Macrolides for use in methods for the prevention or treatment of immune diseases or disorders

Abstract

The present invention relates to a method for preventing or treating an immune disease or disorder, preferably transplant rejection, comprising administering the macrolide at least twice daily, wherein the administration at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, the second time point being a time point within a range from about 1 to about 7 hours, preferably from about 2 to about 5 hours, more preferably from about 2.5 to about 3.5 hours, and even more preferably about 3 hours, from the first time point. The present invention also relates to a tacrolimus mucoadhesive film suitable for buccal administration.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a macrolide for use in a method for the prevention or treatment of an immune disease or disorder, preferably transplant rejection, comprising administering said macrolide at least twice daily, wherein said administering at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, said second time point being a time point within a range of from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, and even more preferably about 3 hours, from said first time point. [Background technology]

[0002] Background of the Invention Poor water solubility of active agents is a challenge in the development of pharmaceutical formulations. In fact, most active drug ingredients exhibit low water solubility. To elicit a therapeutic effect, the active agent must dissolve and reach the bloodstream in sufficient concentrations. Therefore, active agents with low water solubility, such as macrolides, typically have poor bioavailability, i.e., only a small percentage of an administered active agent reaches the bloodstream after ingestion. Low drug bioavailability may be related to incomplete absorption and / or incomplete metabolism in the gastrointestinal tract. Due to poor water solubility, the absorption profiles of oral macrolide formulations, such as tacrolimus formulations, vary widely; for example, the absolute bioavailability of oral tacrolimus formulations ranges from 5 to 93%. Macrolides, such as tacrolimus, undergo extensive presystemic metabolism in the GI tract, limiting their oral bioavailability; for example, tacrolimus averages approximately 25%.

[0003] Tacrolimus is a macrolide lactone with potent immunosuppressive properties, inhibiting both cellular and humoral immune responses through multiple mechanisms of action, including the central effect of calcineurin inhibition. Tacrolimus is approved as a first-line immunosuppressant for the prevention of graft rejection in patients with liver, kidney, or heart allografts and for the treatment of allograft rejection refractory to other immunosuppressive medications. Since the initial approval of Prograf in 1994, oral tacrolimus has become the cornerstone of immunosuppression in adults and children undergoing solid organ transplantation (SOT). Approved oral dosage forms of tacrolimus include immediate-release capsules (e.g., Prograf® hard capsules), extended-release capsules (e.g., Advagraf® extended-release hard capsules), extended-release tablets (e.g., Envarsus® extended-release tablets), and extended-release granules (e.g., Modigraf® granules for oral suspension). Oral tacrolimus is currently considered the world's standard of care for preventing rejection after solid organ transplantation. Tacrolimus therapy is a medication that is administered daily for life.

[0004] Macrolides such as tacrolimus, which have a narrow therapeutic index, exhibit poor oral bioavailability due to their poor solubility and, furthermore, high inter- and intra-individual variability in pharmacokinetic parameters. Therefore, the dose administered to patients must be carefully monitored. Due to the patient-specific dosing (mg / kg body weight / day) of the macrolide tacrolimus, available capsule formulations are often opened, the active ingredient is made into a solution, and then administered to the patient; such a process is called "compounding." Compounding does not meet the high quality requirements of pharmaceutical products and may even jeopardize drug safety by increasing the risk of incorrect dosing. Furthermore, compounding may result in decreased drug stability or microbial contamination of the prepared solution.

[0005] Thus, there remains a need for enhanced macrolide therapy, particularly for macrolides for use with increased efficacy and / or improved bioavailability. In particular, there remains a need for treatments that allow for efficient administration of sufficient doses of macrolides to patients. There also remains a need for dosing regimens and / or formulations that allow for effective delivery of macrolides to patients. In particular, there remains a need for enhanced treatments that allow for sufficient blood concentrations of macrolides to be achieved. There also remains a need for reduced macrolide bioavailability, particularly the variability of macrolide blood concentrations. Furthermore, there remains a need for improved bioavailability of macrolides, such as tacrolimus. Furthermore, there remains a need for enhanced means, such as dosing regimens for administering macrolides, e.g., therapeutic systems that allow for efficient administration of active agents, such as macrolides. There also remains a need for reducing the dose of macrolide required to achieve a therapeutic effect. Furthermore, there remains a need for efficient methods for preventing or treating immune diseases or disorders, such as transplant rejection. Summary of the Invention

[0006] Summary of the Invention Below, the components of the present invention are described. These components are listed with specific embodiments, but it should be understood that they can be combined in any way and in any number to create further embodiments. The various described examples and preferred embodiments should not be construed as limiting the invention to only the specifically described embodiments. This description should be understood to support and encompass embodiments that combine two or more specifically described embodiments, or combine one or more specifically described embodiments with any number of disclosed and / or preferred components. Furthermore, any variations and combinations of all components described in this application should be considered to be disclosed by the description in this application, unless the context dictates otherwise.

[0007] In a first aspect, the present invention relates to a macrolide for use in a method for the prevention or treatment of an immune disease or disorder, preferably transplant rejection, comprising administering said macrolide at least twice daily, wherein said administering at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, said second time point being a time point ranging from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, even more preferably about 3 hours, from said first time point; Preferably, the macrolide is administered by buccal administration.

[0008] In one embodiment, the macrolide is administered in the form of a mucoadhesive layer, preferably a mucoadhesive buccal layer; Preferably, said administering comprises applying said mucoadhesive layer to a body cavity of the patient, preferably the buccal cavity of the patient and / or to a mucosa of the patient.

[0009] The present invention also relates to a method for preventing or treating an immune disease or disorder, preferably transplant rejection, comprising administering a macrolide at least twice daily, The administering at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, the second time point being a time point ranging from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, even more preferably about 3 hours, from the first time point; preferably, the macrolide is administered by buccal administration.

[0010] In one embodiment, said administering comprises administering each of said first dose and said second dose, preferably each in the form of a mucoadhesive layer, for a period ranging from about 1 minute to about 7 hours, preferably from about 10 minutes to about 3 hours, more preferably from about 20 minutes to about 2 hours, even more preferably from about 30 minutes to about 90 minutes, and even more preferably about 60 minutes.

[0011] In one embodiment, the administering comprises administering the first dose over a period ranging from about 30 minutes to about 90 minutes, preferably about 60 minutes; and administering the second dose over a period ranging from about 30 minutes to about 90 minutes, preferably about 60 minutes, at a time ranging from about 2.5 hours to about 3.5 hours, preferably about 3 hours, from the first time point.

[0012] In one embodiment, said first dose and said second dose, preferably prior to said administering, each comprise said macrolide in an amount in the range of from about 0.001 mg to about 250 mg, preferably in the range of from about 0.01 mg to about 50 mg, more preferably in the range of from about 0.05 mg to about 25 mg, even more preferably in the range of from about 0.1 mg to about 15 mg, even more preferably in the range of from about 0.5 mg to about 13 mg, optionally in an amount in the range of from about 8 mg to about 12 mg; and / or Administering the macrolide by buccal administration and administering the macrolide at least twice daily comprises administering a total daily amount of the macrolide that is ≦about 40%, preferably ≦about 30%, more preferably ≦about 25%, and even more preferably ≦about 20% of the total daily amount administered by oral administration.

[0013] In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of 1 cm 2 or more of the mucoadhesive layer prior to administration. 2 and preferably in an amount ranging from about 0.01 mg to about 10 mg per cm of said mucoadhesive layer. 2 and more preferably in the range of about 0.1 mg to about 5 mg per cm of said mucoadhesive layer. 2 and even more preferably in the range of about 1.5 mg to about 3.5 mg per cm of said mucoadhesive layer. 2 The amount of hydroxybenzoates is present in an amount ranging from about 1.8 mg to about 2.5 mg based on the total amount of hydroxybenzoates.

[0014] In one embodiment, - mucoadhesive polymers, preferably selected from a) amphiphilic and hydrophilic polymers, b) poly(methacrylates), c) crosslinked polyacrylic acid polymers, d) copolymers of methyl vinyl ether and maleic anhydride, e) polymers containing polyvinyl acetate and / or polyvinylpyrrolidone, and combinations thereof; administering the macrolide in the form of a mucoadhesive layer comprising Here, preferably, the amphiphilic polymer is selected from polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer, polyoxyl castor oil and D-α-tocopherol-polyethylene glycol-succinate (TPGS), and / or The poly(methacrylate) is selected from neutral protonatable poly(methacrylates) and cationic poly(methacrylates), preferably copolymers of dialkylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trialkylammonioethyl methacrylate and methacrylic acid esters, in particular copolymers of dimethylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trimethylammonioethyl methacrylate and methacrylic acid esters; and wherein, optionally, the mucoadhesive layer comprises: - cellulose derivatives, preferably selected from hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), hydroxypropyl methyl cellulose (HPMC), hydroxyethyl cellulose (HEC), methyl cellulose (MC), hydroxyethyl methyl cellulose (HEMC), ethyl cellulose (EC), and combinations thereof; a plasticizer, preferably selected from glycerol, polyethylene glycol, low molecular weight polyethylene glycol such as PEG 200 or PEG 400, propylene glycol, sorbitol, triacetin, and tributyl citrate; more preferably glycerol; and / or - colorant, preferably TiO2; further comprising Where appropriate, said macrolide is present in said layer as a micronised macrolide, preferably a micronised crystalline macrolide, and / or in molecularly dissolved non-nanoparticulate form.

[0015] In one embodiment, the at least twice-daily administration provides a first peak whole blood concentration of the macrolide at a time within a range from about 2 hours to about 4 hours, preferably from about 2.5 hours to about 3.5 hours, more preferably at about 3 hours, from the first time point, and a second peak whole blood concentration of the macrolide at a time within a range from about 7 hours to about 9 hours, preferably from about 7.5 hours to about 8.5 hours, more preferably at about 8 hours, from the first time point; wherein, optionally, the at least twice-daily administration provides the second peak in whole blood concentration of the macrolide at a time within a range of about 3 hours to about 5 hours, preferably about 3.5 hours to about 4.5 hours, and more preferably about 4 hours, from the second time point.

[0016] In one embodiment, the at least twice daily administration provides a whole blood concentration of the macrolide in the range of about 0.01 ng / mL to about 50 ng / mL, preferably in the range of about 3 ng / mL to about 20 ng / mL, and more preferably in the range of about 5 ng / mL to about 20 ng / mL for a period of about 24 hours.

[0017] In one embodiment, said at least twice daily administration provides a coefficient of variation of less than 40%, preferably less than 35%, more preferably less than 32%, even more preferably less than or equal to 30%, such as in the range of 20% to 30% or in the range of 25% to 30%.

[0018] In one embodiment, the administering at least twice daily - within a range of about 3 hours to about 10 hours from said first time point, preferably within a range of about 7 hours to about 9 hours from said first time point, more preferably within a range of about 7.5 hours to about 8.5 hours from said first time point. maxand / or - within a range of about 1 hour to about 6 hours from said second time point, preferably within a range of about 2 hours to about 5 hours from said second time point, more preferably within a range of about 3.5 hours to about 4.5 hours from said second time point, for example within about 4 hours from said second time point max to provide.

[0019] In one embodiment, the administering at least twice daily - a C in the range of about 0.05 ng / mL to about 50 ng / mL, preferably in the range of about 0.1 ng / mL to about 30 ng / mL, more preferably in the range of about 0.7 ng / mL to about 20 ng / mL, even more preferably in the range of about 5.5 ng / mL to about 20 ng / mL max and / or - a C in the range of about 0.01 ng / mL to about 18 ng / mL, preferably in the range of about 0.5 ng / mL to about 16 ng / mL, more preferably in the range of about 3 ng / mL to about 15 ng / mL, such as in the range of about 3 ng / mL to about 7 ng / mL or in the range of about 5 ng / mL to about 7 ng / mL trough to provide.

[0020] In one embodiment, said administering at least twice daily is performed for a period of at least 2 days, preferably at least 1 week, more preferably at least 1 month, and even more preferably at least 1 year; Where necessary, when the macrolide is used for the prevention or treatment of transplant rejection, the administration at least twice daily is carried out for the lifetime of the patient.

[0021] In one embodiment, the macrolide is administered to a human patient, and preferably, the patient is not a fasting patient.

[0022] In one embodiment, the macrolide is selected from a macrolide immunosuppressant, a macrolide antibiotic, and a macrolide antifungal; Preferably, - the macrolide immunosuppressant is selected from tacrolimus, sirolimus, everolimus, and pimecrolimus; - the macrolide antibiotic is selected from erythromycin, clarithromycin, azithromycin, roxithromycin, josamycin, spiramycin, telithromycin, tylosin and fidaxomicin; and / or - the macrolide antifungal agent is selected from polyenes, in particular nystatin, natamycin and amphotericin B; Here, more preferably, the macrolide is tacrolimus or sirolimus, and even more preferably, it is tacrolimus.

[0023] In one embodiment, the mucoadhesive layer is provided in the form of a mucoadhesive film comprising said mucoadhesive layer, preferably in the form of a mucoadhesive buccal film comprising said mucoadhesive layer.

[0024] In a further aspect, the present invention relates to a method for preventing and / or treating an immune disease or disorder, preferably transplant rejection, said method comprising administering a macrolide, particularly a therapeutically effective amount of a macrolide, to a patient in need thereof at least twice daily, wherein said administering at least twice daily comprises administering a first dose at a first time point and administering a second dose at a second time point, said second time point being at a time point within a range from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, and even more preferably about 3 hours, from said first time point; Preferably, the macrolide is administered bucally.

[0025] In one embodiment, said macrolide is administered in the form of a mucoadhesive layer, preferably in the form of a mucoadhesive buccal layer; preferably said administering comprises attaching said mucoadhesive layer to a body cavity of the patient, preferably to the buccal cavity of the patient and / or to a mucosa of the patient.

[0026] In one embodiment, said administration, said first dose, said second dose, said macrolide, said mucoadhesive layer, and said patient are as defined herein.

[0027] In a further aspect, the present invention relates to the use of a macrolide for the manufacture of a medicament for the prevention or treatment of an immune disease or disorder, preferably transplant rejection, said use comprising administering said macrolide at least twice a day, wherein said administering at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, said second time point being at a time point within a range of from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, and even more preferably about 3 hours, from said first time point; Preferably, the macrolide is administered by buccal administration.

[0028] In one embodiment, the macrolide is administered in the form of a mucoadhesive layer, preferably in the form of a mucoadhesive buccal layer; preferably, said administering comprises attaching said mucoadhesive layer to a body cavity of the patient, preferably to the buccal cavity of the patient and / or to a mucosa of the patient.

[0029] In one embodiment, said administration, said first dose, said second dose, said macrolide, said mucoadhesive layer and said patient are as defined herein.

[0030] Herein, the composition of a film or layer may be exemplarily expressed by reference to the composition and by indicating the presence of individual components therein, as well as by reference to their respective amounts or percentages (e.g., weight percentages) in the film or layer. Such amounts or percentages are often given as ranges where possible. In this context, it should be noted that the actual and specific values ​​of the individual components should be selected from their respective ranges in each case so that the sum of each component is exactly or less than 100%, but does not exceed 100%. That is, for example, components A, B, C, and D, the maximum and minimum values ​​of each component, A, B, C, and D, are given. min From A max , B min From B max , C min From C max , and D min From D max Even when ranges are given, the individual values ​​for each component must be selected from their respective ranges so that the sum of the individual components is either exactly 100% or less than 100%. If the sum of the individual values ​​does not add up to 100% but is <100%, then additional components such as one or more solvents, fillers, extenders, etc. must be present and these must be added in the appropriate amount, i.e., the amount needed to make up 100%.

[0031] The present invention also relates to a tacrolimus mucoadhesive film suitable for buccal administration, comprising a mucoadhesive layer and a backing layer: A) The mucoadhesive layer is: about 12 to about 25 weight percent tacrolimus; about 27 to about 31 weight percent ethylcellulose; about 28 to about 36 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 6 to about 10 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 5 weight percent calcium / sodium copolymer of methyl vinyl ether and maleic anhydride; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; or B) The mucoadhesive layer is: about 18 to about 25 weight percent tacrolimus; about 3 to about 13 weight percent ethylcellulose; about 29 to about 42 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 1 to about 28 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 10 weight percent crosslinked poly(acrylic acid); Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0032] In one embodiment of the tacrolimus mucoadhesive film, particularly in accordance with the preceding embodiments, the backing layer comprises ethylcellulose.

[0033] In one embodiment of the tacrolimus mucoadhesive film, particularly in accordance with the preceding embodiments, the backing layer comprises ethyl cellulose in the range of from about 50 to about 62 weight percent, based on the dry weight of the backing layer.

[0034] In another embodiment of the tacrolimus mucoadhesive film, the backing layer comprises ethyl cellulose in the range of from about 25 to about 27 weight percent, based on the dry weight of the backing layer.

[0035] In one embodiment of the tacrolimus mucoadhesive film, particularly according to any one of the preceding embodiments, the film comprises from 8 mg to 17 mg of tacrolimus.

[0036] In one embodiment of the tacrolimus mucoadhesive film, particularly according to any one of the preceding embodiments, the film tacrolimus film comprises 8.8 mg or 12 mg or 14.4 mg or 16.5 mg of tacrolimus.

[0037] In one embodiment of the tacrolimus mucoadhesive film, particularly according to any one of the preceding embodiments, such film further comprises an adhesive layer disposed between the mucoadhesive layer and the backing layer.

[0038] In another embodiment of the tacrolimus mucoadhesive film, the mucoadhesive layer and the backing layer are in direct contact with each other, and in this tacrolimus mucoadhesive embodiment, there is no layer disposed between the mucoadhesive layer and the backing layer.

[0039] In a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 12 to about 13 weight percent tacrolimus, or about 19 to about 23 weight percent tacrolimus; about 27 to about 30 weight percent ethylcellulose; about 30 to about 35 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 7 to about 8 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 3 to about 4 weight percent of a calcium / sodium copolymer of methyl vinyl ether and maleic anhydride; Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0040] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 19 to about 22 weight percent tacrolimus; about 3 to about 13 weight percent ethylcellulose; about 29 to about 42 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 1 to about 28 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 10 weight percent crosslinked poly(acrylic acid); Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0041] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 5 to about 14 weight percent tacrolimus; about 29 to about 33 weight percent ethylcellulose; about 34 to about 41 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 7 to about 10 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 4 weight percent of a copolymer of methyl vinyl ether and maleic anhydride; Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0042] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 18 to about 20 weight percent tacrolimus; about 11 to about 13 weight percent ethylcellulose; about 30 to about 32 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 11 to about 13 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; Such mucoadhesive layer herein does not comprise any polymer selected from the following mucoadhesive polymers: copolymers of methyl vinyl ether and maleic anhydride; poly(acrylic acid); crosslinked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; polysaccharides, in particular alginate, chitosan, xanthan gum, hyaluronic acid, pectin or pullulan; carboxymethylcellulose; hydroxypropylmethylcellulose; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

[0043] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 9 to about 23 weight percent tacrolimus; about 10 to about 13 weight percent ethylcellulose; about 15 to about 32 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 5 to about 12 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 6 to about 28 weight percent poly(methacrylate); Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0044] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 18 to about 20 weight percent tacrolimus; about 4 to about 6 weight percent ethylcellulose; about 31 to about 32 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 27 to about 28 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; wherein such mucoadhesive layer does not comprise any polymer selected from the following mucoadhesive polymers: copolymers of methyl vinyl ether and maleic anhydride; poly(acrylic acid); crosslinked poly(acrylic acid; poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; polysaccharides, in particular alginate, chitosan, xanthan gum, hyaluronic acid, pectin or pullulan; carboxymethylcellulose; hydroxypropylmethylcellulose; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

[0045] In yet a further aspect, the present invention relates to a tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer: about 18 to about 20 weight percent tacrolimus; about 4 to about 12 weight percent ethylcellulose; about 31 to about 42 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000-Klucel EF] having a viscosity of about 300-600 mPa.s; about 2 to about 23 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000-Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 6 weight percent crosslinked poly(acrylic acid); Here, the weight percentages are based on the dry weight of the mucoadhesive layer.

[0046] In yet a further aspect, the present invention relates to a method of treating a human patient who has received an allogeneic solid organ transplant, the patient requiring prevention of graft rejection, comprising: The method comprises buccal administration to a patient twice daily of a mucoadhesive film described in any of the embodiments and aspects described herein, preferably a mucoadhesive film described in any of the preceding embodiments.

[0047] In one embodiment of the method for treating a human patient who has undergone an allogeneic solid organ transplant, the buccal administration involves administering a first mucoadhesive film described herein for about 30 to 60 minutes, preferably 60 minutes, or equally preferably 30 minutes, followed by about 3-5 hours without administration, followed by buccal administration of a second mucoadhesive film described herein for about 30-60 minutes, preferably 60 minutes, or equally preferably 30 minutes, to the patient.

[0048] In one embodiment of the method for treating a human patient who has undergone an allogeneic solid organ transplant, the human patient's C trough The blood level of tacrolimus is C max This is about 30-40% of the original value. [Brief explanation of the drawings]

[0049] BRIEF DESCRIPTION OF THE DRAWINGS The invention will now be further described by reference to the following drawings. All methods described in the following figure descriptions were performed as detailed in the Examples. [Figure 1] Figure 1 shows the arithmetic mean time profiles of tacrolimus whole blood concentrations (ng / mL; PK Set, N=10). Treatment group 1 (T1): two doses of 8.8 mg tacrolimus MBF; Treatment group 2 (T2): two doses of 12.0 mg tacrolimus MBF. [Figure 2] FIG. 2 shows individual tacrolimus blood levels in 10 healthy volunteers (5 per group), administered bid, 4 hours after the second dose. [Figure 3] FIG. 3 shows the mean blood level profile in 10 healthy volunteers (5 per group), administered bid, 4 hours after the second dose. [Figure 4] Figure 4 shows the individual PK profiles in different groups (5 minipigs per group) using an asymmetric dosing regimen for buccal administration where the second dose was administered 3, 4 or 5 hours after the first dose. [Figure 5]Figure 5 shows a plot of the mean curves. The highest blood concentrations are obtained when the second dosage unit is administered 3 hours after the first dose. [Figure 6] Figure 6 shows the residual drug levels in used films for the low-dose group (8.8 mg tacrolimus loaded = 17.6 mg bid) and the high-dose group (12 mg tacrolimus loaded = 24 mg bid). Each subject was treated with a first dose in the form of a first mucoadhesive film at a first time point and a second dose in the form of a second mucoadhesive film at a second time point approximately 4 hours after the first time point. Each film was removed after a period of approximately 1 hour. The patterned bar indicates the amount of macrolide remaining in the first mucoadhesive film after film removal. The solid bar indicates the amount of macrolide remaining in the second mucoadhesive film after film removal. [Figure 7] FIG. 7 shows the normalized concentration-time curves (tacrolimus blood concentration at each time point as a percentage of maximum concentration) from human studies. [Figure 8] Figure 8 shows the relatively small degree of inter-individual blood level variability (CV) of tacrolimus in both groups over a 24-hour interval for buccal application. Advantageously, buccal administration reduces blood level variability compared to oral application. DETAILED DESCRIPTION OF THE INVENTION

[0050] Detailed Description The present invention aims to provide enhanced administration of macrolides for the prevention and / or treatment of transplant rejection, for example, after organ transplantation, particularly solid organ transplantation. Furthermore, the present invention aims to provide efficient macrolide administration. It is also an object of the present invention to provide macrolides for use in therapies that allow for efficient achievement of therapeutic macrolide blood levels. It is also an object of the present invention to provide dosing regimens that allow for early onset of therapeutic benefit. It is also an object of the present invention to reduce side effects. It is also an object of the present invention to reduce the dosage required to achieve desired macrolide blood levels, such as blood levels within the therapeutic range. The present invention also aims to prevent and treat immune diseases and conditions, such as solid organ transplant rejection. For example, macrolides for use in the present invention can be administered in the form of a mucoadhesive film, e.g., a mucoadhesive, unidirectional, non-dissolving buccal film. It is also an object of the present invention to provide efficient therapies, e.g., therapies that are more effective than standard oral macrolide therapies, such as tacrolimus standard therapy, e.g., standard therapies such as capsules, tablets, or granules. Although macrolides such as tacrolimus typically have very low water solubility, macrolides for use in the present invention, e.g., administered in the form of a layer or film, efficiently achieve therapeutic macrolide blood concentrations. Unexpectedly, clinically relevant blood levels of macrolide are achieved with the macrolides for use in the present invention.

[0051] The term "macrolide," as used herein, refers to a class of compounds of natural products that contain or consist of a large macrocyclic lactone ring, e.g., a 14-, 15-, or 16-membered lactone ring, to which one or more deoxysugars, usually cladinose and desosamine, can be attached; e.g., tacrolimus, sirolimus, everolimus, pimecrolimus, erythromycin, clarithromycin, azithromycin, roxithromycin, josamycin, spiramycin, telithromycin, tylosin, and fidaxomicin, nystatin, natamycin, and amphotericin B. In one embodiment, the macrolide is selected from a macrolide immunosuppressant, a macrolide antibiotic, and a macrolide antifungal. In one embodiment, the macrolide immunosuppressant is selected from tacrolimus, sirolimus, everolimus, and pimecrolimus. In one embodiment, the macrolide antibiotic is selected from erythromycin, clarithromycin, azithromycin, roxithromycin, josamycin, spiramycin, telithromycin, tylosin, and fidaxomicin. In one embodiment, the macrolide antifungal agent is selected from polyenes, particularly nystatin, natamycin, and amphotericin B. In a preferred embodiment, the macrolide is selected from tacrolimus, sirolimus, everolimus, and pimecrolimus, preferably tacrolimus and sirolimus, and more preferably tacrolimus. In one embodiment, the terms "active agent," "active ingredient," and "API" refer to a macrolide. In one embodiment, the macrolide is formulated in the form of a mucoadhesive layer.

[0052] The term "immune disease or disorder" as used herein refers to any immune disease or disorder known to those skilled in the art, particularly diseases, disorders, and / or conditions that can be prevented or treated with a macrolide such as tacrolimus. For example, the immune disease or disorder may be selected from transplant rejection, preferably solid organ transplant rejection, such as liver, kidney, or heart allograft rejection; autoimmune diseases such as systemic lupus erythematosus, psoriasis, vitiligo, or erythema planus; infectious diseases, such as bacterial infections, e.g., Helicobacter pylori infection or Lyme disease; tumors, such as angiofibroma, lymphomas, e.g., T-cell lymphoma, renal cancer, neuroendocrine tumors, or breast cancer; and inflammatory diseases or conditions, such as inflammation associated with atopic eczema, uveitis, dermatitis, Kimura's disease, or coronary artery disease. For example, immune diseases or disorders that can be prevented or treated with a macrolide such as tacrolimus can be selected from transplant rejection, preferably solid organ transplant rejection, e.g., liver, kidney, or heart allograft rejection; autoimmune diseases, e.g., psoriasis; infectious diseases; tumors; and inflammatory diseases, disorders, or conditions, e.g., atopic eczema, uveitis, or Kimura's disease. In one embodiment, the immune disease or disorder is selected from transplant rejection, preferably solid organ transplant rejection, e.g., liver, kidney, or heart allograft rejection; autoimmune diseases, e.g., systemic lupus erythematosus, psoriasis, vitiligo, or erythema planus; infectious diseases, e.g., bacterial infections, e.g., Helicobacter pylori infection or Lyme disease; tumors, e.g., angiofibroma; lymphomas, e.g., T-cell lymphoma, renal cancer, neuroendocrine tumors, or breast cancer; and inflammatory diseases or conditions, e.g., atopic eczema, uveitis, dermatitis, Kimura's disease, or inflammation associated with coronary artery disease. In a preferred embodiment, the immune disease or disorder is transplant rejection. In one embodiment, the immune disease or disorder is transplant rejection, preferably solid organ transplant rejection, such as liver, kidney, or heart allograft rejection. In one embodiment, the terms "immunological disorder" and "immunological condition" are used interchangeably. In a preferred embodiment, the macrolide is for use in the prevention or treatment of transplant rejection.

[0053] In one embodiment, the administration of the macrolide at least twice daily comprises administering the macrolide via mucosal administration, preferably via buccal administration. In one embodiment, the administration at least twice daily comprises administering a first dose and a second dose, and optionally, additional doses via mucosal administration, preferably via buccal administration. For example, mucosal administration can involve administration into a moist cavity, such as the inside of the mouth or nose. Advantageously, mucosal administration, such as buccal administration, can reduce systemic side effects. Advantageously, mucosal routes, particularly buccal routes, can avoid significant drawbacks of oral drug administration, such as first-pass metabolism in the liver, delayed absorption, and drug degradation in the gastrointestinal tract. For example, buccal administration can be a local administration route in which a drug held or applied to the buccal area diffuses through the oral mucosa and directly enters the bloodstream. In a preferred embodiment, the mucosal administration, preferably the buccal administration, comprises administering the macrolide in the form of a mucoadhesive layer, particularly a mucoadhesive layer containing the macrolide. Advantageously, buccal administration avoids first-pass metabolism because the drug does not pass through the digestive system, potentially providing better bioavailability and a more rapid onset of action compared to oral administration. The inventors have found that two doses administered via mucosal administration, such as buccal administration, can efficiently achieve desired macrolide blood levels and significantly reduce the dosage required to achieve the desired macrolide blood levels, e.g., by about 80%, particularly when administered within a 7-hour, preferably 5-hour, more preferably 4-hour, or even more preferably, dosing interval ranging from about 2.5 to about 3.5 hours. Advantageously, buccal administration requires only 20-25% of the dose of standard oral therapy to achieve clinically relevant blood levels. In a preferred embodiment, the macrolide is administered to the buccal mucosa. From the perspective of drug delivery, the buccal mucosa offers advantages, such as a larger surface area compared to other epithelia in the oral cavity, and a highly vascularized system that provides direct access to the systemic circulation via capillaries and venous return, bypassing the first-pass effect of the liver.

[0054] The term "at least twice daily" as used herein refers to the macrolide being administered in the form of at least two doses per day. For example, the macrolide is administered in the form of a first dose and a second dose, and, if necessary, one or more additional doses, such as a third dose. The number of doses can be adjusted to suit the needs of the patient; thus, if necessary, more than two doses per day can be administered, for example, to a patient with a large body weight. In one embodiment, at least twice daily refers to twice, three, or four times per day, preferably twice per day. In a preferred embodiment, the macrolide is administered twice daily. In a preferred embodiment, the administration comprises administering the macrolide twice daily. If necessary, more than two doses per day are administered to the patient. In one embodiment, the at least twice-daily administration comprises administering a first dose at a first time point, a second dose at a second time point, and optionally a third dose at a third time point, the second time point being a time point within a range of about 1 to about 7 hours, preferably about 2 to about 5 hours, more preferably about 2.5 to about 3.5 hours, and even more preferably about 3 hours, from the first time point. In one embodiment, the at least twice-daily administration comprises administering a first dose in the form of a mucoadhesive layer, a second dose in the form of a mucoadhesive layer, and optionally a third dose in the form of a mucoadhesive layer. Advantageously, when the second dose is administered at a time point within a range of about 1 to about 7 hours, preferably about 2 to about 5 hours, more preferably about 2.5 to about 3.5 hours, and even more preferably about 3 hours, from the first time point, i.e., the administration of the first dose, exceptionally high blood levels are achieved, reducing the required dosage. Advantageously, when the second dose is administered at a second time point within a range of about 1 hour to about 7 hours from the first time point, for example about 3 hours from the first time point, the achieved blood levels are much higher than would be expected, particularly the blood levels achieved with the second dose are more than twice the blood levels achieved with the first dose.Therefore, a synergistic effect can be achieved when the second dose is administered at a time point within about 1 to about 7 hours, preferably about 2 to about 5 hours, more preferably about 2.5 to about 3.5 hours, and even more preferably about 3 hours, from the first time point. The inventors have surprisingly discovered that when the second dose is administered within 7 hours, preferably within 5 hours, more preferably within 4 hours, and even more preferably within 3 hours after the administration of the first dose, the dose required to achieve a specific blood level can be significantly reduced compared to when the second dose is administered more than 9 hours, particularly more than 10 hours, for example, about 12 hours, from the administration of the first dose. The inventors have found that the greatest synergistic effect is achieved when the second dose is administered within about 2.5 to about 3.5 hours, for example, about 3 hours, from the first time point. In particular, the inventors have surprisingly found that when the second dose is administered at a second time point within about 2.5 to about 3.5 hours from the first time point, for example at about 3 hours, the blood level achieved by the second dose is significantly greater than the blood level achieved by the first dose, more than twice as much. Furthermore, the inventors have surprisingly found that when the second dose is administered within about 7 hours from the first time point, particularly within about 2.5 to about 3.5 hours, for example at about 3 hours, the dose required to achieve a particular blood level is significantly reduced. The inventors have found that these advantages are strongest when the second dose is administered within 3.5 hours from the first time point, particularly within about 2.5 to about 3.5 hours from the first time point. In one embodiment, the terms "bid," "bid," and "twice daily" are used interchangeably. In one embodiment, a first dose is administered in the form of a first mucoadhesive layer and a second dose is administered in the form of a second mucoadhesive layer, and if desired, further doses can be administered in the form of further mucoadhesive layers.

[0055] The terms "first time point" and "second time point" as used herein preferably refer to the time points at which the first dose and the second dose are first administered, respectively. For example, the first time point refers to the time point at which the mucoadhesive layer is first attached to the body cavity, and the second time point refers to the time point at which the second mucoadhesive layer is first attached to the body cavity. The mucoadhesive layer may then remain attached to the body cavity for a period of, for example, about 1 minute to about 7 hours, e.g., about 60 minutes. Preferably, the second time point is within a range of about 1 hour to about 7 hours from the first time point, and the first time point is relative to the initial administration of the first dose and is independent of the administration period of the first dose. For example, the first time point may be relative to t=0, i.e., the time point at which the first dose is administered, e.g., the time point at which the mucoadhesive layer is attached. The first time point preferably relates to the initiation of prevention or treatment. The term "second time point" as used herein refers to a time point within the range of about 1 hour to about 7 hours, preferably about 2 hours to about 5 hours, more preferably about 2.5 hours to about 3.5 hours, and even more preferably about 3 hours from said first time point.

[0056] The inventors have discovered that asymmetric administration of at least two doses, i.e., administering the second dose within about 7 hours, preferably within about 5 hours, more preferably within about 4 hours, and even more preferably within about 3.5 hours, of the first dose, instead of administering the second dose 10 to about 12 hours after the first dose, makes it possible to achieve higher macrolide blood levels. Advantageously, because the desired blood levels are achieved more efficiently, the applied dose can be reduced. Thus, for example, applying a lower dose can reduce side effects.

[0057] In a preferred embodiment, the macrolide is administered mucosally, preferably bucally. The mucoadhesive layer, e.g., a mucoadhesive layer comprised in a mucoadhesive film, can be attached to a patient's body cavity, e.g., the patient's mouth, nose, or rectum. In a preferred embodiment, the mucoadhesive layer is attached to the mucosa of a patient's body cavity. In one embodiment, the mucoadhesive layer is attached to the buccal cavity, particularly the buccal oral mucosa. Advantageously, by attaching the mucoadhesive layer to the mucosa, the macrolide is released directly to the mucosa, forming a depot within the mucosa as needed, thereby avoiding the first-pass effect. Advantageously, administering the macrolide directly to the mucosa allows a desired blood level to be achieved with a reduced dosage compared to oral administration, e.g., in the form of a tablet or pill. In a preferred embodiment, the body cavity to which the layer is attached is the oral cavity, particularly the buccal cavity. In one embodiment, said administering comprises attaching a mucoadhesive layer, e.g. in the form of a mucoadhesive film, to a body cavity, preferably the buccal cavity, of a patient, said attachment being such that said mucoadhesive film contacts an attachment site within the body cavity, preferably the buccal cavity, in particular the mucosa, via the side of said polymer matrix layer to which the backing layer is not attached.

[0058] In one embodiment, the administration comprises administering each of the first and second doses, and optionally administering the additional doses, for a period ranging from about 1 minute to about 7 hours, preferably from about 10 minutes to about 3 hours, more preferably from about 20 minutes to about 2 hours, even more preferably from about 30 minutes to about 90 minutes, and even more preferably from about 60 minutes. For example, the first dose is administered at a first time point, e.g., in the form of a mucoadhesive layer, and then remains at the administration site, preferably on the mucosa, more preferably in the buccal cavity, for a period ranging from about 1 minute to about 7 hours, preferably from about 10 minutes to about 3 hours, more preferably from about 20 minutes to about 2 hours, even more preferably from about 30 minutes to about 90 minutes, and even more preferably about 60 minutes. For example, a second dose is administered at a second time, e.g., in the form of a mucoadhesive layer, and then remains at the administration site, preferably the mucosa, more preferably the buccal cavity, for a period ranging from about 1 minute to about 7 hours, preferably from about 10 minutes to about 3 hours, more preferably from about 20 minutes to about 2 hours, even more preferably from about 30 minutes to about 90 minutes, and even more preferably about 60 minutes. Advantageously, administering the dose, e.g., in the form of a mucoadhesive layer, for a period ranging from about 1 minute to about 7 hours, e.g., from about 30 minutes to about 90 minutes, allows the macrolide to be released directly to the mucosa, e.g., in the buccal cavity, and efficiently enter the patient's blood system. After the period, the administration device, particularly the mucoadhesive layer, can be removed. In one embodiment, the mucoadhesive layer contains about 50% to about 95%, preferably about 70% to about 93%, of the initial macrolide amount at the end of the period. For example, when the first mucoadhesive layer containing the first dose is removed after the period of time, the first mucoadhesive layer contains about 50% to about 95%, preferably about 70% to about 93%, of the initial amount of macrolide contained in said layer prior to said administration. For example, when the second mucoadhesive layer containing the second dose is removed after the period of time, the second mucoadhesive layer contains about 50% to about 95%, preferably about 70% to about 93%, of the initial amount of macrolide contained in said layer prior to said administration.

[0059] In particular, the present inventors have found that when the administration comprises: administering the first dose for a period of about 30 minutes to about 90 minutes, preferably about 60 minutes; and administering the second dose at a time point from about 2.5 hours to about 3.5 hours, preferably about 3 hours, for a period of about 30 minutes to about 90 minutes, preferably about 60 minutes, from the first time point.Advantageously, by this asymmetric administration, particularly even at a relatively low dose, very high blood level can be achieved.

[0060] In one embodiment, the first dose, the second dose, and optionally, the additional doses each, preferably before administration, each contain the macrolide in an amount ranging from about 0.001 mg to about 250 mg, preferably from about 0.01 mg to about 50 mg, more preferably from about 0.05 mg to about 25 mg, even more preferably from about 0.1 mg to about 15 mg, even more preferably from about 0.5 mg to about 13 mg, and optionally, from about 8 mg to about 12 mg. In one embodiment, the total daily dose contains the macrolide in an amount ranging from 0.001 mg to about 500 mg, preferably from about 0.01 mg to about 100 mg, more preferably from about 0.05 mg to about 50 mg, even more preferably from about 0.1 mg to about 30 mg, even more preferably from about 0.5 mg to about 26 mg, and optionally, from about 8 mg to about 24 mg. The term "total daily dose" as used herein preferably refers to the total amount of macrolide administered per day, such as the total amount of the first and second doses. In one embodiment, the terms "total daily dose" and "total daily amount" are used interchangeably. When two doses are applied per day, the total daily dose preferably refers to the sum of the first and second doses. When more than two doses are applied per day, the total daily dose preferably refers to the sum of the first, second, and further doses. Advantageously, by administering the second dose within 7 hours, preferably within 5 hours, more preferably within 3.5 hours, such as within 3 hours, of the first dose, particularly when the doses are administered in mucoadhesive form, the total daily dose required to achieve the first, second, and / or desired macrolide blood levels is significantly reduced compared to when the second dose is administered at a time within about 10 to about 14 hours, such as about 12 hours, from the first dose.

[0061] In one embodiment, the macrolide is administered mucosally, preferably buccal, and the administration of the macrolide at least twice daily comprises administering a total daily amount of macrolide that is ≦about 40%, preferably ≦about 30%, more preferably ≦about 25%, and even more preferably ≦about 20% of the total daily amount administered orally. For example, if a recommended total daily oral dose is 100 mg, the macrolide for use in the present invention is administered at a total daily amount ≦40 mg, preferably ≦about 30 mg, more preferably ≦about 25 mg, and even more preferably ≦about 20 mg. Those skilled in the art will recognize the recommended total daily oral dose in light of the package insert for oral formulations such as Prograf®. In one embodiment, the macrolide is administered mucosally, preferably buccally, and said administration of the macrolide at least twice daily comprises administering a total daily amount of macrolide that comprises or consists of ≦about 40% of the total recommended daily oral dose, preferably ≦about 30% of the total recommended daily oral dose, more preferably ≦about 25% of the total recommended daily oral dose, and even more preferably ≦about 20% of the total recommended daily oral dose. As used herein, the term "recommended total daily oral dose" preferably relates to the total daily dose approved for marketing and / or as described in the package insert for an oral macrolide formulation. When referring to oral administration, oral formulations, and / or oral doses, the term preferably refers to administration / formulations / dosages that involve swallowing, such as oral administration involving swallowing a tablet, capsule, granules, or pill, whereby the macrolide enters the blood system via the digestive tract; in contrast, "buccal" administration / formulations / dosages do not involve swallowing large amounts of macrolide, but instead administer the macrolide to the blood system via the buccal mucosa. In one embodiment, oral administration refers to peroral administration. In one embodiment, oral dose refers to peroral dose. Advantageously, the macrolides for use in the present invention, in terms of highly efficient administration, allow for the application of much lower amounts of macrolide, e.g., only about 20% of the recommended oral dose, while still achieving therapeutic levels.Advantageously, the bioavailability of macrolides is significantly improved when administered mucosally, particularly buccally, instead of orally, especially when the second dose is administered within 7 hours of the first dose. Without being bound by any theory, the inventors believe that the improved bioavailability of the macrolides used in the present invention is based on the premise that the first-pass effect and pre-systemic degradation after oral administration are not associated with mucosal, particularly buccal, administration. Advantageously, particularly when the first and second doses are provided in the form of a mucoadhesive layer, administering the second dose within 7 hours, preferably within 5 hours, and more preferably within 3.5 hours of the first dose can reduce the patient's drug burden by approximately 80%. This can, for example, reduce the side effects of macrolide therapy.

[0062] The present inventors have determined that the macrolide is present in a layer of the mucoadhesive layer 1 cm thick prior to administration. 2 and preferably in an amount ranging from about 0.01 mg to about 10 mg per cm of said mucoadhesive layer. 2 and more preferably in an amount ranging from about 0.1 mg to about 5 mg per cm of said mucoadhesive layer. 2 and even more preferably in the range of about 1.5 mg to about 3.5 mg per cm of said mucoadhesive layer. 2 It has been found that administration of the macrolide is even further improved when the macrolide is present in the mucoadhesive layer in an amount ranging from about 1.8 mg to about 2.9 mg based on the amount of the macrolide. As used herein, the expressions "before said administration" and "before said administration" in the context of an amount or dose of macrolide are meant to be understood as relating to the mucoadhesive layer, in particular the initial amount / dose contained in the mucoadhesive layer, including the respective amount or dose after preparation and before administration; when the mucoadhesive layer is administered, e.g., attached to the buccal cavity, the amount / dose of the macrolide typically decreases over time, as the macrolide is released from the mucoadhesive layer into the buccal cavity.

[0063] The inventors have also found that administration of the macrolide, e.g., the tacrolimus, is even further enhanced when the macrolide, e.g., the tacrolimus, is present in the mucoadhesive layer in an amount ranging from about 12 to about 25 weight percent, preferably from about 12 to about 13 weight percent or from about 19 to about 23 weight percent, prior to administration (where weight percent is based on the dry weight of the mucoadhesive layer).

[0064] In a preferred embodiment, the at least twice-daily administration provides a first peak in whole blood concentration of the macrolide at a time point within a range of about 2 to about 4 hours, preferably about 2.5 to about 3.5 hours, and more preferably about 3 hours, from the first time point, and a second peak in whole blood concentration of the macrolide at a time point within a range of about 7 to about 9 hours, preferably about 7.5 to about 8.5 hours, and more preferably about 8 hours, from the first time point. In one embodiment, the at least twice-daily administration provides the second peak in whole blood concentration of the macrolide at a time point within a range of about 3 to about 5 hours, preferably about 3.5 to about 4.5 hours, and more preferably about 4 hours, from the second time point. Preferably, the second peak is higher than the first peak. Preferably, the second peak is at least two-fold higher than the first peak. For example, the second peak is two-fold higher than the first peak or more than two-fold higher than the first peak. For example, the sum of the heights of the first and second peaks may be more than twice the height of the first peak. Advantageously, the first and second doses act synergistically when applied within 7 hours or less, preferably 5 hours or less, and particularly 3.5 hours or less. The inventors have surprisingly found that the second peak significantly exceeds the first peak and is higher than expected. In one embodiment, the terms "whole blood concentration" and "blood concentration" are used interchangeably. Preferably, when referring to "whole blood concentration," "blood concentration," or "blood level," it refers to the blood concentration of the macrolide.

[0065] In one embodiment, the at least twice-daily administration provides a whole blood concentration of the macrolide within the range of about 0.01 ng / mL to about 50 ng / mL, preferably within the range of about 3 ng / mL to about 20 ng / mL, and more preferably within the range of about 5 ng / mL to about 20 ng / mL, for a period of about 24 hours. Advantageously, the macrolides for use in the present invention allow for the achievement of desired blood concentrations, such as blood concentrations within the range of about 5 ng / mL to about 20 ng / mL. In one embodiment, the therapeutic range is within the range of about 5 ng / mL to about 20 ng / mL. In a preferred embodiment, the macrolides for use in the present invention are administered to achieve blood concentrations within the therapeutic range.

[0066] The inventors have surprisingly discovered that the coefficient of variation of the macrolides for use in the present invention is significantly reduced compared to standard oral macrolide administration, such as Prograf®, administered twice daily with a dosing interval of approximately 10 to 12 hours. In particular, the coefficient of variation is further improved, i.e., reduced, when the macrolides for use in the present invention are administered in the form of a mucoadhesive layer. Advantageously, reduced intra- and inter-subject coefficients of variation are observed with the macrolides for use in the present invention compared to standard oral macrolide administration, such as Prograf®, administered twice daily with a dosing interval of approximately 10 to 12 hours.

[0067] In one embodiment, the at least twice-daily administration provides a coefficient of variation, preferably a macrolide blood level coefficient of variation, of 30% or less, such as less than 40%, preferably less than 35%, more preferably less than 32%, and even more preferably within the range of 20% to 30% or within the range of 25% to 30%. Patients typically exhibit physiological, particularly biological, variability defined by factors such as hydration, diet, timing of the day, and level of exercise. Furthermore, there is variability between patients in response to a drug. For example, variability within the same subject, particularly intra-subject variability, and / or inter-subject variability, particularly inter-subject variability, can be quantified using the coefficient of variation. The term "coefficient of variation" as used herein preferably relates to intra- and / or inter-subject variability of a pharmacokinetic parameter, such as macrolide blood level. In one embodiment, the variability is intra-subject variability, e.g., intra-subject macrolide blood level variability. In one embodiment, the coefficient of variation is a coefficient of macrolide blood level variability, e.g., inter-subject macrolide blood level variability and / or intra-subject macrolide blood level variability. In one embodiment, the coefficient of variation is the intra-subject coefficient of variation and / or the inter-subject coefficient of variation of the pharmacokinetic index, preferably the intra-subject coefficient of variation of the pharmacokinetic index. The coefficient of variation (CV) can be defined as the ratio of the standard deviation to the mean. Advantageously, the macrolides for use in the present invention have reduced intra- and inter-subject variability compared to standard oral macrolide administration, such as Prograf® administration. For example, inter-individual pharmacokinetic variability can be affected by time since transplantation, patient demographics (age and race), liver and renal function, hematocrit, dietary intake, concomitant medications (such as corticosteroids, antifungals, calcium channel blockers), and metabolic enzyme genotypes.

[0068] In one embodiment, the administering at least twice daily - t within a range of about 3 hours to about 10 hours from the first time point, such as within a range of about 4 hours to about 10 hours from the first time point, preferably within a range of about 7 hours to about 9 hours from the first time point, more preferably within a range of about 7.5 hours to about 8.5 hours from the first time point. max and / or - within a range of about 1 hour to about 6 hours from the second time point, preferably within a range of about 2 hours to about 5 hours from the second time point, more preferably within a range of about 3.5 hours to about 4.5 hours from the second time point, for example, within a range of about 4 hours from the second time point max to provide.

[0069] T max is typically the time to peak drug concentration, particularly the time to maximum drug concentration (Cmax) after drug administration. Advantageously, macrolides for use in the present invention allow very high blood levels to be achieved at relatively low doses, for example, in the range of about 1 hour to about 6 hours from the second time point.

[0070] In one embodiment, the administering at least twice daily - a C in the range of about 0.05 ng / mL to about 50 ng / mL, preferably in the range of about 0.1 ng / mL to about 30 ng / mL, more preferably in the range of about 0.7 ng / mL to about 20 ng / mL, even more preferably in the range of about 5.5 ng / mL to about 20 ng / mL max and / or - a C in the range of about 0.01 ng / mL to about 18 ng / mL, preferably in the range of about 0.5 ng / mL to about 16 ng / mL, more preferably in the range of about 3 ng / mL to about 15 ng / mL, such as in the range of about 3 ng / mL to about 7 ng / mL or in the range of about 5 ng / mL to about 7 ng / mL trough to provide.

[0071] In a preferred embodiment, C max is 20ng / mL or less and / or Ctrough For example, effective and safe prevention and treatment is max is 20 ng / mL or less and C trough This can be achieved when the serum creatine nitrate concentration is 5 ng / mL or higher.

[0072] In one embodiment, the at least twice daily administration provides a whole blood concentration in the range of 0.1 ng / mL to about 20 ng / mL, preferably in the range of 0.2 ng / mL to about 10 ng / mL, and more preferably in the range of 0.3 ng / mL to about 7 ng / mL, at a time point within about 1 hour to about 4 hours, preferably about 1.5 hours to about 3.5 hours, from the first time point. - AUC in the range of about 0.5 h*ng / mL to about 7.0 h*ng / mL, preferably in the range of about 1.0 h*ng / mL to about 6.5 h*ng / mL 0-4 ; - AUC in the range of about 0.7 h*ng / mL to about 15.0 h*ng / mL, preferably in the range of about 1.5 h*ng / mL to about 12.0 h*ng / mL 4-8 ; - AUC in the range of about 4.0 h*ng / mL to about 200.0 h*ng / mL, preferably in the range of about 5.0 h*ng / mL to about 60.0 h*ng / mL, more preferably in the range of about 10.0 h*ng / mL to about 55.0 h*ng / mL 0-24 ; - AUC in the range of about 4.0 h*ng / mL to about 250.0 h*ng / mL, preferably in the range of about 5.0 h*ng / mL to about 70.0 h*ng / mL, more preferably in the range of about 10.0 h*ng / mL to about 60.0 h*ng / mL 0-tlast ; a half-life in the range of about 12 hours to about 16 hours, preferably in the range of about 13.5 hours to about 15.5 hours, more preferably in the range of about 14 hours to about 15 hours; and / or an output rate in the range of about 0.01 1 / h to about 0.08 1 / h, preferably in the range of 0.04 1 / h to about 0.05 1 / h, and even more preferably in the range of 0.045 1 / h to about 0.049 1 / h; to provide.

[0073] In one embodiment, the administration at least twice daily is carried out for a period of at least two days, preferably at least one week, more preferably at least one month, and even more preferably at least one year. Thus, for example, the first and second doses can be administered daily for at least two days, preferably at least one week, more preferably at least one month, and even more preferably at least one year. In one embodiment, when the macrolide is used to prevent or treat a chronic immune disease or disorder, such as transplant rejection, the administration at least twice daily can be carried out for the patient's lifetime. Advantageously, mucosal, particularly buccal, administration of the macrolide for use reduces the dosage required to achieve therapeutic blood levels, thereby reducing side effects and improving the patient's quality of life.

[0074] In one embodiment, the term "patient" relates to a human or animal, preferably a human. In one embodiment, the patient is not a fasting patient. Advantageously, the macrolides for use, particularly when administered buccally, achieve desired blood levels, such as blood levels in the therapeutic range, even when the patient is not a fasting patient. In one embodiment, the patient is not a fasting patient. In one embodiment, the patient is a pediatric patient, a juvenile patient, or an adult patient. An adult patient may be a geriatric patient. The terms "patient" and "subject" may be used interchangeably.

[0075] The term "mucoadhesive layer" as used herein refers to a layer configured to adhere to mucosa and / or to adhere to mucosa, e.g., oral and / or buccal mucosa. In a preferred embodiment, the mucoadhesive layer is provided in the form of a mucoadhesive film, particularly a mucoadhesive film comprising the mucoadhesive layer and, optionally, additional layers. Preferably, the mucoadhesive layer comprises the macrolide. For example, the macrolide may be contained in the layer in the form of a nanoparticulate or non-nanoparticulate macrolide. In one embodiment, the mucoadhesive layer optionally comprises a surfactant, e.g., polysorbate. In one embodiment, the mucoadhesive layer is swellable in aqueous liquid but insoluble and / or has negligible solubility in aqueous liquid. Insolubility or low solubility increases the adhesion time on the mucosa and therefore allows the active ingredient to be released over a prolonged period, e.g., at least 60 minutes. The term "mucoadhesive buccal layer" as used herein refers to a mucoadhesive layer configured for buccal application. In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of at least 2 wt%, preferably at least 3 wt%, e.g., 3.2 wt%, where wt% is based on the dry weight of the mucoadhesive layer. In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of about 12 to about 25 wt%, where wt% is based on the dry weight of the mucoadhesive layer. In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of about 12 to about 13 wt%, or about 19 to about 23 wt%, where wt% is based on the dry weight of the mucoadhesive layer. In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of about 18 to about 25 wt%, where wt% is based on the dry weight of the mucoadhesive layer. In one embodiment, the macrolide is present in the mucoadhesive layer at a concentration of about 19 to about 22 wt%, where wt% is based on the dry weight of the mucoadhesive layer. In one embodiment, the macrolide is administered in the form of a mucoadhesive layer comprising a mucoadhesive polymer. In one embodiment, the mucoadhesive layer comprises a mucoadhesive polymer.In one embodiment, the mucoadhesive layer or film comprises a therapeutically effective amount of the macrolide.

[0076] As used herein, the term "tacrolimus mucoadhesive film" is meant to refer to a mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer, wherein said mucoadhesive layer comprises tacrolimus and optionally a mucoadhesive polymer.

[0077] In one embodiment, the mucoadhesive layer comprises a mucoadhesive polymer and / or a cellulose derivative and further comprises the macrolide, e.g., a nanoparticulate or non-nanoparticulate macrolide. In one embodiment, the mucoadhesive layer comprises a mucoadhesive polymer such as alginate. In one embodiment, the mucoadhesive layer comprises alginate. In one embodiment, the mucoadhesive layer is an alginate layer. In one embodiment, the mucoadhesive layer comprises a non-nanoparticulate macrolide, preferably a micronized crystalline non-nanoparticulate macrolide and / or a molecularly dissolved non-nanoparticulate macrolide. In one embodiment, the non-nanoparticulate macrolide is present in the layer in the form of a micronized macrolide, preferably a micronized crystalline macrolide and / or a molecularly dissolved non-nanoparticulate form. In one embodiment, the micronized macrolide has an average particle size of 1 to 100 μm, preferably 1.5 to 50 μm, more preferably 2 to 25 μm. In one embodiment, the non-nanoparticulate macrolide is present in the mucoadhesive layer in a molecularly dissolved, non-nanoparticulate form; and optionally, the mucoadhesive polymer and / or the cellulose derivative, if present, is a polymer that is soluble in organic solvents.

[0078] In one embodiment, the mucoadhesive layer comprises (in addition to the macrolide described herein) a) a mucoadhesive polymer, preferably an amphiphilic polymer; hydroxypropyl cellulose (HPC); carboxymethyl cellulose (CMC); a plasticizer, preferably glycerol; and, optionally, a colorant, preferably TiO2; Preferably 10 wt% to 30 wt% of a mucoadhesive polymer; 25 wt% to 35 wt% of hydroxypropyl cellulose (HPC); 10 wt% to 25 wt% of carboxymethyl cellulose (CMC); 10 wt% to 16 wt% of said plasticizer; and optionally 1 wt% to 4 wt% of a colorant; or b) a mucoadhesive polymer, preferably selected from cross-linked polyacrylic acid polymers and copolymers of methyl vinyl ether and maleic anhydride; hydroxypropyl cellulose (HPC); ethyl cellulose (EC); a plasticizer, preferably glycerol; and optionally a colorant, preferably TiO2; Preferably, 2 wt% to 17 wt% of a mucoadhesive polymer; 30 wt% to 70 wt% of hydroxypropyl cellulose (HPC); 5 wt% to 40 wt% of said ethyl cellulose; 3 wt% to 20 wt% of said plasticizer; and optionally 1 wt% to 5 wt% of said colorant. Includes.

[0079] In one embodiment, the mucoadhesive layer comprises (in addition to the macrolide described herein) a) a mucoadhesive polymer, preferably selected from crosslinked polyacrylic acid polymers and copolymers of methyl vinyl ether and maleic anhydride; ethyl cellulose; hydroxypropyl cellulose having a viscosity of about 300-600 mPa.s (e.g., Klucel EF); hydroxypropyl cellulose having a viscosity of about 150-400 mPa.s (e.g., Klucel GF); and a plasticizer, preferably glycerol; and optionally, a colorant, preferably TiO2 or yellow iron oxide; Preferably, 2 wt% to 10 wt% of a mucoadhesive polymer; 30 wt% to 70 wt% of hydroxypropyl cellulose (HPC), where optionally the hydroxypropyl cellulose is made of at least two different types of hydroxypropyl cellulose; 3 wt% to 13 wt% of said ethyl cellulose or 27 wt% to 31 wt% of said ethyl cellulose; 4 wt% to 6 wt% of said plasticizer; and optionally 1 wt% to 4 wt% of said colorant. Includes.

[0080] The term "nanoparticulate macrolide" as used herein refers to a macrolide in the form of nanoparticles, particularly nanoparticles having a size ranging from 1 nm to 1000 nanometers. In one embodiment, the mucoadhesive layer comprises the macrolide in the form of nanoparticles, e.g., nanoparticles having a particle size of about 10 to about 400 nm. In one embodiment, the mucoadhesive layer comprises macrolide nanoparticles. For example, the nanoparticles may have a polydispersity index of about 0.4 or less. The nanoparticles may comprise a stabilizer and / or a surfactant. For example, the stabilizer may include polyvinylpyrrolidone, vinylpyrrolidone / vinyl acetate copolymer, polyethylene glycol, and / or a cellulose derivative such as hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose phthalate (HPMCP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), hydroxypropyl cellulose (HPC), and / or carboxymethylcellulose (CMC). In one embodiment, the mucoadhesive layer comprises a nanoparticulate macrolide and / or a non-nanoparticulate macrolide.

[0081] The term "non-nanoparticulate macrolide" as used herein refers to a macrolide that is not in the form of nanoparticles. In one embodiment, the non-nanoparticulate macrolide refers to a macrolide that is not formulated as nanoparticles, particularly nanoparticles having a size ranging from 1 nm to 1000 nanometers. In one embodiment, the non-nanoparticulate macrolide is a macrolide formulated as a molecularly dissolved macrolide and / or a micronized crystalline macrolide. In one embodiment, the molecularly dissolved macrolide is not a molecularly dissolved macrolide in the form of a nanoparticle, e.g., not a molecularly dissolved macrolide present in a micelle or liposome. In one embodiment, the molecularly dissolved macrolide does not refer to nanoparticles containing a molecularly dissolved macrolide. In one embodiment, the molecularly dissolved macrolide is formulated in the form of a solid solution, e.g., embedded and / or dissolved in a polymer matrix. In one embodiment, the micronized crystalline macrolide is not in the form of nanoparticles, e.g., not in the form of nanoparticles containing a micronized crystalline macrolide. In one embodiment, the micronized crystalline macrolide has an average particle size of >1000 nm. In one embodiment, the micronized crystalline macrolide is in the form of microparticles, preferably having an average particle size in the range of 1 μm to 1000 μm, preferably >1000 nm. In one embodiment, the non-nanoparticulate macrolide contained in the mucoadhesive layer and / or mucoadhesive film is not in the form of nanoparticles and / or is not contained in nanoparticles. In one embodiment, the non-nanoparticulate macrolide is present in the layer in the form of a micronized macrolide, preferably in the form of a micronized crystalline macrolide, a molecularly dissolved non-nanoparticulate form, and / or a crystalline macrolide; preferably, the non-nanoparticulate macrolide is present in the layer in the form of a micronized macrolide, preferably in the form of a micronized crystalline macrolide, and / or a molecularly dissolved non-nanoparticulate form. In one embodiment, the mucoadhesive layer comprising a micronized macrolide, preferably in the form of a micronized crystalline macrolide, and / or a molecularly dissolved non-nanoparticulate form of a macrolide, further comprises a crystalline form of the macrolide.

[0082] The term "average particle size" as used herein relates to the average diameter of an analyzed batch of particles, e.g., (micro)particles of a micronized macrolide. For example, the average particle size can be the average diameter of a batch of particles, where if the particles are not spherical, the longest extension of each particle is considered to be the diameter. In one embodiment, the average particle size ranges from 1 to 100 μm, preferably from 1.5 to 50 μm, more preferably from 2 to 25 μm. In one embodiment, the average particle size of, e.g., a micronized macrolide, is measured using any technique known to those skilled in the art, e.g., laser diffraction analysis or dynamic light scattering, preferably laser diffraction analysis. The average particle size is defined as the average diameter of the analyzed particles, determined by a suitable process, e.g., using dynamic light scattering. In one embodiment, the average particle size of the micronized macrolide is 2 μm to 25 μm. In one embodiment, the micronized macrolide has a particle size distribution D 10 2 μm; D 50 10 μm; and D 90 25 μm. In one embodiment, 10% of all particles have a particle size <2 μm, 50% of all particles have a particle size <10 μm, and 90% of all particles have a particle size <10 μm. In one embodiment, all particles have a particle size >1 μm. In one embodiment, the micronized macrolide is not in the form of nanoparticles. In one embodiment, the average particle size is D 50 In one embodiment, the average particle size of the micronized macrolide is >1 μm, preferably it being understood that none of the particles are nanoparticles. In one embodiment, the particle size of the micronized macrolide is >1 μm. In one embodiment, the particle size of each micronized macrolide particle is >1 μm. As used herein, the term "micronized" refers to an average particle diameter reduced to the micrometer range and / or to the micrometer range, preferably reduced by micronization. In one embodiment, "micronized crystalline macrolide" refers to a crystalline macrolide having an average particle diameter in the micrometer range.

[0083] In one embodiment, the macrolide is formulated in the form of a mucoadhesive film; preferably, it is formulated in the form of a mucoadhesive film comprising a mucoadhesive layer containing the macrolide and further layers such as a backing layer. In one embodiment, the mucoadhesive layer is provided in the form of a mucoadhesive film. In one embodiment, the macrolide is administered in the form of a mucoadhesive film, preferably a mucoadhesive buccal film. In one embodiment, the term "mucoadhesive film" as used herein refers to a film that adheres to a mucosa and / or is configured to adhere to a mucosa, e.g., oral and / or buccal mucosa. In one embodiment, the terms "film," "mucoadhesive film," "mucoadhesive buccal film," and "MBF" are used interchangeably. In one embodiment, when referring to a "mucoadhesive buccal film" or "MBF," such abbreviation not only refers to a mucoadhesive buccal film, but also to a mucoadhesive film for other applications besides buccal application. In one embodiment, the mucoadhesive film, preferably a mucoadhesive buccal film, comprises a mucoadhesive layer containing a macrolide, preferably a mucoadhesive layer as defined above; and further comprises a backing layer, preferably impermeable to the macrolide and / or impermeable to water. In one embodiment, the backing layer comprises a plasticizer, preferably selected from glycerol, polyethylene glycol (low molecular weight polyethylene glycol, e.g., PEG 200 or PEG 400), propylene glycol, sorbitol, triacetin, and tributyl citrate, more preferably glycerol; and a water-insoluble polymer and / or cellulose derivative, e.g., hydroxyethyl cellulose, preferably a water-insoluble cellulose derivative, more preferably ethyl cellulose. The mucoadhesive layer or film is advantageous in that the rich blood circulation of the oral and / or buccal mucosa ensures rapid transfer of the active substance into the blood circulation. Advantageously, the active ingredient is largely absorbed through the mucosa, thereby avoiding the "first-pass metabolism" that occurs in conventional active ingredient delivery forms, such as tablet form.Furthermore, the mucoadhesive layer or film has the advantage of protecting the macrolide from degradation by the pH and digestive enzymes of the gastrointestinal tract. The mucoadhesive layer or film also provides a rapid onset of effect, e.g., for the oral route. The mucoadhesive layer or film is a simple method of drug administration and is therefore particularly suitable for use in children and the elderly. Furthermore, the mucoadhesive layer or film avoids the obstacles associated with drug administration via a nasogastric tube and is adaptable in physical shape, condition, size, and surface. The mucoadhesive layer or film has the further advantage of enabling accurate dosing. In a preferred embodiment, the term "mucoadhesive layer" refers to a mucoadhesive layer in the form of a mucoadhesive film, particularly a mucoadhesive film comprising the mucoadhesive layer and, optionally, additional layers. In one embodiment, the mucoadhesive film comprises a mucoadhesive polymer, such as alginate. In one embodiment, the mucoadhesive film comprises alginate. In one embodiment, the mucoadhesive film is an alginate film.

[0084] In one embodiment, the mucoadhesive film further comprises an intermediate layer disposed between the mucoadhesive layer and the backing layer; preferably, said intermediate layer facilitates adhesion between the mucoadhesive layer and the backing layer; optionally, said intermediate layer comprises a plasticizer, preferably selected from glycerol, polyethylene glycol, low molecular weight polyethylene glycol such as PEG200 or PEG400, propylene glycol, sorbitol, triacetin, and tributyl citrate, more preferably glycerol; and a polymer, preferably a polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone, more preferably vinylpyrrolidone-vinyl acetate copolymer.

[0085] In one embodiment, the mucoadhesive film, preferably the mucoadhesive buccal film, comprises a mucoadhesive layer, a backing layer and an intermediate layer; the intermediate layer is disposed between the mucoadhesive layer and the backing layer, the mucoadhesive layer comprising a macrolide, preferably tacrolimus; Preferably, the intermediate layer facilitates adhesion between the mucoadhesive layer and the backing layer; Optionally, the intermediate layer comprises a plasticizer, preferably selected from glycerol, polyethylene glycol, low molecular weight polyethylene glycol such as PEG200 or PEG400, propylene glycol, sorbitol, triacetin and tributyl citrate, more preferably glycerol; Optionally, the intermediate layer comprises a polymer, preferably a polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone, more preferably a vinylpyrrolidone-vinyl acetate copolymer.

[0086] In one embodiment, the mucoadhesive film is a solid formulation comprising one or more layers of a suitable material intended to be applied to a body cavity, preferably the buccal cavity, to obtain a systemic or local effect, preferably a systemic effect; and / or the mucoadhesive film is a flexible single-dose formulation configured to be applied to a body cavity, preferably the oral cavity, to obtain either a systemic or local effect by delivering the macrolide over a period of time, after which it is then removed. In one embodiment, the mucoadhesive film is an oral mucosal patch.

[0087] In one embodiment, the mucoadhesive film comprises the mucoadhesive layer and, optionally, a backing layer. In a preferred embodiment, the mucoadhesive film comprises the mucoadhesive layer and a backing layer. In one embodiment, the mucoadhesive film comprises a mucoadhesive layer; a backing layer, preferably an impermeable backing layer; and, optionally, an intermediate layer. In one embodiment, the mucoadhesive layer comprises a macrolide, for example, in a polymer matrix or nanoparticles, and provides prolonged contact of the macrolide with the buccal mucosa. In one embodiment, the film comprises at least two layers, for example, two layers laminated together; preferably, a mucoadhesive layer comprising an active ingredient and a backing layer. In one embodiment, the back side of the mucoadhesive layer is sealed with a backing layer, preferably a water-insoluble and / or macrolide-impermeable backing layer, to prevent release of the macrolide into the oral cavity. In one embodiment, the back side of the mucoadhesive layer is the side configured to face the oral cavity. In one embodiment, the two layers are attached to each other via an intermediate layer, preferably an adhesive layer. In one embodiment, the mucoadhesive layer is configured to be administered to a patient, for example, by incorporating the mucoadhesive layer into a mucoadhesive film, an oral mucosal patch, or any other application system.

[0088] In one embodiment, the adhesive properties of the mucoadhesive layer and / or film are sufficient for application for at least 30 minutes, preferably at least 60 minutes, i.e., the layer and / or film adheres to the mucosa for at least 30 minutes, preferably at least 60 minutes. In one embodiment, ethylcellulose is included in the mucoadhesive layer and / or film, and its low water solubility allows for an application time of at least 30 minutes, preferably at least 60 minutes. In one embodiment, the backing layer comprises ethylcellulose. In one embodiment, the mucoadhesive film is an insoluble film that must be removed from the mucosa, e.g., the buccal mucosa, after drug release. In one embodiment, the mucoadhesive layer is applied to the patient for a period of at least 30 minutes.

[0089] In one embodiment, the mucoadhesive layer and / or the mucoadhesive film is 0.5 cm 2 10cm from 2 , preferably 2 cm 2 From 8cm 2 In one embodiment, the mucoadhesive film has an area weight of about 20 g / m 2 to approximately 300 g / m 2 , preferably about 50 g / m 2 Approximately 260 g / m 2 In one embodiment, the mucoadhesive film has a film thickness of about 20 μm to about 1000 μm, preferably about 50 μm to about 500 μm, more preferably about 50 μm to about 300 μm, for example about 150 μm to about 260 μm.

[0090] In one embodiment, the area weight of the mucoadhesive layer is from 70 to 180 g / m 2 in the range of 100 to 150 g / m 2 and / or The area weight of the backing layer is 40 to 100 g / m 2 in the range of 40 to 60 g / m 2 in the range up to, for example, 50 g / m 2 and / or If present, the intermediate layer has an areal weight of 40 to 100 g / m 2 in the range of 40 to 70 g / m 2 in the range up to, for example, 60 g / m 2 is.

[0091] In one embodiment, at least one of the layers comprises a colorant, e.g., a white pigment in the mucoadhesive layer and a blue dye in the backing layer. Advantageously, the colorant in at least one of the layers allows the adhesive layer to be distinguished from the backing layer. In one embodiment, the mucoadhesive film further comprises a flavoring and / or sweetening agent, such as an agent selected from citric acid, peppermint oil, saccharin sodium, and citrus flavorings. Advantageously, the flavoring and / or sweetening agent improves the mouthfeel for the patient. In particular, such flavoring and / or sweetening agent effectively masks the unpleasant taste of macrolides or other MBF components. In one embodiment, at least one layer further comprises at least one adjuvant selected from the group including stabilizers, e.g., chelating agents, coloring agents, flavoring agents, sweetening agents, taste masking agents, emulsifiers, enhancers, pH adjusters, humectants, preservatives, and / or antioxidants. In one embodiment, at least one layer comprises a chelating agent, preferably EDTA. In one embodiment, reference to a "layer" refers to a mucoadhesive layer, an intermediate layer, and / or a backing layer. In a preferred embodiment, the mucoadhesive layer is provided in the form of a mucoadhesive film.

[0092] In one embodiment, the dosage strength of the film is determined by the size and / or area of ​​the film, and where, optionally, the quantitative composition per film area is constant. In one embodiment, the film contains a dosage of up to 250 mg, for example up to 50 mg, of a macrolide. In one embodiment, the amount of macrolide is measured using HPLC. In one embodiment, the macrolide is quantified using HPLC. In one embodiment, the mucoadhesive film is administered and / or configured to be administered to adult or pediatric patients, for example, children up to about 18 years of age.

[0093] In one embodiment, the mucoadhesive layer and / or the backing layer comprise a colorant. In one embodiment, the mucoadhesive layer and the backing layer each comprise a colorant, wherein the mucoadhesive layer comprises a first colorant, such as TiO2, and the backing layer comprises a second colorant, such as brilliant blue, and wherein the first colorant and the second colorant are different from each other. In one embodiment, the mucoadhesive layer and / or the mucoadhesive film has a water content of <5 wt%, preferably <3 wt%, more preferably <2 wt%.

[0094] The term "plasticizer" as used herein refers to any plasticizer known to those skilled in the art, preferably a plasticizer suitable for in vivo application. In one embodiment, the plasticizer, for example, contained in the mucoadhesive layer, intermediate layer, and / or backing layer, is selected from glycerol, poly(ethylene glycol), preferably a low molecular weight poly(ethylene glycol) such as PEG200 or PEG400, propylene glycol sorbitol, triacetin, and tributyl citrate. In one embodiment, the low molecular weight poly(ethylene glycol) has a molecular weight of ≦600 Daltons, preferably ≦450 Daltons, for example 420 Daltons. The plasticizer can impart advantageous mechanical properties to the layer.

[0095] The term "backing layer" as used herein refers to a layer of a mucoadhesive film. In one embodiment, the backing layer covers the back side of the mucoadhesive layer in the mucoadhesive film. The back side is preferably the side of the mucoadhesive layer or film configured to face the body cavity, preferably the oral cavity, i.e., the side facing away from the mucosa. In one embodiment, the backing layer is impermeable to the macrolide and / or impermeable to water. Advantageously, the backing layer prevents release of the macrolide from the mucoadhesive layer into the body cavity, e.g., the oral cavity. In one embodiment, the backing layer covers the back side of the mucoadhesive layer and, optionally, further covers the lateral regions of the mucoadhesive layer. In one embodiment, the backing layer is a sealing layer. In one embodiment, the backing layer, preferably an impermeable backing layer, prevents undesired release of the macrolide into the body cavity, preferably the oral cavity. In one embodiment, the backing layer is configured to ensure unidirectional release of the macrolide to the mucosa, preferably the buccal mucosa. In one embodiment, the backing layer prevents release of the macrolide into the buccal oral cavity. Advantageously, preventing release into the oral cavity reduces side effects. In one embodiment, the backing layer is insoluble in aqueous liquids such as water and / or saliva. In one embodiment, the backing layer does not dissolve in aqueous liquids such as water and / or saliva for a period of 30 to 75 minutes, preferably at least 60 minutes. In one embodiment, the backing layer and / or backing layer polymer mixture comprises a water-insoluble polymer and / or a cellulose derivative, such as hydroxyethyl cellulose, preferably a water-insoluble cellulose derivative, more preferably ethyl cellulose, and thus prevents dissolution of the backing layer in the oral cavity. In one embodiment, the backing layer and / or backing layer polymer mixture comprises a water-insoluble polymer and / or a polymer with low water solubility, preferably a water-insoluble polymer such as ethyl cellulose. In one embodiment, the backing layer polymer mixture comprises at least one component, preferably all components of the backing layer. In one embodiment, the backing layer is organic solvent-based, such as ethanol-based, and / or the mucoadhesive layer is water-based.In one embodiment, the backing layer comprises a polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone, preferably a polymer comprising polyvinyl acetate. In one embodiment, the backing layer comprises a colorant, such as brilliant blue. In one embodiment, the backing layer comprises a plasticizer, such as triacetin.

[0096] The term "water-insoluble polymer" as used herein refers to a polymer that is insoluble in water, such as ethyl cellulose and poly(methyl methacrylate). The term "polymer with low water solubility" as used herein refers to a polymer with negligible and / or low water solubility, such as hydroxyethyl cellulose. In one embodiment, the "water-insoluble polymer" and "polymer with low water solubility" are polymers that prevent the dissolution of the mucoadhesive film for at least 30 minutes during application in the body cavity, preferably in the buccal cavity. In one embodiment, the water-insoluble polymer and / or polymer with low water solubility are preferably selected so as to provide a film application time of at least 15 minutes, preferably at least 30 minutes, without film dissolution. In one embodiment, the water-insoluble polymer is ethyl cellulose and / or poly(methyl methacrylate). In one embodiment, the cellulose derivative contained in the backing layer is hydroxyethyl cellulose and / or ethyl cellulose. In one embodiment, the backing layer is configured to be insoluble in aqueous liquids for at least 30 minutes, preferably at least 60 minutes. In one embodiment, the backing layer is configured to be impermeable to macrolides for at least 30 minutes, preferably at least 60 minutes; preferably, configured to be impermeable to macrolides in aqueous liquids, such as saliva, for at least 30 minutes, preferably at least 60 minutes. In one embodiment, the backing layer and / or backing layer polymer blend comprises a polymer that is insoluble in aqueous liquids for at least 30 minutes. In one embodiment, the backing layer comprises ethyl cellulose, hydroxyethyl cellulose, and / or poly(methyl methacrylate), and optionally further comprises a plasticizer. In one embodiment, the backing layer comprises ethyl cellulose and / or poly(methyl methacrylate), and optionally further comprises a plasticizer.

[0097] The term "intermediate layer" as used herein refers to a layer of a mucoadhesive film, preferably a layer disposed between the mucoadhesive layer and the backing layer. In one embodiment, the intermediate layer is an adhesive layer. In one embodiment, the intermediate layer facilitates adhesion between the mucoadhesive layer and the backing layer. In one embodiment, the intermediate layer prevents migration of the macrolide into the backing layer. In one embodiment, the film includes an intermediate layer between the mucoadhesive layer and the backing layer, which prevents migration of the macrolide into the backing layer. In one embodiment, the terms "adhesive intermediate layer" and "intermediate layer" are used interchangeably. In one embodiment, the mucoadhesive layer and the backing layer are bonded via the intermediate layer. For example, in a method, a film including the backing layer and the intermediate layer can be combined with the mucoadhesive layer after drying the layers to prevent migration of the macrolide into the adhesive intermediate layer and / or the backing layer. Advantageously, the incorporation of an intermediate layer into the film makes it possible to prevent dissolution of the macrolide into the backing layer during the method of preparing the film. Advantageously, the intermediate layer prevents the amount of macrolide in the mucoadhesive layer from decreasing and / or the macrolide from decomposing during the process of preparing the backing layer on the mucoadhesive layer. Advantageously, the intermediate layer prevents the macrolide from dissolving in the ethanol contained in the backing layer. In one embodiment, the mucoadhesive film includes an intermediate layer, such as a mucoadhesive film containing a micronized macrolide, preferably a micronized crystalline macrolide, and / or a non-nanoparticulate macrolide in a molecularly dissolved non-nanoparticulate form. In one embodiment, the intermediate layer includes a polymer, optionally an adhesive polymer. In one embodiment, the intermediate layer includes a polymer selected from polyvinyl acetate and / or polyvinylpyrrolidone; for example, poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)], e.g., a copolymer of 6 parts polyvinylpyrrolidone and 4 parts polyvinyl acetate, and a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone), e.g., an 8:2 mixture of poly(vinyl acetate) and poly(vinylpyrrolidone). In one embodiment, the intermediate layer includes a plasticizer, such as glycerol.

[0098] The term "solid solution" as used herein refers to a macrolide molecularly dissolved in a solid-state diluent or matrix, preferably in a polymer matrix. In one embodiment, the term "solid solution" refers to a macrolide dissolved and optionally embedded in a polymer matrix. In one embodiment, a molecularly dissolved non-nanoparticulate form of the macrolide refers to a solid solution of the macrolide. In one embodiment, the terms "solid solution" and "molecularly dissolved non-nanoparticulate form" are used interchangeably. In one embodiment, the solid solution is a molecularly dissolved non-nanoparticulate macrolide provided in a polymer matrix, preferably comprising at least one polymer soluble in an organic solvent. In one embodiment, the molecularly dissolved non-nanoparticulate macrolide is provided in a polymer matrix, preferably comprising at least one polymer soluble in an organic solvent. In one embodiment, a polymer soluble in an organic solvent, such as a copolymer of methyl vinyl ether and maleic anhydride, is used to achieve adequate mucoadhesion of a water-free mucoadhesive film formulation, e.g., for the solid solution. In one embodiment, the molecularly dissolved non-nanoparticulate macrolide is in the form of a solid solution of the macrolide. In one embodiment, the polymer soluble in an organic solvent is a polymer soluble in any organic solvent selected from a) amphiphilic polymers and hydrophilic polymers, b) poly(methacrylates), c) crosslinked polyacrylic acid polymers, d) copolymers of methyl vinyl ether and maleic anhydride, e) polymers containing polyvinyl acetate and / or polyvinylpyrrolidone, f) cellulose derivatives, and combinations thereof. In one embodiment, the polymer soluble in an organic solvent is an amphiphilic or hydrophilic polymer, such as Soluplus®; poly(methacrylates), such as methacrylic acid copolymers; cellulose derivatives, preferably hydroxypropyl cellulose (HPC) or ethyl cellulose (EC), more preferably HPC; crosslinked polyacrylic acid polymers, such as Carbopol® or polycarbophil; copolymers of methyl vinyl ether and maleic anhydride, such as Gantrez®. TMand / or a polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone, such as Kollidon®. In one embodiment, the solid solution of macrolide is HPC; a crosslinked polyacrylic acid polymer, such as Carbopol® or Polycarbophil; a copolymer of methyl vinyl ether and maleic anhydride, such as Gantrez®. TM and / or a polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone, such as Kollidon®. In one embodiment, the solid solution comprises HPC; and a cross-linked polyacrylic acid polymer, such as Carbopol® or polycarbophil, and / or a copolymer of methyl vinyl ether and maleic anhydride, such as Gantrez®. TMIn one embodiment, the non-nanoparticulate macrolide is present in the mucoadhesive layer in the form of a micronized macrolide, preferably a micronized crystalline macrolide, and / or in a molecularly dissolved non-nanoparticulate form, preferably a solid solution of the molecularly dissolved macrolide. Such a mucoadhesive layer allows for achieving a highly advantageous macrolide blood profile. As used herein, the term "molecularly dissolved non-nanoparticulate form" relates to a macrolide that is molecularly dissolved, e.g., in a polymer matrix. In one embodiment, the mucoadhesive layer comprises at least one component, preferably a polymer selected from a) amphiphilic polymers and hydrophilic polymers, b) poly(methacrylate), c) crosslinked polyacrylic acid polymers, d) copolymers of methyl vinyl ether and maleic anhydride, e) polymers containing polyvinyl acetate and / or polyvinylpyrrolidone, f) cellulose derivatives, and combinations thereof; preferably, it comprises at least one component selected from a) to e) and further comprises a cellulose derivative; wherein, preferably, when the layer comprises a molecularly dissolved non-nanoparticulate macrolide, the at least one component is a polymer soluble in organic solvents, preferably ethanol, acetone, and / or isopropanol. In one embodiment, the mucoadhesive layer comprises a plasticizer. In one embodiment, the mucoadhesive layer comprises a non-nanoparticulate micronized crystalline macrolide and / or a molecularly dissolved non-nanoparticulate macrolide.

[0099] In one embodiment, when the non-nanoparticulate macrolide is present in the layer in a molecularly dissolved, non-nanoparticulate form, the mucoadhesive polymer and / or cellulose derivative is a film-forming polymer, preferably a film-forming polymer soluble in an organic solvent such as ethanol. In one embodiment, when the non-nanoparticulate macrolide is present in the layer in a molecularly dissolved, non-nanoparticulate form, the molecularly dissolved, non-nanoparticulate form is solubilized in an organic solvent. In one embodiment, when the non-nanoparticulate macrolide is present in the layer in a molecularly dissolved, non-nanoparticulate form, the cellulose derivative is selected from HPC, EC, and combinations thereof. In one embodiment, when the non-nanoparticulate macrolide is present in the layer in a molecularly dissolved, non-nanoparticulate form, the mucoadhesive polymer is an amphiphilic or hydrophilic polymer, such as Soluplus®; a cross-linked polyacrylic acid polymer, such as Carbopol® or polycarbophil; a copolymer of methyl vinyl ether and maleic anhydride, such as Gantrez®. TM polymers comprising polyvinyl acetate and / or polyvinylpyrrolidone, preferably polymers comprising polyvinyl acetate and / or polyvinylpyrrolidone, such as Kollidon®; and cellulose derivatives, such as HPC, EC, and combinations thereof.

[0100] In one embodiment, the macrolide is dissolved in an organic solvent at a concentration of about 10 mg / ml to about 500 mg / ml to achieve molecular dissolution of the macrolide. As used herein, the term "organic solvent" refers to any organic solvent known to those skilled in the art. In one embodiment, the organic solvent is an ICH Class 3 or ICH Class 2 solvent. In one embodiment, the organic solvent is non-toxic and / or volatile.In one embodiment, the organic solvent is ethanol, methanol, acetone, tetrahydrofuran, acetic acid, acetonitrile, anisole, 1-butanol, 2-butanol, butyl acetate, chloroform, cyclohexane, 1,1-diethoxypropane, 1,1-dimethoxymethane, 1,2-dimethoxyethane, 1,4-dioxane, 2,2-dimethoxypropane, dichloromethane, diethyl ether, diisopropyl ether, dimethyl sulfoxide, dimethylformamide, 2-ethoxyethanol, ethyl acetate, ethyl formate, ethylene glycol (1,2-ethanediol), formic acid, heptane, hexane, isobutyl acetate, isopropyl acetate, 2-methoxyethanol, 2-methyl-1-propanol, 3-methyl-1-butanol, 1-methyl-2-pyrrolidine, methyl acetate, methyl t-butyl ether, methyl buylketone, methylcyclohexane, methyl ethyl ketone (MEK), methyl isobutyl ketone, methyl isopropyl ketone. , methyltetrahydrofuran, n-methylpyrrolidine, 1-pentanol, 1-propanol, 2-propanol, pentane, petroleum ether, propyl acetate, pyridine, sulfolane, t-butyl alcohol, 2,2,4-trimethylpentane (i-octane), toluene, trichloroacetic acid, trichloroethylene, trifluoroacetic acid, xylol, and combinations thereof; preferably acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert-butyl methyl ether, DMSO, ethanol, ethyl acetate, ethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methyl ethyl ketone, 2-methyl-1-propanol, 2-methyltetrahydrofuran, pentane, 1-pentanol, 1-propanol, 2-propanol, propyl acetate, and trimethylamine; more preferably ethanol, acetone, and isopropanol. In one embodiment, the term "micronized macrolide solution" as used herein relates to a solution of a micronized macrolide, particularly a solution of a micronized macrolide dissolved in an organic solvent.In one embodiment, the terms "micronized macrolide solution" and "macrolide solution" are used interchangeably.

[0101] The term "mucoadhesive polymer" as used herein refers to a polymer that facilitates and / or mediates mucoadhesion. In one embodiment, the mucoadhesive polymer is selected from a) amphiphilic polymers and hydrophilic polymers, b) poly(methacrylates), c) cross-linked polyacrylic acid polymers, d) copolymers of methyl vinyl ether and maleic anhydride, e) polymers comprising poly(vinyl acetate) and / or poly(vinylpyrrolidone), and mucoadhesive cellulose derivatives. In one embodiment, the mucoadhesive polymer is selected from poly(acrylic acid), such as Carbopol® 934 or Carbopol® 971 NF; cross-linked poly(acrylic acid), such as polycarbophil, particularly poly(acrylic acid) cross-linked with divinyl glycol, more particularly its calcium salt; amylopectin, such as Proloc® TM 15; poly(methacrylic acid), such as Eudragit® L100 or Eudragit® S100; poly(methacrylate), such as Eudragit® E100, Eudragit® RL, or Eudragit® RS; poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof, such as Gantrez TM AN 119, Gantrez TM AN 139, Gantrez TM AN 149, Gantrez TM AN 169, or Gantrez TMMS-955; gelatin; polysaccharides such as alginate, chitosan, xanthan gum, hyaluronic acid, pectin, or pullulan; cellulose derivatives such as sodium carboxymethylcellulose (CMC), hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), hydroxyethyl methylcellulose (HEMC), hydroxyethyl cellulose (HEC), or methylcellulose (MC); poly(vinylpyrrolidone), such as Kollidon® 30LP or Kollidon® 90F; poly[(vinylpyrrolidone)-co-(vinyl acetate)], such as Kollidon® VA64; a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone), such as Kollidon® SR; poly(vinyl alcohol); and poly(vinyl acetate).

[0102] The term "amphiphilic polymer" as used herein refers to any amphiphilic polymer known to those skilled in the art, preferably a mucoadhesive amphiphilic polymer. Typically, an amphiphilic polymer has a hydrophilic (polar) portion and a hydrophobic (non-polar) portion. In one embodiment, the amphiphilic polymer is selected from polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymers, such as Soluplus®, polyoxyl castor oil, and D-α-tocopherol-polyethylene glycol-succinate (TPGS).

[0103] In one embodiment, hydrophilic polymer is a polymer that contains polar group or charged group.In one embodiment, these groups are nonionic, anionic, or cationic and / or zwitterionic.In one embodiment, hydrophilic polymer is soluble in water.For example, hydrophilic polymer can be selected from cellulose and cellulose derivatives such as carboxymethylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, hydroxypropylethylcellulose, sodium carboxymethylcellulose, ethyl or propylcellulose, polyacrylic acid, polyacrylate, polyvinylpyrrolidone, polyethylene glycol / polyvinyl alcohol copolymer, polyvinyl alcohol, polyethylene oxide polymer, polyethylene oxide / polyethylene glycol copolymer, polyacrylamide, polyethylene glycol, gelatin, collagen, alginate, pectin, pullulan, tragacanth, chitosan, alginic acid, arabinogalactan, galactomannan, agar, agarose, starch and starch derivatives, dextran, carrageenan, shellac, natural gum and / or copolymers thereof.

[0104] In one embodiment, the poly(methacrylate) is selected from neutral protonatable poly(methacrylates) and cationic poly(methacrylates), preferably copolymers of dialkylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trialkylaminoethyl methacrylate and methacrylic acid esters, particularly copolymers of dimethylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trimethylammonioethyl methacrylate and methacrylic acid esters. For example, the poly(methacrylate) can be Eudragit® L100, Eudragit® S100, Eudragit® E100, Eudragit® RL, or Eudragit® RS.

[0105] As used herein, the term "crosslinked polyacrylic acid polymer" refers to polyacrylic acid polymers and crosslinked polyacrylic acid polymers known to those skilled in the art, preferably crosslinked polyacrylic acid polymers, homopolymers, copolymers, and interpolymers, such as acrylic acid crosslinked with allyl sucrose or allyl pentaerythritol, acrylic acid crosslinked with allyl pentaerythritol, and acrylic acid C 10 -C 30 The cross-linked polyacrylic acid polymer may be Carbopol® 934 or Carbopol® 971 NF or polycarbophil.

[0106] The term "copolymer of methyl vinyl ether and maleic anhydride" as used herein refers to any copolymer of methyl vinyl ether and maleic anhydride / maleic acid known to those skilled in the art, such as copolymers of monoalkyl esters of poly(methyl vinyl ether / maleic acid) with various ester groups. In one embodiment, the copolymer of methyl vinyl ether and maleic anhydride is in the anhydride form or in hydrolyzed form, such as mixed sodium and calcium salts of methyl vinyl ether and maleic anhydride, e.g., Gantrez TM MS-955 polymer. In one embodiment, the copolymer of methyl vinyl ether and maleic anhydride is selected from poly(methyl vinyl ether-co-maleic anhydride), poly(methyl vinyl ether-co-maleic acid), the monoethyl ester of poly(methyl vinyl ether / maleic acid), a mixture of the monoethyl ester of poly(methyl vinyl ether / maleic acid) and the monobutyl ester of poly(methyl vinyl ether / maleic acid), and a mixed sodium / calcium salt of poly(methyl vinyl ether / maleic anhydride). In one embodiment, the copolymer of methyl vinyl ether and maleic anhydride, such as Gantrez TMis a mixed sodium and calcium salt of a copolymer of methyl vinyl ether and (hydrolyzed) maleic anhydride. In one embodiment, the copolymer of methyl vinyl ether and maleic anhydride is the complexing agent. For example, the copolymer of methyl vinyl ether and maleic anhydride is available from Gantrez TM AN 119, Gantrez TM AN 139, Gantrez TM AN 149, Gantrez TM AN 169, Gantrez TM AN-903, Gantrez TM S-96, Gantrez TM S-97, Gantrez TM ES-225, Gantrez TM ES-425, or Gantrez TM It could be MS-955.

[0107] As used herein, the terms "polymer comprising polyvinyl acetate and / or polyvinylpyrrolidone" and "polymer comprising polyvinyl acetate and polyvinylpyrrolidone" refer to any polyvinyl acetate and / or polyvinylpyrrolidone-containing polymer known to those skilled in the art, such as poly(vinylpyrrolidone), poly(vinyl acetate), poly[(vinylpyrrolidone)-co-(vinyl acetate)], and mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone). In one embodiment, the polyvinyl acetate and / or polyvinylpyrrolidone-containing polymer is selected from poly(vinylpyrrolidone), poly(vinyl acetate), poly[(vinylpyrrolidone)-co-(vinyl acetate)], such as a copolymer of 6 parts polyvinylpyrrolidone and 4 parts polyvinyl acetate, and mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone), such as an 8:2 mixture of poly(vinyl acetate) and poly(vinylpyrrolidone). In one embodiment, the polyvinyl acetate and / or polyvinylpyrrolidone-containing polymer comprises or consists of poly(vinylpyrrolidone). In one embodiment, the polymer containing polyvinyl acetate and / or polyvinylpyrrolidone comprises or consists of a vinylpyrrolidone-vinyl acetate copolymer, particularly poly[(vinylpyrrolidone)-co-(vinyl acetate)]. In one embodiment, the polymer containing polyvinyl acetate and / or polyvinylpyrrolidone comprises or consists of a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone). In one embodiment, the polymer containing polyvinyl acetate and / or polyvinylpyrrolidone comprises one or more, preferably multiple, vinyl acetate monomers and / or one or more, preferably multiple vinylpyrrolidone monomers. For example, the polymer containing polyvinyl acetate and / or polyvinylpyrrolidone can be Kollidon® 30LP, Kollidon® 90F, Kollidon® VA64, or Kollidon® SR.

[0108] The term "cellulose derivative" as used herein refers to cellulose and its derivatives. In one embodiment, the cellulose derivative is preferably a cellulose derivative that swells rather than dissolves upon contact with water and / or saliva. In one embodiment, the cellulose derivative is selected from hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), such as sodium carboxymethyl cellulose (CMC), hydroxypropyl methyl cellulose (HPMC), hydroxyethyl cellulose (HEC), methyl cellulose (MC), hydroxyethyl methyl cellulose (HEMC), ethyl cellulose (EC), and combinations thereof. In one embodiment, the mucoadhesive layer comprises a cellulose derivative that is a mucoadhesive cellulose derivative, such as a cellulose derivative selected from hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), hydroxypropyl methyl cellulose (HPMC), hydroxyethyl cellulose (HEC), methyl cellulose (MC), and combinations thereof. In one embodiment, the cellulose derivative soluble in organic solvents is HPC and / or EC. In one embodiment, CMC is soluble in water but not in organic solvents.

[0109] In a preferred embodiment, the mucoadhesive layer comprises hydroxypropyl cellulose as the mucoadhesive polymer, alone or in combination with another mucoadhesive polymer selected from: copolymers of methyl vinyl ether and maleic anhydride; poly(acrylic acid); cross-linked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; alginate, chitosan, xanthan gum, hyaluronic acid, pectin, or pullulan; carboxymethylcellulose; hydroxypropyl methylcellulose; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

[0110] In a particularly preferred embodiment, the mucoadhesive layer comprises hydroxypropyl cellulose as the mucoadhesive polymer in combination with another non-cellulosic mucoadhesive polymer selected from: a copolymer of methyl vinyl ether and maleic anhydride; poly(acrylic acid); cross-linked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; alginate, chitosan, xanthan gum, hyaluronic acid, pectin, or pullulan; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

[0111] In a particularly preferred embodiment, which can be combined with any of the other embodiments disclosed herein, particularly the embodiments in the preceding paragraphs, the mucoadhesive layer comprises a combination of two different hydroxypropyl cellulose types, the hydroxypropyl cellulose types differing in molecular weight, preferably a first type of hydroxypropyl cellulose having a weight-average molecular weight of approximately 370,000 and a second type of hydroxypropyl cellulose having a weight-average molecular weight of about 80,000. Preferably, in such a combination of two different hydroxypropyl celluloses, the first type of hydroxypropyl cellulose having a weight-average molecular weight of approximately 80,000 is present in a range of 15 to 43 wt.%, more preferably in a range of 30 to 43 wt.%, and the second type of hydroxypropyl cellulose having a weight-average molecular weight of about 370,000 is present in a range of 2 to 28 wt.%, more preferably in a range of 8 to 13 wt.%, or equally preferably in a range of 12 to 28 wt.%. In the above ranges, the weight percentages are based on the dry weight of each mucoadhesive layer.

[0112] As used herein, the term "hydroxypropyl cellulose having a viscosity of about 300-600 mPa s [weight average molecular weight, about 80,000-Klucel EF]" is meant to refer to hydroxypropyl cellulose having a weight average molecular weight of about 80,000 or a viscosity of about 300-600 mPa s, or both a weight average molecular weight of about 80,000 and a viscosity of about 300-600 mPa s; such type of hydroxypropyl cellulose is commercially available, for example, under the trademark "Klucel EF" (registered trademark).

[0113] As used herein, the term "hydroxypropyl cellulose having a viscosity of about 150-400 mPa s [weight average molecular weight, about 370,000 - Klucel GF]" is meant to refer to hydroxypropyl cellulose having a weight average molecular weight of about 370,000 or a viscosity of about 150-400 mPa s, or both a weight average molecular weight of about 370,000 and a viscosity of about 150-400 mPa s; such type of hydroxypropyl cellulose is commercially available, for example, under the trademark "Klucel GF" (registered trademark).

[0114] In embodiments in which the mucoadhesive layer comprises a combination of two different hydroxypropyl cellulose types, wherein such hydroxypropyl cellulose types differ in terms of their molecular weight, it is particularly preferred that such mucoadhesive layer additionally comprises a further mucoadhesive polymer, more preferably a non-cellulosic polymer. In particularly preferred embodiments, such non-cellulosic polymer is selected from: copolymers of methyl vinyl ether and maleic anhydride; poly(acrylic acid); cross-linked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; alginate, chitosan, xanthan gum, hyaluronic acid, pectin or pullulan; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

[0115] In yet other embodiments, wherein the mucoadhesive layer comprises a combination of two different hydroxypropyl cellulose types as described herein, wherein such hydroxypropyl cellulose types differ in terms of their molecular weight as described herein, it is also preferred, but alternatively preferred, that such mucoadhesive layer does not comprise any additional mucoadhesive polymer; and the mucoadhesive properties of the mucoadhesive film and / or the respective mucoadhesive layer are due solely to the presence of hydroxypropyl cellulose.

[0116] When preparing a mucoadhesive film, the layers of the film can be prepared individually and then bonded together, for example, via an intermediate layer and / or via an adhesive, and / or can be prepared on top of each other, for example, by first preparing a mucoadhesive layer, and then subsequently coating the mucoadhesive layer with a backing layer, optionally coating the mucoadhesive layer with an intermediate layer and a backing layer.For example, preparing a mucoadhesive film can include separately preparing a mucoadhesive layer, a backing layer, and optionally an intermediate layer, and then laminating the layers together, for example, by heat lamination or compression.For example, the mucoadhesive layer is molded, and then, after drying, a second layer, for example, an intermediate layer or a backing layer, is molded on top of the mucoadhesive layer.In one embodiment, the first and second layers, for example, the mucoadhesive layer and the intermediate layer or the backing layer, are attached to each other.In one embodiment, the mucoadhesive layer is not completely dried before applying the intermediate layer and / or the backing layer.In one embodiment, the layers can be compressed onto each other using a roll. In one embodiment, preparing the mucoadhesive film comprises preparing a mucoadhesive layer and a backing layer, and then joining the mucoadhesive layer and the backing layer, e.g., by compression or gluing. In one embodiment, preparing the mucoadhesive film comprises preparing a mucoadhesive layer, an intermediate layer, and a backing layer, and then joining the layers, e.g., by compression or gluing.

[0117] In one embodiment, the steps of preparing a mucoadhesive layer and preparing a backing layer are followed by a step of bonding the mucoadhesive layer and the backing layer together by compression and / or lamination, for example, heat lamination. Optionally, the mucoadhesive layer and / or the backing layer are dried or partially dried before the step of bonding the mucoadhesive layer and the backing layer together. In one embodiment, the steps of preparing a mucoadhesive layer, preparing an intermediate layer, and preparing a backing layer are followed by a step of bonding the mucoadhesive layer, the intermediate layer, and the backing layer together by compression and / or lamination, for example, heat lamination. Optionally, the mucoadhesive layer, the intermediate layer, and / or the backing layer are dried or partially dried before the step of bonding the mucoadhesive layer, the intermediate layer, and the backing layer together. In one embodiment, the bonding is performed by compression and / or lamination, for example, heat lamination.

[0118] In one embodiment, the backing layer is prepared on the surface of the mucoadhesive layer, for example, by coating the polymer mixture on the surface of the mucoadhesive layer, or is prepared separately and then bonded to the mucoadhesive layer, optionally via an intermediate layer. In one embodiment, when the mucoadhesive layer and the backing layer, optionally the intermediate layer, are prepared separately, the layers are prepared on a process liner. In one embodiment, when the mucoadhesive layer contains a micronized macrolide, preferably a micronized crystalline macrolide, the backing layer is prepared on the surface of the mucoadhesive layer, for example, by coating the polymer mixture on the surface of the mucoadhesive layer. In one embodiment, when the mucoadhesive layer contains a molecularly dissolved macrolide, the backing layer is prepared separately from the mucoadhesive layer, and the mucoadhesive layer and the backing layer are then bonded, optionally via an intermediate layer. In one embodiment, the mucoadhesive layer, the backing layer, and optionally the intermediate layer are each prepared on a process liner, and then bonded.

[0119] In one embodiment, the preparation of the mucoadhesive film comprises joining the mucoadhesive layer, the backing layer, and optionally the intermediate layer, for example by compression and / or lamination, for example by heat lamination. In one embodiment, the mucoadhesive layer, the backing layer, and optionally the intermediate layer are dried or partially dried before joining the layers. In one embodiment, the mucoadhesive layer, the backing layer, and optionally the intermediate layer are prepared separately and, after their preparation, are subsequently joined and / or prepared on top of each other.

[0120] The inventors have found that the adverse effects of gastrointestinal transit after oral application can be eliminated by the innovative buccal therapy system presented herein. Another favorable finding from the human kinetic study is the low variability in the pharmacokinetic (PK) profiles of both comparison groups: the PK profiles of the high-dose and low-dose groups varied on average by approximately 25%, which is very small for inter-patient variability for tacrolimus. The inventors have found that advantageously, buccal application can positively impact a central drawback of oral therapy.

[0121] As used herein, the terms "of the present invention," "according to the present invention," "in accordance with the present invention," and the like are intended to refer to all aspects and embodiments of the invention described and / or claimed herein.

[0122] The term "comprising" as used herein should be interpreted as encompassing both "including" and "consisting of," and both meanings are specifically intended, thus including individually disclosed embodiments according to the present invention. As used herein, "and / or" should be interpreted as a specific disclosure of each of the two specified features or components, regardless of the presence or absence of the other. For example, "A and / or B" should be interpreted as a specific disclosure of (i) A, (ii) B, and (iii) each of A and B, as if each were individually described herein. In the context of the present invention, the terms "about" and "approximately" indicate a range of precision that a person skilled in the art can understand to ensure the technical effect of the feature in question. This term typically indicates a deviation of ±20%, ±15%, ±10%, and, for example, ±5% from the indicated numerical value. As will be understood by a person skilled in the art, the specific deviation from the numerical value of a given technical effect will depend on the nature of the technical effect. For example, natural or biological technological effects may generally have greater deviations than man-made or engineered technological effects. Where an indefinite or definite article, e.g., "a," "an," or "the," is used when referring to a singular noun, this includes the plural of that noun unless otherwise specified.

[0123] Reference is now made to the following examples which are offered by way of illustration and not by way of limitation of the invention. [Example]

[0124] Example Example 1: Clinical Human Phase I Kinetic Study (Demonstration in Humans) First-in-human studies have been conducted with a mucoadhesive buccal film formulation of tacrolimus: Research rationale The primary objective of this first-in-human pilot study was to investigate the PK of tacrolimus MBF in healthy male subjects (proof-of-human trial). Additionally, we evaluated the safety and tolerability of tacrolimus MBF and measured the residual amount of tacrolimus in MBF after use. To overcome the PK problems associated with currently marketed oral tacrolimus formulations, a novel tacrolimus formulation in the form of a mucoadhesive buccal film (MBF) was developed.

[0125] [Table 1]

[0126] Methods (study design): The study was conducted as a single-center, open-label, non-randomized, multiple-dose, parallel-group design with two sequential arms. To overcome the pharmacokinetic problems associated with currently available oral tacrolimus formulations, a new formulation of tacrolimus in the form of MBF was developed. The objectives of this study were to investigate the pharmacokinetics of tacrolimus MBF in healthy male subjects. Additionally, we evaluated the safety and tolerability of tacrolimus MBF and measured the residual amount of tacrolimus in MBF after use. The procedure was planned as follows:

[0127] [Table 2]

[0128] The study was structured as follows: Screening Visit: Within 2 to 1 days prior to the first dose of study treatment. Day 1: Subjects are admitted to the study ward and assigned to study treatment. Day 1: Two single doses of the study treatment were administered via the buccal route. The time to start the second dose in a given subject was 4 hours. Each MBF remained attached to the buccal mucosa for 1 hour. Day 2: Subjects will be discharged from the study ward if all protocol procedures are completed and there are no medical objections. Follow-up visit: 1 to 3 days after the last dose of study treatment. Safety and tolerability parameters were collected throughout the study period, from screening to follow-up. Blood samples for PK analysis were collected up to 28 hours after the first dose of tacrolimus.

[0129] Number of subjects (planned and analyzed): There were two sequential treatment groups receiving tacrolimus MBF. At least 10 healthy subjects were planned to be assigned to the study treatment groups, with at least 5 evaluable subjects per treatment group expected to complete the study. Finally, 10 subjects were assigned to treatment groups: 5 in treatment group 1 and 5 in treatment group 2.

[0130] Diagnosis and main inclusion criteria: Weight ≥ 65 kg and ≤ 85 kg and body mass index (BMI) 18.0 to 30.0 kg / m 2 (Inclusive) healthy male subjects (18–40 years old, inclusive) were eligible.

[0131] [Table 3]

[0132] Treatment duration: The study duration was approximately 4 weeks per subject. On Day 1, subjects received two single doses of the study treatment. The time between the start of the two doses for a given subject was 4 hours. Each MBF was left in place on the buccal mucosa for 1 hour.

[0133] Statistical method: Pharmacokinetics: No formal statistical hypotheses were defined in this exploratory study. Appropriate summary tables and graphs were presented. Safety: Safety data were assessed narratively.

[0134] Summary and conclusion: Pharmacokinetics: In treatment group 1 (T1), the whole blood concentration was below the LLOQ in all subjects until 0.5 hours after the start of the first MBF application, but exceeded the LLOQ in all subjects 3 hours after the start of the first MBF application (= 2 hours after the end of the first MBF application).In treatment group 2 (T2), the whole blood concentration was below the LLOQ in all subjects until 0.25 hours after the start of the first MBF application, but exceeded the LLOQ in all subjects 1.5 hours after the start of the first MBF application. The mean whole blood concentration curves for both treatment groups reached a first peak 3 hours after the start of the first MBF application (= 2 hours after the end of the first MBF application), then gradually decreased, and reached a second peak 8 hours after the start of the first MBF application (= 4 hours after the start of the second MBF application = 3 hours after the end of the second MBF application), with mean concentrations of 0.9526 ng / mL (T1) and 6.548 ng / mL (T2). The results are presented in Figure 1. Geometric mean AUC of tacrolimus 0-4 was significantly higher in treatment group 2 (12.0 mg; 6.063 h*ng / mL) than in treatment group 1 (8.8 mg; 1.044 h*ng / mL). 0-tlast( T1: 13.594 h*ng / mL; T2: 56.021 h*ng / mL) and geometric mean C max (T1: 0.878 ng / mL; T2: 3.771 ng / mL) were similar. t max The median values ​​were similar in both treatment groups (T1: 8.083 hours after the start of the first MBF application; = 3.083 hours after the end of the second MBF application; T2: 8.017 hours after the start of the first MBF application; = 3.017 hours after the end of the second MBF application). The geometric mean half-lives and elimination rates were also similar (t1 / 2: T1: 14.475 h and T2: 14.926 h; λz: T1: 0.04789 1 / h and T2: 0.04644 1 / h). Subjects showed individual variability in pharmacokinetic parameters, particularly in treatment group 2. Subject 015 had the highest C maxThe value (17.5ng / ml) was the lowest C max The value was 24.9 times higher than that of subject 012 (0.703 ng / ml). Subject 015 had the highest AUC 0-tlast The value (224.76h*ng / ml) was the lowest AUC 0-tlast This was 19.5 times higher than the value (subject 012; 11.51 h*ng / ml).

[0135] [Table 4]

[0136] Overall, a large amount of tacrolimus remained in the MBF. The mean percentage of tacrolimus remaining in the MBF after each dose was higher in Treatment Group 1 (1st application: 91.076%, 2nd application: 85.650%) than in Treatment Group 2 (1st application: 78.214%, 2nd application: 74.296%).

[0137] First human kinetic data of tacrolimus - MBF Individual tacrolimus blood levels in 10 healthy volunteers (5 per group) following bid administration of tacrolimus, followed by a second dose 4 hours later, are shown in Figure 2. The mean blood level profiles of tacrolimus administered bid in 10 healthy volunteers (5 per group) followed by a second dose 4 hours later are shown in Figure 3.

[0138] conclusion Initial kinetic results in humans demonstrate that systemic exposure is achieved after buccal administration of a unidirectional release MBF formulation, providing proof-of-concept in humans. In clinical practice, tacrolimus dosing is typically adjusted according to body weight, but in this pilot PK study, all subjects in a particular dose group received a fixed nominal dose, which likely contributed to the observed variability in tacrolimus blood levels.

[0139] Example 2: Dosing Regimen for Buccal Application Current recommendations for standard oral tacrolimus therapy (Prograf caps.) for adult patients who have undergone kidney, liver, or heart transplants are as follows:

[0140] [Table 5]

[0141] Tacrolimus TDM is recommended for all patients receiving Prograf. Monitoring tacrolimus blood levels in conjunction with other laboratory and clinical parameters is considered an essential adjunct to patient management for the assessment of rejection, toxicity, dose adjustments, and compliance.

[0142] This novel buccal route of administration will introduce a new dosing regimen for daily tacrolimus therapy, allowing for administration at 24-hour intervals. Based on kinetic data from preclinical animal studies, twice-daily application (bid) with a 4-hour interval between the first and second doses was selected for human kinetic studies. The blood level profile of the high-dose group (n = 5 healthy subjects) loaded with 12 mg of tacrolimus per film unit demonstrates that clinically relevant blood levels are achieved with a twice-daily application system and a 4-hour interval between the first and second doses.

[0143] To validate this particular asymmetric dosing regimen for buccal application, further animal studies were performed under GLP conditions using three different application variants: Study design (GLP study): Intra-subject 3-way crossover Three-group comparative study Bid application (5 naive minipigs per group, GLP)

[0144] [Table 6]

[0145] Results: The individual PK profiles for each group are shown in Figure 4. As shown in Figure 5, the plot of the mean curves demonstrates that in the bid dosing regimen, the highest blood levels are achieved when the second dose unit is administered 3 hours later. The asymmetric dosing scheme tested is highly advantageous. In particular, the asymmetric 3-hour bid application system shows superior results in terms of achieved blood levels.

[0146] Example 3: Dose reduction based on improved bioavailability The standard daily dose of oral tacrolimus (Prograf) in SOT patients is: mg tacrolimus / kg bw / day It is calculated as follows.

[0147] The starting daily dose of oral tacrolimus in a 70 kg kidney transplant patient is: 0.2 mg x 70 kg / day = 14 mg daily dose It is calculated as: This is split into two single doses (morning dose / evening dose). This dosing guideline is part of the international registration document for oral Prograf capsules, the current standard of oral tacrolimus therapy.

[0148] The situation is different for buccal therapy systems: In the human studies presented herein, buccal films were used at two different dose levels: · Low dose: 8.8 mg tacrolimus loading = 17.6 mg (bid). High dose: 12 mg tacrolimus loading = 24 mg (bid).

[0149] The residence time of the film on the buccal mucosa after application is 60 minutes; the film is then actively removed from the oral cavity.

[0150] Only the bioavailable portion of the active ingredient that penetrates the film matrix into the mucosal tissue within 60 minutes and is subsequently released into the systemic circulation by passive diffusion is responsible for achieving therapeutic blood levels. This absorption process typically occurs in three steps: Step 1: Drug release into mucosal tissue Step 2: Formation of API depots in the mucosa Step 3: Diffusion into the bloodstream The amount of residual tacrolimus in the used films after removal from the cheek was analyzed to determine the amount of drug that had penetrated the mucosal tissue to form a depot for diffusion. Figure 6 shows the residual drug levels in the used films for both dose groups.

[0151] result: 1. Remaining tacrolimus after application (average of film) 8.8mg film: 88.4%±5.6% (low dose group) 12mg film: 81.8% ± 8.0% (high dose group) Over 80% of the applied tacrolimus remains within the film. 2. Advantageously, a smaller buccal dose of tacrolimus is required to achieve therapeutic blood levels: Medication example (patient, bw=80kg): Standard oral dose = tacrolimus 16 mg / day Buccal application = tacrolimus 3.2 mg / day

[0152] These data advantageously confirm that the bioavailability of drugs absorbed across the buccal mucosa is higher compared to oral application, as only about 20% of the oral dose is required to achieve therapeutic levels after buccal application.

[0153] Without being bound by any theory, the inventors believe that these effects can be scientifically explained by the low bioavailability of tacrolimus (BCS Class II), the significant first-pass effect, and the fact that pre-systemic degradation of tacrolimus after oral administration is irrelevant for buccal application. Advantageously, the drug burden on patients can be reduced by approximately 80%, which can also significantly alter the side effect profile of tacrolimus therapy.

[0154] Example 4: Blood level fluctuations One of the most challenging aspects of tacrolimus therapy management is the large intra- and inter-patient variability in the concentration-time profile of tacrolimus after oral administration. The reasons for this variation are well known and well documented in the international literature: (1) Low and variable drug bioavailability (4-90%) due to poor water solubility, (2) first-pass effect, (3) degradation before systemic circulation, and (4) food effect. All these effects are not relevant for buccal delivery systems due to better bioavailability and direct access to the systemic circulation.

[0155] This is also demonstrated by the data from this human study (Figure 7), which shows the normalized concentration-time curves (percentage of the maximum tacrolimus blood concentration at each measurement time point) for both dose groups.

[0156] Comparison of the mean normalized concentration-time profiles for both groups illustrates the relatively low inter-individual variability (CV) of tacrolimus blood levels in both groups over a 24-hour interval (Figure 8). Internationally reported variability in tacrolimus blood levels after oral administration is greater than 80%, while the variability in blood levels after buccal administration shown herein is classified as low at 25-30%. Therefore, advantageously, the buccal administration described herein reduces blood level variability compared to oral administration.

[0157] Without being bound by any theory, the inventors believe that one of the reasons for the smaller fluctuation in blood level profile after buccal administration compared to oral administration of tacrolimus is the better bioavailability of the active substance after buccal administration (see Example 2).

[0158] The primary objective of patient-individualized dose titration with tacrolimus is to adjust the blood level profile over a 24-hour treatment interval so that it remains within the therapeutic range of 5 to 20 ng / ml in whole blood on average over 24 hours (mean residence time (MRT)). Of particular clinical importance in this context is the variability of tacrolimus blood levels—the lowest level at the end of the 24-hour dosing interval (C trough ), i.e., the pre-dose level before the next dose.

[0159] There is a direct correlation between time in the therapeutic range and clinical response one year after transplantation. Consequently, dosing regimens (bid) and individual daily doses are preferably designed so that blood levels of tacrolimus are still in the therapeutic range at the end of the 24-hour dosing interval.

[0160] In this human kinetic study, the 24-hour C trough It was shown that the blood level represents 30-40% of the Cmax value. trough Blood levels are important for safe and clinically effective therapy management with tacrolimus.

[0161] Additionally, the lower inter-individual blood level variability compared to oral therapy highlights a further advantage of the innovative buccal therapy system presented herein.

[0162] Example 5: Exemplary Compositions of Mucoadhesive Films Applied tacrolimus mucoadhesive buccal film (8.8 mg / 4.4 cm 2 ) had an exemplary composition for each layer as follows: (The component percentage values ​​used in the tables below are weight percentages and are given based on the dry weight of each layer in which each component is present.)

[0163] [Table 7]

[0164] Applied tacrolimus adhesive buccal film (12.0 mg / 6.0 cm 2 ) had an exemplary composition for each layer as follows:

[0165] [Table 8]

[0166] Additionally, a tacrolimus adhesive buccal film (16.5 mg / 5 cm 2 ) had an exemplary composition for each layer as follows:

[0167] [Table 9]

[0168] Additionally, a tacrolimus adhesive buccal film (14.4 mg / 5 cm 2 ) had an exemplary composition for each layer as follows:

[0169] [Table 10]

[0170] [Table 11]

[0171] Example 6: In vivo testing of mucoadhesive films Pigs were chosen as the animal model because they closely resemble the functional anatomical relationships of humans. The animals were fasted (overnight) for at least 16 hours before treatment and were anesthetized during the procedure. A film having the composition disclosed in the last table of Example 5 was placed with the mucoadhesive layer side on the mucosa of the animal's mouth and gently pressed until the film adhered. The film was left on the buccal mucosa for 30 or 60 minutes and then removed.

[0172] Blood samples were collected via an intravenous catheter. 2 ml of blood was collected from all animals into K2EDTA-coated tubes for determination of tacrolimus blood levels. After blood collection, the blood tubes were cooled on ice and stored in a freezer (temperature ranging from -30°C to -15°C) until analysis. Tacrolimus concentrations were determined by a reliable LC-MS / MS method. Pharmacokinetic parameters were calculated from the time course of blood concentrations. The calculated AUC was normalized to the applied dose and determined in relation to the individual animal weight (AUC_24h / dose).

[0173] [Table 12]

[0174] [Table 13]

[0175] Tacrolimus was detected in the blood of all pigs after buccal administration, with maximum concentrations reached at 2 and 4 hours (t_max), respectively. The films did not cause local irritation to the buccal mucosa. There was no treatment-related mortality, clinical signs, or weight changes.

[0176] The features of the invention disclosed in this specification, in the claims and / or in the accompanying drawings may, both individually and in any combination thereof, be material for realizing the invention in diverse forms thereof.

Claims

1. 1. A macrolide for use in a method for the prevention or treatment of an immune disease or disorder, preferably transplant rejection, comprising administering said macrolide at least twice daily; said administering at least twice daily comprises administering a first dose at a first time point and a second dose at a second time point, said second time point being a time point within a range of from about 1 hour to about 7 hours, preferably from about 2 hours to about 5 hours, more preferably from about 2.5 hours to about 3.5 hours, and even more preferably about 3 hours, from said first time point; Preferably, the macrolide for said use is administered by buccal administration.

2. the macrolide is administered in the form of a mucoadhesive layer, preferably a mucoadhesive buccal layer; 2. A macrolide for use according to claim 1, wherein said administering preferably comprises attaching said mucoadhesive layer to a body cavity of a patient, preferably to the buccal cavity of a patient and / or to a mucosa of a patient.

3. 3. A macrolide for use according to claim 1 or 2, wherein said administering comprises administering each of said first dose and said second dose, preferably each in the form of a mucoadhesive layer, for a period ranging from about 1 minute to about 7 hours, preferably from about 10 minutes to about 3 hours, more preferably from about 20 minutes to about 2 hours, even more preferably from about 30 minutes to about 90 minutes, and even more preferably about 60 minutes.

4. 10. A macrolide for use according to any one of the preceding claims, wherein said administering comprises administering said first dose over a period ranging from about 30 minutes to about 90 minutes, preferably about 60 minutes; and administering said second dose at a time within a range from about 2.5 hours to about 3.5 hours from said first time point, preferably at about 3 hours, over a period ranging from about 30 minutes to about 90 minutes, preferably about 60 minutes.

5. said first dose and said second dose preferably comprise, prior to said administration, said macrolide in an amount in the range of from about 0.001 mg to about 250 mg, preferably in the range of from about 0.01 mg to about 50 mg, more preferably in the range of from about 0.05 mg to about 25 mg, even more preferably in the range of from about 0.1 mg to about 15 mg, even more preferably in the range of from about 0.5 mg to about 13 mg, optionally in an amount in the range of from about 8 mg to about 12 mg; and / or 10. A macrolide for use according to any one of the preceding claims, wherein said macrolide is administered by buccal administration and said administration of said macrolide at least twice daily comprises administering a total daily amount of macrolide that is ≦about 40%, preferably ≦about 30%, more preferably ≦about 25%, even more preferably ≦about 20% of the total daily amount administered by oral administration.

6. The macrolide is present in the mucoadhesive layer at a concentration of 1 cm or more prior to administration. ２ and preferably in an amount ranging from about 0.01 mg to about 10 mg per cm of said mucoadhesive layer. ２ and more preferably in the range of about 0.1 mg to about 5 mg per cm of said mucoadhesive layer. ２ and even more preferably from about 1.5 mg to about 3.5 mg per cm of said mucoadhesive layer. ２ 6. The macrolide for use according to any one of claims 2 to 5, wherein the macrolide is present in an amount ranging from about 1.8 mg to about 2.5 mg based on the total amount of the macrolide.

7. The macrolide is: - administered in the form of a mucoadhesive layer comprising a mucoadhesive polymer, preferably a polymer selected from a) amphiphilic and hydrophilic polymers, b) poly(methacrylates), c) crosslinked polyacrylic acid polymers, d) copolymers of methyl vinyl ether and maleic anhydride, e) polymers comprising polyvinyl acetate and / or polyvinylpyrrolidone, and combinations thereof; Here, preferably, the amphiphilic polymer is selected from polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, polyoxyl castor oil and D-α-tocopherol-polyethylene glycol-succinate (TPGS); and / or the poly(methacrylate) is selected from neutral protonatable poly(methacrylates) and cationic poly(methacrylates), preferably copolymers of dialkylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trialkylammonioethyl methacrylate and methacrylic acid esters, in particular copolymers of dimethylaminoethyl methacrylate and methacrylic acid esters, and copolymers of trimethylammonioethyl methacrylate and methacrylic acid esters; and wherein, optionally, the mucoadhesive layer comprises: cellulose derivatives, preferably selected from hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), hydroxypropyl methyl cellulose (HPMC), hydroxyethyl cellulose (HEC), methyl cellulose (MC), hydroxyethyl methyl cellulose (HEMC), ethyl cellulose (EC), and combinations thereof; a plasticizer, preferably selected from glycerol, polyethylene glycol, low molecular weight polyethylene glycol such as PEG 200 or PEG 400, propylene glycol, sorbitol, triacetin, and tributyl citrate; more preferably glycerol; and / or a colorant, preferably TiO ２ ; further comprising 10. A macrolide for use according to any one of the preceding claims, wherein, optionally, said macrolide is present in said layer in the form of a micronised macrolide, preferably a micronised crystalline macrolide and / or in a molecularly dissolved non-nanoparticulate form.

8. said at least twice-daily administration provides a first peak of whole blood concentrations of the macrolide at a time within a range of from about 2 hours to about 4 hours, preferably from about 2.5 hours to about 3.5 hours, more preferably about 3 hours, from said first time point, and a second peak of whole blood concentrations of the macrolide at a time within a range of from about 7 hours to about 9 hours, preferably from about 7.5 hours to about 8.5 hours, more preferably about 8 hours, from said first time point; 10. A macrolide for use according to any one of the preceding claims, wherein, if necessary, said at least twice daily administration provides a second peak of whole blood concentration of said macrolide at a time within the range of from about 3 hours to about 5 hours, preferably from about 3.5 hours to about 4.5 hours, more preferably at about 4 hours, from said second time point.

9. 10. The macrolide for use according to any one of the preceding claims, wherein said at least twice daily administration provides a whole blood concentration of said macrolide in the range of from about 0.01 ng / mL to about 50 ng / mL, preferably in the range of from about 3 ng / mL to about 20 ng / mL, more preferably in the range of from about 5 ng / mL to about 20 ng / mL, for a period of about 24 hours.

10. 10. The macrolide for use according to any one of the preceding claims, wherein said at least twice daily administration provides a coefficient of variation of less than 40%, preferably less than 35%, more preferably less than 32%, even more preferably less than or equal to 30%, such as in the range of 20% to 30% or in the range of 25% to 30%.

11. said administering at least twice daily - t in the range of about 3 hours to about 10 hours from said first point in time, preferably in the range of about 7 hours to about 9 hours from said first point in time, more preferably in the range of about 7.5 hours to about 8.5 hours from said first point in time ｍａｘ and / or - in the range of about 1 hour to about 6 hours from said second time point, preferably in the range of about 2 hours to about 5 hours from said second time point, more preferably in the range of about 3.5 hours to about 4.5 hours from said second time point, for example about 4 hours from said second time point ｍａｘ 10. A macrolide for use according to any one of the preceding claims, which provides:

12. said administering at least twice daily - C in the range of about 0.05 ng / mL to about 50 ng / mL, preferably in the range of about 0.1 ng / mL to about 30 ng / mL, more preferably in the range of about 0.7 ng / mL to about 20 ng / mL, even more preferably in the range of about 5.5 ng / mL to about 20 ng / mL ｍａｘ and / or - C in the range of about 0.01 ng / mL to about 18 ng / mL, preferably in the range of about 0.5 ng / mL to about 16 ng / mL, more preferably in the range of about 3 ng / mL to about 15 ng / mL, such as in the range of about 3 ng / mL to about 7 ng / mL or in the range of about 5 ng / mL to about 7 ng / mL ｔｒｏｕｇｈ 10. A macrolide for use according to any one of the preceding claims, which provides:

13. wherein said administering at least twice daily is carried out for a period of at least two days, preferably at least one week, more preferably at least one month, and even more preferably at least one year; 10. A macrolide for use according to any one of the preceding claims, wherein, optionally, said administration at least twice daily is carried out for the life of the patient when said macrolide is used for the prevention or treatment of transplant rejection.

14. 10. A macrolide for use according to any one of the preceding claims, wherein said macrolide is administered to a human patient; preferably, said patient is not a fasting patient.

15. the macrolide is selected from a macrolide immunosuppressant, a macrolide antibiotic, and a macrolide antifungal; Preferably, - the macrolide immunosuppressant is selected from tacrolimus, sirolimus, everolimus and pimecrolimus; the macrolide antibiotic is selected from erythromycin, clarithromycin, azithromycin, roxithromycin, josamycin, spiramycin, telithromycin, tylosin and fidaxomicin; and / or the macrolide antifungal agent is selected from polyenes, in particular nystatin, natamycin and amphotericin B; A macrolide for use according to any one of the preceding claims, wherein more preferably said macrolide is tacrolimus or sirolimus, even more preferably tacrolimus.

16. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: A) The mucoadhesive layer comprises: about 12 to about 25 weight percent tacrolimus; about 27 to about 31 weight percent ethylcellulose; about 28 to about 36 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; about 6 to about 10 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 5 weight percent of a calcium / sodium copolymer of methyl vinyl ether and maleic anhydride; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; or B) The mucoadhesive layer comprises: about 18 to about 25 weight percent tacrolimus; about 3 to about 13 weight percent ethylcellulose; about 29 to about 42 weight percent hydroxypropyl cellulose having a viscosity of about 300-600 mPa.s [weight average molecular weight, about 80,000 - Klucel EF]; From about 1 to about 28 weight percent of hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 10 weight percent crosslinked poly(acrylic acid); A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

17. 17. The tacrolimus mucoadhesive film of claim 16, wherein the backing layer comprises ethyl cellulose.

18. 18. The tacrolimus mucoadhesive film of claim 17, wherein the backing layer comprises ethyl cellulose in the range of about 50 to about 62 weight percent, based on the dry weight of the backing layer, or the backing layer comprises ethyl cellulose in the range of about 25 to about 27 weight percent, based on the dry weight of the backing layer.

19. 19. The tacrolimus mucoadhesive film of claim 16, 17 or 18, wherein the tacrolimus film comprises 8 mg to 17 mg of tacrolimus.

20. 20. The tacrolimus mucoadhesive film of claim 19, wherein the tacrolimus film comprises 8.8 mg or 12 mg or 14.4 mg or 16.5 mg of tacrolimus.

21. The tacrolimus mucoadhesive film of any one of claims 16 to 20, further comprising an adhesive layer disposed between the mucoadhesive layer and the backing layer.

22. 21. The tacrolimus mucoadhesive film of any one of claims 16 to 20, wherein the mucoadhesive layer and the backing layer are in direct contact with each other and there is no layer disposed between the mucoadhesive layer and the backing layer.

23. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 12 to about 13 weight percent tacrolimus, or about 19 to about 23 weight percent tacrolimus; about 27 to about 30 weight percent ethylcellulose; about 30 to about 35 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; About 7 to about 8 weight percent of hydroxypropyl cellulose having a viscosity of 150-400 mPa.s [weight average molecular weight, about 370,000 - Klucel GF]; and about 3 to about 4 weight percent of a calcium / sodium copolymer of methyl vinyl ether and maleic anhydride; A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

24. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 19 to about 22 weight percent tacrolimus; about 3 to about 13 weight percent ethylcellulose; about 29 to about 42 weight percent hydroxypropyl cellulose having a viscosity of about 300-600 mPa.s [weight average molecular weight, about 80,000 - Klucel EF]; From about 1 to about 28 weight percent of hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 10 weight percent crosslinked poly(acrylic acid); A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

25. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 5 to about 14 weight percent tacrolimus; about 29 to about 33 weight percent ethylcellulose; about 34 to about 41 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; about 7 to about 10 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 4 weight percent of a copolymer of methyl vinyl ether and maleic anhydride; A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

26. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 18 to about 20 weight percent tacrolimus; about 11 to about 13 weight percent ethylcellulose; about 30 to about 32 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; about 11 to about 13 weight percent of hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa s; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; and The tacrolimus mucoadhesive film does not comprise a mucoadhesive layer comprising a polymer selected from the group consisting of a copolymer of methyl vinyl ether and maleic anhydride; poly(acrylic acid); crosslinked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or a salt thereof; gelatin; polysaccharides, in particular alginate, chitosan, xanthan gum, hyaluronic acid, pectin or pullulan; carboxymethylcellulose; hydroxypropyl methylcellulose; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; a mixture of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

27. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 9 to about 23 weight percent tacrolimus; about 10 to about 13 weight percent ethylcellulose; about 15 to about 32 weight percent hydroxypropyl cellulose having a viscosity of about 300-600 mPa.s [weight average molecular weight, about 80,000 - Klucel EF]; about 5 to about 12 weight percent hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 6 to about 28 weight percent poly(methacrylate); A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

28. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 18 to about 20 weight percent tacrolimus; about 4 to about 6 weight percent ethylcellulose; about 31 to about 32 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; about 27 to about 28 weight percent of hydroxypropyl cellulose having a viscosity of about 150-400 mPa.s [weight average molecular weight, about 370,000 - Klucel GF]; wherein the weight percentage is based on the dry weight of the mucoadhesive layer; and The mucoadhesive layer is a tacrolimus mucoadhesive film that does not comprise a polymer selected from the group consisting of mucoadhesive polymers: copolymers of methyl vinyl ether and maleic anhydride; poly(acrylic acid); crosslinked poly(acrylic acid); poly(methacrylic acid); poly[(maleic anhydride)-co-(vinyl methyl ether)] or salts thereof; gelatin; polysaccharides, in particular alginate, chitosan, xanthan gum, hyaluronic acid, pectin or pullulan; carboxymethylcellulose; hydroxypropyl methylcellulose; poly(vinylpyrrolidone), poly[(vinylpyrrolidone)-co-(vinyl acetate)]; mixtures of poly(vinyl acetate) and poly(vinylpyrrolidone); poly(vinyl alcohol); and poly(vinyl acetate).

29. 1. A tacrolimus mucoadhesive film suitable for buccal administration comprising a mucoadhesive layer and a backing layer: The mucoadhesive layer comprises: about 18 to about 20 weight percent tacrolimus; about 4 to about 12 weight percent ethylcellulose; about 31 to about 42 weight percent hydroxypropyl cellulose [weight average molecular weight, about 80,000 - Klucel EF] having a viscosity of about 300-600 mPa.s; About 2 to about 23 weight percent of hydroxypropyl cellulose [weight average molecular weight, about 370,000 - Klucel GF] having a viscosity of about 150-400 mPa.s; and about 2 to about 6 weight percent crosslinked poly(acrylic acid); A tacrolimus mucoadhesive film, wherein the weight percentage is based on the dry weight of the mucoadhesive layer.

30. 30. A method for treating a human patient who has received an allogeneic solid organ transplant and is in need of prevention of transplant rejection, comprising buccal administration of the mucoadhesive film of any one of claims 16 to 29 to the patient twice daily.

31. The method according to claim 30, wherein the buccal administration comprises administering a first mucoadhesive film according to any one of claims 16 to 29 for about 30 to 60 minutes, preferably 60 minutes, or equally preferably 30 minutes, followed by buccal administration of a second mucoadhesive film according to any one of claims 16 to 29 to a patient for about 30 to 60 minutes, preferably 60 minutes, or equally preferably 30 minutes, after a lapse of about 3 to 5 hours without administration.

32. C in human patients 24 hours after twice-daily buccal administration ｔｒｏｕｇｈ Blood levels of tacrolimus C ｍａｘ 32. The method of claim 30 or 31, wherein the value is about 30-40%.

Images