in question together with e.g. Prograf® or a similar commercially available product containing tacrolimus. The use of a dog model to determine the bioavailability of certain preparations is a general practice in the pharmaceutical industry.

[0061] Studies of relevance to tacrolimus are not radomized, cross-sectional studies where each dog is its own control. Usually four dogs and four treatments are used. Because no intravenous administration is given, the bioavailabilities obtained are relative.

[0062] Furthermore, it has surprisingly been found that the need for simultaneous food intake to provide sufficient uptake of tacrolimus is significantly reduced or even completely abolished.

[0063] Thus, pharmaceutical compositions as disclosed herein provide significant bioavailability of tacrolimus, which can reduce the number of doses administered per day and reduce or eliminate the need for administration in combination with food intake, which provides a higher degree of freedom to recipients of pharmaceutical compositions and thus patient acceptance and / or compliance can be significantly improved. In addition, the compositions cause a significant reduction of side effects, especially side effects associated with high peak concentration (such as nephro- and neurotoxicity, diarrhea, constipation, abdominal pain, nausea, etc.) and predict prolonged release of Tacrolimus, leading to better therapy.

[0064] A further advantage of the invention with a sustained release dosage form is the possibility of obtaining an effective therapeutic response with a reduced dose compared to traditional oral treatment. Similar bioavailability and improved profile following administration at a dose that is at most about 85% w / w, e.g. at most about 80% w / w. At most about 75%,

At most about 70% w / w, at the most about 65% w / w, at most about 60% w / w. At most about 55% w / w. Or at most 50% weight / weight. Doses of tacrolimus Administered in the form of Prograf® or a similar commercial product containing tacrolimus or as a commercially available sustained release product, including Advagraf®.

[0065] The parameters frequently used in bioequivalence studies are t max, cm max, AUC 0 and fineness, AUC 0-t. Other important parameters may be W50, W75 and / or MRT. Accordingly, at least one of these parameters can be used to determine biodiversity. Furthermore, in the present context, the two compositions are considered to be bioequivalent if the value of the parameter used is in the range of 80-125% relative to Prograf® or a similar commercial product containing tacrolimus used in the test.

[0066] In the present context, "tmax" means the time to reach the maximum plasma concentration (cmax) after administration; AUC0-infinity indicates the area under the plasma curve relative to the time curve from time 0 to infinity; AUC0-t is the area under the plasma concentration versus time curve from time 0 to time t; W50 is the time at which the plasma concentration is 50% or more Cmax; Determines the time at which the plasma concentration is 75% or more Cmax; And MRT means the average residence time of tacrolimus.

[0067] Two other major disadvantages associated with the treatment or prophylaxis of tacrolimus are the relative high incidence of adverse events and the relatively high intra-subject variability. It is envisaged that the composition according to the invention leads to a reduction of side effects. Reduction may be associated with reduced frequency or seriousness. The side effects mentioned are e.g. Nephro- and neurotoxicity, diarrhea, constipation, abdominal pain, nausea, etc. The present specification also discloses a particulate pharmaceutical composition containing tacrolimus together with one or more pharmaceutically acceptable excipients,

[0068] Increasing the bioavailability, Area under the curve (AUC), typically decreases within and between the variability associated with the absorption of the drug substance. This is especially true; When low and negative bioavailability is a consequence of poor solubility in water. It is believed that the compositions of the invention provide a CV (coefficient of variance) for the area in the curve data that is significantly less than for the Prograf® products and the like.

[0069] As mentioned above, one of the basic features of the present invention is that it is possible to achieve an improved oral bioavailability by the sustained release formulation. Typically, the low bioavailability of the drug substance after oral administration is a barrier to the design of a composition with controlled or sustained release of drug substance because it is almost impossible to obtain an effective level of the drug over a longer period of time. However, in current technology, it is possible to achieve much better bioavailability and thus possible to design controlled, extended or delayed release compositions.

[0070] Tacrolimus is extensively metabolised by the CYP3A4 isoenzyme in the intestinal and liver walls. Accordingly, a suitable controlled release composition can be a composition that is designed to release tacrolimus in a delayed manner to avoid or reduce CYP3A4 metabolism in the gastrointestinal tract.

[0071] Delayed release is mainly caused by an enteric coating. While the semi-permeable coating exhibits some type of delayed release, it does not have a sufficiently "delayed" release. In addition, a certain amount of time is required to make it available. The coating sought after in this invention is a pH-dependent coating. This type of coating is very resistant to the release of the drug to reach a certain pH. In a very small range of 1/10 pH, the film changes properties and becomes permeable. Examples of pH sensitive polymers that are relatively insoluble and impermeable at the pH of the stomach, but which are more soluble and permeable at the pH of the small intestine and colon include polyacrylamides, phthalate derivatives such as carbohydrate acid phthalates, amylose acetate Phthalates of cellulose phthalate, cellulose phthalate phthalates, other cellulose phthalates, cellulose ether phthalates, hydroxypropyl cellulose phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose phthalate, methyl phthalate, polyvinyl acetate phthalate, polyvinyl phthalate phthalate, cellulose sodium phthalate, starch phthalate, styrene acid - maleic dibutyl Phthalate phthalate copolymer, copolymer of styrene phthalate and maleic acid, copolymers of styrene and maleic acid, polyacrylic acid derivatives such as acrylic acid and copolymers of acrylic ester, polymethacrylic acid and its esters, copolymers of poly (acrylic methacrylic acid), shellac and acetate vinyl D copolymers of crotonic acid. [0072] pH sensitive polymers of particular interest include shellac; Phthalate derivatives, in particular cellulose acetate phthalate, polyvinyl acetate phthalate and hydroxypropyl methylcellulose phthalate; Polyacrylic acid derivatives, in particular polymethacrylate methacrylate mixed with acrylic acid and acrylic ester copolymers; And copolymers of vinyl acetate and crotonic acid.

[0073] The release of the active ingredient from a delayed release composition can also be an enzymatic reaction if, for example, a mixture of zein or mono / diglyceride is used as the coating mixture.

[0074] Following oral administration to mammals, including human, in need thereof, the controlled release pharmaceutical composition of the present invention releases tacrolimus such that a plasma concentration of at least about 5 ng / ml, e.g., at least about 7.5 ng / ml or at least about 10 ng / ml for a period of time of at least about 24 hours. In a particular aspect of the invention the difference between the maximum plasma concentration and the plasma concentration measured 24 hours after administration is at the most about 20 ng / ml, e.g., at the most about 10 ng / ml, at the most about 7.5 Ng / ml or at most 5 ng / ml.

[0075] In a particular aspect, the invention provides a solid dosage form that releases tacrolimus relatively quickly to allow a relatively rapid onset of the therapeutic effect. In one aspect, the invention relates to a pharmaceutical composition in the form of particles comprising tacrolimus together with one or more pharmaceutically acceptable excipients, wherein the composition after oral administration to a mammal in need thereof in a controlled manner releases at least about 50% w / w. The total amount of tacrolimus in about 24 hours, e.g. in about 22 hours, in about 20 hours, in about 18 hours, in about 15 hours or in about 12 hours.

[0076] In addition or alternatively at least about 50% wt / wt. The total amount of tacrolimus is released over about 24 hours, in about 22 hours, in about 20 hours, in about 18 hours, in 15 hours, in about 12 hours, tested in an in vitro dissolution test and using a dissolution medium containing a pH 7.5 buffer. The guidelines for a suitable dissolution test are described in the examples, but differences in the method and ingredients used in the solubilizing agent and the like are within the scope of the present invention. A specialist in this field will be

The skilled person knew how to perform a suitable dissolution test, e.g. with USP guidelines, Ph. Eur. And the like. Suitable conditions for the in vitro dissolution test use the USP dissolution test (paddle method) and pH 7.5 buffer containing 2.5% SDS and 1 g / mL pancreatin as the dissolution medium.

[0077] In other embodiments, the following conditions are met with respect to the in vitro dissolution test:

In other embodiments, the following conditions are met with respect to an in vitro dissolution test performed under acidic conditions:

I) at most about 30% by weight, e.g. at most about 25% wt / wt, at most about 20% wt / wt, at most about 15% wt / wt. Or at most 10% w / w tacrolimus is released within 2 hours in an in vitro dissolution test using a dissolution medium having a pH of at most 5, e.g. At most about 4.5, at most about 4, at the most about 3.5 at most about 3, at most about 2 or at most 1.5;

II) at most about 10% wt / wt, e.g. at most about 7.5% wt / wt, at most about 5% wt / wt. Or at most about 2.5% weight / weight. Tacrolimus is released in 2 hours in an in vitro dissolution test using a dissolution medium having a pH of at most 5, at the most about 4.5, at most about 4, at the most about 3.5, at most about 3, at the most about 2 or at most at most 1.5; Iii) at most about 60% weight / weight, For example, at most about 50% weight / weight, At most about 40% weight / weight. Or at most about 30% w / w. Tacrolimus is released within 15 hours, e.g. in about 12 hours, during an in vitro dissolution test using a dissolution medium having a pH of at most 4.5, such as e.g. At most about 4.0, at the most about 3.5, at most about 3, at most about 2 or at most 1.5;

Iv) at most about 40% weight / weight, e.g. at most about 30% weight / weight, At most about 25% weight / weight. Or at most about 20% w / w. Tacrolimus is released 6 hours after testing in an in vitro dissolution test using a dissolution medium having a pH of at most 4.5, such as, e.g., At most about 4.0, at the most about 3.5, at the most about 3, at the most about 2 or at most around 1.5, and /

V) at most about 30% w / w, e.g. at most about 25% w / w, at most about 20% w / w. Or at most about 15% w / w. Tacrolimus is released 4 hours after testing in an in vitro dissolution test using a dissolution medium having a pH of at most 4.5, such as, e.g., At most about 4.0, at the most about 3.5, at most about 3, at the most about 2 or at most 1.5.

[0078] In addition to tacrolimus, the composition disclosed herein may also contain a further therapeutic active, prophylactic and / or diagnostic. Of particular interest are combinations of tacrolimus with at least one of the following active substances: substances that are indicated for use in connection with organ transplantation, e.g. steroids, calcineurin inhibitors and / or anti-proliferative agents. Specific examples include prednisone, prednisolone, methylprednisone, cyclosporine, mycophenolate mofetil, azathioprine, sirolimus, everolimus, mycophenolate sodium, and FTY720 (Novartis).

[0079] The pharmaceutical compositions may be prepared by any convenient method, such as, e.g., granulating, mixing, spray-drying, etc. A particularly useful method is the method described in WO 03/004001. In the present description, a method for producing a particulate material by a controlled agglomeration method is described, i.e. a method that allows a controlled increase in particle size. Way

The method comprises spraying a first composition comprising e.g. Tacrolimus and a carrier that has been melted onto a second solid carrier substrate. Typically, the melted carrier has a melting point of at least 5 ° C, but less than the melting point of tacrolimus. The melting point of the carrier may range from 10 ° C to 150 ° C, e.g. in the range of 30 ° C to 100 ° C or in the range from 40 ° C to 50 ° C is most preferred.

[0080] It is within the skill of the average practioner to choose the appropriate carrier is pharmaceutically acceptable, capable of dissolving or at least partially dissolving tacrolimus and has a melting point in the desired range, using general knowledge and routine experiments. A suitable carrier candidate is described in WO 03/004001.

[0081] In the present context, suitable carriers are e.g. those listed as oil or an oil-like material (as discussed below) as well as disclosed in WO 03/004001.

[0082] An advantage of using the controlled agglomeration method described in WO 03/004001 is that a relatively large amount of the alloy can be applied to the particulate material without undesirably increasing particle size. Accordingly, in one embodiment, the particulate material of the pharmaceutical composition has a geometric mean diameter of dGw> 10 Pm, for example> 20 Pm, from about 20 to about 2000, from about 30 to about 2000, from about 50 to about 2000, from about 60 to about 2,000, from about 75 to about 2,000, such as from about 100 to about 1500 μm, from about 100 to about 1000 μm, or from about 100 to about 700 μm, and at most about 400 μm or at most 300 μm , for example from about 50 to about 400 μm, e.g. from about 50 to about 350 μm, from about 50 to about 300 μm, from about 50 to about 250 μm, or from about 100 to about 300 μm.

[0083] The particulate material obtained in the above-mentioned method has suitable properties in terms of flowability and / or compressibility and is therefore suitable for further processing into pharmaceutical dosage forms.

Solid dispersion and / or solid solution of tacrolimus The concentration of the active substance in the hydrophilic or water-miscible carrier is at most 15% w / w%, preferably at most 10w / w%, preferably at most 8w / w%, more preferably at most 6w / w%, Even more preferably at most 5w / w%, with a maximum of 4% w / w, and especially at most 3w / w%, in particular at most 2% w / w. And / or 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%.

[0085] Physically, the combination of active substance and carrier may form a solid dispersion, i.e. the active ingredient is dispersed in the vehicle in particulate form or it may form a solid solution, i.e. the active ingredient dissolves in the vehicle at a molecular level. The active substance and the carrier may also form a solid dispersion containing within it a part of the active ingredient dissolved at the molecular level. The physical state of the dispersion and / or solution can be determined using various techniques such as Speed ​​Microscopy (HSM), Differential Scanning Calorimetry (DSC), Scanning Electroscopy (SEM) optionally in combination with X-ray (EDX) and X-ray powder diffraction. In a preferred embodiment, the active ingredient is fully dissolved in the vehicle to form a solid solution at ambient temperature.

[0086] The solid dispersion of the invention exhibits very rapid immediate release of tacrolimus when the composition containing the dispersion or solution is tested in a USP dissolution test, using

An aqueous solution for dissolution, and at least 50% by weight of the active pharmaceutical agent. The component is released in about 30 minutes, preferably within 20 minutes, more preferably within 15 minutes; Such as at least 75% by weight of the active pharmaceutical ingredient is released within about 40 minutes or even better at least 90% by weight of the active pharmaceutical ingredient is released over about 60 minutes, preferably within 45 minutes.

[0087] Examples of useful hydrophilic or water miscible vehicles according to the present invention are selected from the group consisting of polyethylene glycols, polyoxyethylene oxides, poloxamers, polyoxyethylene stearates, poly-epsilon caprolactone, polyglycolized glycerides, such as Gelucire^®And their mixtures.

[0088] In a preferred embodiment of the invention, the vehicle is polyethylene glycol (PEG), in particular PEG with an average molecular weight of at least 1,500, preferably at least 3,000, more preferably at least 4,000, in particular at least 6,000. The polyethylene glycol may preferably be mixed with one or more other hydrophilic or water-miscible vehicles, e.g. a poloxamer, preferably in a ratio (based on weight) between 1: 3 and 10: 1, preferably between 1: 1 and 5: 1, more preferably between 3: 2: 1, especially between 2: 1 and 3: 1, in particular about 7: 3. A specific example of a useful mixture is a mixture of PEG6000 and poloxamer 188 in a ratio of 7: 3.

[0089] For polyethylene glycols (PEG), the melting point (freezing point or melt point) increases with increasing average molecular weight. For example, PEG 400 is in the range of 4-8 ° C, PEG 600 is in the range of 20-25 ° C, PEG1500 is in the range of 44-48 ° C, PEG2000 is about 52 ° C, PEG 4000 Is about 59 ° C, the PEG 6000 is about 65 ° C and the PEG 8000 is about 61 ° C.

[0090] Suitable poloxamers (e.g., polyoxypropylene-polyoxyethylene block copolymers) are, for example, poloxamer 188, poloxamer 237, poloxamer 338 or poloxamer 407 or other block copolymers of ethylene oxide and propylene oxide, such as the Pluronic® and / or Tetronic® series. Suitable block copolymers of the Pluronic® series include polymers having a molecular weight of about 3,000 or more, such as from about 4,000 to about 20,000 and / or a viscosity (Brookfield) of from about 200 to about 4,000 cps, e.g. from about 250 to about 3,000 cps. Suitable 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 copolymers Tetronic® block systems include polymers with a molecular weight of around 8,000 or more,

In a preferred embodiment of the present invention, the poloxamer is poloxamer 188, which has an average molecular weight of about 8,400 and a melting point of about 50-54 ° C.

[0092] Other useful hydrophilic or water-miscible vehicles may be polyvinylpyrrolidones, polyvinyl-polyvinyl acetate copolymers (PVP-PVA), polyvinyl alcohol (PVA), acrylic polymers (Eudragit RS, Eudragit RL, Eudragit NE, Eudragit E), hydroxypropylmethylcellulose ( HPMC) Hydroxypropylcellulose, hydroxypropylcellulose (HPC), methylcellulose, sodium carboxymethylcellulose, hydroxyethylcellulose, pectins, cyclodextrins, galactomannans, alginates, carrageenans, xanthan gums and mixtures thereof.

[0093] "Polyglycolized glycerides" means a mixture of mono- and di-esters of mono-, di- and triglycerides and polyethylene glycol diesters (PEG), preferably with a molecular weight between 200 and 600, where appropriate, free glycerol and free PEG. whose HLB value is corrected by the PEG chain length and whose melting point is regulated by the length of the fatty acid chains, PEG and the degree of saturation of the fatty chains and thus of the starting oil; Examples of such mixtures are Gelucire^®. Gelucire^®The compositions are inert, semi-solid waxy materials having an amphiphilic character and are available with different physical properties. They are surface-active in nature and scatter or dissolve in aqueous micelle-forming media, microscopic beads or vesicles. They are identified by their melting point / HLB value. The melting temperature is expressed in degrees Celsius, and HLB (Hydrophil-Lipofil Balance) is a numerical scale extending from 0 to about 20. Lower HLB values ​​mean more lipophilic and hydrophobic substances, and higher values ​​mean more hydrophilic and lipophobic substances. The affinity of the compound for water or oily substances was determined, and the HLB value was determined experimentally. One or a mixture of different species of Gelucire^® An excipient may be selected to achieve the desired melting point characteristics and / or HLB values. These are mixtures of monoesters, diesters and / or triesters of long-chain glycerides (C12 to C18). Fatty acids and esters of PEG (mono- and / or di) diasterethic acid ester (C12 to C18). Fatty acids and may contain free PEG. Gelucire^® The compositions are generally described as fatty acid esters of glycerol and PEG esters or as polyglycolized glycerides. Gelucire^® The compositions have a wide range of melting points of from about 33 ° C to about 64 ° C, most typically from about 35 ° C to about 55 ° C, and at different HLB values ​​from about 1 to about 14, most often from about 7 to about 14 For example, Gelucire^® 50/13 means a melting temperature of about 50 ° C and an HLB value of about 13 to that of the Gelucire class^®.

Pharmaceutically acceptable excipients [0094] Examples of suitable excipients for use in the composition or solid dosage form of the present invention include fillers, diluents, disintegrants, binders, lubricants, and the like, and mixtures thereof. Because the composition or solid dosage form of the invention may be used for a variety of purposes, the selection of excipients is usually taken into account for various applications. Other pharmaceutically acceptable excipients for appropriate use are e.g. acidic agents, basifying agents, preservatives, antioxidants, buffering agents, chelating agents, coloring agents, complexing agents, emulsifying and / or solubilizing agents, flavoring agents and perfumes, humectants, agents sweetening

[0095] Examples of suitable fillers, diluents and / or binders include lactose (e.g. spray dried lactose, α-lactose, β-lactose, Tabletose®, various Pharmatose® grades, Microse^® Or Fast-Floc^®), Microcrystalline cellulose (various classes of Avicel^®Elcema^®, Vivacel^®, Ming Tai^® Or Solka-Floc^®), Hydroxypropylcellulose, L-hydroxypropylcellulose (low substitution), hydroxypropylmethylcellulose (HPMC) (e.g., Methocel E, F and K, Metolose SH from Shin-Etsu, Ltd, e.g., Class 4,000 cps Methocel E and Metolose 60 SH, 4,000 cps Methocel F and Metolose 65 SH, 4,000, 15,000 and 100,000 cps Methocel K; And 4,000, 15,000, 39,000 and 100,000 species of Metolose 90 SH), methylcellulose polymers (such as, for example, Methocel A, Methocel A4C, Methocel A15C, Methocel A4M), hydroxyethylcellulose, sodium carboxymethylcellulose, carboxymethylene,

Carboxymethyl hydroxyethyl cellulose and other cellulose derivatives, Sucrose, agarose, sorbitol, mannitol, dextrins, maltodextrins, starches or modified starches (including potato starch, corn starch and rice starch), calcium phosphate (e.g. basic calcium phosphate, hydrogen phosphate) calcium, calcium phosphate hydrate), calcium sulphate, calcium carbonate, sodium alginate, collagen, etc.

[0096] Specific examples of diluents are, for example, calcium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, microcrystalline cellulose, powdered cellulose, dextrins, dextrin, dextrose, fructose, kaolin, lactose, mannitol, sorbitol, starch, pre-gelatinized starch, sucrose, sugar, etc.

[0097] Specific examples of disintegrants are e.g. alginic acid or alginates, microcrystalline cellulose, hydroxypropylcellulose and other cellulose derivatives, croscarmellose sodium, crospovidone, potassium polacrilin, sodium starch glycolate, starch, pregelatinized starch, carboxymethyl starch (e.g. Primogel® and Explotab®) etc.

[0098] Specific examples of binding agents are e.g. Gum arabic, alginic acid, agar, calcium carrageenan, sodium carboxymethylcellulose, microcrystalline cellulose, dextrin, ethylcellulose, gelatin, liquid glucose, guar gum, hydroxypropylmethylcellulose, methylcellulose, pectin, PEG, povidone, pre-gelatinized starch, etc.

[0099] Glidants and lubricants may also be included in the composition. Examples include stearic acid, magnesium stearate, calcium stearate or other metallic stearate, talc, waxes and glycerides, light mineral oil, PEG, glyceryl behenate, colloidal silica, hydrogenated vegetable oils, maize starch, sodium stearyl fumarate, polyethylene glycols, alkyl sulphates, Be nium Sodium benzoate, sodium acetate, etc. Other excipients that may be included in the solid dosage form of the invention include, e.g., pH enhancers, buffering agents, preservatives, stabilizers, antioxidants, wetting agents, moisture regulators, agents surface-active agents, suspending agents, absorption enhancers, absorption enhancers, absorption enhancers, extended release, etc.

[0101] Other additives in the solid dosage form of the invention may be antioxidants, e.g. ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphoric acid, monothioglycerol, potassium pyrosulfite, propyl gallate, sodium formaldehyde sulfoxylate, sodium metabisulfite, sodium thiosulphate. , sulfur dioxide, tocopherol, tocopherol acetate, tocopherol toluenesulfonate, 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 typically from about 0.1% w / w. Up to about 5% w / w

[0102] The solid dosage form according to the invention may also contain one or more surfactants or substances with surface-active properties. It is believed that such substances are involved in the wetting of the poorly soluble active principle and thus contribute to the improvement of the solubility of the active ingredient.

[0103] Suitable excipients for use in the solid dosage form of the invention are surfactants such as, e.g., amphiphilic surfactants as disclosed in WO 00/50007 in the name of Lipocine, Inc. Examples of suitable surfactants are

I) polyoxyethylated fatty acids, such as, for example, Mono- or polyethylene glycol fatty acid diesters or mixtures thereof such as, e.g., Mono or polyethylene glycol diesters with lauric acid, oleic acid, stearic acid, myristic acid, ricinoleic acid

The 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 9000, PEG 1000, PEG 10

000, PEG 15,000, PEG 20,000, PEG 3000, PEG 4000, PEG 3500,

II) fatty acid esters of polyethylene glycol glycerol, i.e. Esters as above, but in the form of glyceryl esters of individual fatty acids;

Iii) glycerol, propylene glycol, ethylene glycol, PEG or sorbitol esters, e.g. vegetable oils, such as, for example, hydrogenated castor oil, almond oil, palm kernel oil, castor oil, apricot kernel oil, olive oil, peanut oil , hydrogenated palm kernel oil and the like,

Iv) polyglycerolated fatty acids, e.g. polyglycerol stearate, polyglycerol oleate, polyglycerol ricinoleate, polyglycerol linoleate,

V) propylene glycol fatty acid esters, e.g., propylene glycol monolaurate, propylene glycol ricinoleate and the like,

Vi) mono- and diglycerides, such as, for example, glyceryl monooleinate, glyceryl dioley, glyceryl mono and / or dioleate, glyceryl caprylate, glyceride capsule and the like;

Vii) sterol and sterol derivatives;

Viii) fatty acid esters of polyethylene glycol sorbitan (PEGsorbitan fatty acid esters), such as PEG esters of various molecular weights indicated above and various Tween® series; Ix) polyethylene glycol alkyl ethers, such as, e.g., PEG oleyl ether and PEG lauryl ether;

X) sugar esters, such as, e.g., sucrose monopalmitate and sucrose monolaurate;

Xi) polyethylene glycol polyols, such as e.g. the Triton® X or N series;

Xii) polyoxyethylene-polyoxypropylene block copolymers, such as e.g. Pluronic® series, Synperonic® series, Emkalyx®, Lutrol®, Supronic® etc. The general term for these polymers is "poloxamers" and suitable examples in the present context include: Poloxamer 105, 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 fatty acid esters, such as the Span® series or the Ariacel® series, such as, e.g., sorbinate monolaurate, sorbitan monopalmitate, sorbitan monoleate, sorbitan monostearate, etc .;

XIV) lower alcohol esters of fatty acids, e.g. Oleinate, isopropyl myristate, isopropyl palmitate and the like;

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

[0104] When the surfactant or surfactant mixture is present in the solid dosage form of the invention, the concentration of surfactant (s) is typically from about 0.1 to 80% w / w, 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, 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.

[0105] In a specific aspect of the invention, at least one of the one or more pharmaceutically acceptable excipients is selected from the group consisting of silicic acid or a derivative thereof, or a salt thereof, in

Including silicates, silicon dioxide and polymers thereof; Magnesium aluminate and / or magnesium gluminometasilicate, bentonite, kaolin, magnesium trisilicate, montmorillonite and / or saponite.

[0106] Such materials are particularly useful as a sorption material for oils or oily materials in pharmaceuticals, cosmetics and / or foodstuffs. In a specific embodiment, this material is used as a sorption material for oils or oily substances in pharmaceuticals. A material capable of acting as a sorption material for oils or oil-like materials is also referred to as an "oil sorption material". Furthermore, in the present context, the term "sorption" is used to denote "absorption" and "adsorption". It should be understood that when one of the terms is used, it should include absorption of the phenomenon and adsorption.

[0107] In particular, the pharmaceutically acceptable excipient may comprise silicic acid or a derivative thereof or a 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 may be a lubricant or may be an oil sorption material. The most important are qualifications fulfilling the last function.

[0108] In a specific embodiment, the composition or solid dosage form of the invention comprises a pharmaceutically acceptable excipient that is a silicon dioxide product that has the properties corresponding to Aeroperl® 300 (available from Degussa, Frankfurt, Germany). As can be seen from the examples here, a very suitable material is Aeroperl® 300 (including materials with properties similar or similar to Aeroperl® 300).

[0109] The use of an oil sorption material in the compositions or dosage forms of the invention is very advantageous for the preparation of pharmaceutical, cosmetic, nutritional and / or food compositions, the composition comprising oil or oil similar to the material. One of the advantages is the possibility of incorporating a relatively large amount of oil and oil similar to the material and still has a material that is solid. Thus it is possible to prepare solid compositions with a relatively high oil load or oily material using a sorption oil material according to the invention. In the pharmaceutical field, it is advantageous to include a relatively large amount of oil or oily-like material in a solid composition, especially in these situations, where the active substance does not have adequate properties with respect to water solubility (eg Poor solubility in water), stability in an aqueous environment (eg Degradation occurs in an aqueous environment), oral bioavailability (eg Low bioavailability), etc. Or in situations in which it is desirable to modify the release of the active ingredient from the composition to obtain controlled, delayed, sustained and / or pulsatile delivery of the active ingredient. Thus, in a specific embodiment it is used for the preparation of pharmaceutical compositions. wherein it is desired to modify the release of the active ingredient from the composition to obtain controlled, delayed, sustained and / or pulsatile delivery of the active ingredient. Thus, in a specific embodiment it is used for the preparation of pharmaceutical compositions. wherein it is desired to modify the release of the active ingredient from the composition to obtain controlled, delayed, sustained and / or pulsatile delivery of the active ingredient. Thus, in a specific embodiment it is used for the preparation of pharmaceutical compositions.

[0110] In an important embodiment of the invention, at least part of tacrolimus is present in the composition in the form of a solid solution comprising a molecular dispersion and a solid dispersion. Usually 10% or more, e.g. 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more Such as an example of 95% or more or about 100% wt / wt. Tacrolimus is present in the composition in the form of a solid dispersion.

[0111] The solid dispersion may be obtained in a variety of ways, e.g. by using organic solvents or by dissolving or dissolving the active ingredient in another suitable environment (e.g. oil or an oily material that is in liquid form at room temperature or at elevated temperatures) .

[0112] Solid dispersions (solvent method) can for example be prepared by dissolving a physical mixture of the active substance (e.g. a drug substance) and a carrier in a common organic solvent, followed by evaporation of the solvent. The carrier is often a hydrophilic polymer. Suitable organic solvents include a pharmaceutically acceptable solvent in which the active ingredient is soluble, such as methanol, ethanol, methylene chloride, chloroform, ethyl acetate, acetone or mixtures thereof.

[0113] Suitable water-soluble solvents include polymers such as polyethylene glycol, poloxamers, polyoxyethylene stearates, ρουί-ε-caprolactone, polyvinylpyrrolidone (PVP), polyvinylpyrrolidone-polyvinyl acetate PVP-PVA copolymer (Kollidon VA64), acrylic polymers (Eudragit RS,

Eudragit RL, Eudragit NE, Eudragit E) and polyvinyl alcohol (PVA), hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), methylcellulose and poly (ethylene oxide) (PEO).

[0114] Regarding the amount of active ingredient and polymer in the solid dispersion, the weight ratio of active ingredient to polymer may be from about 3: 1 to about 1:20. However, a narrower ranger may be used from about 3: 1 to about 1: 5, e.g. from about 1: 1 to about 1: 3 or about.

[0115] The solid dispersion is preferably prepared by spray drying techniques, controlled agglomeration, lyophilization or coating of carrier particles or any other solvent removal process. The dried product contains the active substance present in the form of a solid dispersion containing a molecular dispersion and a solid solution.

[0116] As an alternative to the use of organic solvents, the drug and polymer can be co-extruded or extruded at an elevated temperature (melt extrusion).

[0117] Pharmaceutical compositions comprising tacrolimus at least partially in the form of a solid dispersion or solution can be essentially prepared using any suitable procedure for the preparation of pharmaceutical compositions known in the art.

[0118] In addition to using an organic solvent based method, a solid dispersion or solid solutions of tacrolimus can be obtained by dispersing and / or dissolving tacrolimus in a support composition used in a controlled agglomeration process. Stabilizing agents, etc. may be added to ensure the stability of the solid dispersion / solution.

[0119] The present specification also discloses a method of preparing a pharmaceutical composition. In general, any suitable method in the pharmaceutical field can be used. However, in order to allow the introduction of a relatively large amount of oil or oil-like material, especially the method described in WO 03/004001, it has proved useful. The method comprises spraying a first composition in a liquid form, said composition comprising a first carrier and carrier having a melting point above 5 ° C for a second composition comprising a second carrier or carrier material, said second composition, e.g. in a fluidized state and having a temperature below melting temperature of the first vehicle or carrier. The active ingredient may be present in the first carrier or carrier and / or in a second carrier or carrier. However, in those cases where tacrolimus is present, at least in part, in the form of a solid dispersion, it is preferred to include or dissolve tacrolimus in the first carrier or carrier. WO05020993A1 and WO05020994A1. The present inventors further describe tacrolimus compositions using sustained release technology and formulations.

Solid dosage forms

[0120] The pharmaceutical composition of the invention is in particulate form and may be used as such. However, in many cases it is more convenient to present compositions in the form of granules, granules, microspheres, nanoparticles and the like, or in the form of solid dosage forms including tablets, capsules and sachets and the like. The solid dosage form according to the invention may be a single unit dosage form or it may be in the form of a polydipot dosage form comprising a plurality of individual entities, such as for example granules, beads and / or granules. [0121] The solid dosage form of the present invention comprises a particulate pharmaceutical composition as described above. The details and particulars disclosed in this main aspect of the invention apply mutatis mutandis to other aspects of the invention. Accordingly, properties with respect to increased bioavailability, changes in bioavailability parameters, no diurnal effect, as well as the release of tacrolimus described and / or claimed herein to pharmaceutical particulate formulations are analogues for the solid dosage form of the present invention.

[0122] The recommended dosage range of Prograf® is 0.1 to 0.2 mg / kg / day, every 12 hours in two divided doses. More importantly, you need to control your blood levels.

[0123] The typical level for 1-3 months is 7-20 ng / ml, and from 4 to 12 months the level should be from 5 to 15 ng / ml. These are only indicative values ​​and may differ from the types of transplantation and ethnicity.

[0124] In patients with a kidney transplant, it was found that:

<td></td><td colspan="2">Caucasian</td><td colspan="2">black</td>

<td></td><td colspan="2">N = 114</td><td colspan="2">n = 56</td>

<td>Time after transplantation</td><td>Dawka (mg / kg)</td><td>Trough concentrations (ng / ml)</td><td>Dawka (mg / kg)</td><td>Trough concentrations (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>Month 6</td><td>0.14</td><td>11.8</td><td>0.24</td><td>11.5</td>

<td>Month 12</td><td>0.13</td><td>10.1</td><td>0.19</td><td>11.0</td>

[0125] The expected dosage recommendation for the products of the present invention is from 0.02 mg / kg / day to 0.15 mg / kg / day, dosed once a day. Suitable dosage forms (strength) range from 0.1 mg to 15 mg tacrolimus, preferably with a selected strength from 0.5 mg, 1 mg, 2 mg and 5 mg.

[0126] As shown herein, scintigraphic and shown in Figure 2, the release according to the present invention can take place even in the distal part of the colon and continues to be distributed into the mucus and absorbed. Absorption of the sustained release dosage form of the invention is one in which release in vivo after oral administration to a subject occurs substantially in the colon, such as release at one or more locations located within the large intestine, across the colon and within the duodenum.

[0127] Various embodiments of the invention relating to preferred pharmacokinetic parameters obtained with the sustained release dosage forms of the present invention are listed below.

In one embodiment, the invention relates to a sustained release dosage form that, when administered to a patient or a plurality of subjects, provides a subject's intra-body and / or inter-personal variability in TMaks of tacrolimus, which was relatively consistent with the commercial Advagraf® formulation (MR4). ) or sustained release dosage forms are reduced from at least 10%, e.g. at least 15%, e.g. at least 17.5%, such as at least 20%, e.g. at least 22.5%, e.g. at least 25%, e.g. at least 27.5%, e.g. at least 30% under similar conditions and administered at similar molecular doses of tacrolimus.

[0129] In one embodiment, the invention relates to a sustained release dosage form that, when administered to an individual or a number of subjects, provides an individual and / or intra-subject variation in the mean CMaks I / AUC (o-) of tacrolimus, which relative to that obtained from administration the commercial Advagraf® (MR4) or sustained release dosage form is reduced from at least 10%, e.g. at least 15%, e.g. at least 17.5%, e.g. at least 20%, e.g. at least 22, 5%, e.g. at least 25%, e.g. at least 27.5%, e.g. at least 3% under similar conditions and given at similar molecular doses of tacrolimus. [0130] In one embodiment, the invention relates to a sustained release dosage form,

[0131] In one embodiment, the invention relates to a sustained release dosage form that, when administered to a patient or multiple subjects, provides increased bioavailability as compared to administration of Advagraf® (MR4) or sustained release bioactive dose at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45% or at least about 50% Or at least about 55% or at least about 60%, e.g. at least 65%, where bioavailability is defined as AUC (0 &amp; quot;) and under similar conditions and given at similar molecular doses of tacrolimus.

In one embodiment, the invention provides a sustained release dosage form that, when administered to a patient or a plurality of subjects, provides a subject's intra-body and / or inter-personal variability in TMaks of tacrolimus relative to that obtained by administering a commercially available dosage form of Prograf® or The immediate release dosage form of immediate-release tacrolimus is reduced from at least 10%, e.g. at least 15%, e.g. at least 17.5%, e.g. at least 20%, e.g. at least 22.5%, e.g. at least 25%, e.g. at least 27.5%, e.g. at least 30% under similar conditions and given at similar molecular doses of tacrolimus.

[0133] In one embodiment, the invention relates to a sustained release dosage form that, when administered to an individual or the number of subjects, provides individual and / or intra-individual variability in the mean blood CMaks I / or AUC (o-) Tacrolimus, which relative to each other. the dose obtained by administering a commercially available dosage form of Prograf® or an immediate release biodegradable dosage form is reduced from at least 10%, e.g. at least 15%, e.g. at least 17.5%, e.g. at least 20%, for example at least 22.5%, e.g. at least 25%, e.g. at least 27.5%, e.g. at least 30% when determined under similar conditions and given at similar molecular doses of tacrolimus.

In one embodiment, the invention relates to a sustained release dosage form that, when administered to an individual or a number of subjects, provides reduced CMaks relative to that obtained by administering a commercially available Prograf formulation.^® A dosage form or dosage form having a similar biological index of at least about 20%, or at least about 30%, or at least about 35%, or at least about 40% or at least about 45% or at least about 50% Or at least about 55%, or at least about 60%, or at least about 65% when determined under similar conditions and given at similar molecular doses of tacrolimus.

[0135] In one embodiment, the invention provides a sustained release dosage form that, when administered to a patient or multiple subjects, provides greater bioavailability relative to that obtained by administering a commercially available dosage form of Prograf® or immediate release dosage form from at least about 10%, or at least about 15%, or at least about 20%, or at least about 30%, or at least about 35%, or at least about 40 or more, or at least about 45%, or at at least about 50% or at least about 55% of bioavailability is defined as AUC (0 &amp; quot;) and under similar conditions and is administered at similar molecular doses of tacrolimus. [0136] In one embodiment, the invention relates to sustained release dosage forms,

In one embodiment, the invention relates to a sustained release dosage form which, when administered to a patient or more after at least 4 hours on an empty stomach in the evening, provides C (max), which corresponds to the concentration obtained after the dosage Form in the morning after at least 4 hours on an empty stomach is at least 70%, e.g. at least 80%, preferably at least 85%, more preferably at least 9%, and preferably at least 95% measured after administration in the morning.

In one embodiment, the invention relates to a sustained release dosage form, wherein the bioavailability is independent of the dosing time during the day and is therefore suitable for a pre-bedtime regimen.

In one embodiment, the invention relates to a sustained release dosage form that at a dose of 5 mg tacrolimus and after a single dose to at least 6 healthy fasted subjects gives an average maximum concentration (CMaks) of tacrolimus of at most 15 ng / ml, an example of at most 13 ng / ml and an average AUC (0-96 hours) of at least 45 mg • h / l, e.g. at least 55 mg • h / l, e.g. At least 60 mg • h / l.

[0140] In one embodiment, the invention relates to a sustained release dosage form in which the blood concentration after tacrolimus administration is at least 2 ng / ml, e.g. at least 3 ng / ml, e.g. at least 4 ng / ml.

In one embodiment, the invention relates to a sustained release dosage form that when administered once a day at steady-state to a healthy person or a patient, tacrolimus blood swing is measured as (CMaks-CMin) / CMin. It is smaller than the observed swing when administered Advagraf® dosage forms or sustained release dosage forms of tacrolimus sustained release in a once daily dosing schedule and their determination under similar conditions and administration at similar molecular daily doses of tacrolimus. A fall is preferred at least 10%, such as at least 20%, more preferably at least 30%, such as at least 40%, more preferably at least 50%.

[0142] In one embodiment, the invention relates to a sustained release dosage form that when administered once a week at steady-state to a healthy person or a patient, swing the total blood and / or free tacrolimus measured as (CMaks-CMin) / CMin Is less than the observed swing when administering the Prograf® dosage form or bioequivalent immediate-release formulation of tacrolimus twice daily and is determined under similar conditions and administered at similar molecular daily doses of tacrolimus. A fall is preferred at least 10%, such as at least 20%, more preferably at least 30%, such as at least 40%, more preferably at least 50%.

In one embodiment, the invention relates to a sustained release dosage form that is administered once a day at steady-state to a healthy person or patient, fluctuation in whole blood concentration and / or free tacrolimus measured as (CMaks-CMin) / Cd. than the observed fluctuation when administering the Advagraf® dosage forms or sustained release dosage forms of tacrolimus sustained release in the once daily regimen and setting them under similar conditions and administering at similar molecular doses of tacrolimus daily. A fall is preferred at least 10%, such as at least 20%, more preferably at least 30%, such as at least 40%, more preferably at least 50%.

In one embodiment, the invention relates to a sustained release dosage form that when administered once daily in a steady state to a healthy person or patient, fluctuates blood concentration for total tacrolimus measured as (CMaks-CMin) / Aver. It is less than the observed fluctuation during administration of the Prograf® or bioconward bioequivalent immediate-release formulation of tacrolimus in a twice-daily schedule and is determined under similar conditions and administered at similar molecular daily doses of tacrolimus. A fall is preferred at least 10%, such as at least 20%, more preferably at least 30%, such as at least 40%, more preferably at least 50%.

[0145] In one embodiment, the invention relates to a sustained release dosage form that is administered to at least 6 healthy fasted subjects, the average residence time, MRT, of tacrolimus measured in the blood is at least 10% longer than the average residence time measured in bioavailable conditions with a dose of Advagraf® or a similar dosage form of prolonged-release tacrolimus. It is preferred that the MRT is increased by at least 20%, e.g. by 25%.

[0146] In one embodiment, the invention relates to a sustained release dosage form that is administered to at least 6 healthy subjects under the fasting condition, the average residence time of tacrolimus measured in

The blood is at least 35% longer than the average residence time measured under similar conditions with the Prograf® dosage form or immediate-release, equivalent biological immediate release dosage of tacrolimus.

[0147] The present specification also discloses a method of delivering immunosuppressive therapy to a patient in need thereof in a once daily regimen by administering a sustained release formulation described herein providing one or more reduced CMaks, pendulum reduction, decreased fluctuation, increased AUC, increased MRT. , longer time to TMaks, and higher CMin. In addition, methods provide for CMin Co to correlate with bioavailability with a correlation coefficient of at least 0.75 to 1, e.g., at least 0.80, more preferably 0.85, more preferably 0.90, more preferably at least 0.95, and even more preferably at least 0.97.

[0148] In preferred deposition, the difference in bioavailability is substantially independent of the time of day of administration. This gives the possibility of a one-day dosing regimen at bedtime or in the evening, in addition to the normal morning dosing. More importantly, the risk of reducing exposure if the patient actually eats the dosage form at a different time than expected and expected (patient incompatibility), the risk for the patient is reduced to reduce exposure, thereby increasing the risk of graft refusal.

[0149] In a further aspect, the invention provides a recycle of the higher dose delivery method. One method of converting, inter alia, the Prograf formulation to once daily doses reduces the daily dose of immediate-release immediate-release formulation from 25% to 50%, e.g. from 30% to 40%, more preferably from about 33%. Reduction is as much as possible by using a sustained release dose once a day at doses selected from 0.5 mg1 mg, 2 mg and 5 mg doses once a day. Correspondingly, the conversion ratio 1: 0.66 to 0.80 depends on the available strengths as mentioned. In addition, the present invention relates to the conversion of the Advagraf® formulation in a ratio of 1: 0.30 to 0.75, for example in a ratio of 1: 0.33 to 0.7, depending on the dose amounts available selected from 0.5 mg, 1 mg. .

[0150] A particularly important aspect of the invention is the significant lowered peak concentrations that cause a decrease in side effects associated with the peak. This effect may be difficult to measure due to the different and intra-subject variant of current methods of treatment with tacrolimus and the nature of adverse reactions, which are often subjective or may require tissue biopsies. Detailed questionnaires and a large number of patients will be needed in comparative studies to demonstrate this impact with meaning. It is believed, however, that treatment with the sustained release preparation of the present invention may reduce some possible side effects associated with peak concentration, including side effects of neurological origin, such as tremors and headaches. [0151] In a preferred embodiment, the reduction of the side effect is associated with the risk of prolonged QTc interval due to the effect on ventricular repolarization. Other toxicities are thought to include reduction of kidney damage, development of diabetes, and the development of hypertension.

[0152] Particularly the accumulation present in some organs may be associated with organ damage, especially since the accumulation or individual concentration of tacrolimus can not drop during periods with low blood concentrations at later dosing intervals. This lack of organ space can be associated with the high affinity of tacrolimus to this organ, which overcomes otherwise expected gaps in the form of highly-vascular organs. Accordingly, the high peaks present

Available commercial dosage forms can contribute to the accumulation of tacrolimus in those organs that can contain organs up to almost 30 times the steady-state blood concentration using conventional dosage forms.

[0153] The de novo transplanted patient may be very sensitive to high concentrations due to overall poor body condition, low plasma protein levels, still increasing the tacrolimus fraction to freely enter the organs. In particular, these patients avoid high blood levels during titration using the dosage form according to the invention, which can reduce the amount of accumulation or increase the time before reaching high concentrations in organs in which tacrolimus is toxic. This specification also discloses a method of treating patients at risk of toxicity to an organ with tacrolimus. The organs that accumulate tacrolimus include the adrenal glands, lungs, heart, liver, digestive tract and kidneys. According to the present invention, it is furthermore believed that the pancreas, and in particular the islet of Langerhans, it may be sensitive to high concentrations, especially during the start of tacrolimus, with the risk of developing diabetes later being significantly greater. The central nervous system is other organs correlated with toxicity. Many patients withdraw from treatment due to headaches and tremors, side effects that may be reduced as a result of treatment in accordance with the present invention.

[0154] According to a further aspect of the invention, the sustained release preparation of the present invention provides a safety profile for treatment that allows treatment since transplantation as soon as the patient is adequately treated with the oral formulation. Accordingly, the patient does not need the first titration with a conventional twice-daily preparation such as Prograf® and subsequent conversion to treatment according to the present invention, but may initiate oral immunosuppressive therapy with tacrolimus with the formulation once daily in accordance with the present invention.

List A [0155] A list of clinical trials with a sustained release preparation of the present invention, including improved pharmacokinetic parameters obtained in a sustained release formulation of the present invention and claimed in the present application.

Table A

<td>LCP Fast</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>Cmax</td><td>Cmin12</td><td>Cmin24</td><td>kel</td><td>Tmax</td><td>T¹/₂</td>

<td>Study 002</td><td>10.05 ± 3.99</td><td>6.92</td><td>4.32</td><td>0.0217</td><td>8.55 ± 2.72</td><td>31.95</td>

<td>Study 003</td><td>3.62</td><td>ON</td><td>ON</td><td>ON</td><td>6.71 (0.512)</td><td>39.04</td>

<td>Study 004</td><td>2.82 ± 0.96</td><td>2.42 ± 0.87</td><td>1.83 ± 0.72</td><td>ON</td><td>9.69 ± 4.90</td><td>ON</td>

<td>Study 005</td><td>14.31 ± 4.29</td><td>6.504 ± 2.58</td><td>4.09 ± 1.70</td><td>2,07E02</td><td>5.17 ± 1.68</td><td>35.14 ± 8.37</td>

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

EP 2 167 033 B1

<td>Prograf He congratulated</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>Cmax</td><td>Cmin12</td><td>Cmin24</td><td>kel</td><td>Tmax</td><td>T /</td>

<td></td><td>32.75 ±</td><td></td><td></td><td></td><td>1.63 ±</td><td></td>

<td>Test 002</td><td>9.65</td><td>3,508</td><td>2,268</td><td>0.0208</td><td>0.43</td><td>33.16</td>

<td>Test 003</td><td>ON</td><td>ON</td><td>ON</td><td>ON</td><td>ON</td><td>ON</td>

<td></td><td></td><td>0.63</td><td></td><td></td><td>1.38 ±</td><td></td>

<td>Study 004</td><td>5.26 ± 1.28</td><td>± 0.18</td><td>ON</td><td>ON</td><td>0.42</td><td>ON</td>

<td></td><td>31.76 ±</td><td></td><td></td><td>2,05E-</td><td>1.38 ±</td><td>34.86</td>

<td>Test 005</td><td>8.81</td><td>4.24 ± 1.73</td><td>3.13 ± 1.14</td><td>02</td><td>0.34</td><td>± 7.04</td>

[0156] Plasma profiles from the tests shown in Table A and B are shown in Fig. 3 Table B

<td></td><td>items</td><td>Title</td>

<td>LCP-Tacro PK002 Test pilot</td><td>12</td><td>2 Railway crossing; Single dose, on an empty stomach; Tablets LCP-Tacro 2x2mg modified release Vs. Capsules Prograf 4x1 mg (tacrolimus)</td>

<td>LCP-Tacro PK003 Test pilot</td><td>6</td><td>2-made project; One percent, on an empty stomach, in examination of knee absorption in the LCP-Tacro 2 group mg</td>

<td>LCP-Tacro PK004 Test pilot</td><td>13</td><td>2D crossover, multi-dose study, fasting studies, relative relative bioavailability tests; LCP-Tacro 2 mg tablets vs. Capsules Prograf 2x1 mg bid</td>

One-time, comparative PK study (LCP-Tacro PK-002):

[0157] LCP-Tacro PK-002 study, single-dose comparative pharmacokinetic study of LCP-Tacro 4 mg (2x2 mg tablets) vs. Prograf 4 mg (4x1 mg capsules) showed comparable AUC values ​​for both products under fasted conditions. However, they differ in the rate of absorption when administered under fasting conditions, it is slower and more stable for the test preparation, LCP-Tacro. The longer

TMaks For LCP-Tacro tablets (test; HPMC) vs. Prograf capsules (tacrolimus, reference) (8.55 h vs. 1.63h) in combination with lower CMaks (100.05 g / ml vs. 320.75 ng / ml) and lower AUC (0-&gt; 24) (122, 4 ng * h / ml vs. 157.95 ng * h / ml), maintains once daily dosing for LCP-Tacro tablets compared to Prograf capsules (tacrolimus)

LCP-Tacro PK-003 study:

[0158] The LCP-Tacro PK-003 study was designed as a repeated scintigraphic absorption study in 8 healthy volunteers to evaluate transit time, pharmacokinetic profile and release site of LCP-Tacro 2 mg tablets labeled radioactively with a maximum of 1 MBq of 153Sm. IN

In this study, a sustained release profile with TMaks ~ 6.7 hours could be demonstrated. LCP-Tacro was well tolerated. In general, the present study demonstrated the in vivo release profile of LCP-Tacro tablets with release in all parts of the colon and the absorption of tacrolimus from distal parts of the gastrointestinal tract. Scintigraphic images are shown in Figure 2, in which pictures were taken of 0.02, 0.53, 4.32, 4.57, 11.23 and 23.32 hours after dose administration, respectively.

Multiple, stationary, comparative PK study (LCP-Tacro PK-004):

[0159] The LCP-Tacro PK-004 study is a multicenter comparative pharmacokinetic pharmacokinetic LCP-Tacro 2 mg qd vs. Prograf 2x1mg bid in 14 healthy volunteers. Study

1o clearly showed that once a day LCP-Tacro profile versus Prograf bid In addition, the study showed greater bioavailability of LCP-Tacro tablets given once a day compared to Prograf capsules administered twice a day. After administration of LCP-Tacro 2 mg tablets (qd) and Prograf 1 mg (bid) capsules for 1o consecutive days, no significant differences were seen in the morning pre-dose doses between Days 7, 8, 9 and 10, therefore the steady state was maintained from 7. At steady-state, systemic exposure over 24 hours of LCP-Tacro 2 mg tablets (qd) was approximately 50% higher than with Prograf 1 mg (bid) capsules. Time to peak concentration between single and multiple doses of LCP-Tacro 2 mg / d. And Prograf 2x1 mg bid were similar for both treatments.

2o release 1 mg (bid). There was no significant difference observed in Cmax between the two treatments.

<td rowspan="4">Pharmacokinetics Parameters: test summarizing 004</td><td colspan="2">Geometric mean (% CV)</td>

<td colspan="2">The arithmetic mean ± SD</td>

<td>LCPTacro 2 mg tablets</td><td>Prograf® 1 mg capsules</td>

<td>(A, n = 13)</td><td>(B, n = 13)</td>

<td rowspan="2">AUCt (ng · hr / ml)</td><td>(34.98)</td><td>(34.26)</td>

<td>115.07 ± 40.25</td><td>43.65 ± 14.95</td>

<td rowspan="2">AUC0-24 (ng · hr / ml)</td><td rowspan="2">Not applicable</td><td>72.95 (34.40)</td>

<td>76.93 ± 26.46</td>

<td rowspan="2">Cmax (ng / ml)</td><td>6.42 (36.55)</td><td>6.71 (31.30)</td>

<td>6.80 ± 2.49</td><td>7.02 ± 2.20</td>

<td rowspan="2">Cmin (ng / ml)</td><td>3.12 (35.20)</td><td>2.00 (40.51)</td>

<td>3.31 ± 1.17</td><td>2.14 ± 0.87</td>

<td>Tmax (hours) *</td><td>8.00 (6.00 - 10.02)</td><td>1.50 (1.00-2.00)</td>

<td>T1 / 2 (hours)</td><td>32.93 ± 2.66</td><td>32.59 ± 4.08t</td>

3o

EP 2 167 033 B1

<td rowspan="4">Pharmacokinetics Parameters: test summarizing 004</td><td colspan="2">Geometric mean (% CV)</td>

<td colspan="2">The arithmetic mean ± SD</td>

<td>LCPTacro 2 mg tablets</td><td>Prograf® 1 mg capsules</td>

<td>(A, n = 13)</td><td>(B, n = 13)</td>

<td>Cmax / Cmin</td><td>2.12 ± 0.51</td><td>3.43 ± 0.75</td>

<td colspan="3">* Median (min-max); | n = 12.</td>

Table C List of studies in which the results are presented below.

<td>Study nr / phase</td><td>items</td><td>Title</td>

<td>LCP-Tacro 1011 [3330]</td><td>thirty</td><td>A two-way study of a single dose of crossover tablets LCP-Tacro 5 mg Vs. Prograf 5 mg capsules</td>

<td>LCP-Tacro 1012 [3363]</td><td>25</td><td>Bioavailability testing LCP-Tacro2mg qd bilaterally cross, open, multiple dose, bioavailability on fasting. Vs. Prograf 1 mg bid X 10 days</td>

<td>LCP-Tacro 1013 [3386]</td><td>25</td><td>One dose, LCP-Tacro dose-linearity study 5 mg vs 7 mg versus 10 mg</td>

<td>LCP-Tacro 1014 [3385]</td><td>26</td><td>One-time chrono-pharmacokinetic study of tablets LCP-Tacro 2 mg was administered in the morning and in the evening</td>

<td>LCP-Tacro 1015 [3414]</td><td>17</td><td>Homogeneous bioavailability testing of the LCP-Tacro 1 tablet mg vs. Prograf 1 mg capsules</td>

<td>LCP-Tacro 1016 [3423]</td><td>28</td><td>Bioavailability testing LCP-Tacro2mg qd bilaterally cross, open, multiple dose, bioavailability on fasting. Vs. Prograf 1 mg bid X 10 days</td>

Multigrade clinical trial at steady-state, comparative (LCP-Tacro 1012 [3363]):

[0160] In this study, the bioavailability of tacrolimus from the LCP-Tacro 2 mg preparative once-a-day (qd) formulation test in group A was compared and compared to reference capsules.

Prograf 1 mg administered twice a day (bid) in group B, in a multiple dose, On an empty stomach. The study population consisted of 30 healthy volunteers who received tests for a period of 10 days, followed by a two-week rinse and then crossover between study groups. Twenty-five patients were evaluated in this study. The pharmacokinetics of tacrolimus on day 10 are shown in the tables below.

Table: Pharmacokinetic parameters of tacrolimus on day 10 (LCP-Tacro 1012 study [3363])

<td>parameters</td><td>Geometric mean (% CV)</td>

EP 2 167 033 B1

<td rowspan="2">Pharmacokinetic</td><td colspan="2">The arithmetic mean ± SD</td>

<td>LCP-Tacro 2 mg tablets (A; qd; n = 25)</td><td>Prograf® 1 mg Capsules (B; bid; n = 25)</td>

<td rowspan="2">AUC0-12 (ng ^ hr / mL)</td><td>70.71 (27.94)</td><td>48.98 (44.05)</td>

<td>73.65 ± 20.58</td><td>54.22 ± 23.88</td>

<td rowspan="2">AUC12-24 (ng ^ hr / mL)</td><td>54.81 (31/10)</td><td>36.48 (43.15)</td>

<td>57.72 ± 17.95</td><td>40.28 ± 17.38</td>

<td rowspan="2">Cmax (ng / mL)</td><td>7.49 (31.74)</td><td>8.06 (40.91)</td>

<td>7.85 ± 2.49</td><td>8.72 ± 3.57</td>

<td rowspan="2">C12 (ng / mL)</td><td>5.68 (31.88)</td><td>2.47 (46.73)</td>

<td>6.00 ± 1.91</td><td>2.78 ± 1.30</td>

<td rowspan="2">Cavg (ng / mL)</td><td>5.24 (29.04)</td><td>3.57 (42.80)</td>

<td>5.47 ± 1.59</td><td>3.94 ± 1.68</td>

<td>Tmax (hr) *</td><td>6.00 (1.00 - 10.00)</td><td>1.50 (1.00 - 13.00)</td>

<td rowspan="2">AUC0-12 / C12 (hr)</td><td>12.44 (13.44)</td><td>19.82 (11.53)</td>

<td>12.54 ± 1.68</td><td>19.94 ± 2.30</td>

<td rowspan="2">AUCT / Cmin (hr)</td><td>32.00 (6.83)</td><td>32.14 (11.25)</td>

<td>32.07 ± 2.19</td><td>32.34 ± 3.64</td>

<td rowspan="2">AUC12-24 / Cmin (hr)</td><td>13.96 (5.70)</td><td>13.69 (13.13)</td>

<td>13.98 ± 0.80</td><td>13.80 ± 1.81</td>

<td colspan="3">* median (min - max)</td>

Table: Relative assessment of the bioavailability of tacrolimus on day 10 (LCP-Tacro 1o12 study [3363])

<td>Parameter</td><td>90% CI</td><td>Ratio of Means</td><td>Intra-Subject CV</td>

<td>AUCt</td><td>131.49% to 162.84%</td><td>146.33%</td><td>22.31%</td>

<td>C min</td><td>131.27% to 163.97%</td><td>146.71%</td><td>23.22%</td>

Study 1013 (3386) (dose proportionality, NHV, n = 25): [0161]

<td></td><td>Treatment A</td><td>Treatment B</td><td>Treatment C</td>

<td></td><td>1x5 mg</td><td>1x5mg + 1x2mg</td><td>2x5mg</td>

<td>AUC0-12 (ng · hr / ml)</td><td>88.23 ± 32.87</td><td>125.33 ± 44.96</td><td>186.15 ± 56.81</td>

EP 2 167 033 B1

<td></td><td>Treatment A</td><td>Treatment B</td><td>Treatment C</td>

<td></td><td>1x5 mg</td><td>1x5mg + 1x2mg</td><td>2x5mg</td>

<td>AUC0-24 (ng · hr / ml)</td><td>143.00 ± 54.23</td><td>212.69 ± 77.30</td><td>307.05 ± 98.07</td>

<td>AUC0-t (ng · hr / ml)</td><td>303.55 ± 133.60</td><td>449.63 ± 173.51</td><td>648.81 ± 224.20</td>

<td>AUC0-inf (ng · hr / ml)</td><td>324.46 ± 143.52</td><td>473.40 ± 182.86</td><td>67.68 ± 241.22</td>

<td>Cmax (ng / ml)</td><td>12.21 ± 4.84</td><td>16.73 ± 5.22</td><td>24.91 ± 7.15</td>

<td>Tmax (hours)</td><td>5.18 ± 2.36</td><td>5.63 ± 2.53</td><td>5.76 ± 1.39</td>

<td>C24 (ng / ml)</td><td>3.91 ± 1.74</td><td>6.11 ± 2.57</td><td>8.63 ± 3.43</td>

Study 1014 (3385) (daily variation, NHV):

[0162] This study investigated the pharmacokinetic profile of tacrolimus following administration of LCP-Tacro 2 mg in the morning and evening hours in 26 healthy male and female volunteers under fasting conditions. The study included two eight-day periods separated by at least two years of cleaning between treatments. The mean pharmacokinetic parameters are summarized in the table below.

<td></td><td>Evening dose</td><td>The morning dose</td>

<td></td><td>1x2mg, n = 26</td><td>1x2mg, n = 25</td>

<td>AUC0-t (ng · hr / ml)</td><td>61.65 ± 24.31</td><td>113.10 ± 41.89</td>

<td>AUC0-12 (ng · hr / ml)</td><td>28.18 ± 8.28</td><td>29.85 ± 9.20</td>

<td>AUC0-24 (ng · hr / ml)</td><td>50.51 ± 14.74</td><td>54.50 ± 16.80</td>

<td>AUC0-inf (ng · hr / ml)</td><td>106.74 ± 35.11</td><td>124.78 ± 42.82</td>

<td>Cmax (ng / ml)</td><td>4.39 ± 1.21</td><td>4.20 ± 1.46</td>

<td>Tmax (hours)</td><td>6.77 ± 2.69</td><td>6.66 ± 2.79</td>

<td>C24 (ng / ml)</td><td>1.34 ± 0.52</td><td>1.69 ± 0.67</td>

Study 1014 (3385) (daily variation, NHV) [0163]

<td></td><td>The ratio of funds (%)</td><td>Internal CV (%)</td>

<td>AUC0-t</td><td>86.93</td><td>39.48</td>

<td>AUC0-12</td><td>97,95</td><td>26.91</td>

<td>AUC 0-24</td><td>95.67</td><td>26.67</td>

<td>AUC 0-inf</td><td>87.64</td><td>35.56</td>

<td>Cmax</td><td>109.65</td><td>29.49</td>

Exam 1015 (3414)

[0164] In this study, the rate and extent of absorption of tacrolimus after the LCP-Tacro 1 mg tablet versus the Prograf capsule (tacrolimus) at a dose of 1 mg fasted in 17 healthy healthy men or women was compared. The study included two eight-day periods separated by at least two years of cleaning between treatments. Pharmacokinetic results are presented in the table below. The longer the Tmax dose for LCP-Tacro tablets (8.78 h vs. 1.39 h) in combination with the lower Cmax (2.54 ng / mL vs. 7.04ng / ml) and the lower AUC (0-inf) (71.82 ng * h / ml vs 50.18 ng * h / ml), maintains once daily dosing for LCP-Tacro tablets compared to Prograf capsules (tacrolimus).

(LCP-Tacro vs. Prograf, NHV, n = 17)

<td></td><td>LCP-Tacro</td><td>Prograf</td>

<td></td><td>1x1mg tablet</td><td>1x1 mg capsule</td>

<td>AUC0-t (ng ^ hr / mL)</td><td>61.65 ± 24.31</td><td>36.15 ± 22.71</td>

<td>AUC0-12 (ng ^ hr / mL)</td><td>16.43 ± 5.98</td><td>22.93 ± 10.64</td>

<td>AUC0-24 (ng ^ hr / mL)</td><td>31.21 ± 8.90</td><td>28.77 ± 13.75</td>

<td>AUC0-inf (ng ^ hr / mL)</td><td>71.82 ± 26.52</td><td>50.18 ± 28.38</td>

<td>Cmax (ng / mL)</td><td>2.54 ± 1.03</td><td>7.04 ± 3.56</td>

<td>Tmax (hr)</td><td>8.78 ± 5.05</td><td>1.39 ± 0.50</td>

<td>C24 (ng / mL)</td><td>1.04 ± 0.36</td><td>0.46 ± 0.25</td>

Study 1015 (3414):

[0165] (LCP-Tacro vs. Prograf, NHV, n = 17)

<td></td><td>The ratio of funds (%)</td><td>Internal CV (%)</td>

<td>AUC0-t</td><td>187.29</td><td>58.06</td>

<td>AUC0-12</td><td>74.89</td><td>42.87</td>

<td>AUC 0-24</td><td>116.97</td><td>40.10</td>

<td>AUC 0-inf</td><td>154.08</td><td>50.73</td>

<td>Cmax</td><td>37,63</td><td>51,81</td>

[0166] Exposure to tacrolimus is significantly higher after LCP-Tacro tablets than after Prograf capsules, with reduced peak / minimum fluctuation and Tmax delay in NHV. The results are consistent with other PK data obtained in the sustained release preparation of the present invention.

[0167] The release time for the sustained release dosage forms of the invention used in the clinical trials described herein provides a sustained release profile in which less than 63.5% is released within 15 hours. A suitable release profile in this range is shown in Fig. 1, tested according to the USP II dissolution test (blade) in medium at pH 4.5 and containing 0.005% hydroxypropylcellulose and a rotation of 50 rpm.

[0168] The solid dosage form of the invention may also be coated to provide suitable properties, e.g. with respect to the controlled release of the active ingredient. The coating can be applied to individual dosage units (e.g., tablets, capsules) or it can be applied to a poly-depot dosage form or its individual units.

Suitable coating materials are e.g. methylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, acrylic polymers, ethylcellulose, cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, polyvinyl alcohol, carboxymethylcellulose sodium, cellulose acetate, cellulose acetate phthalate, gelatin, methacrylic acid copolymer, glycol polyethylene, shellac, sucrose, titanium dioxide, carnauba wax, microcrystalline wax, glycerol monostearate, zein.

[0170] As plasticizers and other ingredients, they can be added to the coating material. The same or different active ingredients may be added in the coating material.

[0171] In preferred embodiments of the invention, the solid dosage forms aim to release tacrolimus in a controlled manner. In the present context, the term "controlled method" includes all types of release that differ from the release obtained from conventional tablets. Thus, the term includes so-called "controlled release", "sustained release", "sustained release", "pulsed release", "sustained release", release of vesicles "," slow release "," extended release "as well as" Delayed "terms. release &amp; quot; and pH dependent release.] However, a particular aspect of the invention relates to delayed release compositions or dosage forms,

Extended Release Systems [0172] The first sustained release system includes matrix systems in which tacrolimus is embedded or dispersed in a matrix of another material that is intended to delay the release of tacrolimus to the aqueous environment (i.e., Luminescent Gl line fluid). When tacrolimus is dispersed in a matrix of this type, the release of the drug takes place mainly from the surface of the matrix. Thus, the drug is released from the surface of the device that contains the matrix after diffusion through the matrix or when the device surface erodes, exposing the drug. In some embodiments, both mechanisms may operate simultaneously. Matrix systems may be large, i.e. tablets of about 1 cm size or small (&lt;0.3 cm). The system can be uniform (eg Bolus), can be divided by the assembly of several subunits (EXAMPLE, several capsules that form a single dose) that are administered substantially simultaneously or can contain many particles. Also meant multiparticulates. The multimodulant may have many uses of the formulation. In the case of EXAMPLE, it can be used multipartically as powder for filling the capsule shell or used per se to mix with food to facilitate ingestion.

[0173] In a further embodiment, the matrix system is in the form of a hydrophilic matrix tablet comprising tacrolimus (e.g. in the form of a solid dispersion) as a multiparticulate product and an amount of a hydrophilic polymer sufficient to provide a useful degree of tacrolimus control of dissolution. Hydrophilic polymers useful for matrix formation include hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), polyethylene oxide, polyvinyl alcohol, xanthan gum, carbomer, carrageenan and zooglan. The preferred material is HPMC. Other similar polymers may also be used

Hydrophilic. In use, the hydrophilic material is swollen and finally dissolved in water. Tacrolimus is released both by diffusion from the matrix and by matrix erosion. The tacrolimus dissolution matrix of these hydrophilic matrix tablets can be controlled by the amount, molecular weight and gel strength of the hydrophilic polymer. Generally, using more hydrophilic polymer, the rate of dissolution is reduced, as is the case with the higher molecular weight polymer. The use of a low molecular weight polymer normally increases the rate of dissolution. The matrix tablet typically contains about 20 to 90% by weight tacrolimus and about 80 to 10% by weight polymer.

[0174] A preferred matrix tablet contains about 30% by weight to about 80% solid dispersion containing tacrolimus of about 15% to about 35% of matrix comprising a matrix (such as, for example, HPMC), 0% to about 35% lactose, 0% to about 20 % microcrystalline cellulose and about 0.25% to about 2% lubricant (such as, for example, magnesium stearate).

[0175] Matrix systems as a class often show discontinuous release of drug from the matrix. This result may be a consequence of the diffusion mechanism of drug release, and it may be advantageous to modify the geometry of the dosage form so that the drug release rate is more constant.

[0176] The second class of dosage forms of the controlled release tacrolimus of the present invention include those controlled by a membrane or reservoirs. In this class, there is a tank of tacrolimus eg in solid solution / dispersion, as a multiparticulate product is surrounded by a speed limiting membrane. Tacrolimus displaces the membrane with mass transport mechanisms well known in the art, including dissolution in the membrane, followed by diffusion through the membrane or diffusion through the fluid-filled pores within the membrane. These individual dosage forms in a system with resources can be large, as in the case of a tablet containing a single large reservoir or a multiparticulate, as in the case of capsules or tablets containing a plurality of tablets containing a plurality of reservoir particles, each individually coated membrane. The coating may be non-porous but permeable to tacrolimus (in the case of an example, tacrolimus may be dispersed directly through the membrane) or it may be porous. As with other embodiments of the present invention, it is not believed that the particular transport mechanism is critical.

[0177] Sustained release coatings known in the art may be used to form a membrane, especially polymeric coatings, such as a cellulose ester or ether, an acrylic polymer, or a mixture of polymers. Preferred materials include ethylcellulose, cellulose acetate and cellulose acetate butyrate. The polymer can be used as a solution in an organic solvent or as an aqueous dispersion or latex. The coating operation can be carried out in standard equipment, such as a fluid bed spreader, Wurster coater or a rotating bed coater.

[0178] If desired, the permeability of the coating can be adjusted by mixing two or more materials. A particularly useful method of adapting the porosity of the coating includes adding a predetermined amount of a finely divided water-soluble material, such as sugars or salts or water-soluble polymers, to a solution or dispersion (e.g., aqueous latex) of the membrane-forming polymer. When the dosage form is poured into the aqueous medium of the Gl tube, these water-soluble membrane additives are released from the membrane, leaving pores that facilitate release of the drug. The membrane coating can also be modified by adding plasticizers, as is known in the art.

[0179] A particularly useful variation of the membrane coating process is the dissolution of the coating polymer in a mixture of solvents selected such that after the coating dries, a phase inversion takes place in the coating solution used, giving the membrane a porous structure.

[0180] In general, no support for a mechanically reinforcing membrane is required.

[0181] The membrane morphology is not critical as long as the permeability properties listed herein are satisfied. The membrane can be amorphous or crystalline. It may have any morphological category produced in any particular process and may be, for EXAMPLE, a polymerized polymerization membrane (which contains skin at a reduced rate in a porous substrate), a hydrophilic porous membrane, a hydrophobic porous membrane, a hydrogel membrane, an ion membrane and other such materials, which are characterized by controlled tacrolimus permeability.

[0182] In one embodiment of the invention, the aim is to reduce the exposure of the upper GI tract to the high concentration of tacrolimus. Accordingly, suitable dosage forms include those forms that contain a specific delay before starting the controlled release of tacrolimus. An exemplary embodiment may be illustrated by a tablet (or particulate material) comprising a tacrolimus-containing core coated with a first coating of a polymeric material of the type suitable for the sustained release of tacrolimus and a second type coating useful for delaying Drug release when the dose is taken. The first coating is applied and surrounds the tablet or individual particles. The second coating is applied and surrounds the first coating.

[0183] A tablet may be made by techniques well known in the art and contain a therapeutically useful amount of tacrolimus plus such excipients as are necessary to make a tablet using such techniques.

[0184] The first coating may be a sustained release coating known in the art, especially polymeric coatings, to form a membrane as previously discussed for tank systems, or it may be a controlled release matrix core coated for a second time with a delay Release material.

[0185] Materials useful for the preparation of the second coating on the tablet include polymers known in the art as enteric coatings for delaying drug release. These are mostly pH sensitive materials, such as cellulose acetate phthalate, cellulose acetate trimellitate, methylcellulose hydroxypropyl phthalate, polyvinyl acetate phthalate, and acrylic copolymers such as Eudragit L-100 (Rohm Pharma) and associated materials, detailed below under "Released exemption". The thickness of the delayed release coating is adjusted to achieve the desired delay property. In general, thicker coatings are more resistant to erosion and consequently give a longer and more effective delay. Preferred coatings have a thickness of about 30 μm to a thickness of about 3 mm.

[0186] After ingestion, the coated tablet passes through the stomach twice, where the second coating prevents the release of tacrolimus under the acidic conditions prevalent there. When the tablet exits the stomach and into the small intestine where the pH is higher, the second coating erodes or dissolves according to the physicochemical properties of the selected material. After erosion or dissolution of the second coating, the first coating prevents the immediate or rapid release of tacrolimus and modulates the release to prevent the formation of high peak concentrations, thereby minimizing side effects.

[0187] A further preferred embodiment comprises a multiparticulate form in which each particle is double coated, as described above for tablets, first with a polymer intended for

To obtain a sustained release of tacrolimus, and then coated with a polymer designed to delay the initial release in the Gl gl tract environment when the dose is taken.

[0188] The release rate of tacrolimus from extended release coated multiparticulates (i.e., multiparticulates before preparation of the release retarding coating) and methods for modifying the coating are also controlled by the factors previously described for tacrolimus multiparticulates in a collection system.

[0189] The second membrane or coating for double coated multiparticulates is a release retarding coating that overlaps the first sustained release layer as disclosed above for the tablets and may be formed from the same materials. It should be noted that the use of so-called "intestinal" materials for performing this embodiment differs significantly from their use for the manufacture of conventional enteric dosage forms. With conventional intestinal forms, the goal is to delay drug release until the dosage form passes through the stomach and then deliver the dose in the duodenum. Dosing of tacrolimus directly and completely to the duodenum may be undesirable, however, due to side effects, the aim of which is to minimize or avoid using the present invention. Consequently, if conventional intestinal polymers are to be used to implement this embodiment, it may be necessary to use them much more densely than in conventional practice to delay drug release until the dosage form reaches the lower GI tract. However, it is also possible to achieve long-term or controlled administration of tacrolimus after dissolution or erosion of a delayed release coating, and thus the advantages of this embodiment may be accomplished with the proper combination of sustained release release form, and the release retarder itself may or may not correspond USP enteral criteria. The thickness of the delayed release coating is adjusted to achieve the desired delay property. Generally,

[0190] The first delayed release according to the invention is a "coated pH-dependent dosage form", such as for example a tablet or capsule. In the case of a tablet, it contains a tablet core comprising tacrolimus, e.g. in solid solution / dispersion as a multiparticulate product, a controlled release matrix e.g. HPMC, a disintegrating agent, 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 the pH of the stomach and which is more soluble and permeable at the pH of 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 a sufficient amount of polymer to ensure that the substantial release of tacrolimus from the dosage form does not occur until the dosage form leaves the stomach and resides in the small intestine for about 15 minutes or longer, preferably about 30 Minutes or longer, thus providing that the minimum tacrolimus is released in the duodenum. It is also possible to use pH-sensitive polymer mixtures with a water insoluble polymer. The tablets are coated with an amount of polymer containing from about 10% to about 80% of the weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer containing about 15% to about 50% of the weight of the tacrolimus tablet core. that the substantial release of tacrolimus from the dosage form will not occur until the dosage form leaves the stomach and resides in the small intestine for about 15 minutes or longer, preferably about 30 Minutes or longer, thereby ensuring that the minimum tacrolimus is released in the duodenum. It is also possible to use pH-sensitive polymer mixtures with a water insoluble polymer. The tablets are coated with an amount of polymer containing from about 10% to about 80% of the weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer containing about 15% to about 50% of the weight of the tacrolimus tablet core. that the substantial release of tacrolimus from the dosage form will not occur until the dosage form leaves the stomach and resides in the small intestine for about 15 minutes or longer, preferably about 30 Minutes or longer, thereby ensuring that the minimum tacrolimus is released in the duodenum. It is also possible to use pH-sensitive polymer mixtures with a water insoluble polymer. The tablets are coated with an amount of polymer containing from about 10% to about 80% of the weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer containing about 15% to about 50% of the weight of the tacrolimus tablet core. It is also possible to use pH-sensitive polymer mixtures with a water insoluble polymer. The tablets are coated with an amount of polymer containing from about 10% to about 80% of the weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer containing about 15% to about 50% of the weight of the tacrolimus tablet core. It is also possible to use pH-sensitive polymer mixtures with a water insoluble polymer. The tablets are coated with an amount of polymer containing from about 10% to about 80% of the weight of the tacrolimus-containing tablet core. Preferred tablets are coated with an amount of polymer containing about 15% to about 50% of the weight of the tacrolimus tablet core.

[0191] pH-sensitive polymers that are very insoluble and impermeable at pH in the stomach, but which are more soluble and permeable at pH in the small intestine and colon, include polyacrylamides, phthalate derivatives such as carbohydrate acid phthalates, amylose acetate phthalate,

Cellulose acetate phthalate, other phthalate cellulose esters, cellulose ether phthalates, hydroxypropyl cellulose phthalate, hydroxypropyl ethylcellulose phthalate, hydroxypropyl methylcellulose phthalate, polyvinyl acetate phthalate, polyvinyl acetate phthalate, hydrogen phthalate, cellulose acetate phthalate sodium, phthalate acid starch, dibutyl phthalate copolymer styrene - maleic acid, styrene-maleic acid-polyvinyl acetate phthalate, phthalate, copolymers of styrene and maleic acid, polyacrylic acid derivatives such as copolymers of acrylic acid and acrylic acid esters, methacrylic acid and their esters, copolymers of polyacrylic acid and acrylic acid, shellac and acetate COP Olymers vinyl and croton acid.

[0192] Preferred pH-sensitive polymers include shellac; Phthalate derivatives, in particular cellulose acetate phthalate, polyvinyl phthalate and hydroxypropyl methylcellulose phthalate; Derivatives of polyacrylic acid, in particular polymethyl methacrylate mixed with acrylic acid and acrylic ester copolymers; And copolymers of vinyl acetate and crotonic acid.

To delay the release of tacrolimus for about 15 minutes or longer, preferably 30 minutes or longer, after withdrawal of the dose from the stomach, preferred coatings contain from about 9: 1 to about 1: 9 Eudragit-L® / Eudragit-S®, more preferably from about 9: 1 to about 1: 4 Eudragit-L® / Eudragit-S®. The coating may contain from about 3% to about 70% by weight of the uncoated tablet core. Preferably, the coating comprises from about 5% to about 50% by weight of the tablet core.

[0194] For the preparation of a solid sustained release oral dosage form, a solid dispersion and / or solution such as tablets, capsules or sachets may be used; Or for the production of granules, granules of microspheres or nanoparticles.

[0195] A further advantage of the sustained release dosage form according to the present invention is the possibility of obtaining an effective therapeutic response with a reduced dose compared to traditional oral treatment. Thus, it is contemplated that the solid dosage form of the invention is administered orally to a mammal in need thereof at a dose that is at most 85% w / w, e.g. at most about 80% w / w. At most about 75%, at most about 70% weight / weight, At most about 65% weight / weight, At most about 60% weight / weight, At most about 55% weight / weight . Most about 50% wt / wt. Doses of tacrolimus administered in the form of Prograf® or a similar commercial product containing tacrolimus are essentially bioequivalent to Prograf® or a similar commercial product containing tacrolimus.

[0196] Any dosage form, compositions, dispersions or solutions in the form of tacrolimus can improve the treatment of conditions that respond to treatment with tacrolimus.

[0197] Tacrolimus is indicated (or suggested) for the treatment of diseases such as e.g. Rejection reactions by transplanting organs or tissues such as the heart, kidneys, liver, bone marrow, skin,

Cornea, lung, pancreas, small intestine, limbs, muscles, nerves, intervertebral disc, trachea, myoblasts, cartilage, etc.; Graft versus host reactions after bone marrow transplantation; Autoimmune diseases such as rheumatoid arthritis, systemic erythematosus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, etc.; Infections caused by pathogenic microorganisms (e.g. Aspergillus fumigatus, Fusarium oxysporum, asteroids from Trichophyton, etc.); Inflammatory or hyperproliferative diseases of the skin or cutaneous manifestations of immunological diseases (eg psoriasis, atopic dermatitis, contact dermatitis, cystitis, seborrheic dermatitis, lichen planus, pemphigus, vesiculitis, bullous epidermolysis, urticaria, angioedema, vasoconstriction, erythema, eosinophilia with skin, erythematous Erythema, acne, alopecia areata); Eye autoimmune disease (conjunctivitis, keratitis, periodontal keratitis, cone keratitis, corneal epithelial dystrophy, keratoleukoma, orbital eye, Moorena ulcer, multiple sclerosis, Graves' ophthalmic form, Vogt-Koyanagi-Harada syndrome, corneal and conjunctivitis ( dry eye), fliketylene, iridocyclitis, sarcoidosis, endocrine ophthalmopathy, etc.); Reversible obstructive airways disease [asthma (eg bronchial asthma, allergic asthma, asthma itself, external asthma and bronchial asthma), especially bronchial asthma or incurable (eg Late asthma and hypersensitivity to the airways) bronchitis, etc.; Inflammation of the mucous or vascular membrane (e.g. gastric ulcer, ischemic or vascular fracture, intestinal ischemic diseases, colitis, necrotic colitis, intestinal damage associated with thermal burns, diseases transmitted by leukotrienes B4); Inflammation / allergies (e.g. celiac disease, prostatitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis); Allergic diseases related to food with a symptom of symptoms distant from the digestive tract (eg Migrenas, runny nose and eczema); Kidney disease (e.g., intestinal ileitis, Goodpasture syndrome, haemolytic uremic syndrome and diabetic nephropathy); Parkinson's disease, amyotrophic lateral sclerosis (ALS) and radiculopathy); Parkinson's disease, amyotrophic lateral sclerosis (ALS) and degenerative diseases, nervous diseases (e.g. multiple myocarditis, Guillain-Barre syndrome, Meniere's disease, multiple neuritis, one-person neuritis, cerebral infarction, Alzheimer's disease); Ischemic brain disease (e.g. Head injury, cerebral haemorrhage (eg, subarachnoid haemorrhage, haemorrhage), cerebral thrombosis, brain embolism, cardiac arrest, stroke, transient ischemic attack (TIA), hypertensive encephalopathy, cerebral infarction); Endocrine diseases (e.g., Hyperthyroidism and Basedow's disease); Haematological diseases (eg Pure red cell aplasia, aplastic anemia, non-small cell anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, asymptomatic anemia, megaloblastic anorethropplasia); Diseases of bones (e.g., osteoporosis); Respiratory diseases (e.g. sarcoidosis, pulmonary fibrosis and idiopathic interstitial pneumonia); Skin diseases (e.g. dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, hypersensitivity to light and cutaneous T-cell lymphoma); Circulatory diseases (e.g. atherosclerosis, atherosclerosis, aortic inflammatory syndrome, polyangiitis and myocarditis); Collagen diseases (eg hard scleroderma, Wegener's granulomatosis and Sjogren's syndrome); Adipoza; Eosinophilic fasciitis; Periodontal disease (eg Damage to gums, periodontium, alveolar bone or pericardial sacs); Nephrotic syndrome (e.g. glomerulonephritis); Male pattern baldness, senile baldness; muscular dystrophy; Pyoderma and Sezary syndrome; Diseases associated with

Chromosomal abnormalities (e.g., Down's syndrome); Addison's disease; Active diseases carried by oxygen [e.g. Organ damage (eg. Ischemic circulation disorders (eg Heart, liver, kidneys, digestive tract, etc.) Associated with behavior, transplant or ischemic diseases (e.g., thrombosis, myocardial infarction, etc.)); Intestinal diseases (e.g. Endotoxin shock, pseudomembranous colitis and colitis induced by drug or radiation); Kidney disease (eg ischemic acute renal failure, chronic renal failure); Lung diseases (e.g. toxicity caused by tissue or lung drugs (e.g. parakort, bleomycin, etc.), lung cancer and emphysema); Eye diseases (for example, cataracts, iron storage diseases (tuberous siderosis), retinitis, pigmentosis, senile plaques, vitreous scars, keratitis); Dermatitis (e.g. erythema multiforme, Linear immunoglobulin A vesicular dermatitis, cement dermatitis); And other diseases (e.g., Gingivitis, periodontitis, sepsis, pancreatitis, Diseases caused by environmental pollution (e.g., Air pollution), aging, carcinogenesis, cancer metastases and hyperbaropathy); Diseases caused by histamine release or Leukotriene C4 release; Restenosis of the coronary artery after angioplasty and prevention of posturgical adhesions; Autoimmune diseases and inflammations (eg Primary mucosal edema, autoimmune atrophic gastritis, premature menopause, male sterility, juvenile diabetes, pemphigus vulgaris, pemphigoid, sympathetic oftrophites, uveitis caused by the lens, idiopathic leukopenia, active chronic hepatitis, idiopathic cirrhosis, erythematous erythema, autoimmune sclerosis, arthritis (eg arthritis deformed), or polyangiitis); HIV infection (AIDS), AIDS; Allergic conjunctivitis; Obstructive kikatrix and keloid due to injury, inflammation or surgery. [0198] In addition, tricyclic macrolides, such as e.g. Tacrolimus, have liver regeneration activity and / or hepatocyte hyperplasia and hyperplasia effect. Therefore, the sustained release dosage form of the present invention is useful for increasing the effect of therapy and / or prophylaxis of liver diseases [e.g. (Eg liver necrosis due to toxins, viral hepatitis, shock or anoxia),

[0199] In addition, the sustained release dosage form of the present invention is useful to increase the prevention and / or treatment efficacy of various diseases due to the useful pharmacological activity of three-dimensional macrolides, such as increased chemotherapeutic activity, cytomegalovirus activity, infection, anti-inflammatory activity, inhibition of activity against peptidyl-prolyl isomerase or rotamase, antimalarial activity, anti-tumor activity and so on.

Materials and methods

Materials [0200] Tacrolimus (provided by Eurotrade); Lot No. RD 03-111

Lactose monohydrate 200 mesh (with DMV)

EP 2 167 033 B1

Granular Silicon Oxide, Aeroperl® 300, (Degussa)

Polyethylene glycol 6000, Pluracol® E6000 (from BASF)

Poloxamer 188, Pluronic® F-68 (from BASF)

Glyceryl monostearate, Rylo® MD50, (from Danisco Cultor), Ph. Eur.; Lot number 4010056276

Avicel PH200 (microcrystalline cellulose) (with FMC)

Lactose DCL 11 (with DMV)

Magnesium stearate

Croscarmellose sodium, Ac-Di-Sol® (with FMC)

Eudragit® L30D.55 (from Degussa)

Triethyl citrate (Merck)

Anti-foam emulsion (with Unik)

Micro talc

HPMC refers to Metolose 90SH (type 2910, 2208) or Metolose 60SH (type 2910) with ShinEtsu available in various degrees of polymerization (viscosity 3-100,000 cP).

[0201] Tablets, capsules or granules can be enteric coated with various types of polymers, such as hydroxypropyl methylcellulose acetate succinate (Aqoat), cellulose phthalate acetate cap, HPMCP hydroxypropylmethylcellulose or methacrylic acid copolymers such as Eudragit L30D, Eudragit 100 / S, Eudragit 100 / L.

Comparison of the prior art tacrolimus preparation in vivo:

[0202] Prograf gelatin gel capsules, manufactured by Fujisawa Ireland Ltd.

<td>Components</td><td>mg</td>

<td>Tacrolimus, anhydrous.</td><td>1.0</td>

<td>gelatine</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>q, s,</td>

<td>Soya lecithin</td><td>q, s,</td>

<td>Red iron oxide (E172)</td><td>q, s,</td>

<td>Titanium dioxide (E171)</td><td>q, s,</td>

<td>Dimethicone (E900)</td><td>q, s,</td>

methods

Determination of mass deviation [0203] Tablets prepared in EXAMPLES were subjected to the weighting test carried out in accordance with Ph Eur.

EP 2 167 033 B1

Determination of average tablet hardness [0204] Tablets prepared in EXAMPLES were subjected to tablet hardness test using a Schleuniger Model 6D apparatus and were carried out in accordance with the general guidelines for the device.

Determination of disintegration time [0205] The period of disintegration of the tablet, i.e. the degradation into particles or agglomerates, was determined in accordance with Ph Eur. Determination of the average geometric diameter dgw [0206] The average diameter of the geometric diameter was determined by the use of a laser diffraction method to diffuse the obtained material (or starting material) in the air. The measurements were carried out at a leakage pressure of 1 bar in Sympatec Helos, which records the distribution of the equivalent spherical diameter. This distribution is matched to the normal volume distribution.

[0207] As used herein, "geometric mean diameter" means the average diameter of a normal volume distribution.

In vitro dissolution tests [0208] The following test methods related to the compositions and dosage forms of the present invention.

Test 1:

[0209] In vitro release test according to the USP A method, release retardants (USP paddle method, rotational speed 50 rpm, 37 ° C, after 2 hours in an acid medium, the medium is changed to phosphate buffer pH 6.8).

Test 2:

[0210] The in-vitro dissolution test in aqueous solution for dissolution is adjusted to pH 4.5 (900 ml of water with 0.005% HPC (hydroxypropylcellulose) is adjusted to pH 4.5, 37 ° C by Paddy USP, rotation speed 50 rpm. minute.

List of drawings:

[0211]

Figure 1: Dissolution of the sustained release preparation of the invention and tested according to the USP II dissolution test method (stirrer) in an environment of pH 4.5 and containing 0.005% hydroxypropylcellulose and a rotation of 50 rpm.

Figure 2: Scintigraphic assessment of the release location using the sustained release formulation of the present invention. The figure shows that in vivo release with the preparation of the present invention is extended to the extent that the absorption takes place in the intestines of a person. The test is described herein as LCP-Tacro PK-003.

Figure 3: discloses the blood concentration of tacrolimus in a single-dose fasting study of healthy volunteers. The closed triangles represent the concentration of the 5 mg formulation of the present invention, they are Prograf® 5 mg; Open circles indicate a 2 mg formulation according to the present invention; The closed squares denote 2x2 mg treatment in accordance with the present invention; Closed diamonds mean treatment with Prograf® 4x1 mg; The vertical line means

Treatment with Prograf® 2x1 mg. The study was described in List A in this document as LCP-Tacro PK-002.

Figure 4: discloses dissolving a preferred embodiment of the invention having a composition similar to that disclosed in EXAMPLE 20. Triangles treat treatment with 1 mg, squares with a 2 mg preparation. In addition, the dissolution of the commercial Advagraf® product used for comparisons in EXAMPLE 20 occurs where the stars refer to 0.5 mg, to about 1 mg and discs to 5 mg of Advagraf®. The release is measured as a percentage dissolved in time revealed in hours. As the dissolution method, the USP II dissolution test method (agitator) in a medium adjusted to pH 4.5 and containing 0.005% hydroxypropylcellulose and a rotation speed of 50 rpm was used.

Figure 5: discloses steady state blood profiles obtained prior to conversion (steady state of Prograf® 7 in day) and after conversion on days 14 and 21 to the sustained release preparation of the present invention. The squares denote the day of rograph 7, the LCP-Tacro wheels once a day on day 17, and the LCP tacro diamonds once daily on day 21. The details of the study are disclosed in EXAMPLE 19. The profiles reveal actual profiles after conversion to a lower dose with the preparation according to the invention.

Figure 6: discloses the blood profiles in the steady state of Figure 5, however, the corrected dose.

Figure 7: Reveals the results of phase 1 studies in healthy volunteers, LCP-Tacro tablets showed about 50% greater bioavailibility than Prograf® capsules (Astellas Pharma in Japan) and PK profile, supporting once-a-day administration. This test is phase 1, bidirectional crossover, open label, multidose analysis, bioequivalence, comparing pharmacokinetics (Cmax, C24 and AUCtau) and safety of LCP-Tacro tablets compared to Advagraf capsules in fasted state. Twenty healthy male volunteers were randomly assigned to receive one LCP-Tacro 2 mg tablet or two Advagraf® 1 mg capsules per day for 10 days. After two weeks of washing out, each patient received an alternative treatment. The PK profile after 10 days of each treatment is illustrated in Figure 7 and shows

Figure 8: discloses blood plasma concentrations after administration of the same dose of Advagraf® 2x1 mg capsules of tacrolimus in open boxes and as LCT-Tacro 2 mg tablets of the present invention. The study is described in EXAMPLE 20.

[0212] The following EXAMPLES serve to illustrate the invention.

[0213] Pharmaceutical compositions and dosage forms that can be optimized to achieve the desired release profile of the invention are set forth in EXAMPLES 1-16, which preparations are disclosed in patent application WO 2005/020993. The dissolution profiles disclosed therein and in EXAMPLE 17 of the present application may be further optimized for the delivery of release profiles meeting the sustained release parameters of the present invention. Optimization can include changes in the formulation, including changes in the levels of excipients, a change in the proportion of excipients, changes in production, such as a change in stress pressure, change in hardness or disintegration time. As a preferred molecular solution, significant changes include the use of excipients that prevent the degradation of the active substance,

EP 2 167 033 B1

EXAMPLE 1

Modular modified-release poly-depot capsule based on swollen hydrocolloid hydroxypropylcellulose matrix [0214]

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>HPMC</td><td>20.00</td><td>40.00</td>

<td>Lactose 200 mesh</td><td>30.00</td><td>60.00</td>

<td>PEG 6000</td><td>34.65</td><td>69.30</td>

<td>Poloxamer 188</td><td>14.85</td><td>29.70</td>

<td>Total</td><td>100.00</td><td>200.00</td>

[0215] Tacrolimus was dissolved in polyethylene glycol 6000 and poloxamer 188 (70:30 w / w ratio) at 70 ° C. The solution was sprayed with a mixture of 150 g lactose and 100 g HPMC in a fluid bed Strea-1. The granulate was sieved through a 0.7 mm sieve and filled with hard gelatine capsules (200 mg).

EXAMPLE 2

Modular modified-release poly-depot capsule based on swollen hydrocolloid hydroxypropylcellulose matrix [0216]

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>HPMC 2910 3 cp</td><td>20.00</td><td>40.00</td>

<td>Lactose 200 mesh</td><td>30.00</td><td>60.00</td>

<td>Glycerin monostearate</td><td>49.50</td><td>99,00</td>

<td>Total</td><td>100.00</td><td>200.00</td>

[0217] Tacrolimus was dissolved in glyceryl monostearate at 70 ° C. The solution was sprayed with a mixture of 150 g lactose and 100 g HPMC in a fluid bed, Strea-1. The granulate was sieved through a 0.7 mm sieve and filled with hard gelatine capsules (200 mg).

EXAMPLE 3

Extended release matrix tablet based on swollen hydrocolloid hydroxypropylcellulose matrix [0218]

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

EP 2 167 033 B1

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>HPMC</td><td>19,90</td><td>40.00</td>

<td>Lactose 200 mesh</td><td>29.85</td><td>60.00</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>Total</td><td>100.00</td><td>201.01</td>

[0219] Tacrolimus was dissolved in glycol on a polyethylene 6000 and poloxamer 188 (70:30 w / w ratio) at a temperature of 70 ° C. The solution was sprayed onto 250 g lactose in a Strea-1 fluid bed. The resulting granulate was sieved through a 0.7 mm sieve and mixed with HPMC and magnesium stearate for 0.5 min in a Turbula mixer.

The mixture was compressed in 8 mm tablets with 1 mg active ingredient (200 mg tablet) with a molded cup of compound.

[0221] Mean disintegration time: 20 min. Hardness: 45 N EXAMPLE 4 (EXAMPLE OF REFERENCE)

Modified release tablet according to a lipophilic matrix of glyceryl monostearate [0222]

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>Lactose 200 mesh</td><td>49.75</td><td>100.00</td>

<td>Glycerin monostearate</td><td>49.25</td><td>99,00</td>

<td>Magnesium stearate</td><td>0.50</td><td>1.01</td>

<td></td><td>100.00</td><td>201.01</td>

[0223] Tacrolimus was dissolved in glyceryl monostearate at 70 ° C. The solution was sprayed onto 250 g lactose in a Strea-1 fluid bed. The granulate was sieved through a 0.7 mm sieve and mixed with magnesium stearate for 0.5 minutes in a Turbula mixer.

[0224] The resulting mixture was compressed in 8 mm tablets of 1 mg active ingredient (200 mg tablet) cup-shaped compound.

[0225] Mean disintegration time: 20 min. Hardness: 45 N EXAMPLE 5 (EXAMPLE OF REFERENCE)

Modified Release Polydot Capsule Based on a Lipophilic Glyceryl Monostearate Matrix [0226]

EP 2 167 033 B1

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>Lactose 200 mesh</td><td>49.75</td><td>100.00</td>

<td>Glycerin monostearate</td><td>49.25</td><td>99,00</td>

<td>Magnesium stearate</td><td>0.50</td><td>1.01</td>

<td></td><td>100.00</td><td>201.01</td>

[0227] Tacrolimus was dissolved in glyceryl monostearate at 70 ° C. The solution was sprayed onto 250 g lactose in a Strea-1 fluid bed. The granulate was sieved through a 0.7 mm sieve and filled with hard gelatine capsules (200 mg).

EXAMPLE 6 (EXAMPLE OF REFERENCE)

Modified release poly-depot tablet based on the lipophilic matrix Gelucire® 44/14 [0228]

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

<td>tacrolimus</td><td>0.50</td><td>1.00</td>

<td>Aeroperl® 300</td><td>49.75</td><td>100.00</td>

<td>Gelucire® 44/14</td><td>49.25</td><td>99,00</td>

<td>Magnesium stearate</td><td>0.50</td><td>1.01</td>

<td></td><td>100.00</td><td>201.01</td>

[0229] Tacrolimus was dissolved in Gelucire® at 70 ° C. The solution was sprayed onto 250 g Aeroperl® in a fluid bed Strea-1. The granulate was sieved through a 0.7 mm sieve and filled with hard gelatine capsules (200 mg).

[0230] The resulting granules were compressed in 8 mm tablets of 1 mg active ingredient (tablet weight 200 mg).

The tablets were in the shape of a cup.

[0231] Mean disintegration time: 25 minutes. Hardness: 43 N.

EXAMPLE 7 Enteric coating [0232] Capsules and tablets EXAMPLES 1, 2, 3, 5 and 6 were then coated with the following enteric coating to provide a delayed release of the active ingredient after administration.

<td>Components</td><td>%</td>

<td>Eudragit® L30D</td><td>40</td>

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

<td>Triethyl acetylcitrate</td><td>1.8</td>

EP 2 167 033 B1

<td>Components</td><td>%</td>

<td>Anti-foam emulsion</td><td>0.2</td>

<td>Talc</td><td>6</td>

<td>Total</td><td>100</td>

[0233] A coating suspension was prepared by mixing an emulsion of triethylcellulose, an antifoam emulsion and purified water in an Ultra Turrax at a speed of 9,500 rpm. For 30 minutes. After adding 1 minute of talc. The mixture was passed through a No. 300 and mixed using a magnetic stirrer. Eudragit went through sieve no. 300 and a mixture was added, which was stirred for 5 minutes.

[0234] The coating process conditions were as follows: inlet temperature 40 ° C, outlet temperature ° C, air inlet 140 cbm per hour and covering time about 50 minutes (300 g of coating material). About. 400 g tablets or 200 g capsules were coated.

[0235] Tablets coated tablets and capsules were cured for 48 hours at 30 ° C before the dissolution test.

EXAMPLE 8 (EXAMPLE OF REFERENCE)

Modified release tablet according to a lipophilic matrix of glyceryl monostearate [0236]

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

<td>tacrolimus</td><td>0.95</td><td>2.00</td>

<td>HPMC, Pharmacoat 606</td><td>6.75</td><td>14.29</td>

<td>Lactose monohydrate, lactose 125 mesh</td><td>6.75</td><td>14.29</td>

<td>Glyceryl monostearate, Rylo® MD50</td><td>30.56</td><td>64.67</td>

<td>Magnesium stearate</td><td>0.5</td><td>1.06</td>

<td>Talc</td><td>4.5</td><td>9.52</td>

<td>Lactose monohydrate, Pharmacose DCL 14</td><td>50.00</td><td>105.8</td>

<td></td><td>100.00</td><td>211.64</td>

[0237] Tacrolimus was dissolved in glycerol monostearate at a temperature above 80 ° C. The solution was sprayed with a Phast FS1.7 feed unit on 60 g lactose and 60 g HPMC in a Phast FB100 fluid bed.

The granular product was cured in a heating oven for 4 hours at 50 ° C. The resulting granular product was sieved through a 0.71 mm sieve and mixed with lactose for 3 minutes in a Turbula mixer. [0238] Magnesium stearate and talc were sieved through a No. 300 and mixed in a Turbula mixer for 3 minutes. The granulate was mixed with a mixture of magnesium stearate and talc (1: 9) for 0.5 minutes in a Turbula mixer.

The final mixture was compressed in 8 mm tablets with 2 mg active ingredient (210 mg tablet) with a cup shape.

[0240] Mean disintegration time: 2 hours. Hardness: 50 N-EXPOSURE 9

EP 2 167 033 B1

Pill coated tablet with a core based on PEG 6000 / Poloxamer 188 and an enteric coating based on Eudragit L30D 55 [0241]

The composition of the tablet core:

<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>Croscarmellose sodium, Ac-di-sol</td><td>5.00</td><td>5.05</td>

<td></td><td>100.00</td><td>101.01</td>

[0242] The tacrolimus tablet core was prepared by dissolving in PEG 6000 at a temperature above 80 ° C. Poloxamer 188 was added and the solution was heated to above 80 ° C. The solution was sprayed with a Phast FS1.7 feed unit per 200 g of lactose monohydrate in a Phast FB100 fluid bed. . The resulting granulate was sieved through a Comill sieve 1397, 4500 rpm and mixed with croscarmellose sodium for 3 minutes in a Turbula mixer.

[0243] Magnesium stearate and talc were sieved through a No. 300 and mixed in a Turbula mixer for 3 minutes. The granulate was mixed with magnesium stearate and talc (1: 9) for 0.5 minutes in a Turbula mixer.

[0244] The resulting mixture was compressed in 6 mm tablets with 2 mg of active ingredient (100 mg tablets) of cup-shaped compound.

[0245] Mean disintegration time: 7 minutes. Hardness: 65 N

Intestinal coating:

[0246] The enteric coating is based on the Eudragit L30D-55 acrylic polymer. Eudragit L30D is supplied as an aqueous latex suspension forming a water-insoluble film when water is evaporated during coating. The polymer is insoluble at pH below 5.0 and readily soluble at pH values ​​above 6.0. The shell coating composition is:

<td>Substance</td><td>w / in%</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-foam emulsion</td><td>0.2</td>

EP 2 167 033 B1

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

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

<td>Total</td><td>100</td>

[0247] The amount of polymeric film (Eudragit) used was based on the calculation of mg polymer film on the surface of the cm2 tablet. The thickness of the enteric coating was 80 μm. The checking of the applied coating thickness was based on the measurement of the height of the tablet with a digital micrometer. The film coating process was carried out in a Phast FB100 fluid bed equipped with a Wurster cartridge. The process conditions were as follows: air temperature at inlet 50 ° C; Intake air flow 100m3 per hour; Product temperature 38 ° C; Feeding speed 15g / min.

[0248] After coating, proper film formation requires curing of the coated tablets, i.e. at 30 ° C for 48 hours in an oven. Alternatively, the coated tablets may more effectively cure at 40 ° C for 24 hours.

EXAMPLE 10

PEG 6000 / Poloxamer 188 release control tablet based on the HPMC matrix.

[0249]

Composition of tablets:

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

<td>tacrolimus</td><td>1.21</td><td>2.00</td>

<td>Lactose monohydrate, lactose 200 mesh</td><td>24.75</td><td>40.91</td>

<td>PEG 6000</td><td>20.33</td><td>33,60</td>

<td>Poloxamer 188, Lutrol 68</td><td>8.71</td><td>14.40</td>

<td>Magnesium stearate</td><td>0.50</td><td>0.83</td>

<td>Talc</td><td>4.50</td><td>7.44</td>

<td>Hydroxypropylmethylcellulose, Metolose 90SH 15000</td><td>40.00</td><td>66.12</td>

<td></td><td>100.00</td><td>165.29</td>

[0250] Tacrolimus was dissolved in PEG 6000 at a temperature above 80 ° C. Poloxamer 188 is added and the solution is heated to a temperature above 80 ° C. The solution is sprayed through a supply unit

Phast FS1.7 per 200 g lactose monohydrate in a Phast FB100 fluid bed. The granulate is sifted through Comill, sieve 1397, 4500 rpm and mixed with hydroxypropylmethylcellulose for 3 minutes in a Turbula mixer.

[0251] Magnesium stearate and talc are sieved through a 300 sieve and mixed in a Turbola mixer for 3 min. The granulate was mixed with Magnesium Stearate: Talc (1: 9) for 0.5 min in a Turbula mixer. The mixture is compressed in 8 mm 2 mg tablets (165 mg cup-shaped tablet). The average disintegration time: 2 hours 34 minutes, hardness: 50 N

EXAMPLE 11

EP 2 167 033 B1

Intestinal coating preparation. Wet granules and enteric coated tablets [0252]

Composition of tablets:

<td>Component</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

<td>Lactose</td><td>80</td>

<td>Sodium lauryl sulphate</td><td>10</td>

<td>Kollidon VA64</td><td>3</td>

<td>Avicel PH200</td><td>thirty</td>

<td>Magnesium stearate</td><td>0.5</td>

<td>Total</td><td>125.5</td>

[0253] The tablet formulation is based on wet granulation in a high shear mixer Pellmix

1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 80 g sodium lauryl sulfate in a high shear mixer. 15 15 20 25 30 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 20 25 30 35 40 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 And then kneaded for 3 minutes at the same speed. The granulate was dried in a tray drier and sieved through a 0.7 mm sieve.

[0254] The granulate was mixed with 240 g Avicel PH200 for 3 minutes and for and after the addition of 4 g magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine.

[0255] Tablet diameter: 6 mm. Tablet shape: round, cups.

[0256] The tablets were then coated with an enteric coating of the acrylic type as described in EXAMPLE 9.

[0257] The amount of a folic polymer (Eudragit) to be used should be based on a calculation of mg of polymeric film on the surface of a cm2 tablet. The thickness of the enteric coating should be 50-80 μm. The checking of the applied coating thickness is based on the measurement of the height of the tablet with a digital micrometer. The film coating process is performed in a Stre-1 fluid bed equipped with a Wurster cartridge under the following process conditions:

<td>Process parameter</td><td>The value of the process</td>

<td>Product load, g</td><td>400</td>

<td>The temperature of the air at the inlet, ° C</td><td>40</td>

<td>Intake air flow, m3 per hour</td><td>140</td>

<td>Exhaust air temperature, ° C</td><td>31</td>

<td>Feeding speed g / min</td><td>5</td>

[0258] After coating, proper coating preparation requires curing the coated tablets, i.e. 30 ° C in 48 hours in an oven. Alternatively, the coated tablets may more effectively cure at 40 ° C for 24 hours.

EXAMPLE 12

A controlled release tablet formulation based on the eroding HPMC HPMC matrix was added as part of the external phase. Wet granulation.

[0259]

Composition of tablets:

<td>Components</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

<td>Lactose</td><td>80</td>

<td>Sodium lauryl sulphate</td><td>10</td>

<td>Kollidon VA64</td><td>3</td>

<td>Avicel PH200</td><td>thirty</td>

<td>Meteorosis SH 90</td><td>60</td>

<td>Magnesium stearate</td><td>1</td>

<td>Total</td><td>186</td>

[0260] The tablet preparation is based on wet granulation in a high shear mixer Pellmix 1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 80 g sodium lauryl sulfate in a high shear mixer. 15 15 20 25 30 15 15 15 15 15 15 15 15 15 15 15 15 35 35 35

50 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 80 50 The granulate was dried in a tray dryer and sieved through a 0.7 mm sieve.

The granulate was mixed with 240 g Avicel PH200 and 480 g hydroxypropyl methylcellulose Metolose SH 90 15 100 cP for 3 minutes and for and after the addition of 8 g magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine.

[0262] Tablet diameter: 7 mm. Tablet shape: round, cups.

EXAMPLE 13

A controlled release tablet formulation based on the eroding HPMC HPMC matrix was added as part of the intra-granular phase. Wet granulation.

[0263]

Composition of tablets:

<td>Component</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

EP 2 167 033 B1

<td>Component</td><td>mg</td>

<td>Lactose</td><td>80</td>

<td>Sodium lauryl sulphate</td><td>10</td>

<td>Meteorosis SH 90</td><td>80</td>

<td>Avicel PH200</td><td>60</td>

<td>Magnesium stearate</td><td>2</td>

<td>Total</td><td>234</td>

[0264] The tablet formulation is based on wet granulation in a high shear mixer Pellmix 1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 80 g sodium lauryl sulfate and 640 g hydroxypropylmethylcellulose Metolose SH 90 15,000 cP in a high shear mixer. Purified water was pumped to the mixture at a rotor speed of 500 rpm at a feed rate of 20 g / min. And then kneading for 3 minutes. The granulate was dried in a tray drier and sieved through a 0.7 mm sieve.

The granulate was mixed with 480 g Avicel PH200 for 3 minutes, followed by 16 g magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine.

[0266] Tablet diameter: 8 mm. Tablet shape: round, cups.

EXAMPLE 14

A controlled release tablet formulation based on HPMC HPMC eroding was added as part of the intragranular phase. Granulating granulate [0267]

Composition of tablets:

<td>Component</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

<td>Lactose</td><td>80</td>

<td>PEG 6000</td><td>15</td>

<td>Poloxamer 188</td><td>6</td>

<td>Meteorosis SH 90</td><td>80</td>

<td>Avicel PH200</td><td>60</td>

<td>Magnesium stearate</td><td>2</td>

<td>Total</td><td>245</td>

[0268] The tablet formulation is based on melt granulation in a high shear mixer Pellmix 1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 120 g polyethylene glycol 6000, 48 g Poloxamer 188 and 640 g hydroxypropyl methylcellulose Metolose SH 90 15,000 cP in a mixer with

High shear. The jacket of the mixing bowl was heated to 80 ° C and the mixture was heated at a rotational speed of 1000 revolutions per minute until the melting temperature of PEG and Poloxamer. After melting, the kneading was continued for 4 minutes at 800 rpm. The granulate was sieved through a 0.7 mm sieve and cooled on a tray. The granulate was mixed with 480 g of Avicel PH200 for 3 minutes and then 16 g of magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine. Diameter of the tablet: 8 mm. Tablet shape: round, cups.

EXAMPLE 15

A preparation containing a controlled release tablet based on erosion of SR Kollidon matrix added as part of the extra-granular phase.

[0269]

Tablet composition:

<td>Component</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

<td>Lactose</td><td>80</td>

<td>Sodium lauryl sulphate</td><td>10</td>

<td>Kollidon VA64</td><td>3</td>

<td>Lactose DC lac14</td><td>50</td>

<td>Kollidon SR</td><td>60</td>

<td>Magnesium stearate</td><td>1</td>

<td>Total</td><td>206</td>

[0270] The tablet formulation is based on wet granulation in a high shear mixer Pellmix 1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 80 g sodium lauryl sulfate in a high shear mixer. A 15% Kolllidon VA64 aqueous binder solution (Kollidon SR is a polyvinyl acetate mixture and 80:20 polyvinyl pyrrolidone) pumped into the mixture at a rotor speed of 500 rpm at a feed rate of 20 g / min and then kneaded for 3 minutes. The granulate was dried in a tray drier and sieved through a 0.7 mm sieve. The granulation was mixed with 400 g of lactose DC Lac 14 and 480 g of Kollidon SR for 3 minutes followed by the addition of 8 g magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine.

Diameter of the tablet: 8 mm. Tablet shape: round, cups.

EXAMPLE 16

Intestinal coating preparation (melt granules and enteric coated tablets) [0271]

Composition of tablets:

EP 2 167 033 B1

<td>Component</td><td>mg</td>

<td>tacrolimus</td><td>2</td>

<td>Lactose</td><td>80</td>

<td>PEG 6000</td><td>15</td>

<td>Poloxamer 188</td><td>6</td>

<td>Avicel PH200</td><td>60</td>

<td>Magnesium stearate</td><td>2</td>

<td>Total</td><td>165</td>

[0272] The tablet preparation is based on melt granulation in a high shear mixer Pellmix 1/8. 16 g Micronized tacrolimus was mixed with 640 g lactose 125 mesh and 120 g polyethylene glycol 6000, 48g Poloxamer 188 in a high shear mixer. The jacket of the mixing bowl was heated to 80 ° C and the mixture was heated at a rotational speed of 1000 revolutions per minute until the melting point of PEG and

Poloxamer. After melting, the kneading was continued for 4 minutes at 800 rpm. The granulate was sieved through a 0.7 mm sieve and cooled on a tray. The granulate was mixed with 480 g of Avicel PH200 for 3 minutes and then 16 g of magnesium stearate for a further 0.5 minutes. The mixture was compressed into tablets on a single Diaf TM20 tabletting machine. Diameter of the tablet: 7 mm. Tablet shape: round, cups. The intestinal coating of the tablets is carried out according to the procedure described in example 11.

EXAMPLE 17

In Vitro Dissolution Data [0273] The compositions and dosage forms of the previous examples were subjected to in vitro dissolution tests using two different media / dissolution tests.

A. Using a solution / dissolution test: 900 ml of 0.005% HPC (hydroxypropylcellulose) aqueous medium adjusted to pH 4.5 (USP paddle method at 50 rpm) found the following dissolution profiles:

<td></td><td colspan="6">% release</td>

<td>Time</td><td>Ex.</td><td>Ex.</td><td>Ex.</td><td>Ex. 8</td><td>Ex. 9 - EC</td><td>Ex. 10</td>

<td>(Minutes)</td><td>1</td><td>3</td><td>4</td><td>(Uwol%)</td><td>(Uwol%)</td><td>(Uwol%)</td>

<td>0</td><td>0</td><td>0</td><td>0</td><td>0 (0)</td><td>0 (0)</td><td>0 (0)</td>

<td>0.5</td><td></td><td></td><td>2</td><td></td><td></td><td></td>

<td>1</td><td></td><td></td><td>4</td><td></td><td></td><td></td>

<td>1.5</td><td>0</td><td>0</td><td></td><td></td><td></td><td></td>

<td>2</td><td>0</td><td>0</td><td></td><td></td><td></td><td></td>

<td>3</td><td></td><td></td><td>6</td><td></td><td></td><td></td>

EP 2 167 033 B1

<td></td><td colspan="6">% release</td>

<td>Time</td><td>Ex.</td><td>Ex.</td><td>Ex.</td><td>Ex. 8</td><td>Ex. 9 - EC</td><td>Ex. 10</td>

<td>(Minutes)</td><td>1</td><td>3</td><td>4</td><td>(Uwol%)</td><td>(Uwol%)</td><td>(Uwol%)</td>

<td>4</td><td>1</td><td>3</td><td></td><td>7.8 (11.1)</td><td>0.8 (32.3)</td><td>7.4 (9.8)</td>

<td>5</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>6</td><td>3</td><td>4</td><td></td><td></td><td></td><td></td>

<td>8</td><td>5</td><td>7</td><td>17</td><td>17.0 (8.3)</td><td>0.4 (61.1)</td><td>13.3 (16.5)</td>

<td>10</td><td>20</td><td>14</td><td></td><td></td><td></td><td></td>

<td>15</td><td>40</td><td></td><td></td><td>32.2 (4.8)</td><td>11.0 (17,3)</td><td>36.0 (5.8)</td>

<td>16</td><td></td><td>38</td><td></td><td></td><td></td><td></td>

<td>17</td><td></td><td></td><td></td><td>35.1 (9.6)</td><td>13.2 (12.1)</td><td>44.5 (5.4)</td>

<td>20</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>24</td><td></td><td></td><td>37</td><td></td><td></td><td></td>

[0274] Dissolution profile for the tablet cores of example 9 in a dissolution medium: 900 ml, aqueous media with 0.005% HPC (hydroxypropylcellulose) adjusted to pH = 4.5. USP paddle method. Rotational speed: 50 rpm:

<td>Time (minutes)</td><td>% release</td><td>Uwol%</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>

[0275] Dissolution profile for enterally coated tablet-9 tablets in the dissolution medium. 5 For the USP A method, delay release products. The USP Paddle method. Rotational speed: 50 rpm:

<td>Time (minutes)</td><td>% release</td><td>Uwol%</td>

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

<td>120</td><td>0</td><td>ON</td>

<td>155</td><td>84.8</td><td>12.8</td>

EP 2 167 033 B1

<td>Time (minutes)</td><td>% release</td><td>Uwol%</td>

<td>165</td><td>102.9</td><td>ON</td>

<td>175</td><td>101.0</td><td>3.5</td>

EXAMPLE 18 [0276] A list of stability-enhancing agents to improve and optimize tacrolimus sustained release formulation containing tacrolimus in solid solution.

[0277] Tacrolimus in the dissolved form may degrade and several degradation products may be made during storage. Stability tests tested to improve the stability of tacrolimus are disclosed in the list below.

Carriers, antioxidant preparations

Addition

1000 ppm propyl gallate

500 ppm α-tocopherol + 500 ppm Lipoid

500 ppm Ascorbyl palmitate

1000 ppm of α-tocopherol

500 ppm of α-tocopherol ppm α-tocopherol

1000 ppm Ascorbyl palmitate + 1000 ppm α-tocopherol

500 ppm of ascorbyl palmitate + 500 ppm of α-tocopherol

250 ppm ascorbyl palmitate + 250 ppm α-tocopherol ppm ascorbyl palmitate + 50 ppm α-tocopherol

Pollutant removal in poloxamer by filtering with Al2O3 (Compalox)

Dimethicon addition

Addition of BHT

Addition of organic acids, including tartaric acid

0.01% tartaric acid, 0.05% tartaric acid, 0.10% tartaric acid, 0.20% tartaric acid,

0.40% tartaric acid, 0.50% tartaric acid, 0.60% tartaric acid, 0.75% tartaric acid, 1% tartaric acid, 5% tartaric acid

Variability of tartaric acid about 0.15%, at 200 ppm of α-tocopherol

Tablets, dried N2 in ° 4h, dried by N2, low temperature 65 ° C low heating time, ° 24h dried by N2, low temperature 65 ° C low heating time, ° 4h dried by N2, low temperature 65 ° C low heating time, ° 24h dried by N2, low temperature 65 ° C low heating time

Tablets, open storage in various controlled moisture,

11% humidity by LiCl

Humidity 32% using MgCl2,

Humidity 48% with K2CO3, 75% humidity with NaCl

89% humidity by KNO3

EXAMPLE 19

[0278] A clinical trial comparing the sustained release preparation of the present invention with Prograf®. Blood-concentration profiles are disclosed in Figures 5 and 6.

Research title:

[0279] Phase II, Open Label, multicenter study conversion in patients with stable kidney transplants to compare the pharmacokinetics of LCP-Tacro tablets. Disposable to Prograf® capsules twice a day. LCP-Tacro 1 mg, 2 mg, 5 mg tablets

Active ingredient: tacrolimus.

Indications: Tacrolimus (Prograf®) is used to prevent the rejection of liver, kidney and heart transplants.

Research project and development phase: 3-sequential, open, prospective, multi-center, conversion process (phase II).

Study title: Phase II, open label, multi-center conversion study in patients with stable kidney to compare the pharmacokinetics of LCP-Tacro tablets. Disposable to Prograf® capsules. Twice

Objectives: To evaluate exposure of tacrolimus (AUC0-24) and trough levels (C24) in stable kidney transplant patients transformed from Prograf® tablets (tacrolimus, Astellas Pharma US, Inc.) into LCP-Tacro tablets in three sequential studies; To assess the safety of LCP-Tacro compared to Prograf.

Main inclusion criteria: men and women aged 18-65 who have been carrying a kidney transplant at least 6 months before the start of recording. Test products / test products and methods of administration: LCP-Tacro 1 mg, 2 mg, 5 mg tablets , taken orally once a day in the morning.

Reference product, batch number and administration: Prograf® 0.5 mg, 1 mg, 5 mg capsules, given in two divided doses in the morning and evening.

Methods: A third sequence, an open, multicenter, prospective study in kidney transplant patients to evaluate and compare pharmacokinetics (Cmax, C24 and AUC) and the safety of LCP-Tacro tablets (tacrolimus) compared to Prograf® (Tacrolimus).

[0280] Stable kidney transplants that met all inclusion / exclusion (I / E) criteria were saved and stored in the Prograf® program for 7 days. After a 24-hour PK study on day 7 to determine the pharmacokinetics of Prograf®, all patients were converted once daily to LCP-Tacro (Ratio 1: 0.66-0.80) for 7 days without the option of changing the dose. On day 14, a 24-hour LCP-Tacro PK study was conducted. On Day 15, one of the predefined dose changes was allowed if the average score of 3 trough levels was greater than 25% on days 10 ± 1, 12 ± 1 (separated by at least 48 hours from the previous trial) and 14 compared with the Average 3 measured levels on days 0, 7 and 8 for each individual patient.

[0281] Patients remained on a dose determined on Day 14 for a further 7 days without dosing. A further 24-hour LCP-Tacro PK study was performed on day 21. On day 22, patients were converted to the original dose of Prograf® twice daily for 30 days after the end of the safety assessment at 52 days after the safety assessment. A preliminary PK analysis was carried out after 10 patients, finishing day 21, before continuing the recording.

[0282] Points of blood cells collected: The following blood samples were tested during the study:

EP 2 167 033 B1

For LCP-Tacro: blood collection points: 0.00 (before dosing), 0.50, 1.00, 1.50, 2.00, 3.00, 4.00, 6.00, 8.00 , 12.00, 14.00, 16.00, 20.00 and 24.00 hours after the dose, on days 14 and 21.

For Prograf®: blood collection points: 0.00 (initial dose), 0.50, 1.00, 1.50, 2.00, 3.00, 4.00, 6.00, 8.00, 12.00, 12.50, 13.00, 13.50, 14.00, 15.00, 16.00, 20.00 and 24.00 hours after the morning dose, on day 7.

[0283] Evaluation criteria: A pharmacokinetic analysis was carried out in 47 patients. Safety assessment was performed in all patients who received at least one dose during the study.

[0284] Pharmacokinetics (PK): The following pharmacokinetic parameters of tacrolimus were calculated by standard interval methods: AUCt (t = 24), Cmax, Cmin, Cave, Tmax,% fluctuation,% variation and Cmax / Cmin.

[0285] Statistical methods: All demographic data, pharmacokinetic parameters, laboratory data and AE were summarized using descriptive statistics. In the case of continuous data, the average, standard deviation, average, minimum and maximum were recorded. Categorical data are given in percentages and frequencies.

[0286] Non-commercial pharmacokinetic parameters [AUCt (t = 24), Cmax, Cmin, Cave, Tmax] were calculated based on time and blood concentration data.

[0287] Statistical methods: The use of GLM procedures in SAS was carried out ANOVA on natural log transformation parameters (w) of AUCt, Cmax, Cmin and Cavg and on unchanged% fluctuation parameters,% swing and Cmax / Cmin. The model included treatment as a factor. The geometric LSM ratio together with 90% CI was calculated according to the following three comparisons for AUCt, Cmax and Cmin:

Day 14 Prograf versus Day 7 LCP-Tacro Day 21 Prograf versus Day 7 LCP-Tacro Day 21 LCP-Tacro versus Day 14 LCP-Tacro [0288] The Tmax parameter was analyzed using nonparametric methods. The Wilcoxon rank test was used to compare the pairing. The average change between the two treatments (as described above) was estimated using the median unlimited Hodges-Lehmann grade and 90% exact confidence interval.

[0289] The degree of correlation between AUCt and Cmin was quantified by calculating the correlation coefficient and presenting the graphical data displayed on days 7, 14 and 21. Both parameters were subjected to inversion transformation before correlation analysis. Statistical analysis of the subgroup race (Blacks vs. Non-Blacks) was carried out in order to obtain an appropriate division between the two groups.

Summary of pharmacokinetic results:

[0290]

Pharmacokinetic parameters for tacrolimus in all patients:

<td colspan="2"></td><td colspan="2">Geometric mean (% CV)</td>

<td></td><td>parameters Pharmacokinetic</td><td>The arithmetic mean ± SD</td><td></td>

<td></td><td></td><td>Dose Uncorrected Data</td><td>Data corrected in time</td>

EP 2 167 033 B1

<td rowspan="2"></td><td rowspan="2"></td><td>Prograf capsules bid orally</td><td colspan="4">LCP-Tacro tablets qd orally</td><td>Prograf capsules bid orally</td><td colspan="4">LCP-Tacro tablets qd orally</td>

<td>Day 7 (n = 47)</td><td>Day 1 (n = 47)</td><td>4</td><td>Day 2 (n = 46)</td><td>1</td><td>Day 7 (n = 47)</td><td>Day 1 (n = 47)</td><td>4</td><td>Day 2 (n = 46)</td><td>1</td>

<td></td><td></td><td>212.12</td><td>206.79</td><td></td><td>209.05</td><td></td><td>34.81</td><td>47.73</td><td></td><td>48.30</td><td></td>

<td></td><td>AUCt # (ng · hr /</td><td>(25.59)</td><td>(29.27)</td><td></td><td>(31.30)</td><td></td><td>(52.08)</td><td>(57.25)</td><td></td><td>(56.46)</td><td></td>

<td></td><td>ml)</td><td>218.82 ±</td><td>215.71</td><td>±</td><td>218.03</td><td>±</td><td>39.89 ±</td><td>56.83</td><td>±</td><td>56.90</td><td>±</td>

<td></td><td></td><td>55.99</td><td>63.14</td><td></td><td>68.23</td><td></td><td>20.78</td><td>32.54</td><td></td><td>32.13</td><td></td>

<td></td><td></td><td>17.66</td><td>12.64</td><td></td><td>13.05</td><td></td><td></td><td>2.92</td><td></td><td>3.02</td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td>2.90</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>(42.59)</td><td>(36.02)</td><td></td><td>(41.91)</td><td></td><td></td><td>(47.24)</td><td></td><td>(47.92)</td><td></td>

<td></td><td>Cmax (ng / ml)</td><td></td><td></td><td></td><td></td><td></td><td>(41.11)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>19.14 ±</td><td>13.45</td><td>±</td><td>13.94</td><td>±</td><td></td><td>3.29</td><td>±</td><td>3.44</td><td>±</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td>3.18 ± 1.31</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>8.15</td><td>4.84</td><td></td><td>5.84</td><td></td><td></td><td>1.56</td><td></td><td>1.65</td><td></td>

<td></td><td></td><td></td><td>6.59</td><td></td><td>6.64</td><td></td><td></td><td>1.52</td><td></td><td>1.53</td><td></td>

<td></td><td></td><td>6.82</td><td></td><td></td><td></td><td></td><td>1.12</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>(33.41)</td><td></td><td>(31.70)</td><td></td><td></td><td>(64.87)</td><td></td><td>(70.71)</td><td></td>

<td></td><td>Cmin (ng / ml)</td><td>(22.01)</td><td></td><td></td><td></td><td></td><td>(66.74)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>6.96</td><td>±</td><td>6.94</td><td>±</td><td></td><td>1.91</td><td>±</td><td>1.92</td><td>±</td>

<td></td><td></td><td>7.00 ± 1.54</td><td></td><td></td><td></td><td></td><td>1.35 ± 0.90</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>2.32</td><td></td><td>2.20</td><td></td><td></td><td>1.24</td><td></td><td>1.35</td><td></td>

<td></td><td></td><td></td><td>8.62</td><td></td><td>8.71</td><td></td><td></td><td>1.99</td><td></td><td>2.01</td><td></td>

<td></td><td></td><td>8.84</td><td></td><td></td><td></td><td></td><td>1.45</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>(29.27)</td><td></td><td>(31.29)</td><td></td><td></td><td>(57.25)</td><td></td><td>(56.46)</td><td></td>

<td></td><td>Cavg (ng / ml)</td><td>(25.59)</td><td></td><td></td><td></td><td></td><td>(52.08)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>8.99</td><td>±</td><td>9.08</td><td>±</td><td></td><td>2.37</td><td>±</td><td>2.37</td><td>±</td>

<td></td><td></td><td>9.12 ± 2.33</td><td></td><td></td><td></td><td></td><td>1.66 ± 0.87</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>2.63</td><td></td><td>2.84</td><td></td><td></td><td>1.36</td><td></td><td>1.34</td><td></td>

<td></td><td></td><td></td><td>6.00</td><td></td><td>6.00</td><td></td><td></td><td>6.00</td><td></td><td>6.00</td><td></td>

<td></td><td></td><td>1.82 (0.50-</td><td></td><td></td><td></td><td></td><td>1.82 (0.50-</td><td></td><td></td><td></td><td></td>

<td></td><td>Tmax (hours) *</td><td></td><td>(1.00-</td><td></td><td>(1.50-</td><td></td><td></td><td>(1.00-</td><td></td><td>(1.50-</td><td></td>

<td></td><td></td><td>24.00)</td><td></td><td></td><td></td><td></td><td>24.00)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>16.00)</td><td></td><td>16.00)</td><td></td><td></td><td>16.00)</td><td></td><td>16.00)</td><td></td>

<td></td><td>Degree of fluctuation n</td><td>127.41 ±</td><td>73.24</td><td>±</td><td>77.04</td><td>±</td><td>127.41 ±</td><td>73.24</td><td>±</td><td>77.04</td><td>±</td>

<td></td><td>(%)</td><td>57.28</td><td>44.96</td><td></td><td>50.59</td><td></td><td>57.28</td><td>44.96</td><td></td><td>50.59</td><td></td>

<td></td><td></td><td>174.55 ±</td><td>102.80</td><td>±</td><td>110.07</td><td>±</td><td>174.55 ±</td><td>102.80</td><td>±</td><td>110.07</td><td>±</td>

<td></td><td>Swing degree (%)</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>93.72</td><td>75.24</td><td></td><td>89.23</td><td></td><td>93.72</td><td>75.24</td><td></td><td>89.23</td><td></td>

<td></td><td></td><td></td><td>2.03</td><td>±</td><td>2.10</td><td>±</td><td></td><td>2.03</td><td>±</td><td>2.10</td><td>±</td>

<td></td><td>Cmax / Cmin</td><td>2.75 ± 0.94</td><td></td><td></td><td></td><td></td><td>2.75 ± 0.94</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>0.75</td><td></td><td>0.89</td><td></td><td></td><td>0.75</td><td></td><td>0.89</td><td></td>

<td>*</td><td colspan="3">Median (min - max); #: Τ = 24 hours</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

Summary of pharmacokinetic results (continued): [0291]

Pharmacokinetic parameters for tacrolimus in black patients:

<td>parameters</td><td>Geometric mean (% CV)</td>

EP 2 167 033 B1

<td></td><td>Pharmacoki</td><td colspan="11">The arithmetic mean ± SD</td>

<td></td><td rowspan="2">netic</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td colspan="5">Unadjusted dose data</td><td></td><td colspan="4">Data corrected in time</td><td></td>

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

<td></td><td></td><td>capsules</td><td></td><td>tablets</td><td colspan="3">LCP-Tacro</td><td>capsules</td><td colspan="2">tablets</td><td colspan="2">LCP-Tacro</td>

<td></td><td></td><td>bid</td><td></td><td colspan="3">qd orally</td><td></td><td>bid</td><td colspan="3">qd orally</td><td></td>

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

<td></td><td></td><td>Day 7</td><td>(n</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td><td>Day 7 (n</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td>

<td></td><td></td><td>= 20)</td><td></td><td>(n = 20)</td><td></td><td>(n = 19)</td><td></td><td>= 20)</td><td>(n = 20)</td><td></td><td>(n = 19)</td><td></td>

<td></td><td></td><td>242.19</td><td></td><td>204.13</td><td></td><td>216.93</td><td></td><td>26.35</td><td>31.57</td><td></td><td>32.50</td><td></td>

<td></td><td>AUCt # (ng ·</td><td>(24.78)</td><td></td><td>(32.48)</td><td></td><td>(39.32)</td><td></td><td>(74.70)</td><td>(84.23)</td><td></td><td>(84.71)</td><td></td>

<td></td><td>hr / ml)</td><td>250.12</td><td>±</td><td>214.78</td><td>±</td><td>231.41</td><td>±</td><td>31.41 ±</td><td>38.13</td><td>±</td><td>39.17</td><td>±</td>

<td></td><td></td><td>61.98</td><td></td><td>69.77</td><td></td><td>90.99</td><td></td><td>23.47</td><td>32.11</td><td></td><td>33.18</td><td></td>

<td></td><td>Cmax (ng /</td><td>21.72 (41.29)</td><td></td><td>13.89 (41.05)</td><td></td><td>14.48 (48.92)</td><td></td><td rowspan="2">2.36 (46.02)</td><td>2.15 (60.56)</td><td></td><td>2.17 (64.07)</td><td></td>

<td></td><td rowspan="2">ml)</td><td>23.73</td><td>±</td><td>15.20</td><td>±</td><td>15.91</td><td>±</td><td>2.46</td><td>±</td><td>2.53</td><td>±</td>

<td rowspan="2"></td><td rowspan="2">9.80</td><td rowspan="2"></td><td rowspan="2">6.24</td><td rowspan="2"></td><td rowspan="2">7.78</td><td rowspan="2"></td><td rowspan="2">2.61 ± 1.20</td><td rowspan="2">1.49</td><td rowspan="2"></td><td rowspan="2">1.62</td><td rowspan="2"></td>

<td></td>

<td></td><td>Cmin (ng /</td><td rowspan="2">7.43 (21.57)</td><td></td><td>6.21 (37.53)</td><td></td><td>6.68 (38.83)</td><td></td><td rowspan="2">0.81 (107.31)</td><td>0.96 (99.20)</td><td></td><td>1.00 (120.04)</td><td></td>

<td></td><td rowspan="2">ml)</td><td></td><td>6.62</td><td>±</td><td>7.09</td><td>±</td><td>1.26</td><td>±</td><td>1.34</td><td>±</td>

<td></td><td colspan="2">7.62 ± 1.64</td><td></td><td></td><td></td><td></td><td>1.07 ± 1.15</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.49</td><td></td><td>2.75</td><td></td><td></td><td>1.25</td><td></td><td>1.61</td><td></td>

<td></td><td></td><td>10.09</td><td></td><td>8.51</td><td></td><td>9.04</td><td></td><td>1.10</td><td>1.32</td><td></td><td>1.35</td><td></td>

<td></td><td>Cavg (ng /</td><td>(24.78)</td><td></td><td>(32.49)</td><td></td><td>(39.32)</td><td></td><td rowspan="2">(74.70)</td><td>(84.22)</td><td></td><td>(84.70)</td><td></td>

<td></td><td rowspan="2">ml)</td><td>10.42</td><td>±</td><td>8.95</td><td>±</td><td>9.64</td><td>±</td><td>1.59</td><td>±</td><td>1.63</td><td>±</td>

<td rowspan="2"></td><td rowspan="2">2.58</td><td rowspan="2"></td><td rowspan="2">2.91</td><td rowspan="2"></td><td rowspan="2">3.79</td><td rowspan="2"></td><td rowspan="2">1.31 ± 0.98</td><td rowspan="2">1.34</td><td rowspan="2"></td><td rowspan="2">1.38</td><td rowspan="2"></td>

<td></td>

<td></td><td></td><td></td><td></td><td>4.00</td><td></td><td>6.00</td><td></td><td></td><td>4.00</td><td></td><td>6.00</td><td></td>

<td></td><td rowspan="2">Tmax (hours)</td><td colspan="2">1.91 (0.50 -</td><td></td><td></td><td></td><td></td><td rowspan="2">1.91 (0.50 -</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td rowspan="2">(1.00-</td><td></td><td rowspan="2">(1.50-</td><td></td><td rowspan="2">(1.00-</td><td></td><td rowspan="2">(1.50-</td><td></td>

<td></td><td>*</td><td rowspan="2">24.00)</td><td></td><td></td><td></td><td rowspan="2">24.00)</td><td></td><td></td>

<td></td><td></td><td></td><td>12.00)</td><td></td><td>16.00)</td><td></td><td>12.00)</td><td></td><td>16.00)</td><td></td>

<td></td><td>Degree</td><td>145.87</td><td>±</td><td>94.99</td><td>±</td><td>91.42</td><td>±</td><td>145.87 ±</td><td>94.99</td><td>±</td><td>91.42</td><td>±</td>

<td></td><td>fluctuation (%)</td><td>60.72</td><td></td><td>54.11</td><td></td><td>63.47</td><td></td><td>60.72</td><td>54.11</td><td></td><td>63.47</td><td></td>

<td></td><td>Degree</td><td>212.69</td><td>±</td><td>138.16</td><td>±</td><td>135.87</td><td>±</td><td>212.69 ±</td><td>138.16</td><td>±</td><td>135.87</td><td>±</td>

<td></td><td>Swing (%)</td><td>108.38</td><td></td><td>91.69</td><td></td><td>114.34</td><td></td><td>108.38</td><td>91.69</td><td></td><td>114.34</td><td></td>

<td></td><td></td><td></td><td></td><td>2.38</td><td>±</td><td>2.36</td><td>±</td><td></td><td>2.38</td><td>±</td><td>2.36</td><td>±</td>

<td></td><td>Cmax / Cmin</td><td colspan="2">3.13 ± 1.08</td><td></td><td></td><td></td><td></td><td>3.13 ± 1.08</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.92</td><td></td><td>1.14</td><td></td><td></td><td>0.92</td><td></td><td>1.14</td><td></td>

<td>*</td><td>Median (min -</td><td colspan="2">max); #: Τ =</td><td colspan="3">24 hours</td><td></td><td></td><td></td><td></td><td></td><td></td>

Pharmacokinetic parameters for tacrolimus in non-black patients:

<td>parameters</td><td>Geometric mean (% CV)</td>

EP 2 167 033 B1

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

<td></td><td>Pharmacokinetic</td><td colspan="4">The arithmetic mean ± SD</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td colspan="5">Dose Uncorrected Data</td><td colspan="5">Data corrected in time</td>

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

<td></td><td></td><td>Capsules</td><td>tablets</td><td colspan="3">LCP-Tacro</td><td>Capsules</td><td colspan="4">LCP-Tacro tablets</td>

<td></td><td></td><td>bid</td><td colspan="3">qd orally</td><td></td><td>bid</td><td colspan="3">qd orally</td><td></td>

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

<td></td><td></td><td>Day 7 (n</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td><td>Day 7 (n</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td>

<td></td><td></td><td>= 27)</td><td>(n = 27)</td><td></td><td>(n = 27)</td><td></td><td>= 27)</td><td colspan="2">(n = 27)</td><td colspan="2">(n = 27)</td>

<td></td><td></td><td>192.28</td><td>208.78</td><td></td><td>203.68</td><td></td><td>42.79</td><td>64.82</td><td></td><td>63.83</td><td></td>

<td></td><td>AUCt # (ng · hr /</td><td>(19.32)</td><td>(27.32)</td><td></td><td>(21.98)</td><td></td><td>(35.25)</td><td>(36.10)</td><td></td><td>(36.26)</td><td></td>

<td></td><td>ml)</td><td>195.64 ±</td><td>216.40</td><td>±</td><td>208.62</td><td>±</td><td>46.18 ±</td><td>70.69</td><td>±</td><td>69.37</td><td>±</td>

<td></td><td></td><td>37.79</td><td>59.11</td><td></td><td>45.86</td><td></td><td>16.28</td><td>25.52</td><td></td><td>25.15</td><td></td>

<td></td><td></td><td>15.15</td><td>11.79</td><td></td><td>12.14</td><td></td><td></td><td>3.66</td><td></td><td>3.80</td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td>3.37</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>(27.81)</td><td>(24.56)</td><td></td><td>(27.91)</td><td></td><td></td><td>(33.56)</td><td></td><td>(33.35)</td><td></td>

<td></td><td>Cmax (ng / ml)</td><td></td><td></td><td></td><td></td><td></td><td>(34.44)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>15.74 ±</td><td>12.14</td><td>±</td><td>12.55</td><td>±</td><td></td><td>3.92</td><td>±</td><td>4.08</td><td>±</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td>3.60 ± 1.24</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>4.38</td><td>2.98</td><td></td><td>3.50</td><td></td><td></td><td>1.31</td><td></td><td>1.36</td><td></td>

<td></td><td></td><td></td><td>6.88</td><td></td><td>6.60</td><td></td><td></td><td>2.14</td><td></td><td>2.07</td><td></td>

<td></td><td></td><td>6.41</td><td></td><td></td><td></td><td></td><td>1.43</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>(30.70)</td><td></td><td>(25.79)</td><td></td><td></td><td>(41.92)</td><td></td><td>(42.40)</td><td></td>

<td></td><td>Cmin (ng / ml)</td><td>(19.98)</td><td></td><td></td><td></td><td></td><td>(39.02)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>7.20</td><td>±</td><td>6.84</td><td>±</td><td></td><td>2.40</td><td>±</td><td>2.32</td><td>±</td>

<td></td><td></td><td>6.54 ± 1.31</td><td></td><td></td><td></td><td></td><td>1.56 ± 0.61</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>2.21</td><td></td><td>1.76</td><td></td><td></td><td>1.01</td><td></td><td>0.98</td><td></td>

<td></td><td></td><td></td><td>8.70</td><td></td><td>8.49</td><td></td><td></td><td>2.70</td><td></td><td>2.66</td><td></td>

<td></td><td></td><td>8.01</td><td></td><td></td><td></td><td></td><td>1.78</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>(27.32)</td><td></td><td>(21.98)</td><td></td><td></td><td>(36.10)</td><td></td><td>(36.26)</td><td></td>

<td></td><td>Cavg (ng / ml)</td><td>(19.31)</td><td></td><td></td><td></td><td></td><td>(35.25)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>9.02</td><td>±</td><td>8.69</td><td>±</td><td></td><td>2.95</td><td>±</td><td>2.89</td><td>±</td>

<td></td><td></td><td>8.15 ± 1.57</td><td></td><td></td><td></td><td></td><td>1.92 ± 0.68</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>2.46</td><td></td><td>1.91</td><td></td><td></td><td>1.06</td><td></td><td>1.05</td><td></td>

<td></td><td></td><td></td><td>6.00</td><td></td><td>7.87</td><td></td><td></td><td>6.00</td><td></td><td>7.87</td><td></td>

<td></td><td></td><td>1.52 (0.50 -</td><td></td><td></td><td></td><td></td><td>1.52 (0.50 -</td><td></td><td></td><td></td><td></td>

<td></td><td>Tmax (hours) *</td><td></td><td>(2.00</td><td>-</td><td>(1.53-</td><td></td><td></td><td>(2.00</td><td>-</td><td>(1.53-</td><td></td>

<td></td><td></td><td>13.48)</td><td></td><td></td><td></td><td></td><td>13.48)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>16.00)</td><td></td><td>12.05)</td><td></td><td></td><td>16.00)</td><td></td><td>12.05)</td><td></td>

<td></td><td>Degree of fluctuation n</td><td>113.74 ±</td><td>57.13</td><td>±</td><td>66.92</td><td>±</td><td>113.74 ±</td><td>57.13</td><td>±</td><td>66.92</td><td>±</td>

<td></td><td>(%)</td><td>51.53</td><td>28.34</td><td></td><td>37.20</td><td></td><td>51.53</td><td>28.34</td><td></td><td>37.20</td><td></td>

<td></td><td></td><td>146.30 ±</td><td>76.61</td><td>±</td><td>91.92</td><td>±</td><td>146.30 ±</td><td>76.61</td><td>±</td><td>91.92</td><td>±</td>

<td></td><td>Swing degree (%)</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>70.76</td><td>46.88</td><td></td><td>62.46</td><td></td><td>70.76</td><td>46.88</td><td></td><td>62.46</td><td></td>

<td></td><td></td><td></td><td>1.77</td><td>±</td><td>1.92</td><td>±</td><td></td><td>1.77</td><td>±</td><td>1.92</td><td>±</td>

<td></td><td>Cmax / Cmi n</td><td>2.46 ± 0.71</td><td></td><td></td><td></td><td></td><td>2.46 ± 0.71</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td>0.47</td><td></td><td>0.62</td><td></td><td></td><td>0.47</td><td></td><td>0.62</td><td></td>

<td>*</td><td colspan="3">Median (min - max); #: Τ = 24 hours</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

Summary of pharmacokinetic results (continued): [0292]

EP 2 167 033 B1

Relative assessment of bioavailability for day 14 compared to day 7 For tacrolimus in all patients:

<td></td><td></td><td colspan="3">Dose Uncorrected Data</td><td colspan="3">Data corrected in time</td>

<td></td><td>Parameter</td><td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td><td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td>

<td></td><td>AUCt *</td><td>880.63% down 107.23%</td><td>97.49%</td><td>28.44%</td><td>1,110.97% up to 167,84%</td><td>137.09%</td><td>64.84%</td>

<td></td><td>Cmax</td><td>63.09% to 81.23%</td><td>71.59%</td><td>38.29%</td><td>840.52% to 119.91%</td><td>100.67%</td><td>54.74%</td>

<td></td><td>C min</td><td>870.33% down 106,70%</td><td>96.53%</td><td>29.97%</td><td>1,070.43% up to 171.51%</td><td>135.74%</td><td>77.33%</td>

<td colspan="8">*: Τ = 24 hours</td>

Relative assessment of bioavailability for day 21 compared to day 7 For tacrolimus in all patients:

<td rowspan="2"></td><td rowspan="2">Parameter</td><td colspan="3">Dose Uncorrected Data</td><td colspan="3">Data corrected in time</td>

<td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td><td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td>

<td></td><td>AUCt *</td><td>89.55% to 108.46%</td><td>98.55%</td><td>28.44%</td><td>11319% to 170.03%</td><td>138.73%</td><td>64.84%</td>

<td></td><td>Cmax</td><td>65.11% to 83.95%</td><td>73.93%</td><td>38.29%</td><td>87.29% to 124.08%</td><td>104.07%</td><td>54.74%</td>

<td></td><td>C min</td><td>870.91% down 107.53%</td><td>97.23%</td><td>29.97%</td><td>1,080.17% up to 173.12%</td><td>136.84%</td><td>77.33%</td>

<td colspan="8">*: Τ = 24 hours</td>

The relative bioavailability assessment for day 21 compared to day 14 for tacrolimus in all patients:

<td rowspan="2"></td><td rowspan="2">Parameter</td><td colspan="3">Dose Uncorrected Data</td><td colspan="3">Data corrected in time</td>

<td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td><td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td>

<td></td><td>AUCt *</td><td>910.86% down 111.26%</td><td>101.09%</td><td>28.44%</td><td>82From 56% down 124.03%</td><td>101.20%</td><td>64.84%</td>

EP 2 167 033 B1

<td rowspan="2"></td><td rowspan="2">Parameter</td><td colspan="3">Dose Uncorrected Data</td><td colspan="3">Data corrected in time</td>

<td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td><td>90% CI</td><td>Ratio funds</td><td>Intramural CV</td>

<td></td><td>Cmax</td><td>900.96% down 117.27%</td><td>103.28%</td><td>38.29%</td><td>86.71% to 123.25%</td><td>103.38%</td><td>54.74%</td>

<td></td><td>C min</td><td>910.07% down 111.40%</td><td>100.72%</td><td>29.97%</td><td>790,69% down 127.54%</td><td>100.81%</td><td>77.33%</td>

<td colspan="8">*: Τ = 24 hours</td>

Correlation between AUCt and Cmin For tacrolimus:

<td rowspan="3"></td><td rowspan="3">Parameter</td><td colspan="6">Dose Uncorrected Data</td>

<td colspan="2">Day 7</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td>

<td>AUCt (ng · Hr / ml)</td><td>Cmin (ng / ml)</td><td>AUCt (ng · H / m L)</td><td>Cmin (ng / ml)</td><td>AUCt (ng · Hr / ml)</td><td>C min (ng / ml)</td>

<td></td><td>Mean</td><td>5.35716</td><td>1.92049</td><td>5.33171</td><td>1.88517</td><td>5.34260</td><td>1.8923 8</td>

<td></td><td>Deviation standard</td><td>0.25141</td><td>0.23353</td><td>0.29591</td><td>0.33707</td><td>0.28751</td><td>.2998 9</td>

<td></td><td>Correlation</td><td colspan="2">0.78656</td><td colspan="2">0.91380</td><td colspan="2">0.86471</td>

<td></td><td>P-value</td><td colspan="2">&lt;.0001</td><td colspan="2">&lt;.0001</td><td colspan="2">&lt;.0001</td>

<td rowspan="3"></td><td rowspan="3">Parameter</td><td colspan="6">Data corrected in time</td>

<td colspan="2">Day 7</td><td colspan="2">Day 14</td><td colspan="2">Day 21</td>

<td>AUCt (ng · Hr / ml)</td><td>Cmin (ng / ml)</td><td>AUCt (ng · H / m L)</td><td>Cmin (ng / ml)</td><td>AUCt (ng · Hr / ml)</td><td>C min (ng / ml)</td>

<td></td><td>Mean</td><td>3.55001</td><td>0.11346</td><td>3.86547</td><td>0.41903</td><td>3.87736</td><td>.4271 3</td>

<td></td><td>Deviation standard</td><td>0.54311</td><td>0.62557</td><td>0.62680</td><td>0.72963</td><td>0.60442</td><td>.6947 9</td>

<td></td><td>Correlation</td><td colspan="2">0.97266</td><td colspan="2">0.99092</td><td colspan="2">0.98176</td>

<td></td><td>P-value</td><td colspan="2">&lt;.0001</td><td colspan="2">&lt;.0001</td><td colspan="2">&lt;.0001</td>

Proposal:

[0293] The main objective of this study was to estimate steady-state plasma levels of tacrolimus (AUCt) and levels (Cmin) in patients with stable renal

In sequential studies carried out with Prograf® (tacrolimus, Astellas Pharma US, Inc.) on the LCP-Tacro project.

[0294] After systemic dose adjustment, steady-state tacrolimus exposure (AUCt) and tacrolimus trough (Cmin) levels were significantly higher when LCP-Tacro qd tablets were administered compared to therapy with Prograf® Capsules bid in patients with kidney transplants. The systemic exposure (AUCt and Cmin) over 24 hours of LCP-Tacro 2 mg tablets (qd) was ~ 38% and was about 36% greater than that of Prograf® (bid) tablets. The mean drug concentration in the dose range, Cavg, was significantly higher in the LCP-Tacro therapy than Prograf®.

There were no statistically significant differences in the maximal systemic exposure (Cmax) of tacrolimus between Prograf® and LCP-Tacro treatment on days 14 and 21. In addition, treatment with Prograf® showed a significantly higher degree of fluctuation and variation than LCP-Tacro. Compared to Prograf® (Day 7), a greater correlation was observed between AUCt and Cmin on days 14 and 21 (LCP-Tacro treatment); However, the size of the difference was not high. There were no significant differences in total systemic exposure, lower levels of tacrolimus or the extent of variation and fluctuation, comparing LCP-Tacro of day 14 compared to the dose on day 21.

[0296] Analysis of the subgroup of the drug in black and non-melanomas, used in both Prograf®or LCPTacro, indicates that there are statistical peak and systemic differences in tacrolimus exposure, as well as tacrolimus levels indicating the need for caution when administering the black population .

[0297] The results of this study indicate that after Prograf's treatment with LCP-Tacro, LCP-Tacro therapy shows significantly higher systemic exposure and a lower degree of fluctuation and fluctuation of tacrolimus at steady-state compared to the period when Prograf® treatment is stable Patients with kidney transplants. In addition, the highest exposure of tacrolimus is similar after comparing different treatments; However, the mean concentration in the dose range was higher for LCP-Tacro.

[0298] Discussion and general conclusions: The main objective of this study was to assess steady-state plasma levels of tacrolimus (AUCt) and correction levels (Cmin) in stable kidney transplants transplanted from Prograf® (tacrolimus, Astellas Pharma US, Inc.) for LCP-Tacro In three sequential tests.

[0299] After systemic dose adjustment, steady-state tacrolimus exposure (AUCt) and tacrolimus trough (Cmin) levels were significantly higher when LCP-Tacro qd tablets were administered compared to therapy with Prograf® Capsules bid in patients with kidney transplants. The systemic exposure (AUCt and Cmin) over 24 hours of LCP-Tacro 2 mg tablets (qd) was ~ 38% and was about 36% greater than that of Prograf® (bid) tablets. The mean drug concentration in the dose range, Cavg, was significantly higher in the LCP-Tacro therapy than Prograf®. There were no statistically significant differences in the maximum systemic exposure (Cmax) of tacrolimus between Prograf® and LCP-Tacro on days 14 and 21. In addition, treatment with Prograf® showed a significantly higher degree of fluctuation and variation than LCP-Tacro. Compared to Prograf® (Day 7), a greater correlation was observed between AUCt and Cmin on days 14 and 21 (LCP-Tacro treatment); However, the size of the difference was not high. There were no significant differences in total systemic exposure, lower levels of tacrolimus or the extent of variation and fluctuation, comparing LCP-Tacro of day 14 compared to the dose on day 21.

[0300] Analysis of the subgroup of nucleotides compared to kidneys who use the Prograf® or LCPTacro therapy, show statistical differences in the exposure of drugs and tacrolimus in plasma, as well as at the level of the tacrolimus of the channel indicating the need for caution during administration of the black population.

[0301] The results of this study indicate that after the Prograf preparation for treatment with the product

LCP-Tacro LCP-Tacro therapy shows significantly higher systemic exposure and a lower degree of fluctuation and fluctuation of tacrolimus at steady-state compared to the period when treatment with Prograf® is stable. Patients with kidney transplants. In addition, the highest exposure of tacrolimus is similar after comparing different treatments; However, the mean concentration in the dose range was higher for LCP-Tacro.

EXAMPLE 20

Comparison of a formulation according to the invention (LCP-Tacro) against a commercially available prolonged-release single-dose prolol-release tacrolimus formulation, Advagraf® [0302] The main objective of this study is to determine and compare the rate and extent of tacrolimus absorption from a LCP-Tacro 2 mg tablet test preparation administered once daily (qd) versus the reference Advagraf® 2 x 1 mg capsule (qd) for Multiple administration, on an empty stomach.

[0303] Advagraf® is manufactured by Astellas Pharma GmbH in Munich, Germany. Summary study

<td>Experimental design</td><td>Randomized, bi-directional diagonal, open, multiple dose, on an empty stomach.</td>

<td>Population</td><td>Thirty healthy, healthy fingers of male race Caucasian.</td>

<td rowspan="2">Drug research</td><td>LCP-Tacro 2 mg tablets</td>

<td>Advagraf® 1 mg capsules</td>

<td rowspan="3">treatments</td><td>Patients will receive the following 1 treatments on days 1 to 10 each study period, in accordance with the randomization plan:</td>

<td>Treatment A: 1 LCP-Tacro 2 mg tablet (qw) (Daily dose treatment = 2 mg)</td>

<td>Treatment B: 2 Advagraf® 1 mg capsules (qd) (Daily dose treatment = 2 mg)</td>

<td>Duration treatment</td><td>This study consists of two sixteen-day periods (from a follow-up visit lasting 30 to 35 days after the last dose period II) separated by at least a double-period period rinsing from the last dose of the period I to the first dose period II. People will be institutionalized from the day before dosing on Day 1 to 24 hours after dose 10 days for each period of the study. Participants are required to return for the next fate of blood.</td>

EP 2 167 033 B1

<td>Experimental design</td><td>Randomized, bi-directional diagonal, open, multiple dose, on an empty stomach.</td>

<td></td><td>Pharmacokinetics (PK):</td>

<td rowspan="4">Blood collection</td><td>In each period, 50 blood samples are taken (4 ml each), according to the following schedule:</td>

<td>Day 1: 0.00 (before dosing), 0.50, 1.00, 1.50, 2.00, 3.00, 4.00, 5.00, 6.00, 7.00, 8.00, 9.00, 10.00, 12.00, 14.00, 16.00, 20.00 and 24.00 Dosage) after the dose.</td>

<td>Days 5, 6, 7, 8 and 9: 0.00 (before dosing) and 12.00 hours after dose administration.</td>

<td>Day 10: 0.00, 1.50, 2.00, 3.00, 4.00, 5.00, 6.00, 7.00, 8.00, 9.00, 10.00, 12.00, 14.00, 16.00, 20.00, 24.00, 48.00, 72.00, 96.00 and 120.00 hours after the dose.</td>

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

<td></td><td>During this test samples will be taken as follows way:</td>

<td rowspan="5">Blood collection (continued)</td><td>Initial tests: during the control visit (s)</td>

<td>Days 3 and 8: 0.00 hours (initial dose)</td>

<td>End of the test: After the last return of blood</td>

<td>A visit after a study visit after a study visit: between the day 30 a 35 after the last dose of period II.</td>

<td>4570.5 ml of blood will be collected from each patient.</td>

[0304] PK: PK parameters were calculated using extracellular tacrolimus analysis as follows: Day 1: AUC0-24, Cmax, C24 and Tmax.

Day 10: AUCt (t = 24), Cmax, Cmin, Cavg, Tmax, t%, Kel,% variation,% Swing, AUCt / Cmin, Cmax / Cmin and accumulation factor (R).

[0305] Statistics: Statistical data are described for blood concentrations and for all PK parameters.

The Geometric least squares (LSM) and 90% confidence (CI) coefficients (test / reference) were calculated for natural log parameters (Ln): AUC0-24, AUCt, Cmax, C24, Cmin, Cavg and unmodified% Fluctuation parameters,% variation , R, Cmax / Cmin, AUCt / Cmin, Kel and t1 / 2. Tmax were analyzed using nonparametric methods.

[0306] The following results showed a significant higher bioavailability of the product of the invention, while exhibiting significantly higher drug uptake by almost 50%, significantly lower concentration fluctuations during the dosing period, and high concentration. daily and any possible toxicity or side effect associated with periods of high concentration are eliminated to the possibly oral formulation once a day.

EP 2 167 033 B1

The present invention provides a formulation that at a dose of 2 mg orally administered in the morning provides an average concentration over 24 hours that exceeds 4 ng / ml (according to results below Cmin of 4.66 ng / ml), which is significantly higher Than introduced into once a day Advagraf® (Cmin 2.80 ng / ml) with the same administered dose.

The pharmacokinetic parameters of tacrolimus in healthy Caucasian men in the treatment of A, on day 1

<td>Table 20-1</td><td>Cmax (ng / ml)</td><td>AUC (0-24) (Ng.hr/mL)</td><td>C24 (ng / ml)</td><td>Tmax (Hrs.)</td>

<td>Mean</td><td>3.60</td><td>49.79</td><td>1.82</td><td>7.52</td>

<td>SD (±)</td><td>1.04</td><td>13.26</td><td>0.56</td><td>3.08</td>

<td>Median</td><td>3.57</td><td>46.63</td><td>1.69</td><td>7.00</td>

<td>CV (%)</td><td>28.79</td><td>26.63</td><td>30.65</td><td>40.88</td>

<td>Average geometric</td><td>3.46</td><td>48.19</td><td>1.73</td><td>6.99</td>

<td>Range (min)</td><td>2.07</td><td>27.56</td><td>0.66</td><td>3.00</td>

<td>(Max)</td><td>5.47</td><td>79.18</td><td>2.85</td><td>16.00</td>

<td>n =</td><td>21</td><td>21</td><td>21</td><td>21</td>

Treatment A: 1 LCP-Tacro 2 mg tablet (qw) [0307]

Pharmacokinetic parameters of tacrolimus in healthy Caucasian men in the treatment of B, on day 1

<td>Table 20-2</td><td>Cmax (ng / ml)</td><td>AUC (0-24) (Ng.hr/mL)</td><td>C24 (ng / ml)</td><td>Tmax (Hrs.)</td>

<td>Mean</td><td>3.44</td><td>34.00</td><td>0.97</td><td>2.19</td>

<td>SD (±)</td><td>0.99</td><td>9.41</td><td>0.37</td><td>0.77</td>

<td>Median</td><td>3.28</td><td>33.04</td><td>0.83</td><td>2.00</td>

<td>CV (%)</td><td>28.78</td><td>27.67</td><td>38.37</td><td>34.97</td>

<td>Average geometric</td><td>3.31</td><td>32.92</td><td>0.91</td><td>2.07</td>

<td>Range (min)</td><td>2.19</td><td>22.07</td><td>0.58</td><td>1.00</td>

<td>(Max)</td><td>5.48</td><td>61.56</td><td>2.00</td><td>4.00</td>

<td>n =</td><td>21</td><td>21</td><td>21</td><td>21</td>

Treatment e B: 2 Advagraf^® 1 Mg capsule (qd) [0308]

Pharmacokinetic parameters of tacrolimus in healthy Caucasian men in the treatment of A on day 10 68

EP 2 167 033 B1

<td>Table 20-3</td><td>AUCTauNg.hr/mL</td><td>Cmax ng / ml</td><td>C min ng / ml</td><td>Tmax (Hrs.)</td><td>C avg (ng / ml)</td><td>Kel (hr ^Λ (1))</td><td>T / (Hrs.)</td><td>Fluctuation (%)</td><td>Swing</td><td>R</td><td>Cmax / C min</td>

<td>Mean</td><td>142.27</td><td>8.39</td><td>4.66</td><td>7.03</td><td>5.93</td><td>1.87-02</td><td>37.97</td><td>64.72</td><td>85.45</td><td>2.85</td><td>1.85</td>

<td>SD (±)</td><td>49.41</td><td>2.89</td><td>1.71</td><td>2.96</td><td>2.06</td><td>2.76-03</td><td>5.83</td><td>22.97</td><td>37.62</td><td>0.64</td><td>0.38</td>

<td>CV (%)</td><td>34.73</td><td>34.51</td><td>36.60</td><td>42.11</td><td>34.73</td><td>1.84-02</td><td>37.62</td><td>35.48</td><td>44.03</td><td>22.42</td><td>20.29</td>

<td>Median</td><td>136.31</td><td>7.68</td><td>4.51</td><td>8.00</td><td>5.68</td><td>1.48 + 01</td><td>15.36</td><td>64.06</td><td>87.79</td><td>2.91</td><td>1.88</td>

<td>Average geometric</td><td>133.99</td><td>7.93</td><td>4.35</td><td>6.03</td><td>5.58</td><td>1.85-02</td><td>37.56</td><td>60.92</td><td>78.16</td><td>2.77</td><td>1.82</td>

<td>Range (min)</td><td>69.84</td><td>4.53</td><td>1.94</td><td>1.00</td><td>2.91</td><td>1.42-02</td><td>29.03</td><td>31.46</td><td>36.05</td><td>1.52</td><td>1.36</td>

<td>(Max.)</td><td>236.75</td><td>14.30</td><td>7.61</td><td>12.00</td><td>9.86</td><td>2.39-02</td><td>48.94</td><td>123.52</td><td>191.31</td><td>4.29</td><td>2.91</td>

<td>N =</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td>

Tau = m 24 hours

Treatment A: 1 LCP Tacro 2 mg tablet (q.) [0309]

Pharmacokinetic parameters of tacrolimus in healthy Caucasian males in the treatment of B, on day 10

<td>Table 20-4</td><td>AUCTauNg.hr/mL</td><td>Cmax ng / ml</td><td>C min ng / ml</td><td>Tmax (Hrs.)</td><td>C avg ng / ml</td><td>Kel (hr λ (1))</td><td>T / (Hrs.)</td><td>Fluctuation (%)</td><td>Swing</td><td>R</td><td>Cmax / C min</td>

<td>Mean</td><td>94.15</td><td>7.00</td><td>2.80</td><td>2.40</td><td>3.92</td><td>1.83-02</td><td>38.65</td><td>110.22</td><td>158.53</td><td>2.80</td><td>2.59</td>

<td>SD (±)</td><td>28.24</td><td>2.04</td><td>0.98</td><td>1.21</td><td>1.18</td><td>2.85-03</td><td>5.69</td><td>28.20</td><td>48.20</td><td>0.86</td><td>0.48</td>

<td>CV (%)</td><td>29.99</td><td>29.21</td><td>34.79</td><td>50.40</td><td>29.99</td><td>1.77-02</td><td>39.26</td><td>25.58</td><td>30.40</td><td>30.85</td><td>18.64</td>

<td>Median</td><td>89.82</td><td>6.97</td><td>2.52</td><td>2.00</td><td>3.74</td><td>1.55 + 01</td><td>14.71</td><td>116.55</td><td>169.20</td><td>2.59</td><td>2.69</td>

<td>Average geometric</td><td>89.86</td><td>6.71</td><td>2.64</td><td>2.16</td><td>3.74</td><td>1.81-02</td><td>38.24</td><td>106.46</td><td>150.98</td><td>2.70</td><td>2.54</td>

<td>Range (min)</td><td>49.14</td><td>4.32</td><td>1.39</td><td>1.0</td><td>2.05</td><td>1.40-02</td><td>28.10</td><td>54.97</td><td>74.41</td><td>1.82</td><td>1.74</td>

<td>(Max.)</td><td>138.85</td><td>10.50</td><td>4.46</td><td>6.00</td><td>5.79</td><td>2.47-02</td><td>49.39</td><td>161.17</td><td>238.80</td><td>5.56</td><td>3.39</td>

<td>n =</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td><td>20</td>

Treatment B: 2 Advagraf^® 1 Mg capsule (qd)

Tmax comparison of tacrolimus in healthy Caucasian males between 1 and 10 years in the treatment of A and B [0310]

Table 20-5:

EP 2 167 033 B1

<td>Treatment</td><td>Day 1 Median (range)</td><td>Day 10 Median (range)</td><td>Estimation, 90% CI *</td><td>P value **</td>

<td>AND</td><td>7.00 (3.00, 16.00)</td><td>8.00 (1.00, 12.00)</td><td>0.00 (-2.00, 2.00)</td><td>0901</td>

<td>B</td><td>2.00 (1.00, 4.00)</td><td>2.00 (1.00, 6.00)</td><td>0.00 (-0.50, 0.00)</td><td>0.776</td>

<td colspan="5">* Hodges-Lehmann scoring and 90% accurate CI for the difference in treatment. ** p-value for a treatment comparison based on the Wilcoxon-Mann-Whitney test.</td>

Treatment A: 1 LCP Tacro 2 mg tablet (q.)

Treatment B: 2 Advagraf^® 1 Mg capsule (qd) [0311]

Summary of statistics and comparison of pharmacokinetic parameters of tacrolimus in healthy Caucasian men, between treatment A and B, on day 1

<td>Table 20-6</td><td colspan="2">Treatment A</td><td colspan="2">Treatment B</td>

<td>Paramenter</td><td>n</td><td>Average ± SD</td><td>n</td><td>Average ± SD</td>

<td rowspan="2">AUC (0-24) (ng.hr/mL)</td><td>21</td><td>49.79 ± 13.26</td><td>21</td><td>34.00 ± 9.41</td>

<td></td><td>48.19 (26.63) *</td><td></td><td>32.92 (27.67) *</td>

<td rowspan="2">C24 (ng / ml)</td><td>21</td><td>1.82 ± 0.56</td><td>21</td><td>0.97 ± 0.37</td>

<td></td><td>1.73 (30.65) *</td><td></td><td>0.91 (38.37) *</td>

<td rowspan="2">Cmax (ng / ml)</td><td>21</td><td>3.60 ± 1.04</td><td>21</td><td>3.44 ± 0.99</td>

<td></td><td>3.46 (28.79) *</td><td></td><td>3.31 (28.78) *</td>

<td>Tmax (hours)</td><td>21</td><td>7.00 (3.00, 16.00) **</td><td>21</td><td>2.00 (1.00, 4.00) **</td>

<td colspan="5">* Geometric average (% CV) ** Median (range)</td>

Treatment A: 1 LCP Tacro 2 mg tablet (q.)

Treatment B: 2 Advagraf^® 1 Mg capsule (qd) [0312]

Summary of statistics and comparison of pharmacokinetic parameters of tacrolimus in healthy Caucasian men, between treatment A and B, on day 1

<td>Table 20-7</td><td>AUC (0-24)</td><td>C24</td><td>Cmax</td>

<td>90% Geometric</td><td>1330.95% to</td><td>1610.80% to</td><td>93From 92 to</td>

<td>CI1</td><td>163.57%</td><td>222.46%</td><td>118,70%</td>

<td>The ratio of funds2</td><td>148.02%</td><td>189.72%</td><td>105.59%</td>

<td>Integrative CV3</td><td>18.69%</td><td>30.18%</td><td>21.97%</td>

EP 2 167 033 B1

<td>Table 20-7</td><td>AUC (0-24)</td><td>C24</td><td>Cmax</td>

<td>Degree of freedom</td><td>19</td><td>19</td><td>19</td>

<td colspan="4">1. 90% Geometric confidence interval using data transformed with logs 2. Calculated using geometrical agents of the formula: e ((test drug) - (drug reference)) X100% 3. Intense variance coefficient of variability of pharmacokinetic parameters logically transformed</td>

Test: treatment A: 1 tablet LCP Tacro 2 mg (qw)

Reference: Treatment B: 2 Advagraf^® 1 Mg capsule (qd) [0313]

Statistical summary and comparison of pharmacokinetic parameters of tacrolimus in healthy Caucasian men, between treatment A and B, on day 10

<td>Table 20-8</td><td colspan="2">Treatment A</td><td colspan="2">Treatment B</td>

<td>Parameter</td><td>n</td><td>Average ± SD</td><td>n</td><td>Average ± SD</td>

<td>AUCTau ^Λ</td><td rowspan="2">20</td><td>142.27 ± 49.41</td><td rowspan="2">20</td><td>94.15 ± 28.24</td>

<td>(Ng. Hr / mL</td><td>133.99 (34.73) *</td><td>89.86 (29.99) *</td>

<td>Cmax</td><td rowspan="2">20</td><td>8.39 ± 2.89</td><td rowspan="2">20</td><td>7.00 ± 2.04</td>

<td>(Ng / ml)</td><td>7.93 (34.51) *</td><td>6.71 (29.21) *</td>

<td>C min</td><td rowspan="2">20</td><td>4.66 ± 1.71</td><td rowspan="2">20</td><td>2.80 ± 0.98</td>

<td>(Ng / ml)</td><td>4.35 (36.60) *</td><td>2.64 (34.79) *</td>

<td>Tmax</td><td rowspan="2">20</td><td>7.03 ± 2.96</td><td rowspan="2">20</td><td>2.40 ± 1.21</td>

<td>(Hr)</td><td>6.03 (42.11) *</td><td>2.16 (50.40) *</td>

<td>C avg</td><td rowspan="2">20</td><td>5.93 ± 2.06</td><td rowspan="2">20</td><td>3.92 ± 1.18</td>

<td>(Ng / ml)</td><td>5.58 (34.73) *</td><td>3.74 (29.99) *</td>

<td>The degree of fluctuation</td><td rowspan="2">20</td><td>64.72 ± 22.97</td><td rowspan="2">20</td><td>110.22 ± 28.20</td>

<td>(%)</td><td>60.92 (35.48) *</td><td>106.46 (25.58) *</td>

<td>Swing degree (%)</td><td rowspan="2">20</td><td>85.45 ± 37.62</td><td rowspan="2">20</td><td>158.53 ± 48.20</td>

<td></td><td>78.16 (44.03) *</td><td>150.98 (30.40) *</td>

<td>R</td><td rowspan="2">20</td><td>2.85 ± 0.64</td><td rowspan="2">20</td><td>2.80 ± 0.86</td>

<td></td><td>2.77 (22.42) *</td><td>2.70 (30.85) *</td>

<td>Cmax / Cmin</td><td rowspan="2">20</td><td>1.85 ± 0.38</td><td rowspan="2">20</td><td>2.59 ± 0.48</td>

<td></td><td>1.82 (20.29) *</td><td>2.54 (18.64) *</td>

EP 2 167 033 B1

<td>Table 20-8</td><td colspan="2">Treatment A</td><td colspan="2">Treatment B</td>

<td>Parameter</td><td>n</td><td>Average ± SD</td><td>n</td><td>Average ± SD</td>

<td>T /</td><td rowspan="2">20</td><td>37.97 ± 5.83</td><td rowspan="2">20</td><td>38.65 ± 5.69</td>

<td>(Hr)</td><td></td><td></td>

<td>kel</td><td rowspan="2">20</td><td>1.87E-02 ± 2.76E-</td><td rowspan="2">20</td><td>1.83E-02 ± 2.85E-</td>

<td>(hr^AND(-1))</td><td>03</td><td>03</td>

<td colspan="5">* Geometric average (% CV)</td>

Treatment A: 1 LCP Tacro 2 mg tablet (q.)

Treatment B: 2 Advagraf^® 1 Mg capsule (qd) [0314]

Statistical summary and comparison of pharmacokinetic parameters of tacrolimus in healthy Caucasian men, between treatment A and B, on day 10

<td>Table 20-9</td><td>AUCtau λ</td><td>Cmax</td><td>C min</td>

<td>90% Geometric CI *</td><td>1370.43% to 161.12%</td><td>105from 0.33% to 127.73%</td><td>1520,29% to 180.25%</td>

<td>The ratio of funds **</td><td>148,80%</td><td>115.99%</td><td>165.68%</td>

<td>CV internal CV</td><td>13.98%</td><td>16.99%</td><td>14.83%</td>

<td>Degree of freedom</td><td>17</td><td>17</td><td>17</td>

<td colspan="4">^Λ Tau = 24 hours * 90% geometric confidence interval using data transformed with logs ** Calculated using geometric measures in accordance with the formula: e ((tested lek) - (reference medicine)) X100% *** Coefficient of variation for the transformed pharmacokinetic parameter logically.</td>

Treatment A: 1 LCP Tacro 2 mg tablet (q.) Treatment B: 2 Advagraf® 1 mg capsules (qd) [0315]

<td>The sustained release composition has stabilized</td><td>LCP-Tacro 2 mg</td>

<td>Tacrolimus monohydrate (2.00 mg on the anhydrous basis weight)</td><td>20.400 mg</td>

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

EP 2 167 033 B1

<td>The sustained release composition has stabilized</td><td>LCP-Tacro 2 mg</td>

<td>Butylated hydroxytoluene</td><td>10200 μg</td>

<td>Dimetikon 350</td><td>0.25500 μg</td>

<td>Hypromellose 2208 (15,000 cps)</td><td>62866 mg</td>

<td>Lactose monohydrate</td><td>41727 mg</td>

<td>Magnesium stearate</td><td>15757 mg</td>

<td>Opadry II white 85G18490</td><td>4.7232 mg</td>

<td>Poloxamer 188</td><td>14688 mg</td>

<td>Polyethylene glycol 6000</td><td>340.2272 mg</td>

<td>Tartaric acid</td><td>2550.00 μg</td>

EXAMPLE 21 [0316] A table disclosing the dissolution of a preferred embodiment of the invention having the composition set forth above in Example 20 and dissolving the commercial Advagraf® product used for comparison in Example 20.

[0317] Dose as a percentage, dissolution method USP II dissolution test method (agitator) in media adjusted to pH 4.5 and containing 0.005% hydroxylpropyl cellulose and a rotation of 50 rpm. The dissolution is disclosed in Figure 4

<td>Extended dissolution preparation</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Lot number</td><td colspan="2">0988/2006</td><td colspan="2">0989/2006</td><td colspan="2">0990/2006</td>

<td>Time, hours</td><td>1 Mg</td><td></td><td>2 Mg</td><td></td><td>5 Mg</td><td></td>

<td></td><td>analysis</td><td>RSD</td><td>analysis</td><td>RSD</td><td>Analysis</td><td>RSD</td>

<td>1</td><td>1.1</td><td>31.5</td><td>5</td><td>12.9</td><td>6.8</td><td>14.2</td>

<td>2</td><td>7.2</td><td>7.9</td><td>7.9</td><td>10.6</td><td>10.1</td><td>14.5</td>

<td>3</td><td>10.1</td><td>15.4</td><td>10.8</td><td>12.3</td><td>13.4</td><td>18.2</td>

<td>5</td><td>18.1</td><td>10.6</td><td>15.4</td><td>23.5</td><td>20.4</td><td>19.5</td>

<td>9</td><td>30.4</td><td>13.2</td><td>33.8</td><td>33.5</td><td>35.6</td><td>16.1</td>

<td>12</td><td>45.3</td><td>9.9</td><td>48</td><td>18.4</td><td>43.3</td><td>13.2</td>

<td>15</td><td>61.4</td><td>9.1</td><td>59.6</td><td>10.2</td><td>45.3</td><td>10</td>

<td>20</td><td>74</td><td>2.6</td><td>68.4</td><td>5.8</td><td>47</td><td>9.3</td>

<td>24</td><td>80.9</td><td>3.5</td><td>72.2</td><td>3.7</td><td>47.5</td><td>7.6</td>

<td>Formulation of Advagraf</td><td colspan="2"></td><td></td><td colspan="3"></td>

EP 2 167 033 B1

<td>Lot number</td><td>OM4001A</td><td></td><td>1M6002A</td><td></td><td>5M4002A</td><td></td>

<td>Time, hours</td><td>00.5 mg</td><td></td><td>1 Mg</td><td></td><td>5 Mg</td><td></td>

<td></td><td>analysis</td><td>RSD</td><td>analysis</td><td>RSD</td><td>analysis</td><td>RSD</td>

<td>1</td><td>15.9</td><td>4.6</td><td>21.2</td><td>24.5</td><td>13.6</td><td>19.1</td>

<td>2</td><td>27.2</td><td>11.3</td><td>30.9</td><td>29.4</td><td>29.2</td><td>9.6</td>

<td>3</td><td>37.3</td><td>15</td><td>39.2</td><td>24.9</td><td>36.2</td><td>7</td>

<td>5</td><td>48.5</td><td>13</td><td>45.8</td><td>21.5</td><td>46</td><td>7.4</td>

<td>9</td><td>61.2</td><td>7.3</td><td>58</td><td>16.8</td><td>57.1</td><td>7.1</td>

<td>12</td><td>65</td><td>9.3</td><td>63</td><td>15.7</td><td>60.2</td><td>4.4</td>

<td>15</td><td>70.4</td><td>9.9</td><td>66.6</td><td>14.7</td><td>63.6</td><td>7.8</td>

<td>20</td><td>71.3</td><td>9.7</td><td>69.3</td><td>13.4</td><td>67</td><td>6.4</td>

<td>24</td><td>74.1</td><td>7.4</td><td>68.1</td><td>15.3</td><td>69.3</td><td>6.6</td>

[0318] As apparent from the dissolution, the sustained release preparation of the present invention provides a much longer and more expanded profile with much less release initially, e.g. demonstrated by less than 25% release at the 5-hour time point, despite being released in 3 hours every at least 10%. In addition, the curvature profile of the present invention has a significant zero tearing and very long release. The latter is clearly demonstrated by less than 50% release at the 12-hour time point and less than 62% release at the 15-hour time point. As is apparent from EXAMPLE 20, the pharmacokinetic parameters are significantly improved with the sustained release formulation of the invention compared to the commercially available Advagraf® product.