NZ731512B2

NZ731512B2 - Compositions comprising cyclosporin

Info

Publication number: NZ731512B2
Application number: NZ731512A
Authority: NZ
Inventors: Vincenzo Aversa; Ivan Coulter; Donnell Daire O; Of Calhane Wyatt Renaud; Monica Rosa
Original assignee: Sublimity Therapeutics Limited
Priority date: 2014-11-07
Filing date: 2015-11-06
Publication date: 2022-06-28

Abstract

The present invention relates to a formulation comprising a pharmaceutically active ingredient, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, and coating. The invention also relates to the use of the formulation in the treatment and prevention of disorders of the gastrointestinal tract. Also disclosed are methods for preparing the formulations. ent and prevention of disorders of the gastrointestinal tract. Also disclosed are methods for preparing the formulations.

Description

Compositions comprising cyclosporin This invention relates to a composition comprising a pharmaceutically active ingredient and a surfactant. The invention also relates to the use of the composition in the treatment and prevention of disorders, for example disorders of the gastrointestinal tract. Also disclosed are methods for preparing the compositions.

BACKGROUND Cyclosporin A is a cyclic polypeptide which has immunosuppressive and anti-inflammatory properties. The compound has been approved for the prevention of organ rejection following kidney, liver, heart, combined heart-lung, lung or pancreas transplantation, for the prevention of rejection following bone marrow lantation; the treatment and prophylaxis of Graft Versus Host Disease ; psoriasis; atopic dermatitis, rheumatoid arthritis and nephrotic syndrome (Neoral™ Summary of Product Characteristics 24/02/2012). Cyclosporin A may also be useful for the treatment of a range of other diseases including for the treatment of severe recalcitrant plaque psoriasis Bechet's disease, anemia, myasthenia gravis and various conditions affecting the GI tract, including irritable bowel syndrome, Crohn’s disease, s, including ulcerative colitis, diverticulitis, pouchitis, proctitis, Gastro- lntestinal Graft Versus Host Disease (GI-GVHD), colorectal carcinoma and arcinoma as well as ischemia induced disease. A range of other diseases may benefit from ent with cyclosporin A (Landford et al. (1998) Ann Intern Med;128: 1021-1028) the entirety of which is incorporated herein by reference. Cyclosporin A has been used to treat a number of gastrointestinal conditions including inflammatory bowel disease (Sandborn WJ, a critical review of porin therapy in inflammatory bowel disease, Inflamm Bowel Dis. 1995;1:48–63), including ulcerative colitis iger et al, inary report (cyclosporine in the ent of severe ulcerative colitis), Lancet. 1990;336:16–19; Cohen et al, Intravenous cyclosporine in ulcerative colitis (a five-year experience), Am J Gastroenterol. 1999;94:1587–1592).

However porin A has a number of undesirable side effects including hypertension, impaired renal function, and neurotoxicity (Feutren et al, Risk factors for cyclosporine-induced nephropathy in patients with auto-immune diseases, International kidney biopsy ry of cyclosporine for mune diseases, N Engl J Med. 1992;326:1654–1660; ks et al., Neurotoxicity in liver transplant recipients with cyclosporine immunosuppression, ogy. 1995;45:1962–1964; and Porter et al, Cyclosporine-associated hypertension, National High Blood Pressure Education Program. Arch Intern Med. 1990;150:280–283).

Cyclosporin A is available as an intravenous formulation; Sandimmun™, which is a solution of 50 mg/ml of cyclosporin A in ethanol and hoxylated castor oil (for example Kolliphor™ EL). The product is also available as orally administered ations, including a soft gelatin e containing a solution of cyclosporin A in l, corn oil and lineoyl macrogolglycerides (Sandimmune™ Soft Gelatin capsules) and as an orally administered on ning the cyclosporin dissolved in olive oil, ethanol, and labrafil M 1944 CS (polyethoxylated oleic glycerides) (Sandimmune™ Oral Solution). More recently a microemulsion concentrate formulation has been approved containing cyclosporin A dissolved in DL-α-tocopherol, te ethanol, propylene glycol, P213428WO 2 corn no-di-triglycerides, polyoxyl 40 hydrogenated castor oil (Neoral™). Following oral administration the Neoral™ formulation results in the formation of a microemulsion and is stated to have an improved bioavailability compared to orally stered Sandimmune™. These orally administered cyclosporin A compositions are all instant release compositions and cyclosporin A will be t at high concentration in the stomach and small intestine from where it is systemically absorbed.

Sandborn et al.(J Clin Pharmacol. 1991 ; 31 ) determined the relative systemic absorption of cyclosporin following oral and intravenous as well as oil- and water-based enemas.

Based on negligible plasma cyclosporin concentrations observed following enema administration, it was suggested that cyclosporin, even when solubilised, is poorly absorbed from the colon. The enemas however demonstrated considerable efficacy in the treatment of inflammatory bowel disease (Ranzi T, et al, Lancet :97). Intravenous or orally administered cyclosporin efficacy in the treatment of inflammatory bowel disease is dose dependent, requiring high doses to ensure adequate concentration reaches the colon. Systemic ty is known to be dose and duration dependent.

Formulating pharmaceutically active ients into a form suitable for administration to a t is a developed area of science. It is also a key consideration for the efficacy of a drug. There are many examples of methods for formulating drugs and other active ingredients. The aim of these formulations are varied and can range from increasing systemic tion, allowing for a new route of administration, improving bioavilability, reducing metabolism of the active, or avoiding undesirable routes of administration. properties which release cyclosporin in at least the colon. WO2010/133609 discloses compositions comprising a water-soluble polymer matrix in which are dispersed droplets of oil, the compositions comprising a modified release g. The disclosed compositions also contain an active principle.

There remains a need for orally administered cyclosporin A compositions which provide high levels of cyclosporin A in the lower GI tract, particularly in the colon and absorption of the cyclosporin A from the luminal contents into the tissues of the GI tract, particularly into the colonic tissue, for the ent of conditions of the lower GI tract such as ulcerative colitis. Such compositions bly minimise the systemic blood exposure to cyclosporin A thereby minimising the undesirable side effect associated with systemic exposure to porin A. Particularly there is a need for orally administered compositions which have a low exposure/ area under the curve (AUC) and/or low peak blood concentration (Cmax) compared to the orally administered product Neoral™ and/or cyclosporin A administered intravenously as for example Sandimmune™.

Cyclosporin A is oil e; it is hydrophobic. Upon contact of a porin solution with water, the cyclosporin can form a solid by itating or crystalising out of solution. Precipitation or crystallisation of cyclosporin from solution can occur when the porin solution is t in an oilin-water emulsion. Prior art oil-in-water emulsions comprising cyclosporin in solution have been found to have low emulsion stability and to precipitate or crystalise cyclosporin over time. Solid porin formation in an on is undesirable. Therefore, there is a need for emulsions comprising P213428WO 3 porin in solution which have a higher emulsion stability. For example, there is a need for emulsions where the length of time for crystal formation or precipitation to occur is longer.

Formulating an active ingredient into a bead by passing a composition comprising a watersoluble polymer matrix in which are dispersed ts of oil through a single orifice nozzle is disclosed in WO2010/133609. Solid cyclosporin formation in an emulsion is particularly undesirable whilst forming beads from an emulsion comprising solid porin. This is because solid cyclosporin is less active at an intended therapeutic site, for example the gastrointestinal tract, than solubilised porin. The problem of cyclosporin crystallising or precipitating is particularly nt in scale up production of beads from the emulsion. Scaling up production of beads results in batches of the emulsion being kept for longer periods and the sity for crystallisation or precipitation to happen sing. ore, there is a need for an emulsion with a high stability to reduce the amount of solid in the emulsion whilst being processed into beads and consequently to reduce the amount of solid in the bead.

Similarly there is a need for a component of the emulsion to inhibit crystallisation or precipitation.

BRIEF SUMMARY OF THE DISCLOSURE It has surprisingly been found that a surfactant has a stabilising effect on an emulsion sing cyclosporin in solution. The surfactant may comprise or may be a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, optionally n the surfactant does not comprise or is not a polyethyleneglycol ether or ester. The surfactant may have an HLB of up to , preferably of up to 7 or up to 5. Specifically, it has been found that a medium chain or long chain fatty acid mono-or di-glyceride or a combination f, optionally not comprising or being a polyethyleneglycol ether or ester, has a stabilising effect on an emulsion comprising cyclosporin in solution. The emulsion may be an oil-in-water emulsion.

A medium chain or long chain fatty acid mono- or di-glyceride is an ester of such a fatty acid and ol, n there may be one (mono-) or two (di-) fatty acids esterified to glycerol. The glycerol may consist of one glycerol for example mono glycerol (a mono-glycerol ester is also be referred to as a glyceride).

It has been found that the use of certain surfactants during the manufacture of the compositions are particularly effective in stabilising the colloid (for example emulsion), resulting from the mixing of an aqueous solution comprising a hydrogel forming polymer and an oil phase comprising cyclosporin A. When the d comprises an oil-in-water emulsion, it has been found that the presence of a surfactant having an HLB of up to 8 (particularly up to 6 or from 2 to 6) in the oil phase is particularly effective in stabilising the emulsion in particular during the preparation of the composition. The presence of such surfactants has been found to inhibit the ion of cyclosporin A crystals after the formation of the d (oil-in-water emulsion). The presence of a surfactant with an HLB of up to 10 maintains the cyclosporin A in on in the oil phase during manufacture and may also provide favourable release of the cyclosporin A in a solubilised form from the composition P213428WO 4 following oral administration of the composition to a subject. Compositions comprising a tant of the invention with an HLB of up to 8 in at least the oil phase may exhibit high rates of release and/or extent of release of cyclosporin A from the composition compared to the use of surfactants with a higher HLB value in the oil phase. The presence of a surfactant with an HLB of up to 8 in at least the oil phase in the composition may inhibit the precipitation of cyclosporin A after release of the cyclosporin from the composition thereby retaining higher levels of cyclosporin in a solubilised form within the GI tract, for example in the colon. The compositions described herein wherein the composition comprises an oil phase and a surfactant having an HLB of up to 10 form a further independent aspect of the invention.

Accordingly, there is provided a use of a surfactant for stabilising an emulsion. Preferably, the emulsion is an oil in water emulsion. The surfactant preferably is or comprises a medium chain or long chain fatty acid r di-glyceride or a combination f, optionally not comprising or being a hyleneglycol ether or ester.

By the phrase "the surfactant stabilises the emulsion", it is meant that the surfactant ses the length of time for solid particles (for example precipitation or crystallisation) to occur in an emulsion comprising cyclosporin in solution.

It has also been found that a surfactant inhibits llisation of cyclosporin from a cyclosporin solution in an oil phase in an oil-in-water emulsion. The surfactant is or comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, optionally n the surfactant does not comprise or is not a polyethyleneglycol ether or ester. Accordingly, the use of the surfactant provides an emulsion that is free of crystals or precipite for a longer period than prior art emulsions. An emulsion that takes longer for crystallisation or precipitation to occur is beneficial for large-scale bead production. Consequently, the invention contemplates a process ing an emulsion comprising the tant to e beads, ularly in scale up of the bead production process.

Ordinarily surfactants with a low HLB value, for example up to 8, are used as water-in-oil emulsifiers. As part of the invention it has been found that a surfactant is an emulsifier for oil-in-water emulsions comprising dissolved cyclosporin, wherein the surfactant has a HLB value of up to 8 and is or ses a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, optionally not comprising or being a polyethyleneglycol ether or ester.

In an aspect of the invention there is provided a liquid composition sing an aqueous phase, a surfactant and an oil phase in which cyclosporin is ved. The surfactant may comprise or be a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester. The aqueous phase may comprise a hydrogel forming polymer. The oil phase may be dispersed in the s phase. The oil phase may be dispersed in the aqueous phase in the form of a colloid, for example a -liquid colloid. The oil phase may be dispersed in the aqueous phase in the form of an emulsion. Accordingly, the liquid composition may be a liquid emulsion composition.

P213428WO 5 The liquid composition may be converted into a solid form by allowing the hydrogel forming polymer to form a el matrix. There is also provided a process for converting the liquid composition into a bead, wherein the liquid composition is ejected through a single orifice nozzle.

In an aspect of the invention there is provided a composition comprising cyclosporin, a hydrogel forming r matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix. The tant may be or may comprise a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and may not comprise or may not be a polyethyleneglycol ether or ester. The composition may be a solid composition. The composition may be in the form of a dried bead. The composition may be in the form of a dried colloid.

Advantageously an enhanced release profile is provided by the presence of the surfactant being or comprising a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof that does not comprise or is not a polyethyleneglycol ether or ester in compositions of the invention compared to itions with a different surfactant. A solid ition of the ion exhibits a release e with higher release of cyclosporin and maintenance of high porin levels in solution when compared to compositions with a different surfactant (Kolliphor EL, a polyethoxylated castor oil). The dissolution may be measured in deionised water.

Optionally, the porin, the hydrogel g polymer matrix, the surfactant and the oil phase are comprised within a core. Thus, the ition may comprise a core. Accordingly, the composition may comprise a core, wherein the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, wherein the surfactant may be or comprise a medium chain or long chain fatty acid mono- or diglyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester.

The liquid composition may be a colloid, i.e. it may be a colloidal liquid composition. The composition may be a solid colloid or the composition may be in the form of a solid colloid, i.e. it may be a solid colloidal composition. The colloidal liquid composition of the invention may comprise a continuous phase which is or comprises a hydrogel-forming polymer and a disperse phase which is or ses cyclosporin A and an oil phase, wherein the dal liquid composition or the solid colloidal composition further comprise a surfactant (also ed to as a first surfactant) comprising or being a medium chain or long chain fatty acid mono- or ceride or a combination thereof and not comprising or not being a polyethyleneglycol ether or ester.

The solid colloidal composition of the invention may comprise a continuous phase which is or comprises a hydrogel-forming polymer matrix and a disperse phase which is or ses cyclosporin A and an oil phase, wherein the colloidal liquid composition or the solid dal composition further comprise a surfactant (also referred to as a first surfactant) comprising or being a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and not comprising or not being a polyethyleneglycol ether or ester.

Throughout the specification both the liquid composition and the composition are referred to by "composition". Furthermore, where an ment or aspect is referred to as a "composition" this P213428WO 6 may optionally be referring to a liquid composition (for example a dal liquid composition) and/or to a solid ition (for example a solid colloidal composition).

In an embodiment the oil phase comprises a solution of the cyclosporin. As such, the cyclosporin may be dissolved in the oil phase, for example tely ved, substantially completely dissolved, or lly dissolved. Thus, the oil phase may comprise a solution of cyclosporin and some undissolved cyclosporin.

Throughout this specification the term cyclosporin may be referring to the class of compounds or to porin A. Preferably, the use of cyclosporin is in nce to cyclosporin A.

The cyclosporin is suitably present in the composition in an amount of from about 5% to about 20%, from about 8% to about 15%, or from about 9% to about 14% by weight based upon the dry weight of the core or of the composition.

The cyclosporin is suitably present in the liquid composition in an amount of up to 10%, optionally from about 1% to about 10%, from about 2% to about 8%, from about 3% to about 6%, from about 3% to about 5% by weight of the liquid ition. Optionally the cyclosporin may be present in the liquid composition in about 4% by weight of the liquid composition.

A medium chain fatty acid mono-ester or di-ester comprises a fatty acid having 8 to 12 in chain carbon atoms. A long chain fatty acid mono-ester or di-ester comprises a fatty acid having at least 13 in chain carbon atoms, preferably 13 to 26 in chain carbon atoms. The long chain fatty acid may optionally have from 14 to 22 in chain carbon atoms or 16 to 20 in chain carbon atoms.

Preferably, the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof that does not comprise or is not a polyethyleneglycol ether or ester. Where the surfactant comprises a medium chain or long chain fatty acid mono- or di-glyceride or a ation thereof that does not comprise or is not a polyethyleneglycol ether or ester, the medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof is substantially all of the surfactant. For example, the surfactant may comprise medium chain or long chain fatty acid mono -or ceride or a combination f that does not comprise or is not a polyethyleneglycol ether or ester in an amount of greater than 80% of the surfactant, optionally greater than 85%, 90%, 95%, 97%, 98% or 99%. ly, the surfactant is substantially free of a triglyceride. For example, the surfactant may comprise less than 10%, 8%, 5%, 3%, 2% or 1% of a triglyceride.

The presence of the surfactant may enhance the rate and or extent of release of cyclosporin from the composition following oral administration. The presence of the surfactant may act to maintain a high proportion of the cyclosporin in a solubilised form after it has been released from the composition into an aqueous medium such as that found in the lower GI tract, particularly the colon.

The surfactant may have an HLB value of up to 8, up to 6, or up to 5. Alternatively the surfactant may have an HLB value selected from: up to 7, 1-8, 1-7, 2-6, 1-5, 2-5, 1-4, 1-3, 1-2, 2-4, 3- 4, 3-6, 5-8, 6-8 and 6-7. Preferably, the surfactant has an HLB value of up to 6, 2-6 or 3-6.

P213428WO 7 The cyclosporin A may be soluble in the surfactant, for example the cyclosporin A may have a solubility of more than 200mg/g in the surfactant. Thus, the tant may have a cyclosporin lity of more than 200 mg/g. The surfactant may have a cyclosporin lity of from 200 mg/g to 500 mg/g, optionally from 250 mg/g to 500 mg/g.

The tant may have a cyclosporin solubility of from 200 mg/g to 400 mg/g, from 225 mg/g to 375 mg/g, from 200 mg/g to 300 mg/g, from 300 mg/g to 400 mg/g, from 225 mg/g to 275 mg/g, from 350 mg/g to 400 mg/g. Preferably, the surfactant has a cyclosporin solubility of from 200 mg/g to 400 mg/g or from 225 mg/g to 375 mg/g. The surfactant may have a porin solubility of from 250 mg/g to 400 mg/g, from 250 mg/g to 375 mg/g, from 250 mg/g to 300 mg/g, from 300 mg/g to 400 mg/g, from 250mg/g to 275 mg/g, from 350 mg/g to 400 mg/g. Preferably, the surfactant has a cyclosporin solubility of from 250 mg/g to 400 mg/g or from 250 mg/g to 375 mg/g. The solubility of cyclosporin in a surfactant may be determined by techniques known to those skilled in the art, for example by following the protocol described in Development of a Self Micro-Emulsifying Tablet of Cyclosporine- A by the Liquisolid Compact Technique, Zhao et al (International Jpurnal of Pharmaceutical Sciences and Research, 2011, Vol. 2(9), 308) which is incorporated herein by reference.

The surfactant may have an HLB of up to 6 and a cyclosporin solubility of from 200 mg/g to 400 mg/g. The surfactant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 200 mg/g to 400 mg/g. The surfactant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 225 mg/g to 275 mg/g. The surfactant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 250 mg/g to 300 mg/g.

The surfactant may have an HLB of up to 6 and a porin solubility of from 250 mg/g to 400 mg/g. The surfactant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 250 mg/g to 400 mg/g. The tant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 250 mg/g to 375 mg/g. The surfactant may have an HLB value of 2-6 (optionally 3-6) and a cyclosporin solubility of from 250 mg/g to 300 mg/g.

The surfactant may be or comprise a medium chain or long chain fatty acid mono- or diglyceride or a combination thereof and may not comprise or may not be a polyethyleneglycol ether or ester, wherein the fatty acid ester is saturated or unsaturated. ably, the fatty acid is unsaturated.

The unsaturated fatty acid may n only one or only two double bonds.

Where the surfactant is a medium chain or long chain fatty acid di-glyceride (by which it is meant that there are two fatty acids esterified to a glycerol) the surfactant may comprise two fatty acids which are the same or ent. For example the two fatty acids may both be unsaturated or may both be saturated. Alternatively, one of the two fatty acids may be saturated and the other fatty acid may be unsaturated.

Preferably the surfactant is a long chain mono- or di-glyceride or a combination f and does not comprise or is not a polyethyleneglycol ether or ester. A further preferred surfactant is a long chain mono- or di-glyceride or a combination thereof and does not comprise or is not a P213428WO 8 polyethyleneglycol ether or ester, wheren the fatty acid has a chain length of 13 to 22 carbon atoms, optionally 16 to 20 carbon atoms. In ular the fatty acid may have a chain length of 18 carbon atoms.

In an embodiment the tant is selected from: glyceryl monocaprate, glyceryl dicaprate, glyceryl monocaprylate, glyceryl dicaprylate, glyceryl caprate, glyceryl monocaprylate/caprate, glyceryl caprylate/caprate glyceryl dicaprylate/caprate, glyceryl monooleate/dioleate, glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl lmitostearate, glyceryl dipalmitostearate, glyceryl monobehenate, glyceryl dibehenate, glycerol noleate, glyceryl dilinoleate, polyglyceryl te, ene glycol monoheptanoate, and a combination thereof.

A red surfactant may be or comprise a surfactant selected from: glyceryl monocaprylate/caprate, glyceryl dicaprylate/caprate, glyceryl monooleate, ol monolinoleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitostearate, glyceryl dipalmitostearate, glyceryl monobehenate, glyceryl dibehenate, yl monolinoleate, glyceryl dilinoleate, polyglyceryl dioleate and a combination f.

Accordingly, there is provided a composition comprising porin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming r matrix, wherein the surfactant may be or may comprise a surfactant ed from: yl monocaprylate/caprate, glyceryl dicaprylate/caprate, glyceryl monooleate, glycerol monolinoleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitostearate, glyceryl dipalmitostearate, glyceryl monobehenate, glyceryl dibehenate, glyceryl monolinoleate, glyceryl dilinoleate, polyglyceryl dioleate and a combination thereof.

The surfactant may comprise or be a surfactant selected from: glyceryl caprylate, glyceryl caprate, glyceryl monooleate, glyceryl dioleate, glycerol monolinoleate or a combination thereof.

A particularly red surfactant may be or comprise a surfactant selected from: glyceryl caprylate/caprate (Capmul MCM), glyceryl monooleate/dioleate (Capmul ) and glycerol monolinoleate (Maisine 35-1).

Optionally, the surfactant is not a mixture of glyceryl monostearate EP/NF and PEG-75 palmitostearate (for example GeltoTM 64). Suitably, the surfactant may not be or comprise a mixture of glyceryl monostearate.

In an ment the oil phase comprises an oil or liquid lipid and the surfactant is present in an amount greater than the oil or liquid lipid. ally, the surfactant may be present in an amount of more than 6 wt% of the dry weight of the composition. This refers to the ed composition or the core. The surfactant may comprise more than 12 wt% of the oil phase, for example in the liquid composition. The surfactant may be present in the composition in an amount of from about 5% to about 20%, from about 8% to about 20%, from about 8% to about 15%, or from about 10% to about 14% by weight based upon the dry weight of the core. It is to be understood that reference to the "dry weight of the core" means the weight of the components present in the uncoated core other than water.

P213428WO 9 The weight ratio of the surfactant : oil may be from about 5:1 to about 1:5, from about 3:1 to about 1:2, from about 3:1 to about 1:1 or from about 2.5:1 to 1.5:1. Suitably the weight ratio may be about 1:1, about 2:1, about 2.5:1, about 3:1, about 1:1.5 or about 1:2.

Accordingly, in a red embodiment there is provided a liquid composition comprising an aqueous phase, a tant and an oil phase in which cyclosporin is dissolved, wherein the aqueous phase comprises a hydrogel forming polymer, the oil phase is dispersed in the s phase and the surfactant comprises or is a surfactant selected from: glyceryl caprylate/caprate (Capmul MCM), glyceryl monooleate/dioleate l GMO-50), glycerol monolinoleate (Maisine 35-1) and a combination thereof. The oil phase may be dispersed in the aqueous phase in the form of a colloid, for e a -liquid colloid. The oil phase may be dispersed in the aqueous phase in the form of an on. ingly, the liquid composition may be a liquid emulsion composition.

There is provided a composition comprising cyclosporin, a hydrogel g polymer matrix, a surfactant and an oil phase being dispersed in the el forming polymer matrix, wherein the surfactant comprises or is a surfactant selected from: glyceryl caprylate/caprate (Capmul MCM), glyceryl monooleate/dioleate (Capmul GMO-50), glycerol monolinoleate (Maisine 35-1) and a combination thereof. The composition may be a solid ition. The composition may be in the form of a dried bead. The composition may be in the form of a dried colloid.

Optionally, the surfactant is or comprises glyceryl monooleate, glyceryl te or a combination thereof. Capmul GMO -50 is an example of a commercially available surfactant that comprises a combination of glyceryl monooleate and glyceryl dioleate. Thus, the surfactant may be Capmul GMO-50. Where Capmul GMO-50 is ned in the specification it will be understood that it is referring to a mixture of glyceryl monooleate and glyceryl te. Capmul GMO-50 may also refer to glyceryl monooleate alone.

Similarly, the skilled person would understand that a surfactant that is described as, for example glyceryl monooleate/dioleate, contemplates a combination of glyceryl monooleate and glyceryl te. In other words a "/" in a surfactant name indicates that the surfactant is a mixture of two components.

The composition may comprise a coating to control or te release of the cyclosporin from the composition. Advantageously the c oating is a polymeric coating to provide delayed and/or sustained release of the cyclosporin from the composition. Suitably such coatings are described in more detail below and include a coating which is or comprises a coating selected from a controlled e polymer, a sustained release polymer, an enteric polymer, a pH independent polymer, a pH dependent r and a polymer specifically tible to degradation by bacterial enzymes in the gastrointestinal tract, or a combination of two or more such polymers. In a particular embodiment the coating is or comprises a pH-independent polymer, for example a coating which is or comprises ethyl cellulose. In a further specific embodiment the coating is or comprises a pH-independent polymer, for example ethyl cellulose, and optionally a water-soluble polysaccharide, for example pectin or an, or a combination thereof, particularly pectin. 8WO 10 In an ment the coating that is referred to in the preceding paragraph is an outer coating, also referred to as a second coating. The composition may optionally comprise a further coating, ed to as a sub-coat or a first coating. The respective polymers of the first g and the second coating are ent. Often the second coating does not have any polymer found in the first coating; for example, if the first coating comprises (e.g. is) a hydroxypropylmethyl cellulose, then the second coating will not also comprise a hydroxypropylmethyl cellulose. In addition the situation is contemplated where the first coating is or comprises a water-soluble ether or ester of a cellulose ether, the major component(s) (e.g. more than 50%) of the second coating is or comprises a different polymer to that of the first coating. Accordingly, the first and second coatings suitably provide two layers of material as part of the composition. It is to be understood that when the second coating comprises a mixture of components, minor components of the outer second coating may be the same as the material of the sub-coating. By way of example, when the first g is or comprises HPMC and the second g ses ethyl cellulose, the ethyl cellulose may optionally further comprise a minor amount (e.g. less than 50%,40%, 30% or 20%) of the first g material, HPMC in this example. In such embodiments the first coating and the second coating are considered to be different.

The ition of the invention may comprise cyclosporin, a hydrogel forming polymer , a surfactant and an oil phase being dispersed in the hydrogel g polymer matrix, wherein the surfactant may be a medium chain or long chain fatty acid mono- or di-glyceride or a ation f and may not comprise or may not be a hyleneglycol ether or ester. Optionally, the composition may further comprise a first coating, wherein the first coating is or comprises a watersoluble cellulose ether as described above and elsewhere herein. In addition to the first coating or alternatively to the first coating the composition may comprise a second coating. Optionally, the second coating is or comprises a coating, suitably a polymeric coating, to control or modulate release of the active ingredient from the composition. The polymeric coating may be as further described elsewhere in this specification.

Where the composition comprises a first coating and a second coating the second coating may be outside the first coating.

The composition may comprise: a core, wherein the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer ; a first coating outside the core, wherein the first coating is a water-soluble cellulose ether as described above and elsewhere herein; and a second coating outside the first coating, wherein the surfactant is as described herein, the surfactant being, for example, a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and not comprising or not being a polyethyleneglycol ether or ester. Throughout this specification "core" may refer to a core comprising cyclosporin, a hydrogel forming polymer , a surfactant, as described herein, and an oil phase being dispersed in the hydrogel forming polymer matrix.

According to an embodiment of the invention, the tant optionally is a medium chain or long chain fatty acid mono-glyceride, di-glyceride or a combination f, the first coating is or comprises a soluble cellulose ether, and the composition further comprises a second coating P213428WO 11 outside the first coating wherein the second coating is or ses a coating, suitably a polymeric coating, to control or modulate release of the active ingredient from the composition. The polymeric coating may be as further described elsewhere in this specification.

The first g suitably may be or comprise a water-soluble ose ether. The watersoluble cellulose ether may be any cellulose ether or derivative of a cellulose ether, for example an ester of a cellulose ether that is soluble in water. ore, the water-soluble cellulose ether may be selected from: an alkyl cellulose; a hydroxyalkyl ose; a hydroxyalkyl alkyl cellulose; and a carboxyalkyl cellulose. Suitably the first coating is or comprises one or more water-soluble cellulose ethers selected from: methyl cellulose, hydroxyethyl cellulose, ypropyl cellulose and hydroxypropylmethyl cellulose, and ations thereof. In particular embodiments the first coating is or comprises a water-soluble hydroxypropyl methylcellulose. The water-soluble cellulose ethers and water-soluble tives thereof ( e.g. water-soluble esters of a cellulose ether) present in the first coating oat) suitably form at least 20%, 40%, 50%,60%, 70%, 80%, 85% or 90% by weight of the dry weight of the first coating.

In accordance with the present invention there is provided a pharmaceutical composition sing a core and a first coating, n the core comprises cyclosporin, a hydrogel forming polymer matrix, a tant and an oil phase being dispersed in the hydrogel forming polymer matrix and the first coating comprises or is a water soluble cellulose ether and the first coating is present in an amount corresponding to a weight gain due to the first coating of from 0.5% to 20% by weight of the core, wherein the surfactant is as described herein, for example a medium chain or long chain fatty acid mono- or di-glyceride or a ation thereof and not comprising or not being a polyethyleneglycol ether or ester.

The first coating of the present invention modifies the release of the active ingredient from the composition. There would be an expectation that a g on a composition would slow the rate of release of the active ingredient within a composition. One might reasonably expect this as coating the composition with onal material would provide an additional barrier to a dissolution medium coming into contact with the active ient in the composition. In contrast to this expected outcome, the compositions of the present invention se a coating comprising or being a water soluble cellulose ether that increases the rate of release of the active ingredient compared to a composition without the coating. In addition the coating of the present invention has the beneficial effect of maintaining the active ingredient in solution, whereas a comparable composition lacking the coating of the invention provides less of the active ingredient in solution as time progresses. Without wishing to be bound by theory, it is believed that the coating prevents precipitation of the active ingredient from solution, thereby maintaining a higher amount of the active in on.

Throughout the present application active ingredient, active, and pharmaceutically active ient are used interchangeably and all refer to cyclosporin, preferably cyclosporin A.

The composition of the present invention may take any form known to the person skilled in the art. Preferably, the composition is an oral composition. The composition may be in the form of a single minibead or a multiplicity of minibeads. Accordingly the invention provides a minibead P213428WO 12 sing porin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, wherein the surfactant is or comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester. The invention also provides a composition comprising a multiplicity of minibeads. Similarly, the invention provides a multiple minibead formulation comprising a unit dosage form comprising a licity of minibeads.

The invention also provides for a pharmaceutical composition sing a core and a first coating, wherein the core comprises porin, a hydrogel forming polymer , a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix and the first coating comprises or is a water-soluble cellulose ether and the first coating has a thickness of from 1 µm to 1 mm wherein the surfactant is as described herein, for example a medium chain or long chain fatty acid mono- or diglyceride or a combination thereof and not comprising or not being a polyethyleneglycol ether or ester.

Any of the pharmaceutical compositions of the invention may comprise a further coating, referred to herein as a second coating. The second coating may be outside the first coating. The second coating may be or comprise a delayed release polymer. In any embodiment and any aspect of the invention the first and second g may be different.

The invention ore, contemplates a pharmaceutical composition comprising a core, a first coating and a second coating outside of the first g, wherein the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, the first coating comprises or is a water soluble cellulose ether (for example HPMC), and the second coating comprises or is a delayed release polymer (for example ethylcellulose), wherein the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester.

The composition of any aspect or ment of the invention may be in the form of a solid colloid. Furthermore, the core of a composition may be in the form of a solid colloid. The colloid comprises a continuous phase and a disperse phase. Suitable continuous phases and disperse phases which may be used to form the core are defined in more detail below and in the detailed description of the invention. The uous phase may comprise or be the hydrogel g polymer matrix. Hence, where the continuous phase is the hydrogel forming polymer matrix, the composition of the invention may take the form of a solid unit of the el forming polymer comprising a disperse phase. The disperse phase may be droplets dispersed in the uous phase, or the el forming polymer matrix. The disperse phase may comprise or be the oil phase.

Thus, the invention provides a composition in the form of a solid colloid comprising a continuous phase and a sed phase, wherein the continuous phase ses or is a hydrogel forming polymer matrix and the continuous phase is or comprises oil phase, wherein the composition r comprises cyclosporin and a surfactant. Preferably, the tant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not se or is not a polyethyleneglycol ether or ester. The oil phase may se the cyclosporin in solution.

P213428WO 13 The composition may comprise a core in the form of a solid colloid comprising a continuous phase and a dispersed phase, wherein the continuous phase comprises or is a hydrogel forming polymer matrix and the continuous phase is or comprises oil phase, wherein the core further comprises cyclosporin and a surfactant, wherein the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not se or is not a polyethyleneglycol ether or ester. The oil phase may comprise the cyclosporin in solution.

The continuous phase of a solid colloid composition or core is or comprises a hydrogelforming polymer matrix. In embodiments the hydrogel-forming polymer matrix is or comprises a hydrocolloid, a non-hydrocolloid gum or an. In a particular ment the hydrogel-forming polymer matrix is or comprises gelatin, agar, a polyethylene glycol, starch, casein, chitosan, soya bean protein, safflower protein, alginates, gellan gum, eenan, xanthan gum, phthalated gelatin, ated gelatin, cellulosephthalate-acetate, oleoresin, polyvinylacetate, polymerisates of acrylic or methacrylic esters and polyvinylacetate-phthalate and any derivative of any of the foregoing; or a mixture of two or more such polymers. In a further ment the hydrogel-forming polymer matrix is or comprises a hydrocolloid selected from carrageenan, gelatin, agar and pectin, or a combination thereof optionally selected from gelatin and agar or a combination thereof. Particularly, the r of the hydrogel-forming polymer matrix is or comprises gelatin. In an embodiment, the hydrogel-forming r does not comprise a cellulose or a cellulose derivative, e.g. does not comprise a cellulose ether.

In this aspect of the ion the composition may be in the form of a solid colloid the colloid comprising a continuous phase and a disperse phase and the cyclosporin may be in solution or suspended in the disperse phase. For example, the cyclosporin may be in solution in the se phase.

It is to be understood that the individual ments described above may be combined with one or more of the other embodiments described to provide further embodiments of the invention.

The first coating may be in contact with the core. The second coating may be on the first coating. In embodiments the first coating is in contact with the core and the second coating is on the first coating.

The second coating may be or may comprise a delayed release r and the delayed release polymer may be selected from an enteric polymer, a pH independent polymer, a pH dependent r and a polymer specifically susceptible to ation by bacterial enzymes in the gastrointestinal tract, or a combination of two or more such polymers. Hence, the second coating may be any of the aforementioned delayed release polymers or any may be or s the characteristics mentioned in relation to the delayed release polymer mentioned below.

In ments the delayed e polymer may be water-soluble or water-permeable in an aqueous medium with a pH greater than 6.5. The delayed release polymer may be or comprise a pH-independent polymer, for example ethyl cellulose. 8WO 14 In any aspect and any embodiment of the invention the water-soluble cellulose ether may be selected from any one or a combination of: methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose and ypropyl methylcellulose. The water-soluble cellulose ether may preferably be hydroxylpropyl methylcellulose (HPMC).

In embodiments the first coating may be or comprise hydroxypropyl methyl cellulose and the second coating may be or comprise ethyl cellulose.

The disclosure of the weight gain of the first g is given as a % by weight of the core.

Similarly, the weight gain of the second coating is given as a % by weight of the core, where there is no first coating (sub-coat) on the core. Where the composition comprises a first coating, the weight gain of the second coating is given as a % by weight of the composition that is coated by the second coating, for e the core and the first g.

The hydrogel forming r or the hydrogel forming polymer matrix may be or comprise a hydrocolloid, a non-hydrocolloid gum or chitosan. The hydrogel forming polymer or the hydrogel forming polymer matrix may be a reversible hydrocolloid, for example a thermoreversible hydrocolloid or a thermoreversible hydrogel forming polymer. atively, the hydrogel forming polymer or the hydrogel forming polymer matrix may be or comprise an irreversible hydrocolloid. The hydrogel forming polymer or the el forming polymer matrix may be or comprise gelatin, agar, a polyethylene , starch, casein, chitosan, soya bean protein, safflower protein, alginates, gellan gum, carrageenan, xanthan gum, phthalated gelatin, succinated gelatin, cellulosephthalate-acetate, oleoresin, polyvinylacetate, polymerisates of c or methacrylic esters and polyvinylacetatephthalate and any derivative of any of the foregoing; or a mixture of one or more such a hydrogel forming polymers. The hydrogel forming r or the hydrogel forming polymer matrix may be or comprise a hydrocolloid selected from carrageenan, gelatin, agar and , or a combination thereof optionally selected from gelatin and agar or a combination f, more optionally the hydrogel forming polymer or the or the hydrogel forming polymer matrix forming polymer matrix is or comprises gelatin. The hydrogel forming polymer matrix is or comprises a non-hydrocolloid gum optionally selected from a cross-linked salt of alginic acid. In preferred embodiments the hydrogel forming polymer or the hydrogel forming polymer matrix is or comprises gelatin.

In embodiments the el forming polymer or the hydrogel forming polymer matrix further sing a plasticiser, optionally a plasticiser selected from glycerin, a polyol for example sorbitol, polyethylene glycol and triethyl citrate or a e thereof, particularly sorbitol.

The hydrogel forming polymer matrix may encapsulate the cyclosporin. The cyclosporin may be encapsulated in solution. The cyclosporin may be in solution or suspended in another component, for example the oil phase or the disperse phase sed elsewhere, of the composition that is also encapsulated by the hydrogel forming polymer matrix.

The disperse phase may be solid, olid or liquid. In particular, the disperse phase may be . In other particular instances the disperse phase may be semi-solid, for example it may be waxy.

P213428WO 15 The disperse phase may be or comprise the oil phase, for example the oil phase may be a solid, a semi-solid or a liquid. Suitably the disperse phase or the oil phase is or comprises a liquid lipid and optionally a solvent miscible ith. The liquid lipid is optionally a medium chain mono- di- or triglyceride (particularly a medium chain triglyceride).

Suitably, porin is soluble in the solvent. The solvent may be an alcohol (for example ethanol or isopropanol), a glycol (for example propylene glycol or a hylene glycol) or a glycol ether. The solvent may be a glycol ether, for example an ethylene glycol ether, more particularly an alkyl, aryl or aralkyl ethylene glycol ether. The solvent may be a glycol ether selected from 2- methoxyethanol; xyethanol; 2-propoxyethanol; 2-isopropoxyethanol; 2-butoxyethanol; 2- phenoxyethanol; 2-benzyloxyethanol; 2-(2-methoxyethoxy)ethanol; 2-(2-ethoxyethoxy)ethanol; and 2- (2-butoxyethoxy)ethanol. More particularly the solvent is 2-(2-ethoxyethoxy)ethanol or 2- phenoxyethanol. A particular solvent is 2-(2-ethoxyethoxy)ethanol.

The cyclosporin may be dissolved in the disperse phase. The cyclosporin may be suspended in the disperse phase. The disperse phase may be as described ere herein, for example it may be as described in the immediately preceding two paragraphs.

The oil phase or disperse phase may be or may comprise a liquid lipid. Particularly, the oil phase or disperse phase may comprise or be a , medium- or long- chain triglyceride formulation, or a combination thereof, for example a caprylic/capric triglyceride, i.e. a ic/capric ceride formulation.

Accordingly, in an embodiment the liquid composition comprises an aqueous phase, a tant and an oil phase in which cyclosporin is dissolved, wherein the tant may comprise or be a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise is not a polyethyleneglycol ether or ester, the aqueous phase may se a hydrogel forming polymer, and the oil phase comprises a short-, medium- or long- chain triglyceride formulation, or a combination thereof (optionally a caprylic/capric triglyceride, i.e. a caprylic/capric triglyceride formulation) and is dispersed in the aqueous phase. The oil phase may be dispersed in the aqueous phase in the form of a colloid, for example a liquid-liquid colloid. The oil phase may be sed in the aqueous phase in the form of an emulsion. Accordingly, the liquid composition may be a liquid emulsion composition.

Additionally, in an embodiment the composition comprises cyclosporin, a hydrogel forming polymer , a surfactant and an oil phase comprising a short-, medium- or long- chain triglyceride formulation, or a combination thereof (optionally a caprylic/capric triglyceride, i.e. a caprylic/capric triglyceride formulation) and being dispersed in the hydrogel forming polymer matrix, n the tant may be or may comprise a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester. The composition may be in the form of a dried colloid. The composition may be in the form of a bead.

In a particular ment the disperse phase or the oil phase further ses a solvent, thus ally the disperse phase or the oil phase may be or comprise a liquid lipid and a solvent. The P213428WO 16 solvent may be miscible with the liquid lipid and water, optionally wherein the solvent is ed from 2-(2-ethoxyethoxy)ethanol and a poly(ethylene ), particularly wherein the solvent is 2-(2- ethoxyethoxy)ethanol. In a further embodiment the disperse phase or oil phase is or comprises a medium chain mono- di- or triglyceride (particularly a medium chain triglyceride), 2- (ethoxyethoxy)ethanol and the surfactant. The disperse phase or oil phase as described in this aph may n the cyclosporin, the cyclosporin may optionally be in solution.

Preferably, the oil phase or disperse phase comprises a short-, medium- or long- chain ceride formulation, or a combination f (optionally a caprylic/capric triglyceride, i.e. a caprylic/capric triglyceride formulation). Where the oil phase or disperse phase comprises a short-, medium- or long- chain triglyceride formulation, or a combination thereof, the triglyceride is substantially all of the disperse phase or oil phase (optionally the liquid lipid). For example, the oil phase or disperse phase may se short-, medium- or long- chain triglyceride formulation in an amount of greater than 80% of the oil phase or disperse phase (optionally the liquid , optionally greater than 85%, 90%, 95%, 97%, 98% or 99%. Suitably, the short-, medium- or long- chain triglyceride formulation is substantially free of mono- or di-glycerides. For example, the surfactant may comprise less than 10%, 8%, 5%, 3%, 2% or 1% of a mono- or di-glycerides.

In embodiments the composition further comprises one or more additional surfactants, preferably one additional surfactant. The additional surfactant may be ed to as a second surfactant or further surfactant throughout the specification and these terms are used interchangeably.

Where the compositions of the invention comprise a second surfactant the tant that may be or may comprise a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a hyleneglycol ether or ester is referred to as a first tant.

Suitable surfactants for the second surfactant are described in more detail in the detailed description of the invention. The second surfactant may be an anionic or non-ionic surfactant. The second surfactant may be a sucrose monoester, an alkyl e or a polyethylene glycol alkyl ether.

The second surfactant may be sucrose laurate, sucrose palmitate, sodium octyl sulfate, sodium cyl sulfate, sodium dodecyl sulphate, polyethylene glycol hexadecyl ether, polyoxyethylene glycol octadecyl ether, or polyethylene glycol dodecyl ether. Optionally, the second surfactant may be sodium octyl sulfate, sodium octadecyl sulfate, sodium dodecyl sulphate or polyethylene glycol dodecyl ether.

Preferably the second surfactant is an anionic surfactant. For example, the second surfactant may be an alkyl sulphate, for example sodium octyl sulfate, sodium octadecyl sulfate, or sodium dodecyl sulphate (preferably sodium l te).

In those ments where the liquid composition is in the form of a colloid, the composition is in the form of a solid colloid or the composition comprises a core in the form of a solid colloid, the colloid comprises a continuous phase and a disperse phase, wherein the continuous phase comprises the hydrogel-forming polymer matrix and the second surfactant may be present in the continuous phase, the disperse phase or both. ably the second surfactant is present in the continuous phase and the first surfactant is present in the disperse phase. Accordingly, the aqueous P213428WO 17 phase of the liquid composition may comprise the second surfactant and the oil phase may comprise the first surfactant. In one embodiment the core further comprises one additional surfactant present in at least the continuous phase, the surfactant having an HLB value of greater than 10, for example greater than 20.

The ition may have the characteristics of a composition formed by mixing a disperse phase with a continuous phase to form a colloid, wherein the continuous phase is an aqueous phase sing hydrogel forming r and the disperse phase is a oil phase, wherein the pharmaceutically active ingredient is in the uous phase or the se phase, wherein the colloid is gelled to form the composition. The composition is thus in the form of a solid colloid. rmore, the composition may comprise a core having the characteristics of a core formed by mixing a disperse phase with a continuous phase to form a d, wherein the continuous phase is an aqueous phase comprising hydrogel forming polymer and the disperse phase is a oil phase, wherein the pharmaceutically active ingredient is in the continuous phase or the disperse phase, wherein the colloid is gelled to form the core. The core is thus in the form of a solid colloid.

The cyclosporin may be present in the composition in solution or in suspension. In the aspect where the invention provides a liquid composition the cyclosporin is in solution.

The liquid composition comprises an aqueous phase, a surfactant and an oil phase in which cyclosporin is ved and may have the characteristics of a liquid composition obtained by a process comprising: (i) dissolving a hydrogel-forming polymer in an aqueous liquid to form an aqueous phase solution; (ii) dissolving the cyclosporin in the oil phase to form a solution; and (iii) mixing the aqueous phase solution (i) and the oil phase solution (ii) to form a colloid (optionally an on).

The composition or core of the ion may have the characteristics of a composition obtained by a process comprising: (a) ejecting the liquid composition through a nozzle to form ts; (b) causing or allowing the a hydrogel-forming polymer to gel or solidify to form a hydrogelforming polymer matrix; and (c) drying the solid.

The composition or core comprises cyclosporin, a hydrogel forming polymer matrix, a tant and an oil phase and may have the characteristics of a composition obtained by a process comprising: (i) dissolving a hydrogel-forming r in an aqueous liquid to form an aqueous phase solution; (ii) dissolving the cyclosporin in the oil phase to form a solution; (iii) mixing the aqueous phase solution (i) and the oil phase solution (ii) to form a colloid (optionally an emulsion); P213428WO 18 (iv) ejecting the colloid through a nozzle to form droplets; (v) causing or allowing the a el-forming r to gel or solidify to form a elforming r matrix; and (vi) drying the solid.

The aqueous phase and oil phase may be mixed (for example in step (iii)) in an oil phase to aqueous phase ratio of from 1:4 to 1:10, optionally from 1:4 to 1:8, from 1:5 to 1:7. For example, the oil phase to aqueous phase ratio may be 1:4, 1:5, 1:6 or 1:7.

The oil phase solution (ii) may be prepared by dissolving or sing the porin A in a suitable hydrophobic liquid. The hobic liquid may be for example, any of the oils or liquid lipids described herein. By way of example the hydrophobic liquid may be, or se, ted or unsaturated fatty acids or a triglyceride, or an ester or ether thereof with polyethylene glycols. A particular oil for the oil phase is or comprises a triglyceride, for example an oil comprising a medium chain triglyceride, optionally wherein the oil comprises a triglyceride of at least one fatty acid ed from fatty acids having 6, 7, 8, 9, 10, 11 or 12 carbon atoms, e.g. C8-C10 fatty acids.

The aqueous phase solution (i) may further comprise a surfactant selected from: sucrose monoester, an alkyl sulfate and a polyethylene glycol alkyl ether, optionally the sselected from: sucrose laurate, sucrose palmitate, sodium octyl sulfate, sodium octadecyl sulfate, sodium dodecyl sulphate, polyethylene glycol hexadecyl ether, polyoxyethylene glycol octadecyl ether, and polyethylene glycol dodecyl ether. The aqueous phase solution (i) may further comprise a surfactant selected from: sodium octyl sulfate, sodium cyl sulfate, sodium dodecyl sulphate or polyethylene glycol dodecyl ether. Preferably, the aqueous phase solution (i) further comprises an anionic surfactant, e.g. as described elsewhere herein, for example sodium l sulphate (SDS).

In one embodiment the liquid composition or composition having the characteristics of a liquid composition or composition obtained by the process above is a composition or liquid composition comprising an oil phase dispersed in the aqueous phase solution, wherein the liquid composition or composition is or comprises cyclosporin, glyceryl monooleate/dioleate, gelatin, SDS, ol, caprylic/capric triglyceride, 2-(ethoxyethoxy)ethanol; wherein the aqueous phase solution (i) is or comprises gelatin, sorbitol and SDS; and the oil phase solution (ii) is or comprises cyclosporin, glyceryl monooleate/dioleate, caprylic/capric triglyceride, 2-(ethoxyethoxy)ethanol and the active ingredient.

Cores having the characteristics of cores obtained by the above-described processes, for example cores obtained by the processes, may be coated to provide a coating that comprises or is a soluble cellulose ether, optionally with a second coating to control or modify release, preferably a polymeric coating as described above and herein. The coated composition may be obtained by applying to the core the g, e.g. applying to the core first and second gs as described.

Before the coating is applied, the core may be made by a process having steps (i) to (vi) or (i) to (v) described above. Suitable methods for applying the coating(s) are described below and include ng the coatings by spray g a coating composition onto the core. The processes having steps (i) to (vi) or (i) to (v) themselves form aspects of the invention.

P213428WO 19 The ition or core may further se a second tant (also referred to as a further surfactant), optionally wherein the second surfactant is an anionic surfactant, optionally selected from alkyl sulphates, carboxylates or phospholipids, or a non-ionic surfactant, optionally selected from sorbitan-based surfactants, PEG-fatty acids, fatty alcohol ethoxylates, alkylphenol ethoxylate, fatty acid ethoxylates, fatty amide ethoxylates, alkyl glucosides or glyceryl fatty acids, or poloxamers, or a combination thereof. Hence the liquid composition of the invention may comprise at least the following features, an aqueous phase comprising a hydrogel forming polymer, a first surfactant and an oil phase being dispersed in the aqueous phase in which cyclosporin is dissolved and a second surfactant. Similarly, the composition of the invention may comprise at least the following features, cyclosporin, a hydrogel forming polymer matrix, a first surfactant and an oil phase being dispersed in the el forming polymer matrix and a second surfactant.

In embodiments where the composition is in the form of a solid colloid, the second surfactant may be in the se phase or the continuous phase. The second surfactant may be in the uous phase and may be an anionic surfactant, for example at least one surfactant selected from fatty acid salts and bile salts, particularly an alkyl sulphate, for example sodium dodecyl sulphate. The surfactant in the disperse phase may be a non-ionic surfactant.

In embodiments the composition comprises a second surfactant which is or comprises an c surfactant, for e sodium dodecyl sulphate, which is in the continuous phase.

In embodiments the composition further comprises a combination of excipients selected from: an anionic tant and a solvent; an anionic surfactant and an oil; and an anionic surfactant, a t and an oil. ably, the c surfactant is an alkyl sulphate, for example sodium dodecyl te, the oil is a medium chain mono-, di- and/or tri-glyceride (optionally a medium chain triglyceride, for example caprylic/capric triglyceride, and the solvent is 2-(ethoxyethoxy)ethanol.

The ition may further se an excipient selected from: a surfactant, a solubiliser, a permeability enhancer, a disintegrant, a crystallisation inhibitor, a pH modifier, a stabiliser, or a combination thereof.

The composition of the invention or, where the composition comprises a core, the core may comprise a disperse phase or oil phase, wherein the disperse phase or oil phase is or comprises: cyclosporin; a medium or long chain fatty acid mono- or di-ester or a combination f which does not comprise is not a polyethyleneglycol ether or ester, such as a medium or long chain fatty acid monoor di-glyceride or a combination f, for example glyceryl monooleate/dioleate; a medium chain mono- di- or tri-glyceride, for example caprylic/capric triglyceride; and a solvent, for example 2-(ethoxyethoxy)ethanol and the composition or the core may further comprise a continuous phase or aqueous phase being or comprising: an anionic surfactant, for e at least one surfactant selected from fatty acid salts and bile salts, particularly an alkyl sulphate, for e sodium dodecyl sulphate P213428WO 20 a hydrogel forming polymer matrix which is or comprises a hydrocolloid selected from carrageenan, gelatin, agar and pectin, or a combination thereof optionally selected from gelatin and agar or a combination thereof, more optionally the polymer of the a hydrogel forming polymer matrix is or comprises gelatin; and optionally a plasticiser, for example a plasticiser selected from glycerin, a polyol for example sorbitol, polyethylene glycol and triethyl citrate or a mixture thereof, particularly ol.

In one embodiment the composition comprises a core and a coating outside the core, wherein the core is in the form of a solid d, the colloid comprising a continuous phase and a disperse phase, wherein the disperse phase is or comprises: cyclosporin A; a medium or long chain fatty acid mono- or di-glyceride or a combination thereof which does not comprise is not a hyleneglycol ether or ester, for example glyceryl monooleate/dioleate; a medium chain mono- di- and/or tri-glyceride, for example caprylic/capric triglyceride; and a co-solvent, for example 2-(ethoxyethoxy)ethanol; and wherein the uous phase is or ses: a hydrogel-forming polymer matrix which is or comprises a hydrocolloid selected from carrageenan, gelatin, agar and pectin, or a ation thereof optionally selected from gelatin and agar or a combination thereof, more optionally the polymer of the water-soluble polymer matrix is or comprises gelatin; optionally a plasticiser, optionally a plasticiser selected from glycerin, a polyol for example sorbitol, polyethylene glycol and triethyl citrate or a mixture thereof, particularly sorbitol; and an anionic surfactant, for example at least one surfactant selected from fatty acid salts and bile salts, particularly an alkyl te, for e sodium dodecyl sulphate; and wherein the coating on the core is a first coating or a second coating, as bed herein.

Suitably the g comprises a first coating and a second coating outside the first coating; and wherein the first coating is the coating which is or comprises a water-soluble ose ether as described above; and the second coating is or comprises a coating, suitably a polymeric g, as defined above to control or modulate release of cyclosporin A from the composition.

In ments comprising a first coating and/or a second g, for example as mentioned in the immediately preceding paragraph, a ular first coating is or comprises hydroxypropylmethyl cellulose and a ular second coating outside the first coating is or comprises a pH independent r, for example ethyl cellulose; more particularly the second coating is or comprises ethyl cellulose and ally a polysaccharide selected from water soluble and naturally occurring ccharides, for example pectin or another water-soluble naturally occurring polysaccharide. The second coating may therefore contain pectin or another said polysaccharide or it may be substantially free of pectin and other said ccharides. There are therefore disclosed second coatings which comprise ethylcellulose as a controlled release polymer and which further comprise pectin or another said polysaccharide as well as second coatings which comprise P213428WO 21 ethylcellulose as a lled release polymer and which do not further comprise pectin or another said polysaccharide.

The hydrogel forming polymer, optionally comprising gelatin, may be present in an amount of 300 to 700 mg/g (optionally 380 to 500 mg/g). The medium chain mono, di and/or tri-glycerides, may be present in an amount of 20 to 200 mg/g (optionally 40 to 80 mg/g). The solvent, for example 2- (ethoxyethoxy)ethanol, may be present in an amount of 100 to 250 mg/g nally 160 to 200 mg/g).

The medium or long chain fatty acid mono- or di-ester or a combination f which does not comprise or is not a polyethyleneglycol ether or ester, for example glyceryl monooleate/dioleate, may be present in an amount of 80 to 200 mg/g (optionally 100 to 150 mg/g). The anionic surfactant, for example sodium dodecyl sulphate, may be t in an amount of up to 100 mg/g or up to 50 mg/g (optionally 10 - 70 mg/g, 15 - 60 mgm/g or 15 – 50 mg/g, preferably 25 – 50 mg/g or 25 – 45 mg/g).

The composition or the core may comprise a el forming polymer comprising gelatin, optionally in an amount of 300 to 700 mg/g, the core further comprising medium chain mono, di and/or tri-glycerides, optionally in an amount of 20 to 200 mg/g, wherein the composition or core further ses the following components: t, for example 2-(ethoxyethoxy)ethanol, optionally in an amount of 100 to 250 mg/g; a medium or long chain fatty acid mono- or di-ester or a combination thereof which does not comprise or is not a polyethyleneglycol ether or ester, for example glyceryl monooleate/dioleate, optionally in an amount of 80 to 200 mg/g; and anionic surfactant, for example sodium dodecyl sulphate, ally in an amount of up to 70 mg/g or up to 50 mg/g.

As will be recognised the composition or core further comprises cyclosporin.

The ition or the core may se: a hydrogel forming polymer, for example which is, or comprises, gelatin in an amount of 300 to 700 mg/g; porin in an amount of up to about 250 mg/g, for example 50 to 250 mg/g; medium chain triglycerides, for example Miglyol 810 in an amount of 20 to 200 mg/g, optionally a solvent, for example 2-(ethoxyethoxy)ethanol, which when present is in an amount of 100 to 250 mg/g; a surfactant comprising a medium or long chain fatty acid mono- or di-ester or a combination thereof which does not comprise or is not a polyethyleneglycol ether or ester, for e glyceryl monooleate/dioleate, in an amount of 80 to 200 mg/g; and anionic surfactant, for example sodium dodecyl sulphate, in an amount of up to 60 mg/g or up to 50 mg/g, for example 10 to 50 mg/g, or optionally 20 to 45 mg/g.

The composition or the core may comprise: gelatin in an amount of 380 – 500 mg/g; cyclosporin in an amount of 90 – 250 mg/g nally 90 – 200 mg/g or 90 – 160 mg/g); and caprylic/capric triglyceride in an amount of 40 – 80 mg/g; 2-(2-ethoxyethoxy) ethanol in an amount of 160 – 200 mg/g; P213428WO 22 glyceryl monooleate and/or glyceryl te in an amount of 100 – 150 mg/g; and SDS in an amount of 15 – 60 mg/g or 15 – 50 mg/g (optionally 25 – 50 mg/g or 25 – 45 mg/g); ally D-sorbitol in an amount of 30 – 80 mg/g.

The composition or the core may comprise: gelatin in an amount of 380 – 500 mg/g; cyclosporin in an amount of 90 – 140 mg/g; and caprylic/capric triglyceride in an amount of 40 – 80 mg/g; 2-(2-ethoxyethoxy) ethanol in an amount of 160 – 200 mg/g; glyceryl monooleate and/or glyceryl dioleate in an amount of 100 – 150 mg/g; and SDS in an amount of 15 – 50 mg/g nally 25 – 50 mg/g or 25 – 45 mg/g); and optionally D-sorbitol in an amount of 30 – 80 mg/g.

] The composition or core may be a colloid. Where the composition or the core is a colloid, the cyclosporin may be dissolved in the disperse phase of the colloid.

The composition or core may be a colloid; thus, the composition or core may comprise a continuous phase and a disperse phase wherein the uous phase comprises: gelatin in an amount of 380 – 500 mg/g; and optionally D-sorbitol in an amount of 30 – 80 mg/g; the disperse phase comprises: cyclosporin in an amount of 90 – 140 mg/g; and caprylic/capric triglyceride in an amount of 40 – 80 mg/g; and the composition further comprises: 2-(2-ethoxyethoxy)ethanol in an amount of 160 – 200 mg/g; glyceryl monooleate and/or glyceryl dioleate in an amount of 100 – 150 mg/g; and SDS in an amount of 15 – 50 mg/g.

A colloidal composition or core comprising a uous phase comprising: a hydrogel forming polymer matrix comprising n in an amount of 300 to 700 mg/g; a disperse phase comprising: cyclosporin in an amount of up to 200 mg/g; and a medium chain tri-glyceride in an amount of 20 to 200 mg/g; and the composition further comprising: solvent in an amount of 100 to 250 mg/g; surfactant (a first surfactant) being or comprising a medium or long chain fatty acid mono- or di-ester or a combination thereof which does not comprise is not a polyethyleneglycol ether or ester, for example glyceryl monooleate/dioleate; and anionic tant (a second surfactant) in an amount of up to 50 mg/g.

In the ments above which refer to mg/g of a component, the concentration is based upon the dry weight of the composition.

P213428WO 23 Suitably in the six compositions or cores described immediately above, the composition is a colloid comprising a disperse phase and a continuous phase; wherein the disperse phase comprises porin, medium-chain triglyceride and medium or long chain fatty acid mono- or di-ester surfactant; and the continuous phase comprises the el forming polymer (e.g. n) and an anionic surfactant (e.g. SDS).

The invention includes within its scope compositions wherein the core is a colloid having a disperse phase and the uous phase (matrix phase) of the d further includes dispersed particles of a pharmaceutically active ingredient, for example microparticles or nanoparticles. The disperse phase and uous phase may otherwise be as described elsewhere in this specification.

The composition of the invention and / or the core may be in the form of a minibead. It may be that the core is a minibead and the first coating and, where able, the second coating in ction with the core are in the form of a minibead. However, it may be possible for the core to be a minibead and the composition not to be a minibead. The composition may additionally comprise a multiplicity of minibeads. Hence the invention contemplates a minibead with the features of the pharmaceutical compositions disclosed herein.

The composition or the minibead may have a largest cross sectional dimension of a core of from about 0.01 mm to about 5 mm, for example from 1 mm to 5 mm, as in the case of from 1mm to 3mm or 1mm to 2mm. The minibead may be spheroidal. The spheroidal minibeads may have an aspect ratio of no more than 1.5, for e from 1.1 to 1.5.

The composition of the ion may be for oral administration. The composition may be formulated into a unit dosage form for oral administration comprising from 0.1 mg to 1000 mg, optionally from 1mg to 500 mg, for example 10mg to 300 mg, or 25 to 250 mg suitably about 25mg, about 35 mg, about 37.5 mg, about 75mg, about 150 mg, about 180 mg, about 210 mg, about 250 mg or about 300 mg of cyclosporin. Suitably the composition is in a multiple minibead unit dosage form selected from multiple minibeads in, for example, soft or hard gel capsules, gelatin capsules, HPMC capsules, compressed tablets or sachets. The ads may be as described elsewhere herein.

The compositions described herein may be used to deliver cyclosporin A y to specific locations in the GIT, for example the solid compositions described , may be adapted to provide release of cyclosporin A in at least the colon. The compositions may be used to provide the cyclosporin A locally in the GIT in a solubilised form, y providing high concentrations of cyclosporin in an active (available) form within the GIT where it acts to provide a therapeutic benefit in a number of medical conditions, particularly conditions affecting the GIT as described in more detail herein, such as ulcerative colitis. The release of cyclosporin A in an active form, for example a solubilised form, enables high concentrations of cyclosporin A to be ed directly into the local tissues of the GIT, such as the colon. However, as described above, systemic exposure cyclosporin A has a number of undesirable side effects. Therefore, a cyclosporin A composition which minimises systemic exposure to cyclosporin whilst ining therapeutically cial concentrations in the tissues of the GIT would be desirable.

P213428WO 24 The major pathways of cyclosporin A metabolism in humans are via cytochrome P450 3A4 (CYP 3A4) and cytochrome P450 2J2 (CYP 2J2), with three major metabolites being formed (two hydroxylated metabolites AM1, AM9 and one N-demethylated metabolite, AM4N). These metabolites have minimal, if any immunosuppressive activity. ore minimising the lism of cyclosporin is desirable, because this minimises the formation of inactive metabolites and maximises the quantity of cyclosporin available to interact y with the tissues in the GIT. The primary metabolism of cyclosporin is via CYP 3A4, which is mainly found in the liver and the small intestine.

The total mass of CYP3A in the entire small intestine has been estimated to be_1% of that in the liver. However, despite the relatively low mass of CYP3A in the small intestine, enteric CYP3A can bute significantly, and in some cases equally, with hepatic CYP3A, to the overall first-pass metabolism of several drugs including cyclosporin (Paine et al; Drug Metabolism and Disposition, Vol 34(5), 2006, 880-885). CYP 3A4 expression in the colon is lower than in the small ine.

Compositions which control the release of cyclosporin to limit or inhibit release in the upper GI tract, for example, by using one or more modified release coatings, may reduce enteric (i.e. stemic or "pre-systemic") P450 metabolism. However, some metabolism would still be expected resulting from the P450 expressed in the tissues in the lower GIT.

It has been found that n itions described herein, particularly compositions which release cyclosporin in the lower GI tract, ally in the colon, provide very low levels of cyclosporin metabolism following oral administration of the composition. The compositions therefore maximise the amount of active (solubilised) cyclosporin available to interact with the tissues of the GI tract following release of the cyclosporin from the composition. Without wishing to be bound by theory, it is thought that certain components present in the composition, for example the medium chain or long chain fatty acid mono- or di-glyceride surfactant t in the composition, may act to inhibit cyclosporin metabolism by CYP 3A4 present in the s of the GI tract. When the composition is provided in a modified release format which prevents or inhibits e of cyclosporin in the upper GI tract, systemic absorption and metabolism of cyclosporin in liver is also minimised. Therefore, the modified e compositions described herein minimise both systemic and enteric metabolism of cyclosporin.

Low levels of cyclosporin metabolism may enable a lower dose of cyclosporin to be administered whilst maintaining a therapeutic t, thereby, widening the therapeutic window of the drug.

The relative degree of porin metabolism ing oral administration of a composition may be assessed by, for e, measuring the tration of cyclosporin and the concentration of cyclosporin metabolites present in a faecal sample collected from a patient following oral administration of a composition comprising cyclosporin. As will be illustrated in the Examples, modified release compositions sing cyclosporin and the surfactant (for example Capmul GMO- 50) resulted in very low levels of cyclosporin metabolism compared to a similar composition sing a different surfactant (Cremophor). Compositions, particularly orally administered compositions, which exhibit low cyclosporin metabolism following release of porin from the composition, form a further aspect of the invention.

P213428WO 25 Accordingly, there is provided a composition comprising cyclosporin A, wherein after oral administration of the composition to a human, the mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in a faecal sample from the human is greater than 12:1.

The mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in the faecal sample may be selected from: greater than 19:1; r than 24:1; greater than 31:1 and greater than 50:1. The mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in the faecal sample may be selected from: from 20:1 to 30:1; from 20:1 to 35:1; from 20:1 to 40:1; from 20:1 to 60:1; from 30:1 to 50:1; and from 20:1 to 100:1. T he mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in the faecal sample may be selected from 12.5:1 to 90:1; from 13:1 to 85:1; from 15:1 to 85:1; from 16:1 to 85:1; from 20:1 to 83:1; and from 65:1 to 79:1; optionally wherein the ratio is about 76:1.

The mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in the faecal sample may be determined from a faecal sample collected from 12 to 28 hours after oral administration of a single dose of the composition to the human. Alternatively the faecal sample may be collected after a more prolonged period of regular oral administration of the composition to the human, after which the cyclosporin and metabolite concentrations in the faeces may have reached a steady state, thereby reducing the variability in the measured ratio of cyclosporin : metabolites. For example, the faeces may be collected 4 to 6 hours after the oral administration of the last dose of a dosage regimen wherein the cyclosporin composition is orally stered once or twice per day for 2, 3 4, 5, 6 or 7 days. The faeces may, for example, be collected after oral administration of 75mg of cyclosporin once or twice per day. Suitably, the faecal sample is collected 4 to 6 hours after oral administration of the last dose of a dosing regimen of the composition; the dosing n comprising once or twice daily oral administration of the composition to the human for seven days; optionally wherein the dosing n comprises once daily stration of the composition comprising 75mg of cyclosporin A for seven days. In a further embodiment the composition comprising cyclosporin is orally administered once per day for two days (for example as a single 75 mg daily dose of cyclosporin) and the faecal sample is collected 4 to 6 hours after the last dose of the composition on the second day.

The main metabolites of cyclosporin are the AM1, AM4N and AM9 cyclosporin metabolites.

The ratio of cyclosporin : cyclosporin lites in the faecal sample is suitably the ratio of cyclosporin: the total concentration of AM4N and AM9 cyclosporin metabolites. The ratio of cyclosporin: cyclosporin metabolites in the faecal sample may be the ratio of cyclosporin: the total concentration ofAM1, AM4N and AM9 cyclosporin metabolites.

The concentration of cyclosporin and its metabolites in the faeces may be measured using any suitable analytical method, for e chromatography and mass spectrometry as illustrated in the es section.

Suitably the concentrations of porin and lites in the faeces are ed in faecal samples obtained from healthy male subjects so as to se the inter-patient variability in the ed . The faecal samples are suitably obtained from healthy male subjects aged P213428WO 26 between 20 and 50 years, preferably weighing between 60 and 100 kg. Suitably the ratio is the arithmetic mean of the measured ratio of cyclosporin : porin metabolites in a representative number of subjects, for example at least 4, 5, 6, 7, 8, 9 or 10 subjects. lly at least 4 ts would be a sufficient number to provide a representative mean ratio.

The composition comprising cyclosporin A may be any composition comprising cyclosporin A which provides a ratio of porin : cyclosporin metabolites which is greater than 12:1 (or within any of the ranges described above in relation to this aspect of the invention). The composition is suitably a composition sing cyclosporin A, wherein the composition releases cyclosporin A in a solubilised form when the composition is placed in an aqueous dissolution medium. By "solubilised" in meant that the cyclosporin in released in an active form, for example in a dissolved form such as a solution, when the ition is placed in an aqueous dissolution medium, for example an s environment encountered in the lower GIT tract, particularly in the colon, following oral administration of the composition.

The composition may be, or comprise, cyclosporin A that is partially or completely ved in dissolved in a lipophilic substance. The ilic substance may be or comprise an oil, or a surfactant in which the cyclosporin is at least partially, or preferably fully dissolved. Suitable oils and surfactants which may be used include, but are not limited to any of the the oils and surfactants described herein.

] In one embodiment the cyclosporin may be dissolved or sed in a low melting point lipophilic substance, ly a substance with a melting point in the range of 30 to 70oC. Suitably the hydrophobic material is a wax like solid with a melting point in the range of 30 to 60oC, ularly suitable are lipophilic waxy material which are solid at room temperature, but which melt or soften at temperatures in the range of 30 to 50oC, or more preferably 30 to 40oC. The lipophilic material may be selected from one or more of unsaturated alcohols, hydrogenated alcohols, fatty acids, fatty acid esters, fatty acid amides, fatty acid mono- di- or triglycerides, polyethoxylated fatty acids and polyethyoxylated fatty acid esters, terol derivatives and waxes. The wax may be a suitable animal or plant d wax, for example carnauba wax or a tic wax such as paraffin wax. The lipophilic material may comprise a wax, a saturated ot rated fatty acid (for example palmitic, stearic, myristic, lauric, laurylic, or oleic acid), or a derivative thereof for example a mono-, di-, or triglyceride or a polyethylene glycol ester thereof. The lipophilic material comprising the dissolved or dispersed cyclosporin is suitably used in the form of a particulate composition, for example as a granule composition. Suitably the lipophilic substance comprising the dissolved or dispersed cyclosporin is itself dispersed, in a suitable carrier matrix. For example, granules sing the the lipophilic substance and dissolved or dispersed porin are dispersed in a suitable carrier matrix.

The carrier matrix may be a modified release matrix material, particularly a modified release polymer matrix. A modified release matrix may provide d or sustained release of the cyclosporin from the matrix, thereby ing a modified release composition. The carrier matrix is suitably a hydrophilic material. The matrix al may be, or comprise, acrylic or methacrylic acid polymers or copolymers, alkylvinyl polymers, hydroxyalkyl celluloses, carboxyalkyl celluloses, polysaccharides, dextrins, pectins, starches, starch derivatives, or natural or tic gums for example an alginate.

P213428WO 27 The carrier matrix may be a el forming polymer such as those described herein, including The composition may comprise cyclosporin A and a surfactant. Suitably the surfactant is or ses a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof as described herein. Suitably, the in this ment the weight ratio of cyclosporin to medium chain or long chain fatty acid mono- or di-glyceride or a combination is from about 3:1 to about 1:3, for example about 3:1 to about 1:2, or about 2.5:1 to about 1:1.8, about 1.5:1 to 1:1.5, about 1.2:1 to 1:1.2, about 1.2:1, about 1:1, or about 1:1.2. Suitably the cyclosporin composition in this embodiment r comprises an oil phase. The oil phase may be any suitable hydrophobic oil, for example an oil having a low HLB value (for example HLB less than 10). Particularly the oil phase may se any of the oil phases described herein, for example the oil phase may be or comprise cerides, for example a medium chain triglyceride. The weight ratio of oil to surfactant may be for example 12:1 to 1:5, for example from about 5:1 to about 1:5, from about 3:1 to about 1:2, from about 3:1 to about 1:1 or from about 2.5:1 to 1.5:1. The ition may further comprise a solvent. The solvent is suitably an organic solvent in which cyclosporin is soluble. More particularly suitable solvents e those in which both cyclosporin and the oil phase (when present) are soluble. For example the solvent may comprise 2-(ethoxyethoxy)ethanol. The cyclosporin may be partially or fully dissolved in the composition. Accordingly, the cyclosporin may be ntially dissolved in the composition. Suitably the porin is fully dissolved in the composition.

The ition is suitably formulated such that e of cyclosporin in the upper GI Tract, for example in the duodenum and jejunum, is minimised so as to se the systemic absorption of cyclosporin and both hepatic and enteric P450 metabolism. Accordingly a particular composition is a modified release composition. Suitably the release of cyclosporin from the composition is minimised for the first 4 hours after oral administration such that the composition can pass through the duodenum and jejunum and into the ileum before releasing large amounts of porin. Preferably the composition releases the majority (for example at least 50 %) of the cyclosporin into the colon.

Suitably the composition releases less than 40% (for example less than 35%, or less than 30%) of the cyclosporin from the composition 4 hours when ed in the two stage dissolution test described . Suitably the composition releases less than 15% (for example 0 to 10%) of the cyclosporin A after 2 hours; and es 10% to 40% (for example 10% to 35%, or suitably 15% to 35%) of the cyclosporin A at 4 hours, when ed in the two stage dissolution test.

The composition may be formulated to provide the desired modified release profile by, for example, use of any of the coatings described herein, in particular coatings which are adapted to release cyclosporin in at least the colon. Suitable coatings include, for example, a modified release coating comprising a pH independent polymer such as ethyl cellulose. The coating may also comprise a first coating comprising a water soluble cellulose ether such as HPMC, as described herein. Other modified release coatings are also contemplated including but not limited to enteric coating systems and other delayed release coatings. Generally the modified e coatings comprise a polymeric coating. 8WO 28 In this aspect of the invention, when the composition comprises cyclosporin A and a surfactant which is or comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof as described herein, the cyclosporin and surfactant are suitably dispersed within in matrix. The cyclosporin is released from the matrix when the composition is placed in an aqueous environment, for example as would be found in the lower GI tract such as in the colon. le matrix materials which may be used to disperse the cyclosporin and surfactant may be any of the matrix materials described herein, for example those described under "Composition" in the detailed description. In n ments the matri x material may be selected such that the matrix itself modifies the release of cyclosporin from the composition as described in more detail in the detailed description of the invention. In such itions it may be possible to achieve the desired inhibition of release of cyclosporin in the first 4 hours following oral administration without the need for additional modified release coating(s). In other embodiments the matrix material may be coated with one or more of the modified release coatings, such as those described herein to provide the required cyclosporin release profile. In a particular, in this aspect of the invention the matrix is a hydrogel polymer, as described herein, more particularly the matrix is or comprises n.

In a preferred embodiment in this aspect of the invention, the composition sing cyclosporin may be any of the cyclosporin compositions described herein which comprise a surfactant wherein the surfactant is, or ses, a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof.

The cyclosporin compositions described , more particularly the modified release compositions described herein, provide pharmacokinetic (PK) properties which minimise systemic exposure to cyclosporin compared to, for example, intravenous administration of cyclosporin and/or oral administration of instant e cyclosporin compositions such as ™. The following paragraphs describing suitable PK properties of the compositions are applicable to any of the cyclosporin compositions described .

The ition may provide a low systemic whole blood exposure to cyclosporin following oral administration of the composition. The composition may provide a mean whole blood cyclosporin A AUC0-inf of less than about 450 ng.hr/ml, less than about 350 ng.hr/ml, or less than about 300 ng.hr/ml after oral stration of the ition as a single dose containing 75 mg cyclosporin A to a human in a fasted state, or an AUC0-inf directly proportional thereto for a total dose other than 75mg. For e, the composition may provide a mean whole blood cyclosporin A AUC0-inf of from about 140 to about 420 ng.hr/ml, for example from about 150 to about 300 ng.hr/ml after oral administration of the composition as a single dose containing 75 mg cyclosporin A to a human in a fasted state, or an AUC0-inf directly proportional thereto for a total dose other than 75mg.

A high peak blood concentration of cyclosporin A (Cmax) may result in undesirable side s and potentially reduce the therapeutic window available for a composition ning cyclosporin A. Accordingly, the compositions ly provide a low Cmax. The composition comprising cyclosporin A, may provide a mean maximum whole blood concentration of cyclosporin A (Cmax) of less than 100 ng/ml. The composition may for example provide a Cmax of from about 15 to about 60 P213428WO 29 ng/ml, for example about 20 to 50 ng/ml, wherein in each case the Cmax is that measured after oral administration of the composition as a single dose ning 75 mg of porin A to a human in a fasted state, or a Cmax ly proportional thereto for a total dose other than 75mg.

The time taken to reach maximum whole blood concentration (Tmax) of the cyclosporin A suitably occurs n about 3 and about 10 hours after oral administration of the composition as a single dose to a human in a fasted state. The Tmax may occur between about 4 hours and about 10 hours, or between about 4 hours and about 8 hours, or between about 5 and about 6 hours after oral administration of the composition. For example a Tmax at about 5 hours, about 5.5 hours or about 6 hours.

An IV dose of 2 to 4 mg/kg/day cyclosporin is known to be efficacious in the treatment of ulcerative colitis patients (Lichtiger et al N. Engl J Med 1994; 330: 1841-1845). An IV dose of 2 mg/kg is approximates to a cyclosporin dose of approximately 150 mg (assuming an average weight of about 75 kg). As illustrated in the examples section it has been found that the AUC resulting from IV stration of cyclosporin as Sandimmun™ is significantly higher than the AUC resulting from the ed release compositions comprising the surfactant. IV administration of cyclosporin results in effectively 100% ic bioavailability. Accordingly, a comparison of the AUC for IV administration with the orally stered cyclosporin compositions described herein enable the absolute oral bioavailability (F%) to be determined. As illustrated in the examples, the cyclosporin compositions described herein provide a low absolute oral bioavailability. The F% is calculated by calculating the relative % of the AUC following oral administration relative to the AUC observed following IV administration of 2 mg/kg Sandimmun™. As will be ed compensation for the actual dose of cyclosporin needs to be accounted for when ating the F%. For example if the AUC for oral administration is that measured following a single dose of 75 mg of cyclosporin, the relative % needs to be multiplied by 2 to compensate for the fact that the effective total IV dose was 150 mg porin. Similarly, if a 37.5 mg dose of cyclosporin is administered , the relative % needs to be multiplied by 4.

The composition comprising cyclosporin A may provide a cyclosporin A absolute bioavailability following oral administration of the composition is less than 15%, for example less than %; ally wherein the absolute bioavailability is from 0.5% to 15% suitably from 1% to 10%.

Suitably the composition releases the cyclosporin in at least the colon. The composition may also release cyclosporin in other parts of the GI tract for example in the duodenum, jejunum and/or ileum. Suitably however, release of cyclosporin in the upper GI tract such as the duodenum and jejunum is minimised so as to reduce systemic exposure to cyclosporin A and/or reduce P450 metabolism of the drug. The release profile of porin A from the composition may be assessed by measuring the release in an in vitro dissolution test. The composition comprising cyclosporin may e less than 15% (for example 0 to 10%) of the cyclosporin A after 2 hours; releases 10% to 40% (for example 10% to 35%, or ly 15% to 35%) of the cyclosporin A at 4 hours; and releases from about 30% to 70% (for example 40% to 70%) of the cyclosporin A between 4 hours and 12 hours, when measured in a two stage dissolution test using a USP Apparatus II with a paddle speed of 75 P213428WO 30 rpm and a dissolution medium temperature of 37oC; wherein for the first 2 hours of the dissolution test the dissolution medium is 750 ml of 0.1 N HCl, and at 2 hours 250 ml of 0.2M tribasic sodium phosphate containing 2% SDS is added to the dissolution medium and the pH is adjusted to pH 6.8 (herein referred to as "the age dissolution test).

The composition may e less than 20% of the cyclosporin A after 2 hours; releases 10 to 40% of the cyclosporin A at 4 hours; and releases at least 60% of the cyclosporin A at 12 hours, when measured in the two stage dissolution test. The composition may release less than 10% of the porin A after 2 hours; releases 10 to 30% of the cyclosporin A at 4 hours; and releases at least 50% of the cyclosporin A at 12 hours, when measured in the two stage dissolution test. The composition may release from about 30 to about 75% of the porin A between 4 hours and 12 hours in the two stage dissolution test, for example the composition releases from about 40 to about 75%, particularly from about 45 to 70% of the cyclosporin A between 4 hours and 12 hours in the two stage dissolution test. The composition may release less than 15% (for example 0 to 10%) of the cyclosporin A after 2 hours; releases 10% to 40% (for example 10% to 35%, or suitably 15% to 35%) of the cyclosporin A at 4 hours; and es from about 25% to 70% (for example 40% to 70%) of the cyclosporin A between 4 hours and 12 hours in the two stage dissolution test.

It is to be understood that any of the individual PK parameters and/or in-vitro or other release es described herein may be ed with the compositional features of the cyclosporin compositions described herein, for example relating to any one or a ation of AUC; Cmax; Tmax; cyclosporin A concentration in the luminal contents; cyclosporin A tration in the GI tract tissue; the ratio of cyclosporin A in the luminal contents : cyclosporin A in GI tract tissues; the ratio of the concentration of cyclosporin A: the concentration of cyclosporin A metabolites in collected faeces; the concentration of cyclosporin A in intracolonic faeces: the concentration of cyclosporin A in colonic tissue; or the concentration of cyclosporin A in colonic tissue. By way of a non -limiting example of such a combination of es composition comprising porin A, provides a mean concentration of cyclosporin A: the concentration of cyclosporin A metabolites in a faecal sample from the human of greater than 12:1 after oral administration of the composition to the human; and wherein the composition provides an nf of less than about 450 ng.hr/ml, (e.g. from about 140 to about 420 ng.hr/ml) after oral administration of the composition as a single dose containing 75 mg cyclosporin A to a human in a fasted state, or an AUC0-inf directly proportional thereto for a total dose other than 75mg. Optionally this composition may release 0 to 10% of the cyclosporin A after 2 hours; 10% to % of the cyclosporin A at 4 hours; and releases from about 40% to 70% of the cyclosporin A between 4 hours and 12 hours, when ed in the two stage dissolution test.

In another embodiment the composition releases 0 to 10% of the cyclosporin A after 2 hours; and releases from 50 to 100% of the cyclosporin A after 12 hours, when measured in the two stage dissolution test. In another embodiment the composition releases less than 20% of the cyclosporin A after 2 hours; es 5 to 40% of the cyclosporin A at 4 hours and releases at least 50% of the cyclosporin A at 12 hours, when measured in the two stage dissolution test.

P213428WO 31 The cyclosporin compositions described herein are expected to provide similar or higher levels of cyclosporin A in the colonic tissue compared to IV administration of Sandimmun™, but with a higher intracolonic faecal concentration of cyclosporin A as a result of the local release of the cyclosporin directly into the colon. The vely high local concentration of cyclosporin in the colon is expected to provide beneficial therapeutic effects.

The composition comprising cyclosporin A described herein, may provide a ratio of the mean concentration of cyclosporin A present in intracolonic faeces : the mean concentration of cyclosporin A present in colonic tissue in an adult human patient after oral stration of the composition of from about 50:1 to about 500:1, optionally from about 80:1 to about 300:1, or optionally about 100:1 to about 250:1; wherein the concentration of the cyclosporin A is measured in samples of the intracolonic faeces and the colonic tissue taken substantially simultaneously 4 to 6 hours after oral administration of the last dose of a once daily oral dosing regimen of the composition, the dosing regimen comprising once daily oral administration of the composition for seven days. Optionally the once daily oral dosing regimen of the composition provides a single daily dose of 75 mg cyclosporin A. However, other doses may be administered for e any of the cyclosporin doses described herein including but not limted to 37.5 mg or 150 mg once per day. Optionally the dosage regimen may be a twice daily dosage regimen for seven days, for example 37.5 mg twice per day, 75 mg twice per day or 150 mg twice per day.

Reference herein to a sample being taken "substantially simultaneously" means that the s are obtained close to the same time point, for example the colonic tissue and/or intracolonic faeces and/or blood samples are taken within about 2 hours, 1 hour or 30 minutes of each other, suitably the samples are all taken at the same time point.

In contrast to the compositions according to the invention, IV administration porin as Sandimmun™ s in a lower ratio of the mean concentration of cyclosporin A present in intracolonic faeces : the mean tration of cyclosporin A present in c tissue. As illustrated in the Examples, when patients were treated with mun® IV (2mg/kg) stered as an infusion over 24 hours (2mg/kg/day) a ratio of about 3:1 was observed.

Accordingly, the orally administered cyclosporin compositions described herein are expected to provide a relatively high colonic tissue concentrations ed to IV administration of an equivalent dose of cyclosporin (e.g. as Sandimmun™ IV).

The composition comprising cyclosporin A may provide a concentration of cyclosporin A in c tissue of at least 250 ng/g following oral administration of the composition to a human, for e at least 300, 350, 400 or 450 ng/g. Accordingly, the composition may provide a cyclosporin A concentration of from about 250 to 6000 ng/g, for example from 400 ng/g to 6000 ng/g, from 500 to 5000 ng/g, from 600 to 4000 ng/g, or from 600 to 2000 ng/g. Particularly the composition provides a porin A concentration in colonic tissue of from about 1000 to about 1500 ng/g, for example about 1200 ng/g. Suitably the compositon is orally administered to provide a daily dose of cyclosporin A within the ranges described herein, suitably a total daily dose in the range of 15 to 300 mg P213428WO 32 cyclosporin. Optionally the composition provides a dose may be 37.5 mg, 75 mg or 150 mg cyclosporin A once or twice per day.

The colonic tissue samples described above may be obtained using conventional methods, for example by taking a tissue sample during endoscopy as described in the examples herein.

As described above, compositions comprising cyclosporin A and a surfactant, wherein the surfactant comprises, or is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof may inhibit P450 metabolism of porin A. Such compositions may be useful for the preparation of modified release of porin in the lower GI tract as bed above. Also contemplated are such compositions for administration of cyclosporin to any part of the GI tract, for example duodenum or jejunum as, for example, an t release ition. The compositions may reduce the rate and or extent of cyclosporin metabolism and thereby maximise the amount of cyclosporin in the GI tract.

According to a further feature of the invention there is provided a composition comprising cyclosporin A and a surfactant, wherein the surfactant comprises, or is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof. Suit ably the ition does not comprise or is not a polyethyleneglycol ether or ester. Suitably the surfactant is present in an amount of at least 6% by weight of the composition, for example at least 10%, at least 15% or at least 20% by weight of the composition. Optionally the surfactant is present in an amount of from 10 to about 50% by weight.

The composition according to this aspect of the invention may further comprise an oil phase, for example any of the oil phases bed herein.

The porin may be partially or completely dissolved in the composition. Suitably the cyclosporin is completely dissolved in the composition.

A particular composition comprises: (i) 10 to 60 parts cyclosporin A; (ii) 5 to 40 parts of a medium chain fatty acid triglyceride, for example a caprylic/capric triglyceride; (iii) 10 to 50 parts of the surfactant; and (iv) 0 to 60 parts solvent; n all parts are parts by weight and the sum of the parts (i) + (ii) + (iii) + (iv) = 100.

Another composition ses: (i) 10 to 40 parts cyclosporin A; (ii) 5 to 25 parts of a medium chain fatty acid triglyceride, a ic/capric triglyceride; (iii) 15 to 30 parts of the surfactant; and (iv) 10 to 60 parts solvent nally 20 to 40 parts or 25 – 30 parts solvent), for example 2- (2-ethoxyethoxy)ethanol); wherein all parts are parts by weight and the sum of the parts (i) + (ii) + (iii) + (iv) = 100.

] Optionally in this aspect the surfactant is selected from glyceryl caprylate, glyceryl caprate, glyceryl monooleate, glyceryl dioleate and glycerol monolinoleate, or a combination thereof. 8WO 33 The cyclosporin compositions according to this aspect may be stered orally, for example to provide an instant release composition. Also plated is the administration of the composition to the GI tract rectally, for example in the form of an enema or suppository. Other routes of administration of the composition are also contemplated, for example the composition may be administered directly to the GIT, by for example intra-duodenal administration, intra-jejunal or intraileal administration. Such routes of administration enable the compositon to bypass the h (and optionally other parts of the GI tract) for delivery to specific points in the lower GI tract. These routes of administration may be achieved using for example suitable tubing with an exit at the desired location within the GI tract. Suitably the tubing is inserted orally or nasally into the GI Tract.

Alternatively, administration may be achieved by gastric tubing, or continuous or discontinuous p ercutaneous endoscopic stomy (PEG) tubing. PEG is an endoscopic medical ure in which a tube (PEG tube) is passed into a patient's stomach through the abdominal wall. This method of administration may be ularly suitable for patients that cannot take the drug orally due to for example gia or sedation.

A further aspect of the ion provides a composition described herein for use as a medicament. The composition may comprise at least one further active ingredient, for example at least one further immunosuppressant. In particular there is provided a composition for use in the treatment, e.g. prevention, of a condition of the GIT. The composition may be for use in the treatment of an inflammatory bowel disease, irritable bowel syndrome, Crohn’s disease, ulcerative colitis, celiac disease, graft-versus-host disease, intestinal graft-versus-host disease, gastroenteritis, duodenitis, jejunitis, s, peptic ulcer, Curling’s ulcer, appendicitis, colitis, pseudomembraneous s, diverticulosis, diverticulitis, pouchitis, collagenous colitis, macorscopic colitis, diarrheal colitis, triosis, colorectal carcinoma and adenocarcinoma. The composition may also be for use in the treatment of proctitis. The composition may be for use in the prevention or treatment of primary sclerosing gitis, familial adenomatous polyposis, or perinanal Crohn’s, including perianal fistulae.

] In embodiments where the pharmaceutical composition does not comprise a second coating, the composition may be for use in the treatment of conditions that affect the small intestine. Such compositions may be able to treat conditions selected from celiac disease, GVHD or s disease.

The invention onally provides a method for administering cyclosporin to a subject, comprising orally administering to the subject a composition described herein. The method may be performed in the treatment, e.g. prevention, of disease. The subject may be a mammal, in particular a human. Also provided is a method for treating a condition of the GI tract in a subject, preferably a human, in need thereof comprising orally administering to the mammal a therapeutically effective amount of a composition described . Conditions of the GI tract which may be treated or ted include the conditions disclosed herein.

A further aspect of the ion provides the use of a composition described herein for use in the manufacture of a medicament for the treatment, e.g. prevention, of a condition of the GIT.

Conditions of the GI tract include those disclosed herein.

P213428WO 34 The invention also contemplates a method of treating a condition selected from inflammatory bowel disease, irritable bowel e, s disease, tive colitis, celiac disease, graft vs host disease, gastrointestinal graft-versus-host disease, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, Curling’s ulcer, appendicitis, colitis, pseudomembraneous colitis, diverticulosis, diverticulitis, collagenous colitis, endometriosis, colorectal carcinoma and adenocarcinoma, n the method comprises administering a ceutical composition of the invention.

In r aspect the invention provides a method of treating conditions that affect the small intestine, wherein the method comprises stering a composition of the ion which does not comprise a second coating. The conditions of the small intestine may be selected from celiac disease, GVHD or Crohn’s disease.

In an aspect of the invention there is provided a process for making a liquid composition, the process comprising mixing an oil phase with an aqueous phase comprising a hydrogel forming polymer, wherein the oil phase has cyclosporin in solution and comprises a surfactant which is medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, wherein the surfactant does not comprise or is not a polyethyleneglycol ether or ester.

Optionally, the oil phase and the aqueous phase are mixed in an oil phase to aqueous phase ratio of from 1:2 to 1:12, optionally 1:4 to 1:10, 1:4 to 1:8, for example 1:5 or 1:7.

The process may further comprise the step of causing the emulsion to solidify.

The process may further comprise the step of: coating a core with a coating comprising HPMC wherein the weight gain due to the coating is from 0.5% to 20% of the weight of the pharmaceutical composition. The core may compris e a ceutically active ingredient and may be a core as described in this ication.

] An additional advantage of the present application may be that a composition ved in a dissolution medium yields a uniform droplet size with low polydispersivity compared to ations with a different first surfactant.

Accordingly, there is provided a ition comprising cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, wherein the surfactant is or comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not se or is not a polyethyleneglycol ether or ester, wherein the composition releases droplets with a uniform size. Optionally the droplets may have low polydispersity. Optionally the uniform size may be a droplet size of from about 1 nm to about 350 nm.

The droplet size may be selected from: from about 20 nm to about 350 nm; from about 20 nm to about 300 nm; from about 20 nm to about 250 nm from about 100 nm to about 350 nm; from about 100 nm to about 300 nm; from about 100 nm to about 250 nm; from about 100 nm to about 200 nm; from about 150 nm to about 250 nm; from about 150 nm to about 200 nm; from about 150 nm to about 350 nm; and from about 150 nm to about 300 nm. Preferably, the t size may be selected P213428WO 35 from: from about 20 nm to about 250 nm; from about 100 nm to about 250 nm; and from about 100 nm to about 200 nm.

The size of the droplets may be measured using c light scattering. The dynamic light scattering experiments were carried out by analysis of a liquid medium arrived at as follows. Minibeads of the invention (0.5 g) comprising Capmul GMO-50 as the first surfactant were added to a beaker ning 50 g of deionised water. The beaker contents were mixed at 250 rpm and at 37 °C throughout the study. Samples of the beaker contents were taken at 0, 1, 2, 3, 4, 5, 6 and 24 hours.

Samples of the beaker contents were filtered 0.65µm pore size filters (Merck Millipore Ultrafree-CL Centrifugal Filter). The particle size and zeta potential was measured using a Malvern Nano-Zetasiser.

For certain active ingredients it may be desirable to limit or delay release of the active from the composition until the composition has passed through the stomach and upper GI tract. The compositions of the invention comprising a second coat may be particularly suitable for such ations. The second coat acts to delay release from the composition, whilst the presence of the coating of the invention (e.g. HPMC) increases the amount of active released when the composition releases the active in the lower GI tract. The period of delay to the release of the active as a result of the ce of the second coating can be tailored by appropriate selection of the nature or amount of second g used. For a given second coating material a higher weight gain of coating will generally increase the time period between administration of the composition and e of the active. The compositions of the invention can therefore be used to provide high levels of release of active agent at very specific parts of the GI tract to provide, for example, topical treatment to diseased tissue within the GI tract. Such delayed release compositions may be particularly beneficial when the active has undesirable side effects which may arise from systemic absorption higher in the GI tract. ed in this description by reference are the subject matters of the appended claims.

The description is therefore to be read together with the claims and features mentioned in the claims are applicable to the subject matters of the ption. For example, a feature described in a s claim is applicable also to products mentioned in the description, where the feature is manifested in the product. For example, a e mentioned in a product claim is applicable also to relevant process t s contained in this description. rly, a feature mentioned in the description in the context of a process is applicable also to products mentioned in the description, where the feature is manifested in the product. Also, a e mentioned in the description in the context of a product is applicable also to relevant s subject matters contained in this ption.

BRIEF DESCRIPTION OF THE GS Embodiments of the invention are further described hereinafter with reference to the anying drawings, in which: Figure 1 is an image showing crystal ion over time of a comparative composition.

Figure 2 is an image showing crystal formation over time of a composition of the invention comprising Capmul GMO-50 (glyceryl monooleate/dioleate) as the surfactant (the first surfactant).

Figure 3 is an image showing crystal formation over time of a composition of the invention comprising Capmul MCM (glyceryl caprylate/caprate) as the surfactant (the first surfactant).

P213428WO 36 Figure 4 is an image showing l formation over time of a composition of the invention comprising Maisine 35-1 (glycerol monolinoleate) as the surfactant (the first surfactant).

Figure 5 is a graph ng % of cyclosporin released t time over 24 hours and showing the e profiles of ads of Example 2.

Figure 6 is a graph ng % of cyclosporin released against time over 24 hours and showing the release profiles of minibeads of Comparative Example 1.

Figure 7 is a graph plotting % of cyclosporin released against time over 24 hours in deionised water and showing the release profiles of minibeads of Example 6a.

Figure 8 is a graph plotting % of porin released against time over 24 hours in the twostage dissolution test g the release profiles of beads of Example 4a, specifically those of Table Figure 9 is a graph showing the droplet size and zeta potential of a coated composition of the invention when the composition has been dissolved in deionised water.

Figure 10 shows the dosing schedule used in the clinical trial described in Example 9.

Figure 11 shows the median whole blood cyclosporin concentration time profiles r linear) on day 1 in those subjects treated with CyCol® in the clinical trial described in Example 9.

Figure 12 the median whole blood cyclosporin concentration time profiles (linear linear) on day 7 in those subjects treated with CyCol® in the clinical trial described in Example 9. In Figures 11 and 12, BID=twice daily; OD=once daily. Values below the lower limit of quantification (<0.2 ng/mL) are presented as equal to zero.

Figure 13 shows the mean whole blood tration of cyclosporin A obtained from the comparative clinical trial of Comparative Example 10 on linear and log scales.

Figure 14 shows the ratio of cyclosporin A to the concentration of the (AM4N + AM9) cyclosporin metabolites measured in the faecal samples for each of the tested formulations in Example 9 and Comparative Example 10.

Figure 15 shows the in-vitro release es of the compositions used in Example 9 and Comparative Example 10. In Figure 14 "PK fast", "PK " and "PK slow" refer to the Fast Release Formulation, Formulation I and Formulation II used in Comparative Example 10. "CyCol 2014" refers to the formulation used in Example 9.

Figures 16 to 22 show photomicrographs of the emulsions described in Example 11 sing surfactants in an aqueous phase at specified time points.

DETAILED DESCRIPTION A mono-glyceride or di-glyceride of the present invention may comprise one glycerol fied to one fatty acid or one glycerol esterified to two fatty acids the fatty acids may be the same or ent, ordinarily the fatty acids will be the same. The surfactant of the invention is a surfactant that does not comprise or is not a polyethyleneglycol ether or ester; by this it is meant that there is no polyethyleneglycol component bonded to the surfactant molecule by an ether or ester linkage. For example a pegylated fatty acid glyceride such as oleoyl macrogol-6 ides (commercially available as Labrafil M1944CS). It is possible that a commercial surfactant of the invention is supplied with a small amount of polyethyleneglycol (PEG) contained within the supplied tant composition.

P213428WO 37 The use of such commercial formulations of surfactants which contain non-bonded PEG, put another way free PEG) are not excluded by the limitation that the tant does not comprise or is not a hyleneglycol ether or ester.

Reference to "cyclosporin" herein is a reference to cyclosporin-A (also known as porine and the INN ciclosporin. It is plated that other forms of cyclosporin may be used in the compositions described herein, for example cyclosporin –B, -C, -D or –G and derivatives or pro drugs of any thereof.

The term "treatment", and the therapies encompassed by this invention, include the following and combinations thereof: (1) reducing the risk of or inhibiting, e.g. delaying, initiation and/or progression of, a state, disorder or condition; (2) preventing, e.g. reducing the risk of, or delaying the appearance of clinical symptoms of a state, disorder or condition developing in a patient (e.g. human or animal) that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display al or subclinical symptoms of the state, disorder or ion; (3) inhibiting the state, disorder or condition (e.g., arresting, reducing or delaying the development of the disease, or a relapse f in case of maintenance treatment, of at least one clinical or subclinical symptom thereof); and/or (4) relieving the condition (e.g. causing sion of the state, disorder or condition or at least one of its clinical or subclinical symptoms). Where the composition of the invention is used in the treatment of a patient, treatment contemplates any one or more of: maintaining the health of the patient; restoring or improving the health of the patient; and delaying the ssion of the disorder.

The benefit to a patient to be treated may be either statistically significant or at least perceptible to the patient or to the physician. It will be understood that a medicament will not necessarily produce a clinical effect in every patient to whom it is administered, and this paragraph is to be understood accordingly. The itions and s described herein are of use for therapy and/or prophylaxis of disease.

The treatments may include maintenance therapy of patients who have ed a disorder and whose condition has subsequently improved, e.g. because of treatment. Such ts may or may not suffer a symptomatic disorder. nance therapy aims to arrest, reduce or delay (re- )occurrence or progression of a disorder.

"Effective amount" means an amount sufficient to achieve the desired treatment, e.g. result in the desired therapeutic or prophylactic response. The therapeutic or prophylactic response can be any response that a user (e.g., a clinician) will recognise as an ive response to the therapy. It is further within the skill of one of ordinary skill in the art to determine appropriate treatment on, appropriate doses, and any potential combination treatments, based upon an evaluation of therapeutic or prophylactic response.

The terms "dry" and "dried" as applied to compositions or compositions of the disclosure may each include reference to compositions or compositions containing less than 5% free water by weight, e.g. less than 1% free water by weight. Primarily, however, "dry" and "dried" as applied to itions of the sure mean that the hydrogel present in the initial solidified composition has P213428WO 38 dried sufficiently to form a rigid ition. Where a solid colloid is referred to this also refers to a dried colloid according to the definition herein.

Ingredients and excipients of the described compositions are suitable for the intended purpose. For example, pharmaceutical compositions comprise pharmaceutically acceptable ingredients.

If not otherwise stated, ingredients, components, excipients etc. of the compositions of the ion are suitable for one or more of the intended purposes sed elsewhere herein.

For the avoidance of doubt, it is hereby stated that the information disclosed r in this specification under the heading "Background" is nt to the invention and is to be read as part of the disclosure of the invention.

Where the invention is referred to as a formulation it takes the same meaning as the composition of the invention. Accordingly, formulation and composition are used interchangeably.

Throughout the description and claims of this specification, the words "comprise" and "contain" and variations of them mean "including but not limited to", and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this ication, the ar encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the t requires otherwise.

Features, rs, characteristics, compounds, chemical es or groups bed in conjunction with a particular , embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments.

The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying , ct and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

The reader’s attention is directed to all papers and documents which are filed rently with or previous to this ication in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference. ition The liquid composition comprises cyclosporin, a polymer capable of forming a matrix (a hydrogel forming polymer), a tant and an oil phase. The oil phase, cyclosporin and the surfactant are contained within the polymer capable of creating a matrix. The cyclosporin is dissolved P213428WO 39 in the oil phase. When the polymer forms a matrix the liquid composition is formed into a composition of the invention.

The composition comprises a matrix and cyclosporin. The matrix may be formed with a hydrogel-forming polymer, and may contain additional excipient(s) to the polymer. The active ingredient is contained within the matrix. The active ingredient may be in solution or in suspension, or in a combination thereof; however the invention is not limited to compositions sing a solution or sion of the active and it includes, for example, active ingredients encapsulated in liposomes or extrin. The matrix may contain inclusions in which the active ingredient is comprised; for example, the inclusions may comprise a hydrophobic medium in which the active ingredient is dissolved or suspended. An active ingredient may ore be directly dissolved or suspended in the matrix, or it may be dissolved or suspended indirectly in the matrix by way of inclusions in which the active ingredient is dissolved or suspended.

The composition, therefore, ses a matrix-forming polymer, in particular a hydrogelforming polymer. The matrix of the composition may be or comprise a polymer matrix comprising a polymer selected from a water-permeable r, a water-swellable polymer and a biodegradable polymer. In particular, the matrix is or comprises a hydrogel-forming polymer described in more detail below.

] Modified release of the active ingredient from the composition may be achieved by virtue of the ties of the matrix material. For example the matrix may be a permeable or erodible polymer within which the active ient is contained, e.g. dissolved or suspended; following oral stration the matrix is gradually dissolved or eroded thereby releasing the active ingredient from the matrix. Erosion may be ed by biodegradation of a biodegradable polymer matrix. Where the matrix is permeable, water permeates the matrix enabling the drug to diffuse from the matrix. A matrix formed with a hydrogel-forming polymer may therefore include a modified release polymer. As such modified release rs may be mentioned cellulose tives, for example hydroxypropylmethyl cellulose, poly(lactic acid), poly(glycoloic)acid, poly(lactic –co glycolic acid copolymers), polyethylene glycol block co-polymers, polyorthoesters, polyanhydrides, polyanhydride esters, hydride imides, ides and polyphosphazines.

Water Soluble ose Ether Coating The invention provides pharmaceutical compositions that may have a first coating which is or ses a water-soluble cellulose ether. The invention provides pharmaceutical compositions that have a first polymer coating, wherein the polymer is or comprises a water-soluble ose ether. The water-soluble cellulose ether may be, for e selected from methyl cellulose, hydroxyethyl cellulose, hydroxylpropyl cellulose and hydroxypropylmethyl cellulose.

Suitably the material of the first coating (i.e. the sub-coating) is different to the second coating on the composition. For example, where the first coating is or comprises a water-soluble ester of a cellulose ether, the major component(s) (e.g. more than 50%) of the second coating is or ses a different polymer to that of the first coating. Accordingly, the first and second coatings P213428WO 40 suitably provide two layers of material as part of the composition. It is to be understood that when the second coating comprises a mixture of components, minor ents of the outer second g may the same as the material of the first coating. By way of example, when the first coating is or ses HPMC and the second coating ses ethyl cellulose, the ethyl cellulose may optionally r comprise a minor amount (e.g. less than 50%,40%, 30% or 20%) of the first coating material, HPMC in this example. In such embodiments the sub-coat and the second coating are considered to be ent.

The water-soluble ose ether may be a water-soluble cellulose ether selected from an alkyl cellulose, for example methyl cellulose, ethyl methyl cellulose; a hydroxyalkyl cellulose, for example hydroxyethyl ose able as Cellosize™ and Natrosol™), hydroxypropyl cellulose (available as Klucel™) or hydroxymethyl cellulose; a hydroxyalkyl alkyl cellulose, for example hydroxyethyl methyl cellulose (HEMC), hydroxypropyl methyl cellulose (available as Methocel™, Pharmacoat™, Benecel™) or ethyl hydroxyethyl cellulose (EHEC); and a carboxyalkyl cellulose, for example carboxymethyl cellulose (CMC). Suitably the water-soluble cellulose ether may, for example be selected from methyl cellulose, hydroxyethyl cellulose, hydroxylpropyl cellulose and hydroxypopylmethyl cellulose.

The water-soluble ose ether may be a low viscosity polymer which is suitable for ation as a film or coating to the composition. The viscosity of the polymer may be from about 2 to about 60 mPa.s, for example a viscosity of: about 2 to about 20 mPa.s; about to 2 to about 8 mPa.s; more suitably a viscosity of about 4 to about 10 mPa.s, for example about 4 to about 6 mPa.s.

Alternatively, the viscosity of the polymer may fall outside any or all of the just-mentioned ranges, for example be above 20 mPa.s. Alternatively, the viscosity of the polymer may fall outside any or all of the just-mentioned ranges, for example be above 20 mPa.s. The viscosity of the polymer may be determined by ing the viscosity of a 2% solution of the polymer in water at 20oC using a Ubbelode eter using ASTM standard methods (D1347 and D2363).

The water soluble ose ether may be a water-soluble hydroxypopylmethyl cellulose (HPMC or hypromellose). HPMC is prepared by modifying ose to substitute hydroxy groups with methoxy and hydroxypropyl groups. Each anhydroglucose unit in the cellulose chain has three hydroxyl groups. The amount of substituent groups on the anhydroglucose units may be expressed as the degree of substitution. If all three hydroxyl groups on each unit are substituted, the degree of substitution is 3. The number of substituent groups on the ring determines the properties of the HPMC. The degree of substitution may also be sed as the weight % of the methoxy and hydroxypropyl groups present. Suitably the HPMC has from about 19 to about 30% methoxy substitution and from about 7 to about 12% hydroxypropyl substitution. ularly the HPMC has 25 to 30% methoxy substitution and 7 to 12% hydroxypropyl substitution. Suitably the HPMC is a low viscosity HPMC which is suitable for application as a film or coating to the composition. The viscosity of the HPMC is suitably from about 2 to 60 mPa.s, for e about 2 to about 20 mPa.s, more suitably a viscosity of about 4 to about 10 mPa.s.. The viscosity of the HPMC is determined by measuring the viscosity of a 2% solution of the HPMC in water at 20oC using a Ubbelode viscometer 8WO 41 using ASTM rd methods (D1347 and D2363). Such HPMC is available as for example Methocel™, for example Methocel™ E, including Methocel™ E5.

] When the first g is or comprises a soluble derivative of a cellulose ether, the derivative may, for example be a water-soluble ester of a cellulose ether. Water-soluble esters of cellulose ethers are well known and may comprise esters of a cellulose ether, formed with one or more le acylating s). Acylation agents may be, for example suitable acids or acid anhydrides or acyl halides. Accordingly the ester of a cellulose ether may contain a single ester moiety or two or more ester moieties to give a mixed ester. Examples of water-soluble esters of cellulose ethers may be water-soluble phthalate, acetate, succinate, propionate or butyrate esters of a cellulose ether (for example HPMC). Suitably the water-soluble ester of a cellulose ether is a soluble phthalate, acetate-succinate, propionate, acetate-propionate or acetate-butyrate ester of a cellulose ether (for example HPMC).

In one embodiment the water-soluble ester of a ose ether may be or comprise a watersoluble ester of any of the water-soluble cellulose ethers described above in relation to the ting.

Particular water-soluble esters of cellulose ethers are water-soluble esters of HPMC. Esters of HPMC which are soluble in water at a pH greater than 5.5 may be or comprise hydroxypropyl methylcellulose phthalate (HPMCP), or hydroxypropyl methylcellulose acetate succinate (HPMCAS) in which the presence of ionisable carboxyl groups causes the polymer to solubilize at high pH (> 5.5 for the LF grade and > 6.8 for the HF grade). These polymers are commercially available from Shin-Etsu Chemical Co. Ltd.

The cellulose ether-containing g may comprise or be hypromellose, e.g. it may be made of a e of hypromellose, titanium dioxide and polyethylene glycol; the coating may se at least 20wt% hypromellose and optionally at least 50% or at least 75wt% hypromellose, e.g. at least 80wt% or at least 85wt% or 90wt% hypromellose. The coating material used to form the coating may therefore se a dry weight percentage of hypromellose mentioned in the preceding sentence.

If it is desired for the coating to use a mixture of hypromellose, titanium dioxide and polyethylene glycol, commercial products ponding to such mixtures are available including Opadry White, a product commercialised by Colorcon. More generally, there may be mentioned various products commercialised under the trade name Opadry and Opadry II. Further non limiting examples include Opadry YS7706-G white, Opadry Yellow 03B92357, Opadry Blue 03B90842).

These formulations are available as dry film coating formulations that can be diluted in water shortly before use. Opadry and Opadry II formulations comprise a cellulosic film forming polymer (e.g., HPMC and/or HPC), and may contain polydextrose, maltodextrin, a plasticizer (e.g., tin, polyethylene glycol), polysorbate 80, a colorant (e.g., titanium dioxide, one or more dyes or lakes), and/or other suitable film-forming rs (e.g., acrylate-methacrylate copolymers). Suitable OPADRY or OPADRY II formulations may comprise a plasticizer and one or more of maltodextrin, and polydextrose (including but not limited to a) triacetin and polydextrose or maltodextrin or lactose, or b) P213428WO 42 polyethylene glycol and polydextrose or maltodextrin). Particularly preferred commercial products are Opadry White (HPMC/HPC-based) and Opadry II White (PVA/PEG-based).

The cellulose ether-containing coating may also be d as a simple solution comprising water and the polymer of the first coating. For example when the polymer is an HPMC, for example such as Methocel, the first g may be applied to the core as an aqueous solution or dispersion of the HPMC. ally the coating solution may include other solvents such as an alcohol. atively the g may be applied as a solution or dispersion in a volatile organic solvent.

Suitably the first coating that contains a water e ose ether is present in an amount corresponding to a weight gain of the composition due to the coating of from 0.5% to 40% (for example from 0.5% to 30%; from 0.5% to 20%; from 1% to 25%; from 1% to 15%; from 1% to 6%; from 1% to 4%; from 4% to 6%; from 6% to 10%; from 9% to 15%; or from 12% to 15%) by weight based upon the weight of the composition prior to applying the g. The first g that contains a water soluble cellulose ether is present in an amount corresponding to a weight gain of the composition due to the coating of from 1% to 10%; from 4% to 10%; from 4% to 8%; and from 5% to 8% by weight based upon the weight of the core or the composition prior to applying the coating.

In r embodiment the first coating that contains a water-soluble cellulose ether is present in an amount corresponding to a weight gain due to the first coating in a range selected from 9 to 30%, suitably 9% to 20%, or particularly 10% to 15% by weight based upon the weight of the composition prior to applying the coating.

Suitably the first coating that contains a water soluble ose ether provides a coating thickness on the composition of at least 5 µm, suitably from about 5 µm to about 1 mm, for example from about 10µm to about 1mm, , from about 10 µm to about 500 µm, from about 50 µm to about 1 mm, or about from about 50 µm to about 500 µm. The thickness may therefore be from about 100 µm to about 1 mm, e.g. 100 µm to about 750 µm or about 100 µm to about 500 µm. The thickness may be from about 250 µm to about 1 mm, e.g. about 250 µm to about 750 µm or 250 µm to about 500 µm.

The thickness may be from about 500 µm to about 1 mm, e.g. about 750 µm to about 1 mm or about 500 µm to about 750 µm. The thickness may therefore be from about 10 µm to about 100 µm, e.g. from about 10 µm to about 50 µm or about 50 µm to about 100 µm.

When the first coating comprises a water-soluble cellulose ether the cellulose ether(s) suitably forms at least 40%, 50%, 60%, 70%, 80%, 85% or 90% by weight of the dry weight of the first coating. Alternatively the water-soluble cellulose ether is the first coating.

It is preferred to dry the composition of the invention before the first coating that ns a water-soluble cellulose ether is applied, as is described in more detail below in relation to the coating process.

It has been found that applying to a core sing a pharmaceutically active ingredient a sub-coating, referred to ere in the application as the subcoat (hence the t and the first coating are equivalent), that contains a water soluble cellulose ether prior to applying a delayed release coating provides unexpected advantages. The presence of such a sub-coating has been P213428WO 43 found to enhance the dissolution properties of the delayed release compositions according to the ion. In particular the presence of such a sub-coating has been found to increase the rate of release of the active ingredient from the composition and also to increase the amount of the active ingredient released in a set time period ed to compositions prepared without using such a ting.

These findings are unexpected, because it would have been expected that the presence of a sub-coating in on to a delayed release outer coating would act to delay or inhibit release of drug from the composition and, at a given time, for there to be less drug released, e there is a r coating present. However, as illustrated in the Examples, contrary to these expectations both the extent and rate of release of active ingredient are increased compared to compositions without such a sub-coating. Accordingly, delayed e compositions according to the ion which comprise a sub-coat that comprises or is a water-soluble cellulose ether and a delayed e coating outside the at, provide a unique dissolution e. The presence of such a ating has also been found to reduce batch-to-batch variability, particularly when the core is in the form of a minibead. A sub-coating that comprises or is a soluble cellulose ether may therefore also reduce intra- and inter-patient variability as a result of a more tent dissolution profile. The unique properties of sub-coated compositions according to the invention (particularly the dissolution profile) are expected to contribute to favourable pharmacokinetic properties of the compositions ing to the invention.

Accordingly in an embodiment there is provided a composition comprising cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, wherein the surfactant is a medium chain or long chain fatty acid mono- or diglyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester, the composition r comprising a first coating; and wherein the first coating is or comprises a water-soluble cellulose ether.

The composition may have a second coating comprising or being a delayed release polymer.

Accordingly in an embodiment there is provided a composition comprising cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, wherein the surfactant is a medium chain or long chain fatty acid mono- or diglyceride or a combination thereof and does not comprise or is not a hyleneglycol ether or ester, the ition further comprising a first coating and a second coating outside the first coating; and wherein the first coating is or comprises a water-soluble cellulose ether; and the second coating is or comprises a delayed release coating, e.g. is or comprises a delayed release polymer.

An aspect of the invention resides in a multiple minibead composition comprising at least two populations of active ingredient-containing minibeads, wherein s of at least one minibead tion are minibeads as described herein (i.e. compositions of the invention in minibead format).

It will be understood that the two populations are different. Such a plural minibead tion composition may comprise or consist of the following two populations: P213428WO 44 a first population having a coating that is or comprises a water-soluble cellulose ether but having no outer coating, e.g. as bed herein; and a second population having a first coating that is or comprises a water-soluble cellulose ether and a second coating that is or comprises a delayed release coating, for e as described herein e.g. a coating that is or comprises a delayed e polymer.

The tive minibeads of each population of a plural minibead composition may contain cyclosporin as the minibeads of some or all of the other populations, or one population may contain cyclosporin and another population may contain a different active ingredient(s) thereto, e.g. a different combination.

A multiple population composition may be for use in treating a disorder of the GI tract, for example as described herein. Such a composition may be for use in treating a disorder affecting multiple regions of the GIT, e.g. the upper intestine and the lower intestine, and may se an active ient selected from immunosuppressants (e.g. cyclosporin), hydroxylase inhibitors (e.g. hydralazine) and anti-inflammatories (e.g. mesalazine).

The minibeads of a multiple population ition may by way of example be contained in a gel capsule or a sachet.

The second coating is outside the first coating and may be any of the delayed release coatings described herein. In particular, the second coating is or comprises a pH independent polymer modified release coating described above. For e the second coating may be or comprise an enteric coating or a pH independent coating. The second coating may comprise a mixture of polymers including a polymer degradable by bacterial or other enzymes. In a particular embodiment the second coating comprises ethyl cellulose and optionally a water-soluble ccharide, in particular one susceptible to degradation by colonic bacteria, suitably .

Accordingly the second coating may comprise the Surelease-pectin mixture described above.

It is not a requirement that both the first and second coatings are present in the composition at the same time. For example, the composition may comprise second coating (outer coating) in the absence of a first coating. Conversely, the composition may se a first coating in the absence of a second coating.

] The first and second coating may independently be aqueous-based coatings or may be solvent-based gs. By this it is meant that the first and/or second coating may be formulated prior to being applied to the core or composition and/or applied to the core or composition as an aqueousbased composition or as a solvent-based (non-aqueous solvent-based) ition. The aqueousbased or solvent-based coating compositions may be a sion or a solution of the coating material in water or in a solvent.

In an ment the ition comprises a core and an outer coating (also referred to as a second coating herein), the core sing cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming r matrix, wherein the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof P213428WO 45 and does not comprise or is not a polyethyleneglycol ether or ester. The composition may optionally further comprise a at.

In one embodiment of the invention there is provided a composition comprising a core, a first coating and a second coating outside the first coating; and wherein: the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer , wherein the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester; the first coating is or comprises a water-soluble cellulose ether, particularly hydroxypropylmethyl cellulose; the second coating is or comprises a modified release coating or delayed release coating, particularly a pH independent modified release coating; the first coating is present in an amount ponding to a weight gain due to the first g in a range selected from : (i) 8% to 15%; (ii) from 8% to 12%, for example about 10%; or (iii) from 2.5% to 6%, for e about 5% by weight based upon the weight of the composition prior to applying the first coating; and wherein the second coating is present in an amount corresponding to a weight gain of the composition due to the second coating selected from (a) from 4% to 20%; (b) from 7.5% to 20%; (C) from 10% to 12%, for example about 11% or about 11.5%; or (d) from 16% to 18%, for e about 17% by weight based upon the weight of the composition prior to applying the second coating.

The first and second coatings in the embodiment of the immediately preceding paragraph are suitably any of the first and second coatings described above or below. Accordingly it is intended that the coatings described in this section may be applied to any of the compositions described herein to e a delayed release coating if required. The coatings are particularly useful to provide a modified e coating to the cores comprising a polymer matrix and pharmaceutically active ingredient described in this application.

Outer Barrier or Protective Coating The compositions described herein may comprise a protective coating outside the first and/or second coating, for example outside the second coating, the modified release coating. The protective coating may help to protect the ed release coating from damage resulting from, for example ating the ition into a final dosage form, or during the handling, transport or storage of the composition. The protective coating is suitably applied to the outer e of the composition. The protective coating may be applied directly to the second coating (the modified release coating) such that the protective g is in contact with the second coating (the modified release coating). The protective coating is suitably a water soluble coating which does not adversely affect the e of the porin A from the composition when in use. Suitably the protective coating is or comprises a water-soluble polymer. The protective coating may comprise a water-soluble osic or PVA orming polymer. ly the protective coating may be or comprise Opadry (HPMC/HPC-based), Opadry II (PVA/PEG-based) or polyvinyl alcohol-polyethylene glycol graft P213428WO 46 copolymers coat IR) as described herein. The tive coating may be present as a layer of from about 2 to about 50 µm. Suitably the protective coating is applied to give a weight-gain of from about 0.5 to about 10 %, based upon the weight of the ition prior to applying the protective Polymer Matrix The composition of the invention ses cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix. In addition, in certain embodiments of the invention the composition of the invention comprises a core wherein the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix. The composition or the core comprises a continuous phase or matrix phase, which may be or comprise the hydrogel forming polymer matrix, to provide mechanical strength. In embodiments the cyclosporin is comprised within a disperse phase or oil phase within the continuous phase or matrix. The cyclosporin may be present as a disperse phase within the hydrogel-forming polymer matrix (continuous phase or aqueous phase) of the core or composition. The disperse phase may be or comprise the oil phase. For example the disperse phase may comprise a lipid and cyclosporin A. The core or the composition may be prepared by dispersing the cyclosporin, dissolved in the oil phase within an aqueous phase comprising the hydrogel forming polymer matrix to form a d and then causing the composition to solidify (gel), thereby immobilising the cyclosporin within the hydrogel-forming polymer .

The core may have the form of a solid colloid, the colloid comprising a continuous phase and a disperse phase, wherein the continuous phase is or comprises the hydrogel-forming polymer matrix and the disperse phase is or comprises an oil phase optionally comprising the cyclosporin. The disperse phase may comprise a vehicle containing the cyclosporin, for example ning it as a solution or a suspension or a combination of both. The vehicle may be an oil phase as described herein.

Such cores comprising a hydrogel-forming polymer and a disperse phase comprising cyclosporin A are described in more detail below. d Release Coatings The invention provides compositions having a g that comprises, or is, a coatingforming r, n the coating-forming polymer is a hydrogel-forming r; the coating may be a first coating outside which is a second coating. The second coating may be a d release coating, although the invention does not require that the second coating be a d release coating.

The second coating may comprise or be a delayed release polymer.

] The first g may be t in an amount described elsewhere in this specification.

The first coating may be present in an amount corresponding to a weight gain due to the first coating of from 0.5% to 20% by weight of the core.

P213428WO 47 Furthermore, the composition may comprise a first coating present in an amount corresponding to a weight gain due to the coating selected from ranges of from: 0.5% to 15%; 1% to %; 1% to 12%; 1% to 10%; 1% to 8%; 1% to 6%; 1% to 4%, 2% to 10%; 2% to 8%; 2% to 6%; 2% to 7%; 2% to 4%; 4% to 8%; 4% to 7%, 4% to 6%, 5% to 7%; 7% to 20%; 7% to 16%; 9% to 20%; 9% to 16%; 10% to 15%; and 12% to 16%.

The invention provides for a pharmaceutical composition sing a core, a first coating and a second coating outside of the first coating, wherein the core comprises cyclosporin, a hydrogel forming r , a surfactant and an oil phase being sed in the hydrogel forming polymer matrix, the first coating comprises or is a water soluble cellulose ether, and the second coating comprises or is a delayed e polymer, and the first coating may be present in an amount ponding to a weight gain due to the first coating of from 0.5% to 20% by weight of the core, wherein the surfactant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester.

The composition of the invention may comprise a first coating with a thickness of 1 µm to 1mm. Thus, the % weight gain due to the coating specified above may correspond to a ess of 1 µm to 1mm.

The first coating may have a thickness selected from ranges of from: 1 µm to 500 µm; 10 µm to 250 µm; 10 µm to 100 µm; 10 µm to 50 µm; 10 µm to 20 µm; 50 µm to 100 µm; 100 µm to 250 µm; 100 µm to 500 µm; 50 µm to 500 µm; 50 µm to 250 µm; 100 µm to 1 mm; 500 µm to 1 mm. The coating having the thicknesses disclosed in this paragraph may be any of the coatings in the application. In particular the coating referred to in this paragraph may be the water-soluble cellulose ether coating.

The first coating may be present in a weight gain selected from a range of from: 1% to 20%, 4% to 7%, 5% to 7%, 4% to 15%, 8% to 15%, 4% to 12% and 8% to 12%. The second coating may be present in a weight gain selected from a range of from: 8% to 15% or 8% to 12%.

In addition, the invention provides for a pharmaceutical composition comprising a core, a first coating and a second coating outside of the first coating, wherein the core comprises cyclosporin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel g polymer matrix, the first coating ses or is a water soluble cellulose ether, and the second coating comprises or is a d release polymer, and the first coating has a thickness of from 1 µm to 1 mm. The core may optionally r se a hydrogel forming polymer, wherein the tant is a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester.

The second coating may be present in an amount described elsewhere herein. Suitably the second coating provides a coating thickness on the composition of from about 10µm to about 1mm, for example, from about 10 µm to about 500 µm, from about 50 µm to about 1 mm, or about from about 50 µm to about 500 µm. The thickness may therefore be from about 100 µm to about 1 mm, e.g. 100 µm to about 750 µm or about 100 µm to about 500 µm. The thickness may be from about 250 µm to P213428WO 48 about 1 mm, e.g. about 250 µm to about 750 µm or 250 µm to about 500 µm. The thickness may be from about 500 µm to about 1 mm, e.g. about 750 µm to about 1 mm or about 500 µm to about 750 µm. The thickness may therefore be from about 10 µm to about 100 µm, e.g. from about 10 µm to about 50 µm or about 50 µm to about 100 µm.

] It is contemplated within any aspect or embodiment where there is a second coating (also referred to as an outer coating) that the second coating may be present in a % weight gain of from 2% to 40%. In addition the second coating may be present in an amount corresponding to a weight gain due to the coating selected from ranges of from: 4% to 30%, 4% to 7%, 7% to 40%, 7% to 30%, 8% to %, 8% to 20%, 2% to 25%, 2% to 20%, 4% to 25%, 4% to 20%, 4% to 15%, 4% to 13%, 7% to %, 7% to 13%, 8% to 12%, 9% to 12% and 20% to 25%.

In any aspect and embodiment of the invention the first coating may be present in a % weight gain relative to the core of from 0.5% to 20%, preferably 1% to 16% or 4% to 16%, and the second coating may be present in a % weight gain of 4% to 24%, 7% to 24%, 22% to 24%, 7% to %, or 8% to 12%, ably 22% to 24%, 7% to 15%, or 8% to 12%.

The core is preferably in the form of a minibead, for example as bed hereafter in more detail, for example in the form of a solid colloid. The second coat may be a film or it may be a membrane. The second coat, e.g. film or ne, may serve to delay release until after the stomach; the coat may therefore be an enteric coat. The delayed release coat may comprise one or more delayed release substances, preferably of a polymeric nature (e.g. rylates etc; ccharides etc as described in more detail below), or combination of more than one such substance, optionally including other excipients, for example, plasticizers. Preferred plasticizers, if they are used, include hydrophilic plasticizers for example triethyl citrate (TEC) which is particularly preferred when using the Eudragit® family of polymers as coatings as described below. Another preferred plasticiser, described in more detail below in on to coating with ethyl cellulose, is dibutyl sebacate (DBS). Alternative or additional optionally included excipients are glidants. A glidant is a substance that is added to a powder or other medium to improve its flowability. A typical glidant is talc which is preferred when using the Eudragit® family of rs as coatings.

The delayed release coating (the second coating) may be applied as described below and may vary as to thickness and density. The amount of coat is defined by the onal weight added to (gained by) the dry composition (e.g. the core) to which it is applied. Weight gain is preferably in the range 0.1% to 50%, preferably from 1% to 15% of the dry weight of the core, more preferably in the range 3% to 10% or in the range 5-12% or in the range 8-12%.

Polymeric coating al of a d release coating may comprise rylic acid copolymers , ammonio methacrylate co-polymers, or es thereof. Methacrylic acid co-polymers such as, for example, EUDRAGIT™ S and EUDRAGIT™ L (Evonik) are particularly suitable. These polymers are gastroresistant and enterosoluble polymers. Their polymer films are insoluble in pure water and d acids. They may dissolve at higher pHs, depending on their content of carboxylic acid. EUDRAGIT™ S and EUDRAGIT™ L can be used as single components in the polymer coating or in ation in any ratio. By using a combination of the polymers, the polymeric material can P213428WO 49 exhibit solubility at a variety of pH levels, e.g. between the pHs at which EUDRAGIT™ L and EUDRAGIT™ S are separately e. In particular, the coating may be an enteric coating comprising one or more co-polymers described in this paragraph. A particular coating material to be mentioned is Eudragit L 30 D-55.

The trade mark "EUDRAGIT" is used hereinafter to refer to rylic acid copolymers, in particular those sold under the trade mark EUDRAGIT by Evonik.

The delayed release coating, where t, can comprise a polymeric material sing a major proportion (e.g., greater than 50% of the total ric coating content) of at least one pharmaceutically acceptable water-soluble polymer, and ally a minor proportion (e.g., less than 50% of the total polymeric content) of at least one pharmaceutically acceptable water insoluble polymer. Alternatively, the membrane coating can comprise a polymeric material comprising a major proportion (e.g., greater than 50% of the total ric content) of at least one pharmaceutically acceptable water insoluble polymer, and optionally a minor proportion (e.g., less than 50% of the total polymeric content) of at least one pharmaceutically acceptable water-soluble polymer.

] Ammonio methacrylate co-polymers such as, for example, IT™ RS and EUDRAGIT™ RL (Evonik) are suitable for use in the present invention. These polymers are insoluble in pure water, dilute acids, buffer solutions, and/or digestive fluids over the entire logical pH range. The polymers swell in water and digestive fluids independently of pH. In the swollen state, they are then permeable to water and dissolved active agents. The permeability of the rs depends on the ratio of ethylacrylate (EA), methyl methacrylate (MMA), and hylammonioethyl methacrylate chloride (TAMCl) groups in the polymer. For example, those polymers having EA:MMA:TAMCl ratios of 1:2:0.2 GIT™ RL) are more permeable than those with ratios of 1:2:0.1 (EUDRAGIT™ RS). Polymers of EUDRAGIT™ RL are insoluble polymers of high permeability.

Polymers of EUDRAGIT™ RS are insoluble films of low permeability. A diffusion-controlled pH- independent polymer in this family is RS 30 D which is a copolymer of ethyl acrylate, methyl methacrylate and a low content of methacrylic acid ester with quaternary ammonium groups present as salts to make the polymer ble. RS 30 D is available as an aqueous dispersion.

The amino methacrylate co-polymers can be combined in any desired ratio, and the ratio can be modified to modify the rate of drug release. For example, a ratio of EUDRAGIT™ RS: EUDRAGIT™ RL of 90:10 can be used. Alternatively, the ratio of EUDRAGIT™ RS: EUDRAGIT™ RL can be about 100:0 to about 80:20, or about 100:0 to about 90:10, or any ratio in between. In such compositions, the less ble polymer EUDRAGIT™ RS generally comprises the majority of the polymeric material with the more soluble RL, when it dissolves, permitting gaps to be formed through which solutes can come into contact with the core allowing for the active to escape in a lled manner.

] The amino rylate co-polymers can be combined with the methacrylic acid copolymers within the polymeric material in order to achieve the d delay in the release of the drug and/or poration of the coating and/or exposure of the composition within the coating to allow egress of drug and/or dissolution of the immobilization or water-soluble r matrix. Ratios of ammonio P213428WO 50 methacrylate co-polymer (e.g., EUDRAGIT™ RS) to methacrylic acid co-polymer in the range of about 99:1 to about 20:80 can be used. The two types of polymers can also be combined into the same polymeric material, or provided as separate coats that are applied to the beads.

Eudragit™ FS 30 D is an anionic aqueous-based acrylic ric dispersion ting of methacrylic acid, methyl acrylate, and methyl methacrylate and is pH sensitive. This polymer contains fewer carboxyl groups and thus ves at a higher pH (> 6.5). The advantage of such a system is that it can be easily manufactured on a large scale in a reasonable processing time using conventional powder layering and fluidized bed coating techniques. A further e is EUDRAGIT® L 30D-55 which is an aqueous dispersion of anionic polymers with methacrylic acid as a functional group. It is available as a 30% aqueous dispersion.

In addition to the EUDRAGIT™ polymers described above, a number of other such mers can be used to control drug release. These include methacrylate ester co-polymers such as, for example, the EUDRAGIT™ NE and EUDRAGIT™ NM ranges. r information on the EUDRAGIT™ polymers can be found in "Chemistry and Application ties of Polymethacrylate Coating Systems," in Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms, ed. James McGinity, Marcel Dekker Inc., New York, pg 109-114 the entirety of which is incorporated herein by Several derivatives of hydroxypropyl methylcellulose (HPMC) also exhibit pH dependent solubility and may be used in the invention for the delayed release coating. As examples of such derivatives may be mentioned HPMC esters, for example hydroxypropyl methylcellulose ate (HPMCP), which rapidly dissolves in the upper inal tract and hydroxypropyl methylcellulose acetate succinate (HPMCAS) in which the presence of ble carboxyl groups causes the polymer to solubilize at high pH (> 5.5 for the LF grade and > 6.8 for the HF grade). These polymers are commercially available from tsu Chemical Co. Ltd. As with other polymers described herein as useful for delayed release coatings, HPMC and tives (e.g. esters) may be combined with other polymers e.g. EUDRAGIT RL-30 D.

Other polymers may be used to provide a coating in particular enteric, or pH-dependent, polymers. Such polymers can include phthalate, butyrate, succinate, and/or mellitate groups. Such polymers include, but are not limited to, cellulose acetate phthalate, cellulose acetate succinate, cellulose hydrogen phthalate, cellulose acetate trimellitate, hydroxypropyl-methylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, starch acetate ate, amylose acetate phthalate, polyvinyl acetate phthalate, and polyvinyl butyrate phthalate. pH Independent Polymer Delayed Release Coatings In a ular embodiment the second coating, where present, is or comprises a polymeric g which is pH-independent in its dissolution profile and/or in its y to e the active ingredient incorporated in the itions of the invention. A pH-independent r delayed e coating comprises a delayed release polymer, optionally a plurality of delayed release polymers, and one or more other optional components. The other components may serve to modulate P213428WO 51 the properties of the composition. Examples have already been given (e.g., Eudragit RS and RL).

Another example of a ependent polymeric coating is a coating that comprises or is ethylcellulose; a pH-independent polymeric coating may have a delayed e polymer that is ethylcellulose, therefore. It will be understood that an ethylcellulose formulation for use in coating a dosage form may comprise, in addition to ethylcellulose and - in the case of a liquid formulation - a liquid e, one or more other components. The other components may serve to modulate the properties of the composition, e.g. stability or the al properties of the coating such as the flexibility of the film coating. The ethylcellulose may be the sole delayed release polymer in such a composition. The et hylcellulose may be in an amount of at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95% by weight of the dry weight of a coating composition for use in g a dosage form. Accordingly, an ethylcellulose coating may include other components in addition to the ethylcellulose. The ethylcellulose may be in an amount of at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95% by weight of the ethylcellulose g.

Consequently, ethylcellulose may be in an amount of at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95% by weight of the dry weight of the second g. ly the ethyl cellulose coating r comprises a plasticizer as described below to improve the flexibility of the film and to e the film-forming properties of the coating composition during application of the coating.

A particular ethylcellulose coating composition which may be applied to the composition, optionally to the core (i.e. in the absence of a first coating) or to the first coating is a dispersion of ethylcellulose in a sub-micron to micron particle size range, e.g. from about 0.1 to 10 µm in size, homogeneously suspended in water with the aid of an emulsification agent, e.g. ammonium oleate.

The ethylcellulose dispersion may optionally and preferably contain a plasticizer. Suitably plasticisers include for example dibutyl sebacate (DBS), diethylphthalate, triethyl citrate, tributyl citrate, triacetin, or medium chain triglycerides. The amount of plasticizer present in the coating ition will vary depending upon the desired properties of the coating. Typically the plasticizer ses from 1 to 50%, for example about 8 to about 50% of the ed weight of the plasticizer and ethyl ose.

Such ethylcellulose dispersions may, for example, be ctured according to U.S. Pat. No. 4,502,888, which is incorporated herein by reference. One such ellulose dispersion suitable for use in the present invention and available commercially is marketed under the trademark Surelease®, by Colorcon of West Point, Pa. USA. In this marketed product, the ethylcellulose les are, e.g., blended with oleic acid and a plasticizer, then optionally extruded and melted. The molten plasticized ethylcellulose is then directly emulsified, for example in ammoniated water optionally in a high shear mixing device, e.g. under pressure. Ammonium oleate can be formed in situ, for instance to stabilize and form the dispersion of plasticized ethylcellulose particles. Additional purified water can then be added to achieve the final solids content. See also U.S. Pat. No. 4,123,403, which is incorporated herein by nce.

The trademark "Surelease®" is used hereinafter to refer to ethylcellulose coating materials, for example a dispersion of ethylcellulose in a cron to micron particle size range, e.g. from P213428WO 52 about 0.1 to 10 µm in size, homogeneously suspended in water with the aid of an emulsification agent, e.g. ammonium oleate. In particular, the trademark "Surelease®" is used herein to refer to the t marketed by Colorcon under the Surelease® trademark.

] Surelease® dispersion is an example of a combination of film-forming polymer, plasticizer and stabilizers which may be used as a second g to adjust rates of active principle release with reproducible profiles that are relatively insensitive to pH. The pal means of drug release is by diffusion through the Surelease® dispersion membrane and is directly controlled by film thickness. Use of ase® is particularly preferred and it is possible to increase or se the quantity of Surelease® applied as coating in order to modify the dissolution of the coated composition.

Unless ise stipulated, use of the term "Surelease" may apply to ase E19020, E 19030, E19040 or E19050. An ethylcellulose coating formulation, for example Surelease E 19020, may comprise ethylcellulose blended with oleic acid and dibutyl sebacate, then extruded and melted. The molten plasticized ethylcellulose is then directly emulsified in ammoniated water in a high shear mixing device under pressure. Ammonium oleate is formed in situ to stabilize and form the dispersion of plasticized ethylcellulose particles. Additional purified water is then added to achieve the final solids content. An ethylcellulose coating formulation, for example Surelease E19030, may additionally comprise dal anhydrous silica dispersed into the material. An ethylcellulose coating formulation, for e Surelease E19040, may comprise medium chain triglycerides instead of dibutyl sebacate, in particular in a formulation comprising colloidal anhydrous silica and oleic acid. An ethylcellulose coating formulation, for example Surelease E19050, may derive from blending ethylcellulose with oleic acid before melting and extrusion. The molten plasticized ethylcellulose is then directly emulsified in ammoniated water in a high shear mixing device under pressure.

Ammonium oleate is formed in situ to stabilize and form the dispersion of cized ethylcellulose particles. However, ations that comprise medium chain triglycerides, colloidal anhydrous silica and oleic acid are red. Surelease E19040 is particularly preferred.

The invention also contemplates using combinations of ethylcellulose, e.g. a Surelease formulation, with other coating components, for example sodium alginate, e.g. sodium alginate ble under the trade name Nutrateric™.

In addition to the EUDRAGIT™ and ase® polymers discussed above, where compatible, any combination of g polymers disclosed herein may be blended to provide additional delayed-release profiles.

] The delayed release coating can further comprise at least one soluble excipient to increase the permeability of the polymeric material. These soluble excipients can also be referred to or are pore formers. Suitably, the at least one soluble excipient or pore former is selected from among a soluble polymer, a surfactant, an alkali metal salt, an c acid, a sugar, a polysaccharide, and a sugar alcohol. Such soluble excipients include, but are not limited to, nyl pyrrolidone, polyvinyl alcohol (PVA), polyethylene glycol, a water-soluble hydroxypropyl methyl cellulose, sodium chloride, surfactants such as, for example, sodium lauryl sulfate and polysorbates, organic acids such as, for example, acetic acid, adipic acid, citric acid, fumaric acid, glutaric acid, malic acid, succinic acid, and P213428WO 53 tartaric acid, sugars such as, for example, dextrose, fructose, glucose, lactose, and sucrose, sugar alcohols such as, for example, lactitol, ol, mannitol, sorbitol, and xylitol, xanthan gum, dextrins, and maltodextrins; and a polysaccharide susceptible of degradation by a bacterial enzyme normally found in the colon, for example polysaccharides include chondroitin sulphate, pectin, dextran, guar gum and amylase, chitosan etc. and derivatives of any of the foregoing. In some embodiments, polyvinyl pyrrolidone, mannitol, and/or polyethylene glycol can be used as soluble excipients. The at least one e excipient can be used in an amount ranging from about 0.1% to about 15% by weight, based on the total dry weight of the polymer g, for example from about 0.5% to about %, about 0.5% to about 5%, about 1% to about 3%, suitably about 2% based on the total dry weight of the polymer coating. The delayed release coating may be free from HPMC.

The modifications in the rates of release, such as to create a delay or extension in release, can be achieved in any number of ways. Mechanisms can be dependent or independent of local pH in the intestine, and can also rely on local enzymatic ty to achieve the desired effect. Examples of modified-release compositions are known in the art and are described, for example, in U.S. Pat. Nos. 3,845,770; 3,916,899; 809; 123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; ,073,543; 5,639,476; 5,354,556; and 5,733,566 all of which are incorporated herein by reference in their entirety.

The addition to Surelease® or other pH-independent polymer nce of a second polymer (e.g. a ccharide, especially a heteropolysaccharide) which is susceptible to degradation by colonic bacterial enzymes (and optionally or atively by pancreatic or other relevant enzymes), helps provide targeted release of the active ient to a site or sites within the GI tract where the second r is degraded. By varying the amount of second polymer added present in the coating the dissolution profile may be optimized to provide the required release of cyclosporin A from the composition.

In a particular embodiments the delayed release coating provides for release of the active agent in at least the colon. ingly in one embodiment the coating comprises a combination of ethylcellulose (preferably a described above, and ularly formulated with an emulsification agent such as, for example, ammonium oleate and/or a plasticizer such as, for example, dibutyl sebacate or medium chain cerides) and a polysaccharide susceptible of degradation by a bacterial enzyme normally found in the colon. Such polysaccharides include chondroitin sulphate, pectin, dextran, guar gum and amylase, chitosan etc. and derivatives of any of the foregoing. Chitosan may be used in connection with obtaining a specific release profile; additionally or alternatively, pectin may be so used.

The use of polysaccharides by lves for delayed release coating purposes has been tried with limited success. Most of the arch polysaccharides suffer from the ck of g good film forming properties. Also, they tend to swell in the GI tract and become porous, resulting in the early release of the drug. Even amorphous amylose, which is resistant to degradation by pancreatic alpha amylase but e of degradation by colonic bacterial enzymes, has the disadvantage of swelling in aqueous media although this can be controlled by incorporating insoluble P213428WO 54 polymer, for example ethyl ose and / or acrylate, into the amylose film. Amylose r is not water-soluble and although water-insoluble ccharides are not excluded, use of a soluble polysaccharide (WSP) susceptible to bacterial enzymatic ation brings particularly advantageous results when used as a coating in accordance with this embodiment of the present invention. A particularly preferred polysaccharide in this embodiment of the present ion is pectin.

Various kinds of pectin may be used including pectin of different grades ble i.e. with differing degrees of methylation (DM), i.e. percentage of carbonyl groups esterified with methanol, for example pectins with a DM of more than 50%, known as High Methoxy (HM) Pectins or Low Methoxy (LM) pectins, or a pectin combination comprising an HM pectin and an LM pectin. It is also possible in this embodiment to use pectins having various degrees of acetylation (Dac). Taken together, the DM and Dac or the degree of substitution is known as Degree of fication (DE). Pectins of various DE’s may be used according to the invention. As an alternative to pectin, sodium te may be used as a ccharide according to an embodiment of the invention. However, other embodiments may conveniently include amylose and/or starch which contains amylose. Various grades of starch, containing different percentages of amylose may be used including for example Hylon V (National Starch Food tion) which has an e percentage of 56% or Hylon VII which has an amylose percentage of 70%. The remaining percentage is amylopectin. The polysaccharides pectin, amylose and sodium alginate are particularly preferred for achieving colon delivery of the active ingredient.

It has been found that water-soluble polysaccharide, suitably pectin, can act as a former of pores in the coating ise provided by ethylcellulose (preferably Surelease). By "pores" is not meant shaft-like holes from the surface to the core of the composition, rather areas of weakness or absence of coating occurring stochastically on and within the coating of the invention.

Pore formers have been described before in connection with Surelease (see e.g. US 2005/0220878).

] According to a particular embodiment of the invention the delayed e coating comprises ethylcellulose, e.g. Surelease™, and a water-soluble polysaccharide (WSP) n the proportion of ethylcellulose (in particular Surelease™) to WSP is ideally in the range 90:10 to 99:1, preferably, 95:5 to 99:1, more preferably 97:3 to 99:1, for example about 98:2 based upon the dry weight of the coating. Suitably in this embodiment the weight gain of the composition due to application of the coating comprising ethylcellulose, e.g. Surelease™, and the WSP is in the range of from 1 to 30% (for example from: 3% to 25%; 5% to 15%; 8% to 14%; 10% to 12%; 12% to 18%; or 16% to 18%, suitably the weight gain is about 11%, about 11.5%, or about 17%). It is particularly preferred that the WSP in this embodiment is pectin. ularly favoured weight gains using coatings comprising ethylcellulose, e.g. Surelease™, are those in the range 5-12% or in the range 8-12%.

Accordingly in an embodiment the second coating comprises ethyl ose and a water soluble polysaccharide (particularly pectin) wherein the water-soluble polysaccharide (WSP) is present in an amount of 0.1% to about 10% by weight, based on the dry weight of the second coating.

Suitably the WSP is present in an amount of from about 0.5% to about 10%, for example about 0.5% to about 5%, about 1% to about 3%, suitably about 2% based on the total dry weight of the second P213428WO 55 coating. In this ment the WSP is preferably pectin. In this embodiment the second composition ly further comprises a plasticizer. Suitable plasticizers include these described above in on to Surelease™. Suitably the weight gain of the composition due to application of the second coating in this embodiment is in the range of from 1 to 30% (for example from: 3% to 25%; 5% to 15%; 8% to 14%; 10% to 12%; 12% to 18%; or 16% to 18%, suitably the weight gain is about 11%, about 11.5%, or about 17%).

In an embodiment the delayed release polymer is not a water-soluble cellulose ether. Where the second coating comprises or is a delayed e polymer the delayed release polymer may not be the same as the water-soluble cellulose ether of the first coating. Accordingly the second coating may not be the same as the first coating.

Continuous Phase r Matrix (Aqueous Phase) This section of the specification refers to a polymer matrix and continuous phase both of which concern the hydrogel forming polymer matrix. Therefore, reference to a polymer matrix or uous phase can be equated to the hydrogel forming polymer matrix. Furthermore, this n of the ication relating to the polymer matrix recites amounts of constituents in terms of percent by weight of the ition. In the context of this section of the ication, what is meant is percent by weight of the dry weight of the composition or core ing coating(s).

The composition or the core may comprise a matrix or continuous phase and also a disperse phase or oil phase. Similarly the liquid composition of the invention comprises an aqueous phase comprising a hydrogel forming polymer. Suitably the uous phase or matrix phase of the composition or core is or comprises a hydrogel-forming polymer. A hydrogel-forming polymer is a polymer capable of forming a hydrogel. A hydrogel may be described as a solid or semi-solid al, which ts no flow when at rest, comprising a k (matrix) of hydrophilic polymer chains that span the volume of an aqueous liquid medium. A hydrogel forming polymer matrix is a network of hydrogel forming polymer chains, thus a hydrogel forming polymer matrix is a hydrogel forming polymer that has been allowed or caused to form a matrix.

The ition or core may comprise a hydrogel-forming polymer matrix and the liquid composition may select a hydrogel-forming polymer selected from the group consisting of: gelatin; agar; agarose; pectin; carrageenan; chitosan; alginate; starch; xanthan gum; gum Arabic; guar gum; locust bean gum; polyurethane; polyether polyurethane; cellulose; cellulose ester, cellulose acetate, cellulose triacetate; cross-bonded polyvinyl alcohol; polymers and copolymers of acrylic acid, hydroxyalkyl acrylates, hydroxyethyl acrylate, diethylene glycol monoacrylate, 2- hydroxypropylacrylate, 3-hydroxypropyl acrylate; polymers and copolymers of methacrylic acid, yethyl methacrylate, leneglycol monomethacrylate, 2-hydroxypropyl methacrylate, 3- hydroxypropyl methacrylate, dipropylene glycol monomethylacrylate; vinylpyrrolidone; mide polymers and copolymers, N-methylacrylamide, N-propylacrylamide; methacrylamide polymers and copolymers, N-isopropylmethacrylamide, Nhydroxyethylmethacrylamide; and vinyl pyrrolidone; and combinations thereof. In specific embodiments binary or tertiary etc ations of any of the above substances are foreseen. 8WO 56 In a further embodiment the hydrogel-forming polymer or the hydrogel forming polymer matrix is selected from the group consisting of gelatin, agar, a polyethylene glycol, starch, casein, chitosan, soya bean protein, safflower protein, alginates, gellan gum, carrageenan, xanthan gum, ated gelatin, succinated gelatin, cellulosephthalate-acetate, oleoresin, polyvinylacetate, polymerisates of c or methacrylic esters and polyvinylacetate-phthalate and any derivative of any of the foregoing; or a mixture of one or more such hydrogel-forming polymers.

The hydrogel-forming polymer or the hydrogel forming r matrix may also be referred to as a hydrocolloid i.e. a d system wherein the colloid particles are disperse in water and the quantity of water ble allows for the formation of a gel. In embodiments it is red to use reversible hydrocolloids preferably thermo-reversible hydrocolloids (e.g. agar, agarose, gelatin etc) as opposed to rsible (single-state) hydrocolloids. Thermo-reversible hydrocolloids can exist in a gel and sol state, and alternate between states with the addition or elimination of heat. n, agar and agarose are thermo-reversible, rehydratable ds and are particularly preferred. Gelatin derivatives such as, for example, succinated or phthalated gelatins are also contemplated. reversible hydrocolloids which may be used according to the invention, whether individually or in combination, include those derived from natural sources such as, for example, carrageenan (extracted from seaweed), gelatin (extracted from bovine, porcine, fish or vegetal sources), agar (from seaweed), agarose (a polysaccharide obtained from agar) and pectin (extracted from citrus peel, apple and other ). A non-animal based hydrocolloid may be preferred for certain applications e.g. administration to vegetarians or to duals not wishing to ingest animal products for religious or health reasons. In relation to the use of carrageenan, reference is made to US patent ation 029660 A1 (Fonkwe et al), the entirety of which is incorporated herein by nce. The hydrogel-forming polymer may comprise or be a combination of gelatin with one or more other thermoreversible hydrocolloids, e.g. with one or more other of the thermoreversible hydrocolloids just listed. The hydrogel-forming polymer may comprise or be a combination of gelatin with agar; optionally, at least one further thermoreversible hydrocolloid may be included in the combination, for example one just Thermo-reversible colloids present a benefit over other hydrogel-forming polymers. on or hardening of thermo-reversible colloids occurs by cooling the colloid, e.g. in a liquid g bath or by air flow. Gelation of other hydrogel-forming polymers, which is chemically driven, can lead to leakage of the composition contents into the gelation medium as the hardening s can take time to occur. Leakage of the content of the composition may lead to an inaccurate quantity of the active ingredient within the composition. Thermo -reversible colloids are also known as thermo-reversible gels, and it is therefore preferred that the hydrogel former be a -reversible gelling agent.

Another term which may be applied to hydrogel formers which are advantageous is "thermotropic": a thermotropic gelling agent (which the reader will infer is red as a hydrogel former used in the invention) is one caused to gel by a change in temperature and such gelling agents are able to gel more rapidly than those whose gelling is chemically induced, e.g. ionotropic gelling agents whose gelling is induced by ions, for example chitosan. In embodiments of the invention, P213428WO 57 therefore, the hydrogel former is a thermotropic gel-forming polymer or a combination of such polymers.

The cture of the ition to prepare a core may require that the hydrogel-forming polymer be present as a solution, which is preferably an s solution. The hydrogel-forming polymer represents between 5% and 50%, preferably between 10% and 30%, still more preferably between 15% and 20% by weight of the aqueous phase during manufacture as described herein. In addition the hydrogel-forming polymer may comprise 8 to 35%, (for example 15-25%, preferably 17- 18%) hydro-gel forming r; 65%-85% (preferably 77-82%) of water plus, ally, from 1-5% (preferably 1.5 to 3%) sorbitol. When present surfactant (e.g. anionic tant) in the aqueous phase pre-mix may be present in an amount of 0.1 to 5% (preferably 0.5 to 4%) wherein all parts are by weight of the aqueous phase.

In the aspect of the invention where the liquid composition is provided the hydrogel forming polymer may be present as an aqueous solution in the aqueous phase. The amounts of hydrogel forming polymer recited in the immediately preceding paragraph are equally relevant to embodiments of the liquid composition.

In embodiments the composition comprises at least 25%, suitably at least 40% by weight based upon the dry weight of the composition of the hydrogel-forming polymer matrix. For example the hydrogel-forming polymer matrix is present from 25 to 70%, for example 40 to 70% suitably 45 to 60% of the composition, wherein the % is by weight based upon the dry weight of the composition.

In embodiments the el-forming polymer is a pharmaceutically acceptable r.

In certain ments the hydrogel-forming r is gelatin. In certain embodiments the el-forming polymer matrix is gelatin. In certain embodiments the hydrogel-forming polymer comprises gelatin. In certain ments the gelatin comprises at least 30%, for example 30 to 70% or 40 to 70% suitably 40 to 60% of the composition, wherein the % is by weight based upon the dry weight of the composition.

The hydrogel-forming polymer may optionally comprise a plasticiser for example sorbitol or ine, or a combination f. In particular one or more plasticisers may be combined with gelatin.

In embodiments in which the hydrogel-forming polymer comprises or is gelatin, reference is hereby made to "Bloom strength", a measure of the strength of a gel or gelatin developed in 1925 by O. T. Bloom. The test determines the weight (in grams) needed by a probe (normally with a diameter of 0.5 inch) to deflect the surface of the gel 4 mm without breaking it. The result is expressed in Bloom (grades) and usually ranges between 30 and 300 Bloom. To perform the Bloom test on gelatin, a 6.67% gelatin solution is kept for 17-18 hours at 10°C prior to being .

When the hydrogel-forming polymer ses or is gelatin the bloom strength of the gelatin may be in the range of 125 Bloom to 300 Bloom, 200 Bloom to 300 Bloom and preferably 225 Bloom to 300 Bloom. It should be appreciated that higher bloom strength gelatin can be replaced by lower bloom strength n at higher concentrations.

P213428WO 58 According to the invention, in embodiments in which the hydrogel g polymer or hydrogel-forming polymer matrix comprises or is gelatin, the gelatin may be sourced by a variety of means. For example, it can be obtained by the partial ysis of collagenous material, such as the skin, white connective s, or bones of animals. Type A gelatin is derived mainly from porcine skins by acid processing, and exhibits an isoelectric point between pH 7 and pH 9, while Type B gelatin is derived from ne processing of bones and animal (bovine) skins and exhibits an isoelectric point between pH 4.7 and pH 5.2. Type A gelatin is somewhat red. Gelatin for use in the invention may also be derived from the skin of cold water fish. Blends of Type A and Type B gelatins can be used in the invention to obtain a gelatin with the ite viscosity and bloom strength characteristics for bead manufacture.

Lower ature gelatin (or gelatin derivatives or mixtures of gelatins with melting point reducers) or other polymer matrices able to be solidified at lower temperatures (e.g. sodium te) may also be used. It is therefore believed that polymer which ses or is low temperature gelatin is a preferred matrix polymer.

According to the invention, in embodiments in which the hydrogel forming r or hydrogel forming polymer matrix comprises or is gelatin, the starting gelatin al is preferably ed before cture to produce "soft gelatin" by the addition of a plasticizer or softener to the n to adjust the hardness of the composition of the invention. The addition of cizer es enhanced softness and flexibility as may be desirable to optimise dissolution and/or further sing such as, for example, coating. Useful plasticizers of the present invention for combination with gelatin or another hydrogel-forming polymer include glycerine (1,2,3-propanetriol), itol (D-glucitol), sorbitol BP (a non-crystallizing sorbitol solution) or an aqueous solution of D-sorbitol, sorbitans (e.g. iborb 85/70), mannitol, maltitol, gum arabic, triethyl citrate, tri-n-butyl citrate, dibutylsebacate.

Other or similar low molecular weight polyols are also contemplated for example ethylene glycol and propylene glycol. Polyethylene glycol and polypropylene glycol may also be used although these are less preferred. Glycerine and D-sorbitol may be obtained from the Sigma Chemical Company, St.

Louis, Mo. USA or Roquette, France. Some active agents and excipients included for other functions may act as plasticisers.

Softeners or plasticisers, if utilized, can be ideally incorporated in a proportion rising to 30%, preferably up to 20% and more preferably up to 10% by dry weight of the composition of the invention, even more preferably between 3 and 8%, and most preferably between 4% and 6%.

Although not essential, the hydrogel-forming polymer matrix may also optionally contain a disintegrant where it is particularly desired to enhance the rate of disintegration of the composition of the invention. Examples of disintegrants which may be included are c acid, croscarmellose , crospovidone, low-substituted hydroxypropyl cellulose and sodium starch glycolate.

A crystallisation inhibitor (e.g. approximately 1% by dry weight of the composition) may also be included in the composition of the ion. An example is hydroxy propyl/methyl cellulose (HPC or HPMC, hypromellose etc) which may play other roles such as, for example, emulsifier.

P213428WO 59 In r embodiment, the hydrogel-forming polymer matrix is chitosan which can exist in the form of s with or without additives as described e.g. in United States Patent 4,659,700 (Johnson & Johnson); by Kumar Majeti N.V. Ravi in Reactive and Functional Polymers, 46, 1, 2000; and by Paul et al. in ST.P. Pharma Science, 10, 5, 2000 the entirety of all 3 of which is orated herein by reference. Chitosan derivatives e.g. thiolated entities are also contemplated.

The hydrogel-forming polymer matrix may be a non-hydrocolloid gum. es are the cross-linked salts of alginic acid. For example, aqueous ons of sodium alginate gums extracted from the walls of brown algae have the well known property of gelling when exposed to di- and trivalent cations. A typical nt cation is calcium, often in the form of aqueous calcium chloride solution. It is preferred in this embodiment that the cross-linking or gelling have arisen through reaction with such a multivalent cation, particularly m.

The hydrogel-forming polymer matrix may have a low water content, therefore the composition may have a low water content. As described below, during manufacture of a core the disperse phase or oil phase, optionally comprising cyclosporin, is mixed with an s solution of the hydrogel-forming polymer and the composition is gelled, for example to provide a composition or a core which are minibeads. Suitably the composition or cores are dried following formation to reduce the water content t therein.

In certain embodiments the ition does not se compounds containing a disulphide bond. In ments the hydrogel-forming polymer does not comprise compounds containing a disulphide bond.

The hydrogel-forming polymer matrix forming the continuous phase of the core (aqueous phase) may further comprise a surfactant. tants which may be used in the composition are described in the n "surfactants" below.

Surfactant which may be present in the continuous phase, aqueous phase or the el forming polymer matrix of the composition or core include, for example a tant selected from the group consisting of: cationic; amphoteric (zwitterionic); anionic surfactants, for example perfluorooctanoate (PFOA or PFO), perfluoro-octanesulfonate (PFOS), sodium dodecyl sulfate (SDS), ammonium lauryl sulfate, and other alkyl sulfate salts, sodium laureth sulfate, also known as sodium lauryl ether sulfate (SLES) and alkyl benzene sulphonate; and non-ionic surfactants for example perfluorocarbons, polyoxyethyleneglycol dodecyl ether (e.g. Brij such as, for example, Brij 35), Myrj (e.g. Myrj 49, 52 or 59), fatty alcohol ethoxylates, alkylphenol ethoxylate, fatty acid ethoxylates, fatty amide ethoxylates, alkyl glucosides, Tween 20 or 80 (also known as Polysorbate) (Brij, Myrj and Tween products are available commercially from Croda), poloxamers which are nonionic triblock copolymers composed of a central hydrophobic chain of ypropylene (poly(propylene oxide)) flanked by two hydrophilic chains of yethylene (poly(ethylene oxide)), or a combination of the foregoing. In particular, the surfactant may be selected from, or comprise, anionic surfactants and combinations thereof, the anionic surfactants optionally being those mentioned in this paragraph. A particular class of tant comprises sulfate salts. A preferred c surfactant in the aqueous P213428WO 60 phase is SDS. Mixtures of c surfactants may be used. Mixtures of r surfactants are also contemplated, e.g. mixtures comprising orocarbons.

In embodiments of the invention, the core comprises a hydrophilic tant which, without being bound by theory, is believed at least partially to partition the aqueous phase (polymer matrix).

Such surfactants intended for such inclusion in the aqueous phase of the core are preferably readily diffusing or diffusible surfactants to facilitate cturing and processing of the composition of the invention.

The surfactant may have an HLB of at least 10 and optionally of at least 15, e.g. at least 20, or at least 30 and optionally of 38-42, e.g. 40. Such surfactants can be of any particular type (ionic, non-ionic, zwitterionic) and may comprise as a proportion of dry weight of the composition from 0.1% to 6%, e.g. 0.1% to 5%. 0.1% to 4% or 0.1% to 3%, more preferably in a proportion of at least 1% and in particular between 1.0 and 4.5 or 5%, ideally within or just outside the 2-4% range, for example from 2 to 3% or approximately 2% or approximately 4%.

Unless otherwise stated or required, all percentages and ratios are by weight.

] In one embodiment the anionic surfactant which may be present in the continuous phase, aqueous phase or the hydrogel forming polymer matrix of the composition or core may be an anionic surfactant selected from alkyl sulphates, carboxylates or phospholipids, or combinations f.

The physical form of the surfactant at the point of introduction into the aqueous phase during preparation of the composition or core plays a role in the ease of manufacture of the composition or core. As such, gh liquid surfactants can be employed, it is preferred to utilize a surfactant which is in solid form (e.g. crystalline, granules or powder) at room ature, particularly when the aqueous phase comprises gelatin.

Disperse Phase The polymer matrix or the continuous phase of the composition, or in embodiments where a core is present, the core described above (for example the hydrogel-forming polymer) may comprise a se phase. Similarly the aqueous phase of the liquid composition comprises a disperse phase which is or comprises the oil phase. Suitably the disperse phase, where present, may comprise the cyclosporin. In such ments the cyclosporin is preferably e in the disperse phase, i.e. the se phase comprises a vehicle in which the active is dissolved. Embodiments wherein the cyclosporin is solubilised in the se phase are preferred, because such compositions release the cyclosporin in a solubilised form, which may enhance the therapeutic effect of the drug at the site of release, for example by enhancing absorption into the colonic mucosa.

In embodiments the porin is or is comprised in the disperse phase. The disperse phase is or comprises the oil phase. Preferably, the disperse phase is the oil phase.

The disperse phase may se a water immiscible phase (also referred to herein as an oil phase). The water ible phase may be solid, semi-solid or liquid at ambient temperature (e.g.

° C), and ore the oil phase may for example be waxy at ambient temperature. The oil phase P213428WO 61 may be or may comprise a liquid lipid and optionally a t miscible therewith. The cyclosporin may be present in the oil phase. Suitably the cyclosporin is soluble in the oil phase.

The se phase may comprise a combination of oils, for example liquid lipids. The liquid lipid may be a short-, medium- or long- chain triglyceride formulation, or a combination thereof. A medium chain triglyceride(s) (MCT) comprises one or more triglycerides of at least one fatty acid selected from C6, C7, C8, C9, C10, C11 and C12 fatty acids. It will be understood that commercially available triglyceride, in particular MCT, formulations useful in the invention are mixtures derived from natural products and usually or always contain minor amounts of compounds which are not MCTs; the term "medium chain triglyceride formulation" is therefore to be interpreted to include such formulations.

A short chain triglyceride(s) comprises one or more triglycerides of at least one short chain fatty acid selected from C2-C5 fatty acids. A long chain triglyceride(s) comprises one or more triglycerides of at least one long chain fatty acid having at least 13 carbon atoms.

The liquid lipid may comprise or be cerides and/or erides. Such glycerides may be selected from medium chain triglycerides or short chain triglycerides or a combination thereof.

The liquid lipid may be a caprylic/capric triglyceride, i.e. a caprylic/capric triglyceride ation (which it will be understood may contain minor amounts of compounds which are not caprylic/capric triglycerides).

The disperse phase may optionally se a solvent. Accordingly the oil phase may comprise a solvent. Said solvent which is optionally ed in an oil phase may be le with both the liquid lipid and with water. Examples of le ts are 2-(2-ethoxyethoxy)ethanol available commercially under trade names Carbitol™, Carbitol cellosolve, Transcutol™, Dioxitol™, Poly-solv DE™, and Dowanal DE™; or the purer Transcutol™ HP . Transcutol P or HP, which are available commercially from Gattefosse, are preferred. Another possible co-solvent is poly(ethylene ). PEGs of molecular weight 0 (e.g. PEG 200) or 380-420 (e.g. PEG 400) are preferred in this embodiment. Suitable PEGs can be obtained commercially under the name "Carbowax" manufactured by Union Carbide Corporation gh many alternative manufacturers or suppliers are possible.

The disperse phase may represent from 10-85% by dry weight of the core.

As sed above the disperse phase may be an oil phase comprising any pharmaceutically suitable oil, e.g. a liquid lipid. The oil phase may be t as oil drops. In terms of dry weight of the core, the oil phase may comprise a proportion from 10% to 85%, e.g. 15% to 50%, for example 20% to 30% or from 35% to 45%. The term "oil" means any substance that is wholly or partially liquid at ambient temperature or close-to-ambient temperature e.g. between 10°C and 40°C or between 15°C and 35°C, and which is hydrophobic but soluble in at least one organic solvent. Oils include vegetable oils (e.g. neem oil) and petrochemical oils.

The oil may be present in the composition in an amount of from about 2% to about 25%, from about 3% to about 20%, from about 3% to about 10% or from about 5% to about 10% by weight based upon the dry weight of the core.

P213428WO 62 Oils which may be included in the oil phase include poly-unsaturated fatty acids such as, for example, 3 oils for example eicosapentanoic acid (EPA), docosohexaenoic acid (DHA), alphalinoleic acid (ALA), conjugated linoleic acid (CLA). Preferably ultrapure EPA, DHA or ALA or CLA are used e.g. purity up to or above 98%. Omega oils may be sourced e.g. from any appropriate plant e.g. sacha inchi. Such oils may be used singly e.g. EPA or DHA or ALA or CLA or in any combination.

Combinations of such components including binary, ry etc combinations in any ratio are also contemplated e.g. a binary mixture of EPA and DHA in a ratio of 1:5 available commercially under the trade name Epax 6000. The oil part of the oil phase may comprise or be an oil mentioned in this paragraph.

Oils which may be included in the oil phase are ularly natural triglyceride-based oils which include olive oil, sesame oil, coconut oil, palm kernel oil, neem oil. The oil may be or may comprise saturated coconut and palm kernel oil-derived caprylic and capric fatty acids and glycerin e.g. as ed under the trade name Miglyol™ a range of which are available and from which one or more components of the oil phase of the invention may be selected including Miglyol™ 810, 812 lic/capric triglyceride); Miglyol™ 818: (caprylic/capric/linoleic triglyceride); Miglyol™ 829: (caprylic/capric/succinic triglyceride; Miglyol™ 840: lene glycol ylate/dicaprate). Note that Miglyol™ 2 are MCT ations which differ only in C8/C10-ratio and because of its low C10- t, the viscosity and cloud point of Miglyol™ 810 are lower. The Miglyol™ range is available commercially from Sasol Industries. As noted above, oils which may be included in the oil phase need not necessarily be liquid or fully liquid at room temperature. Waxy-type oils are also possible: these are liquid at manufacturing temperatures but solid or semi-solid at normal ambient temperatures. The oil part of the oil phase may comprise or be an oil mentioned in this paragraph.

Alternative or onal oils which may be included in the oil phase according to the invention are other medium chain triglyceride formulations such as for example Labrafac™ Lipophile manufactured by osse in particular product number WL1349. Miglyol™ 810, 812 are also medium chain triglyceride formulations.

Accordingly the oil phase may be or comprise medium chain mono-di- or tri rides.

] The medium chain glyceride(s) (eg mono- di- or tri- glyceride(s)) mentioned herein are those which comprise one or more triglycerides of at least one fatty acid selected from fatty acids having 6, 7, 8, 9, 10, 11 or 12 carbon atoms, e.g. C8-C10 fatty acids.

The oil phase may further comprise the surfactant as described above and elsewhere herein.

The ce of the surfactant in the oil phase may also provide a stabilising effect on the liquid composition when the oil phase is dispersed in the aqueous phase. In addition the presence of the surfactant in the oil phase may inhibit crystallisation of cyclosporin from a cyclosporin solution in the oil phase. The surfactant may also provide enhanced emulsification when the disperse phase is mixed with the aqueous phase during preparation of the liquid composition, composition or core (i.e act as an emulsifier).

P213428WO 63 The liquid lipid or oil of the oil phase or disperse phase is suitably not a surfactant. However, certain oils, particularly those derived from natural sources will comprise components which may have surface active ties. For example many triglyceride oils also comprise mono and diglyceride components and may therefore t some surfactant like properties. Accordingly the oil suitably has an HLB value of 0-10, however ly the oil has an HLB which is close to 0 for example an HLB of 0 to 3, optionally about 0, about 1 or about 2.

Surfactant in the oil phase may for e be or comprise a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, n the surfactant does not comprise or is not a polyethyleneglycol ether or ester. Optionally the surfactant is a medium chain or long chain fatty acid mono-glyceride, di-glyceride or a combination thereof, optionally wherein the surfactant does not comprise or is not a polyethyleneglycol ether or ester. Two particular surfactants contemplated by the invention are glyceryl caprylate/caprate and yl monooleate/dioleate. Commercial preparations may also be used as a tant e.g. those commercial preparations which contain minor components. Preferred examples are Capmul GMO-50 (glyceryl monooleate/dioleate) and Capmul MCM (glyceryl caprylate/caprate).

Within embodiments, the HLB of the oil may be in the range 0-10 (optionally 1-8, e.g. 1-6 and sometimes 1-5).

In another embodiment the oil phase comprises an oil with an HLB in the range 0-10 (preferably 1-5) and the has an HLB of up to 10 and optionally up to 7, 1-8, 1-7, 1-5, 2-5, 1-4, 1-3, 1-2, 2-4, 3-4, 5-8, 6-8 and 6-7..

In another ment the oil phase comprises an oil and the surfactant wherein the oil and the surfactant both have an HLB in the range 0-10. For example the oil has an HLB of 1-5, for example 1 to 4 or 1-2 and the tant has an HLB 2 -8, for example 3-7, 2-6, or 3-4).

Suitable oils which may comprise or be the oil phase or disperse phase with a low HLB (HLB less than 10) include medium chain triglycerides, caprylocaproyl macrogolglycerides and caprylic/capric triglyceride. In terms of commercial products, particularly preferred oils in the lower HLB range are Labrafac™ Lipophile (e.g. 1349 WL), Captex 355 and Miglyol 810.

It is to be understood that the oil phase or se phase in the ments above may further comprise one or more solvents, for example 2-(2-ethoxyethoxy)ethanol or low molecular weight PEG as mentioned above. The solvent may be present in the composition in an amount of form about 1% to 30%, for about 5% to about 30% , for about 10% to about 25%, or from about 12% to about 22% by weight based upon the dry weight of the uncoated composition or upon the dry weight of the core.

A particular oil phase comprises an oil (low HLB), the surfactant and a co-solvent. For example the ing three commercial ts: Transcutol P (as co-solvent), Myglyol 810 (as oil) and Capmul GMO-50 (surfactant). An oil phase may ore comprise or consist of a ation of the following: 2-ethoxyethanol, an MCT and particularly a caprylic/capric triglyceride formulation, and glyceryl monooleate/dioleate. The oil phase may further comprise the cyclosporin.

P213428WO 64 Preferably, cyclosporin is soluble in the oil phase. As discussed below in relation to preparation of the composition, the cyclosporin is suitably ved in the oil phase and the oil phase is mixed with an aqueous phase comprising the hydrogel-forming polymer.

The disperse phase (oil phase) may be or comprise a glyceride formulation, optionally wherein the disperse phase is or comprises a fatty acid yceride, diglyceride or triglyceride or a combination thereof, or the disperse phase is or comprises a ic/capric triglyceride formulation.

The disperse phase of the colloidal core may comprise self-assembly structures, for example micelles, vesicles, liposomes or nanoparticles, or at least the structures which result from drying aqueous colloids of such types (have the characteristics of structures which result from drying aqueous colloids of such types). The invention in particular includes formulations in which the disperse phase is micellar, i.e. formed of micelles and/or promicelles. The term "promicelle" refers to a part of a formulation which will form a micelle upon contact with water, e.g. gastrointestinal contents.

The following sion for convenience refers to es but is applicable in general to other self-assembly ures. A micelle-forming surfactant is t as micelles dispersed within the hydrogel-forming polymer in a "wet" (not yet dried) composition made as an intermediate in the manufacturing process described herein. It is believed also to be present as micelles in the dried ition but observability of micelles or micelle-like structures in the dried composition is not a requirement of the invention. It is mentioned at this point that the presence of a surfactant in micelle form does not require that the entire surfactant content of a composition is in micelle form as it is considered more probable that a portion of the tant will be outside the micelles. Thus in the "wet" composition, whether the hydrogel-forming polymer is in the gel state or the sol (liquid) state may comprise the micelle-forming surfactant at a tration above the critical micelle concentration.

The diameter of the sed micelles may be between 0.5 nm and 200 nm, 1 nm and 50 nm, or 5 nm and 25 nm. The size of the micelles may be determined by dynamic light scattering or diffusion NMR techniques known within the art. Although the size of the es is given as a diameter this does not imply that the micelles must be purely spherical species only that they may possess some approximately circular dimension.

The surfactant may be a non-ionic surfactant. The surfactant may be a yethylated tant. The surfactant has a hilic head which may be a hydrophilic chain, for example a polyoxyethylene chain or a polyhydroxylated chain.

The surfactant of course has a hydrophobic part and in particular a hydrophobic chain. The hydrophobic chain may be a hydrocarbon chain, for example having at least 6 carbon atoms and optionally at least 10 carbon atoms, and particularly of at least 12 carbon atoms; some hydrocarbon chains have no more than 22 carbon atoms, for example C10-C20, C12-C20 or C15-C20 arbon chains. It may be an alkyl chain, e.g. having a number of carbon atoms just mentioned. It may be an alkenyl chain comprising one or more carbon-carbon double bonds, e.g. having a number of carbon atoms just mentioned. The surfactant may comprise a hydrocarbon chain, e.g. alkyl chain or alkenyl chain that is substituted provided that it ins a hobic characteristic. There may for P213428WO 65 example be one or two substituents, for example a single substituent, e.g. ed from halogen (e.g.

F or Cl), y, thiol oxo, nitro, cyano; hydroxy or thiol substituents may be esterified by for example a fatty acid. One class of surfactants se a hydrocarbon bstituted by y; optionally, at least a portion of the hydroxy groups of an aliquot of surfactant, e.g. of the surfactant in a bead, may be esterified by a fatty acid or mono-hydroxy fatty acid as disclosed herein or etherified by a fatty alcohol for example having at least 6 carbon atoms and optionally at least 10 carbon atoms, and particularly of at least 12 carbon atoms; some hydrocarbon chains have no more than 22 carbon atoms, for example C10-C20, C12-C20 or C15-C20 fatty alcohols.

The hydrophobic chain may be part of an esterified fatty acid R1-COOH or of an etherified or esterified fatty ether R1-COH where R1 is the hydrophobic chain, e.g. as mentioned in the preceding paragraph. The ester-forming or, as the case may be, forming group will typically comprise a hydrophilic chain.

As mentioned, the tant may have a hydrophilic chain and may be a non-ionic surfactant, and may satisfy both requirements. The hydrophilic chain may be a poly(ethyleneglycol), also known as poly(oxyethylene) or macrogol. The hydrophilic chain may be of the formula 2- CH2)n-OR where n is 5 or 6 to 50 and R is H or alkyl, e.g. ethyl or methyl. The ion includes implementations in which n is from 6 to 40, e.g. from 6 to 35. In some embodiments, n is from 6 to 25 and optionally is from 8 to 25 or from 8 to 15. In other embodiments, n is from 8 to 50 or from 8 to 40, e.g. is from 10 to 50, 10 to 40 or 10 to 35. In a particular ment, n is 15. For all hydrophilic chains of the formula -(O-CH2-CH2)n-OR, in one class of embodiments R is H.

The hydrophilic chain may be a polyhydroxylated chain (for example a C5-C20 e.g. C5-C10 chain), e.g. having a hydroxy group on the carbon atoms of the chain, for example a glucamide.

The micelle-forming surfactant may comprise a combination of a hydrophobic chain as bed above and a hydrophilic chain as described above. It may ore be, or comprise, a macrogol ester of a fatty acid as described herein or a macrogol ether of a fatty alcohol as described herein.

Micelle-forming surfactants comprising a hydrophobic chain and a hydrophilic chain can be selected from the group consisting of: macrogol esters; macrogol ethers; diblock copolymers; triblock copolymers; and amphiphilic polymers. In certain embodiments of the invention any combinations of the group are included within the invention.

Examples of macrogol esters which are suitable for use in the present invention are macrogol esters of fatty acids having at least 6 carbon atoms and optionally at least 10 carbon atoms, and particularly of at least 12 carbon atoms; some fatty acids have no more than 22 carbon atoms, for example C10-C20, C12-C20 or C15-C20 fatty acids. The fatty acids may be ted or unsaturated but are in particular saturated. To be mentioned are macrogol 25 earyl ether (Cremophor® A25); macrogol 6 cetostearyl ether (Cremophor® A6); ol glycerol ricinoleate 35 (Cremophor® EL); macrogol-glycerol hydroxystearate 40 (Cremophor® RH 40); macrogolhydroxystearate (Solutol® HS 15). Examples of macrogol ethers which are suitable for use in the present invention are macrogol P213428WO 66 ethers of fatty alcohols having at least 6 carbon atoms and optionally at least 10 carbon atoms, and particularly of at least 12 carbon atoms; some fatty alcohols have no more than 22 carbon atoms, for example C10-C20, 0 or C15-C20 fatty alcohols. The fatty alcohols may be saturate or rated but are in one embodiment saturated.

Examples of amphiphilic polymers which are suitable for use in the present invention are : alkyl glucamides; fatty alcohol poly(ethoxyl)ates also known as polyethoxylated alkyl ethers; poly(ethoxyl)ated fatty acid esters (Myrj or Solutol); fatty amide polyethoxylate; fatty amine ethoxylate; alkylphenol ethoxylate; polyethoxylated an esters (polysorbates); polyethoxylated glycerides; or poly-glycerol esters.

Examples of mers, which are le for use in the present invention are: pluronics(poloxamers); polyvinylpyrollidone-polyvinylacetate (Plasdone S630); aminoalkyl methacrylate copolymer (Eudragit EPO); methacrylic acid – methyl methacrylate copolymer (Eudragit S100, L100); prolactone-PEG; prolactone-methoxy – PEG; poly(aspartic acid)-PEG; poly(benzyl-L-glutamate)-PEG; poly(D,L-lactide)methoxy-PEG; poly(benzyl-L-aspartate-PEG; or poly(L-lysine)-PEG In a preferred embodiment the micelle-forming surfactant cis a ol ester, more preferably a macrogol ester that conforms to the European Pharmacopoeia monograph number 2052 macrogolhydroxystearate, such as Kolliphor® HS 15 marketed by BASF.

Kolliphor® HS 15 consists of polyglycol mono- and di-esters of 12-hydroxystearic acid and about 30% of free polyethylene glycol. The main components of the ester part have the following chemical structures: H O H O O O where x and y are integers and a small part of the 12-hydroxy group can be etherified with hylene glycol.

Suitable surfactants comprise those which during manufacture e with the aqueous phase (including hydrogel-forming polymer) in an amount above their CMC to form a clear liquid.

Kolliphor® HS 15 is such a surfactant.

P213428WO 67 In certain embodiments the weight ratio of the micelle-forming surfactant to the antigen is from 10:1 to 100:1, optionally from 50:1 to 100:1. In some embodiments, the ratio is from 80:1 to 90:1.

In ular embodiments, the ratio is from 50:1 to 60:1.

In particular embodiments, the compositions of the invention comprise a combination of micelle-forming compounds. Such a combination of micelle-forming compounds may consist of two or more surfactants as mentioned in the ing section of this specification. Alternatively, a surfactant may be combined with one or more other compounds at least ially able to form micelles with the surfactant, optionally selected from cationic lipids and glycolipids, amongst others.

As an additional option, a composition may comprise a plurality of surfactants as mentioned in the preceding section of this specification and one or more other nds at least potentially able to form micelles with the surfactant, optionally selected from cationic lipids and glycolipids, amongst others.

The ion therefore includes itions as described herein which comprise: two or more micelle-forming surfactants, e.g. two or more surfactants having a hydrophobic chain and a hydrophilic chain; a compound, e.g. a single compound or two or more compounds, selected from cationic lipids and glycolipids; two or more micelle-forming surfactants and a compound, e.g. a single compound or two or more nds, ed from cationic lipids and ipids.

A disperse phase which is or comprises a surfactant may enhance the absorption of an active ingredient, for example cyclosporin A, into the tissue of the GIT, for example by forming selfassembly structures, such as micelles, which are ated with the active ingredient and thus t the drug to the mucosa tissue of the GI tract in a form which es uptake/absorption into the tissue.

The oil phase may also include one or more volatile or non-volatile ts, which may be the same or different from the solvent or co-solvent previously mentioned. Such solvents may for example remain in the formulation of the invention following processing e.g. initial dissolution of the components present in the core, and have no particular function in the core formulation. Alternatively, such solvents if present may function to maintain the cyclosporin a ved state (in solution) within the oil phase or to facilitate dispersion, egress etc. In other embodiments, the solvent may have partly or fully evaporated during processing and therefore be t in only minor quantities if at all. In a related embodiment, the t, ularly when a solvent which is both oil and water-soluble is used, may be partly or completely present in the aqueous phase of the core. An example of such a solvent is ethanol. Another example is transcutol which is already mentioned as a co-solvent.

Accordingly, the composition may comprise a hydrogel-forming polymer matrix which forms a continuous phase and a disperse phase comprising cyclosporin, a low HLB medium or long chain mono- or di-ester surfactant, a low HLB oil, and optionally a co-solvent. Optionally, the medium or long-chain mono- or di-ester tant is a medium- or long-chain mono- or di-glyceride surfactant.

P213428WO 68 In a particular embodiment the composition or the core is in the form of a solid colloid, the d comprising a continuous phase and a disperse phase, wherein the disperse phase is or comprises: cyclosporin; a medium chain mono-, di- and/or yceride, for example a medium chain triglyceride, ularly caprylic/capric triglyceride; a medium- or long-chain mono- or di-glyceride, particularly glyceryl monooleate/dioleate; and a co-solvent (for example 2-(ethoxyethoxy)ethanol); and wherein the continuous phase is or ses: a hydrogel-forming polymer matrix which is or comprises a hydrocolloid selected from carrageenan, gelatin, agar and pectin, or a combination thereof optionally selected from gelatin and agar or a combination thereof, more particularly the polymer of the a hydrogel-forming polymer matrix is or comprises gelatin; a plasticiser, optionally a plasticiser selected from glycerin, a polyol for example sorbitol, polyethylene glycol and triethyl citrate or a mixture thereof, particularly sorbitol; and an anionic surfactant, for example at least one surfactant selected from fatty acid salts, alkyl sulphates and bile salts, ularly an alkyl sulphate, for example sodium dodecyl sulphate.

] In a further specific embodiment the disperse phase comprises: cyclosporin in an amount of 60 – 180 mg/g; caprylic/capric triglyceride in an amount of 40 – 80 mg/g; 2-(2-ethoxyethoxy)ethanol in an amount of 100 – 200 mg/g; and glyceryl monooleate and/or glyceryl te in an amount of 100 – 150 mg/g, wherein weights are based upon the dry weight of the composition.

The oil phase or disperse phase may comprise: cyclosporin in an amount of 120 – 360 mg/g; ic/capric ceride in an amount of 80 – 160 mg/g; 2-(2-ethoxyethoxy) ethanol in an amount of 200 – 400 mg/g; and glyceryl monooleate and/or glyceryl dioleate in an amount of 200 – 300 mg/g, wherein the weights are based on the weight of the wet composition.

The liquid composition may comprise an oil phase comprising: cyclosporin in an amount of 20 – 60 mg/g; caprylic/capric triglyceride in an amount of 13 – 27 mg/g; thoxyethoxy) ethanol in an amount of 50 – 70 mg/g; and glyceryl eate and/or glyceryl dioleate in an amount of 30 – 55 mg/g, wherein weights are based upon the wet weight of the composition, i.e. the liquid composition, optionally wherein the oil phase to aqueous phase ratio may be 1:5.

In an embodiment the aqueous phase or continuous phase comprises a hydrogel-forming r matrix comprising gelatin in an amount of 300 to 700 mg/g, and SDS in an amount of 15 – 50 mg/g, wherein s are based upon the dry weight of the composition.

P213428WO 69 In an embodiment the aqueous phase may comprise a hydrogel-forming polymer matrix comprising gelatin in an amount of 120 to 280mg/g and SDS in an amount of 6 – 20 mg/g wherein the weights are based upon the weight of the s phase. The s phase may comprise a hydrogel-forming polymer matrix comprising gelatin in an amount of 100 to 230mg/g and SDS in an amount of 5 – 16 mg/g, n the weights are based on the weight of the composition, i.e. the liquid ition, optionally n the oil phase to aqueous phase ratio may be 1:5.

Suitably in the ment of the immediately preceding two paragraphs the cyclosporin may be present in an amount of 90 to 140 mg/g, for example of 60 to 150 mg/g, 80 to 120 mg/g or particularly 80 to 100 mg/g. The anionic surfactants are as defined , for example an anionic surfactant selected from alkyl sulphates, carboxylates or phospholipids (particularly SDS).

The composition or the cores described above comprising hydrogel-forming polymer matrix may be coated as described herein. A particular coating for these embodiments is a coating comprising: a first coating (sub-coating) which is or comprises a water-soluble cellulose ether, particularly hydroxypropylmethyl cellulose; a second coating outside the first coating which is or comprises a modified e coating, particularly a pH independent modified release coating, more especially a coating comprising ethyl cellulose (e.g. Surelease) still more particularly a coating comprising ethyl cellulose and a watersoluble polysaccharide such as pectin (e.g. a Surelease-pectin coating as described herein); and wherein the first coating is present in an amount corresponding to a weight gain due to the first coating in a range selected from : (i) from 8% to 12%, for e about 10%; or (ii) from 4% to 6%, for example about 5% by weight based upon the weight of the formulation prior to applying the first coating; and wherein the second coating is present in an amount corresponding to a weight gain of the formulation due to the second coating ed from (a) from 10% to 12%, for example about 11% or about 11.5%; or (b) from 16% to 18%, for example about 17% by weight based upon the weight of the formulation prior to ng the second coating. y, the composition or the cores described above comprising el-forming polymer matrix may be coated with a coating comprising: a second coating which is or comprises a modified release coating, particularly a pH independent modified release coating, more especially a coating comprising ethyl cellulose (e.g.

Surelease) still more particularly a coating comprising ethyl ose and a water-soluble polysaccharide such as pectin (e.g. a Surelease-pectin coating as described herein); and wherein the second coating is present in an amount corresponding to a weight gain of the formulation due to the second coating selected from (a) from 10% to 12%, for example about 11% or about 11.5%; or (b) from 16% to 18%, for e about 17% by weight based upon the weight of the formulation prior to applying the second coating.

Surfactant P213428WO 70 The composition comprises a surfactant, as described above. The surfactant may be t in the composition or the core, for example in the el-forming polymer matrix, or in the disperse phase or both. The tant may also be present in one or more of the coatings comprised in the composition or applied to the core.

The composition may comprise a further surfactant. Where the composition comprises a further surfactant this surfactant can be referred to as a second surfactant and the surfactant present in the composition of the invention can be referred to as a first tant. ingly, the first surfactant is or comprises the medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof, which does not comprise or is not a polyethyleneglycol ether or ester. The further surfactant may be present in the composition or the core, for example in the hydrogel-forming polymer matrix, or in the disperse phase or both. The further surfactant may also be t in one or more of the coatings comprised in the composition or applied to the core. Suitable r surfactants can be anionic, cationic, zwitterionic, or non-ionic.

In the description and claims of this specification, the term ctant" is employed as a contraction for "surface active agent". For the purposes of this description and claims, it is assumed that there are four major classifications of surfactants; therefore the further surfactant may be: anionic, ic, non-ionic, and amphoteric (zwitterionic). The nic surfactant remains whole, has no charge in aqueous solutions, and does not dissociate into positive and negative ions. Anionic surfactants are water-soluble, have a negative charge and dissociate into positive and negative ions when placed in water. The negative charge lowers the surface tension of water and acts as the surface-active agent. Cationic surfactants have a positive charge, and also dissociate into positive and negative ions when placed in water. In this case, the positive ions lower the surface tension of the water and act as the surfactant. The amphoteric (zwitterionic) surfactant assumes a positive charge in acidic ons and performs as a cationic surfactant, or it assumes a negative charge in an alkaline solution and acts as an anionic surfactant.

The further surfactant(s) may be selected from: anionic tants and combinations thereof; from non-ionic surfactants and combinations thereof; and from combination of an anionic surfactant (e.g. a single such surfactant or a plurality thereof) and a non-ionic surfactant (e.g. a single such tant or a plurality thereof). Preferably the second surfactant is an anionic surfactant.

Accordingly, in an embodiment the liquid composition comprises an aqueous phase comprising a el forming polymer, a first surfactant and an oil phase being sed in the aqueous phase in which cyclosporin is dissolved, wherein the first surfactant is or comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and the first surfactant does not comprise or is not a polyethyleneglycol ether or ester, the liquid composition, r sing a second surfactant, preferably wherein the second surfactant is an anionic surfactant. The second tant may be present in an amount of 10 to 70mg/g or 15 to 60 mg/g.

Furthermore, in an ment the composition comprises cyclosporin, a hydrogel forming polymer matrix, a first surfactant and an oil phase being sed in the hydrogel forming polymer matrix, wherein the first surfactant is or comprises a medium chain or long chain fatty acid mono- or di- P213428WO 71 glyceride or a combination thereof and does not comprise or is not a polyethyleneglycol ether or ester, the composition further comprising a second surfactant, preferably wherein the second surfactant is an anionic surfactant.

Surfactants can also be classified according to their hydrophilic-lipophilic balance (HLB) which is a e of the degree to which the surfactant is hydrophilic or lipophilic, determined by calculating values for the different regions of the molecule, as described nally for non-ionic surfactants) by Griffin in 1949 and 1954 and later by Davies. The s apply a formula to the molecular weight of the whole molecule and of the hydrophilic and ilic portions to give an arbitrary (semi-empirical) scale up to 40 although the usual range is between 0 and 20. An HLB value of 0 corresponds to a completely hydrophobic molecule, and a value of 20 would correspond to a molecule made up completely of hilic components. The HLB value can be used to t the surfactant properties of a molecule: HLB Value ed properties 0 to 3 antifoaming agent from 4 to 6 W/O emulsifier from 7 to 9 wetting agent from 8 to 18 an O/W emulsifier from 13 to 15 typical of detergents to 18 solubiliser or hydrotrope Although HLB numbers are assigned to surfactants other than the non-ionic, for which the system was invented, HLB numbers for anionic, cationic, non-ionic, and amphoteric (zwitterionic) surfactants can have less significance and often ent a relative or comparative number and not the result of a mathematical ation. This is why it is possible to have tants above the "maximum" of 20. HLB numbers can however be useful to describe the HLB requirement of a desired application for a given emulsion system in order to achieve good performance.

Non-ionic Surfactants The further surfactant (second surfactant) may be or comprise at least one tant selected from the following non-ionic surfactants. tty acid ter surfactants, PEG-fatty acid diester surfactants, PEG-fatty acid monoester and diester surfactant mixtures, PEG glycerol fatty acid esters, transesterified products of oils and alcohols, lower alcohol fatty acid esters, polyglycerised fatty acids, propylene glycol fatty acid esters, mono and diglyceride tants, sterol and sterol derivative surfactants, PEG-sorbitan fatty acid , sorbitan fatty acid esters, polyethylene glycol alkyl ethers, sugar ester surfactants, polyethylene glycol alkyl phenol surfactants, POE-POP block copolymers, phospholipids,.

A PEG-fatty acid mono ester tant for example PEG 4-100 monolaurate, PEG 4-100 monooleate, PEG 4-100 monostearate, PEG-laurate, PEG-oleate, PEG stearate, and PEG ricinoleate.

A tty acid diester surfactant for example PEG dilaurate; PEG dioleate, PEG distearate, PEG dipalmitate. A mixture of PEG-fatty acid mono- and diesters.

P213428WO 72 A PEG glycerol fatty acid ester for example PEG glyceryl laurate, PEG glyceryl stearate, PEG glyceryl oleate.

PEG-sorbitan fatty acid esters for example PEG an laurate, PEG sorbitan monolaurate, PEG sorbitan lmitate, PEG sorbitan monostearate, PEG sorbitan arate, PEG sorbitan tearate, PEG sorbitan monooleate, PEG sorbitan oleate, PEG sorbitan trioleate, PEG sorbitan tetraoleate, PEG an monoisostearate, PEG sorbitol hexaoleate, PEG sorbitol hexastearate.

Propylene glycol fatty acid esters for example propylene glycol monocaprylate, propylene glycol monolaurate, propylene glycol oleate, propylene glycol myristate, ene glycol monostearate, propylene glycol 72ydroxyl stearate, propylene glycol ricinoleate, ene glycol isostearate, propylene glycol monooleate, propylene glycol dicaprylate/dicaprate, propylene glycol dioctanoate, ene glycon caprylate/caprate, propylene glycol dilaurate, propylene glycol distearate, propylene glycol dicaprylate, propylene glycol dicaprate.

A sorbitan fatty acid ester for example sorbitan monolaurate, sorbitan lmitate, sorbitan monooleate, sorbitan monostearate, sorbitan trioleate, sorbitan sesquioleate, sorbitan tristearate, sorbitan monoisostearate, sorbitan sesquistearate.

] Lower l fatty acid esters for example ethyl oleate, isopropy myristate, isopropyl palmitate, ethyl linoleate, pyl linoleate. ] yethylene-polyoxypropylene block copolymers for example poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, mer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, mer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, mer 401, poloxamer 402, poloxamer 403, poloxamer 407.

Polyglycerised fatty acids for example polyglyceryl stearate, polyglyceryl , polyglyceryl isostearate, polyglyceryl laurate, polyglyceryl ricinoleate, polyglyceryl linoleate, polyglyceryl pentaoleate, polyglyceryl dioleate, polyglyceryl distearate, polyglyceryl trioleate, polyglyceryl septaoleate, polyglyceryl tetraoleate, polyglyceryl decaisostearate, polyglyceryl decaoleate, polyglyceryl monooleate, dioleate, polyglyceryl polyricinoleate.

PEG alkyl ethers for example PEG oleyl ether, PEG lauryl ether, PEG cetyl ether, PEG stearyl ether.

PEG alkyl phenols for example PEG nonyl phenol, PEG octyl phenol ether.

] Transesterification products of alcohol or polyalcohol with natural or hydrogenated oils for example PEG castor oil, PEG hydrogenated castor oil, PEG corn oil, PEG almond oil, PEG apricot kernel oil, PEG olive oil, PEG-6 peanut oil, PEG hydrogenated palm kernel oil, PEG palm kernel oil, PEG triolein, PEG corn glycerides, PEG almond glycerides, PEG ate, PEG caprylic/capric triglyceride, lauroyl macrogol glyceride, stearoyl macrogol glyceride, mono, di, tri, tetra esters of vegetable oils and sorbitol, rythrityl tetraisostearate, pentaerythrityl distearate, pentaerythrityl P213428WO 73 tetraoleate, pentaerythrityl tetrastearate, pentaerythrityl tetracaprylate/tetracaprate, pentaerythrityl tetraoctanoate.

Oil-soluble ns for example vitamins A, D, E, K, and isomers, analogues, and derivatives thereof. The derivatives include, for example, c acid esters of these oil-soluble vitamin substances, for example the esters of vitamin E or vitamin A with succinic acid. Derivatives of these vitamins include tocopheryl PEG-1000 succinate in E TPGS) and other tocopheryl PEG succinate tives with various lar weights of the PEG moiety, for example PEG 100-8000.

Sterols or sterol derivatives (e.g. esterified or etherified sterols as for example PEGylated sterols) for example cholesterol, sitosterol, lanosterol, PEG cholesterol ether, PEG cholestanol, phytosterol, PEG phytosterol.

Sugar esters for example sucrose distearate, sucrose distearate/monostearate, sucrose dipalmitate, sucrose monostearate, sucrose monopalmitate, sucrose monolaurate, alkyl glucoside, alkyl maltoside, alkyl maltotrioside, alkyl glycosides, derivatives and other sugar types: glucamides.

] Carboxylates (in particular carboxylate esters) for e ether carboxylates, succinylated monoglycerides, sodium stearyl fumarate, stearoyl propylene glycol hydrogen succinated, mono/diacetylated tartaric acid esters of mono- and diglycerides, citric acid esters of mono-, diglycerides, glyceryl-lacto esters of fatty acids; acyl lactylates: lactylic esters of fatty acids, calcium/sodium stearoyllactylate calcium/sodium stearoyl lactylate, alginate salts, propylene glycol alginate.

A fatty acid monoglyceride, diglyceride or triglyceride or a combination f.

Anionic Surfactants The further surfactant d tant) may be or comprise at least one anionic surfactant.

] The second tant may be a fatty acid salt or bile salt for e sodium caproate, sodium caprylate, sodium caprate, sodium laurate, sodium ate, sodium myristolate, sodium ate, sodium palmitoleate, sodium oleate, sodium ricinoleate, sodium linoleate, sodium linolenate, sodium stearate, sodium lauryl sulfate, sodium tetradecyl sulfate, sodium lauryl sarcosinate, sodium dioctyl sulfosuccinate; sodium cholate, sodium taurocholate, sodium glycocholate, sodium deoxycholate, sodium taurodeoxycholate, sodium glycodeoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, sodium taurochenodeoxycholate, sodium glyco chenodeoxycholate, sodium cholylsarcosinate and sodium N-methyl taurocholate. Preferably the second surfactant is sodium lauryl sulphate.

Phospholipids for example egg/soy lecithin, cardiolipin, sphingomyelin, phosphatidylcholine, phosphatidyl lamine, phosphatidic acid, phosphatidyl glycerol, atidyl serine.

Phosphoric acid esters having the general formula RO-PO3-M+ where the R group is an ester forming group, e.g. an alkyl, alkenyl or aryl group ally tuted by a PEG moiety through which the alkyl, alkenyl or aryl group is coupled to the phosphate moiety. R may be a residue of a P213428WO 74 long chain (e.g. >C9) alcohol or a phenol. Specific examples include diethanolammonium polyoxyethylene-10 oleyl ether phosphate, esterification products of fatty alcohols or fatty alcohol ethoxylates with phosphoric acid or anhydride.

Sulfates and sulfonates (in particular esters thereof) for example ethoxylated alkyl sulfates, alkyl benzene sulfones, -olefin sulfonates, acyl onates, acyl taurates, alkly glyceryl ether sulfonates, octyl sulfosuccinate disodium, disodium undecylenamideo-MEA-sulfosuccinate, alkyl phosphates and alkyl ether phosphates.

Cationic Surfactants The further surfactant (second surfactant) may be or se at least one cationic surfactant selected from the following cationic surfactants.

] Hexadecyl triammonium e, dodecyl ammonium chloride, alkyl benzyldimethylammonium salts, utyl phenoxyethoxydimethyl benzylammonium salts, alkylpyridinium salts; betains kylglycine): lauryl betaine (N-lauryl,N,N-dimethylglycine); ethoxylated amines: yethylene-15 coconut amine, alkyl –amines/ diamines/ quaternaty amines and alkyl ester.

Emulsifiers ] The surfactant may act as an emulsifier such surfactants include non-ionic fiers, for example selected from: a mixture of triceteareth-4 phosphate, ethylene glycol palmitostearate and diethylene glycol palmitostearate (for e sold under the trade mark SEDFOSTM 75); sorbitan , e.g. sorbitan monooleate, sorbitan monolaurate, sorbitan monpalmitate, sorbitan monostearate (for example products sold under the trade mark Span®), PEG-8 beeswax e.g. sold under the trade mark Apifil; a mixture of cetyl alcohol, ceteth-20 and steareth-20 (for example EmulcireTM 61 WL 2659); a mixture of PEG-6 stearate and PEG-32 stearate (for example Tefose 1500); a mixture of PEG-6 palmitostearate, ethylene glycol palmitostearate, and PEG-32 ostearate (e.g. Tefose 63); triglycerol diisostearate (for example products sold under the trade mark Plurol Diisostearique); polyglyceryl-3 dioleate (for example products sold under the trade mark Plurol Oleique).

Preferred Second tants.

] Preferably the second surfactant is an anionic surfactant. For example, the second surfactant may be an alkyl sulphate, for e sodium dodecyl sulphate. The second surfactant may be present in an amount of 10 to 70mg/g or 15 to 60 mg/g.

The second tant may be a fatty acid salt or bile salt for example sodium caproate, sodium caprylate, sodium caprate, sodium e, sodium ate, sodium myristolate, sodium palmitate, sodium palmitoleate, sodium oleate, sodium ricinoleate, sodium linoleate, sodium linolenate, sodium stearate, sodium lauryl sulfate, sodium tetradecyl sulfate, sodium lauryl sarcosinate, sodium dioctyl sulfosuccinate; sodium cholate, sodium taurocholate, sodium glycocholate, sodium deoxycholate, sodium taurodeoxycholate, sodium glycodeoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, sodium taurochenodeoxycholate, sodium glyco chenodeoxycholate, P213428WO 75 sodium cholylsarcosinate and sodium N-methyl taurocholate. Preferably the second surfactant is sodium lauryl sulphate.

Other Excipients ] The composition optionally contains one or more of the following additional substances or categories of substances. For example, the composition may contain a protectant such as, for example, a proteolytic enzyme inhibitor or a protector t acid degradation or both (e.g. an alkali for example sodium hydroxide); an adhesive entity such as, for e, a muco- or bio-adhesive; excipients to maximize solubility of the active ingredient; excipients to maximize permeability of the active ingredient in the GIT. Typical excipients for enhancing the permeability of the epithelial r include but are not d to sodium e, sodium dodecanoate, sodium palmitate, SNAC, an and derivatives thereof, fatty acids, fatty acid esters, polyethers, bile salts, phospholipids, alkyl polyglucosides, sugar esters, ylase inhibitors, antioxidants (e.g. ascorbic acid) and/or nitric oxide donors. The preceding list is of particular interest to e permeability in the ileum.

To enhance permeability in the colon, typical ents include, but not limited to sodium caprate, sodium dodecanoate, sodium palmitate, SNAC, chitosan and derivatives f, fatty acids, fatty acid esters, polyethers, bile salts, phospholipids, alkyl polyglucosides, ylase inhibitors, antioxidants (optionally selected from curcuminoids, flavonoids, curcumin, beta-carotene, αtocopherol , ascorbic acid, ascorbate, lazaroid, carvedilol, butylated hydroxytoluene, propyl gallate, hydralazine, carnosic acid, vitamin E, lecithin ithin (vitelin), vegilecithin, fumaric acid or citric acid) and/or nitric oxide , including nitric oxide donor groups covalently attached to various pharmaceutically active ingredients. The composition may further comprise excipients or other active pharmaceutical or other ingredients to enhance local tissue bioavailability in the GIT, such as the small intestine or colon, ing efflux pump inhibitors, including, but not limited to PgP pump inhibitors (optionally selected from NSAIDs, dine , omeprazole, Vitamin E TPGS, verapimil, quinidine, PSC833, avir (APV), indinavir (IDV), nelfinavir (NFV), ritonavir (RTV) and saquinavir (SQV)), and metabolism inhibitors, including, but not limited to, cytochrome P450 inhibitors, ally selected from: essential oils, cimetidine, surfactants (for example cremophor), oils, omeprazole, verapamil, ritonavir and carbamazepine as well as plant extracts, e.g, from citrus fruits. The composition may therefore further comprise a P450 tor to r reduce metabolism of cyclosporin following administration of the composition. The P450 inhibitor may act to inhibit enteric and/or c metabolism of the cyclosporin. The composition may further comprise a PgP inhibitor. Optionally the ition may comprise a P450 inhibitor and a PgP inhibitor.

] The composition may further comprise excipients to enhance the therapeutic potential of an active ingredient, for example cyclosporin A or another immunosuppressant, throughout the gastrointestinal tract, including in the ileum and colon including, but not limited to absorption limiters, essential oils such as, for example, omega 3 oils, natural plant extracts such as, for example, neem, ion-exchange resins, bacteria degradable conjugation linkers such as, for example, azo bonds, polysaccharides such as, for example, amylose, guar gum, pectin, chitosan, inulin, cyclodextrins, P213428WO 76 chondroitin sulphate, dextrans, guar gum and locust bean gum, nuclear factor kappa B inhibitors, acids such as, for example, fumaric acid, citric acid and others, as well as modifications f.

] The composition may further comprise excipients to reduce systemic side effects associated with absorption of certain active, for example cyclosporin or other immunosuppressants, in the GIT, such as the small intestine, including, but not limited to, antioxidants, such as, for example, curcuminoids, flavanoids or more specifically including curcumin, beta-carotene, α-tocopherol, ascorbate or lazaroid.

The composition may further or separately comprise antioxidants (such as, for example, ascorbic acid or BHT - butyl hydroxy toluene) taste-masking or photosensitive components or rotective components. Antioxidants may be incorporated in the aqueous phase (e.g. hydrophilic antioxidants) or in the disperse phase of the core (e.g. hydrophobic antioxidants such as, for example, vitamin E) for example up to 1% by weight, preferably between 0.01 and 0.50% by weight, more preferably n 0.10 to 0.20% by weight.

The composition may further comprise immune-enhancing nutrients such as ns A/B/C/E; carotenoids/beta-carotene and iron, manganese, selenium, zinc, especially when the composition contains an immunosuppressant, as in the case of an immunosuppressant targeted to the ileum and/or colon, e.g. the colon. Such nutrients may be present in composition, or if the composition has a coating, for example if it is the form of a bead, the nutrients may be included in the coating.

The composition may also include other well know excipients used in pharmaceutical itions including colorants, taste masking agents, diluents, fillers, binders etc. The presence of such optional additional components will of course depend upon the particular dosage form adopted.

Shape, Size and Geometry ] The composition of the invention can be formed into a limitless number of shapes and sizes.

In the n below describing the process for making the composition, s methods are given including pouring or introducing a fluid dispersion into a mould where it hardens or can be caused to harden. Thus the composition can be created in whichever form is desired by creating an appropriate mould (e.g. in the shape of a disc, pill or tablet). However, it is not essential to use a mould. For example, the ition may be formed into a sheet e.g. resulting from pouring a fluid dispersion onto a flat surface where it hardens or can be caused to harden.

] Preferably, the composition may be in the form of spheres or cal-like shapes made as described below. ably, the composition of the invention is in the form of ntially spherical, seamless minibeads. The absence of seams on the minibead e is an advantage e.g. in further processing, for example coating, since it allows more tent coating, flowability etc. The absence of seams on the minbeads also enhances consistency of dissolution of the beads.

The preferred size or diameter range of minibeads according to the invention can be chosen to avoid retention in the stomach upon oral administration of the minibeads. Larger dosage forms are ed for variable periods in the stomach and pass the pyloric ter only with food whereas smaller particles pass the pylorus independently of food. Selection of the appropriate size range (see P213428WO 77 below) thus makes the therapeutic effect post-dosing more consistent. Compared to a single large monolithic oral format such as, for example, a traditional compressed pill, a population of beads released into the GI tract (as foreseen by the dosage form of the present invention) permits greater intestinal lumen dispersion so ing absorption via exposure to greater epithelial area, and achieves greater topical coating in certain parts of the GI tract for example the colon). Reduction of residence time in the ileo-caecal junction is r potential advantage.

The composition of the invention is preferably monolithic meaning internally (i.e. crosssectionally ) homogeneous, excluding a possible thin skin of matrix material and excluding any coating layers.

The minibeads provided for by the ation of the present ion generally range in diameter from 0.5 mm to 10 mm with the upper limit preferably 5 mm, e.g. 2.5 mm A particularly convenient upper limit is 2mm or 1.7mm. The lower limit can preferably be 1mm, e.g. 1.2mm, more preferably from 1.3mm, most preferably from 1.4mm. In one embodiment the diameter is from 0.5 to 2.5mm, for example from 1mm to 3mm, 1mm to 2mm, 1.2mm to 3mm or 1.2mm to 2mm. The ads may have a diameter of no more than 2.5mm, irrespective of their minimum size. The beads may have a diameter of no more than 2mm, ective of their minimum size.

A minibead as described herein may have an aspect ratio of no more than 1.5, e.g. of no more than 1.3, for example of no more than 1.2 and, in particular, of from 1.1 to 1.5, 1.1 to 1.3 or, 1.1 to 1.2. A population of minibeads as described herein, e.g. at least 10 beads, may have an e aspect ratio of no more than 1.5, e.g. of no more than 1.3, for example of no more than 1.2 and, in particular, of from 1 to 1.5, 1 to 1.3 or 1 to 1.2. The aspect ratios mentioned in this paragraph optionally apply to coated minibeads and optionally apply to uncoated minibeads. Average aspect ratio is suitably determined for a population of minibeads, e.g. at least 10 minibeads, using a particle size analyser, for example an Eyecon™ particle characteriser of Innopharma Labs, Dublin 18, Ireland.

The minibeads of the disclosure may, therefore, have a size as disclosed above and an aspect ratio of from 1 to 1.5. The beads of the disclosure may have a size as disclosed above and an aspect ratio of no more than 1.3, for example of no more than 1.2 and, in particular, of from 1.1 to 1.5, 1.1 to 1.3 or, 1.1 to 1.2.

Bead size (diameter) may be measured by any suitable technique, for example microscopy, sieving, sedimentation, optical g zone method, ical sensing zone method or laser light ring. For the purposes of this specification, bead size is measured by analytical sieving in accordance with USP General Test <786> Method I (USP 24–NF 18, (U.S. Pharmacopeial Convention, lle, MD, 2000), pp. 1965–1967).

In embodiments, minibeads of the invention are sperse. In other embodiments, ads of the invention are not monodisperse. By "monodisperse" is meant that for a population of beads (e. g. at least 100, more preferably at least 1000) the minibeads have a cient of variation (CV) of their diameters of 35% or less, ally 25% or less, for example 15% or less, such as e.g. of % or less and optionally of 8% or less, e.g. 5% or less. A particular class of polymer beads has a P213428WO 78 CV of 25% or less. CV when referred to in this specification is defined as 100 times ard deviation) divided by average where "average" is mean particle diameter and standard deviation is rd ion in particle size. Such a ination of CV is performable using a sieve.

The invention includes minibeads having a CV of 35% and a mean diameter of 1 mm to 2 mm, e.g. 1.5 mm. The invention also includes minibeads having a CV of 20% and a mean diameter of 1 mm to 2 mm, e.g. 1.5 mm, as well as minibeads having a CV of 10% and a mean diameter of 1 mm to 2 mm, e.g. 1.5 mm. In one class of embodiments, 90% of ads have a diameter of from 0.5 mm to 2.5 mm, e.g. of from 1 mm to 2 mm.

Dosage Forms The composition of the invention may be ed as an orally administrable dosage form suitable for pharmaceutical use. In those embodiments where the ation is in the form of a minibead, the present invention provides for a dosage form comprising a plurality of the minibeads for example as a capsule, a tablet, a sprinkle or a sachet. The minibeads may also be administered rectally or vaginally administered compositon, for example as an enema or suppository. The composition, for example in the form of minibeads may be d in a suitable medium to provide a suppository or enema compositons. Suitable media for suppositories and enemas are well known and include for example, a low g point wax for a suppository or a suitable aqueous or oil based medium for an enema compositon.

The liquid composition of the invention may be formulated as an orally, rectally or vaginally administrable dosage form suitable for pharmaceutical use. The liquid composition may be formulated into a hard or soft gelatin e, a suppository or an enema. Deliverly of the liquid compositon to the stomach may also be achieved via a gastric feeding tube located in the stomach or by means of a percutaneous endoscopic gastrostomy tube (PEG tubing) as hereinabove described. The liquid composition may also be administered directly to specific points in the GI tract, for example the duodenum, jejunem or ileum via oral or intranasal tubing with an exit at the desired point in the GI tract. Delivery of the liquid composition via tubing may be under ty flow or under positive pressure using a pump or syringe drive etc.

In embodiments the dosage form comprising a population of beads may be presented in a single unit dosage form e.g. contained in a single hard gel capsule which releases the beads e.g. in the stomach. Alternatively the beads may be presented in a sachet or other container which s the beads to be sprinkled onto food or into a drink or to be administered via a feeding tube for example a naso-gastric tube or a duodenal feeding tube. Alternatively, the beads may be administered as a tablet for example if a population of beads is compressed into a single tablet as described below.

Alternatively, the beads may be filled e.g. compressed into a specialist bottle cap or otherwise fill a space in a specialised bottle cap or other element of a sealed container (or container to be sealed) such that e.g. on ng the bottle cap, the beads are released into a fluid or other contents of the bottle or vial such that the beads are disperse (or dissolve) with or t agitation in such ts.

The fluid or other ts of the bottle or vial may optionally contain one of more additional active agent(s) to tate the conventiaent co-administration of the cyclosporin composition with other P213428WO 79 active agents. An example is the Smart Delivery Cap manufactured by Humana Pharma International (HPI) S.p.A, Milan, Italy.

The dosage form may be formulated in such a way so that the beads of the ion can be further developed to create a larger mass of beads e.g. via compression (with appropriate oil or powder-based binder and/or filler known to persons skilled in the art. The larger (e.g. ssed) mass may itself take a y of shapes including pill shapes, tablet shapes, capsule shapes etc. A particular problem which this version of the bead embodiment solves is the "dead space" (above the settled particulate ts) and/or "void space" en the particulate t elements) typically found in hard gel capsules filled with powders or pellets. In such pellet- or powder-filled es with dead/void space, a patient is required to swallow a larger e than would be ary if the capsules contained no such dead space. The beads of this embodiment of the invention may readily be compressed into a capsule to adopt the inner form of whichever capsule or shell may be desired leaving much reduced, e.g. essentially no, dead/void space. Alternatively the dead or void space can be used to age by suspending beads in a e such as, for example, an oil which may be inert or may have functional properties such as, for example, permeability enhancement or ed dissolution or may comprise an active ingredient being the same or different from any active ingredients in the bead. For example, hard gelatin or HPMC capsules may be filled with a liquid medium combined with ed and/or coated beads. The liquid medium may be one or more of the surfactant phase constituents described herein or it may be one or more surfactants. Particularly preferred but non-limiting examples are corn oil, sorbitane trioleate (sold under the trade mark SPAN 85), propylene glycol dicaprylocaprate (sold under the trade mark Labrafac), 2-(2- ethoxyethoxy)ethanol (sold under the trade mark Transcutol P) and polysorbate 80 (sold under the trade mark Tween 80).

In a representative embodiment the bead of the dosage form is prepared as described herein for example by mixing together at least the following materials: a hydrogel-forming polymer; an oil phase, a surfactant being or comprising a medium chain or long chain fatty acid mono- or diglyceride or a combination f, wherein the surfactant does not se or is not a polyethyleneglycol ether or ester, and cyclosporin A, suitably cyclosporin A being dissolved in the oil phase, such as a liquid lipid to form a dispersion of the cyclosporin A in the hydrogel-forming polymer.

The dispersion is immobilized within the solidified bead by ejection from a single orifice nozzle into a suitable cooling liquid. Following removal of the cooling liquid the bead is coated with a modified release coating (the second coating) bly with a sub-coat under the modified release coating), the coated bead is then optionally filled into a gelatin or HPMC capsule suitable for pharmaceutical use.

Suitably the dosage form is prepared as a unit dosage form containing from for oral administration comprising from 0.1 mg to 1000 mg, optionally from 1mg to 500 mg, for example 10mg to 300 mg, 15 mg to 300 mg, or 25 to 250 mg, suitably about 15 mg, about 25mg, about 35 mg, about 50mg, about 75mg, about 100 mg, about 150 mg, about 180 mg, about 200 mg, about 210 mg or about 250 mg cyclosporin A Determination of Contents and Distribution of Formulations P213428WO 80 The ty and/or distribution of one or more of the components of a composition according to the invention can be determined by any method known to those skilled in the art. The distribution of one or more components of a composition can, for example, be determined by near-infrared (NIR) chemical imaging technology. NIR chemical imaging technology can be used to generate images of the surface or cross section of a composition, for example a minibead. The image produced by this technique shows the distribution of one or more components of the composition. In addition to NIR chemical imaging technology, the distribution of one or more components of a composition such as minibead, for example, be determined by time-of-flight secondary ion mass spectrometry (ToFSIMS).

ToFSIMS imaging can reveal the distribution of one or more components within the ition. The images produced by ToFSIMS analysis or NIR analysis can show the distribution of ents across a surface of the ition or a cross section of the composition. The methods described in this paragraph are applicable, for example, to composition comprising a polymer , e.g. a dried, colloid, on or dispersion.

Pharmacokinetics The orally administered compositions comprising cyclosporin A described herein may provide, amongst other features, a favourable pharmacokinetic profile compared to orally administered Neoral and/or to intravenously administered cyclosporin A as for example Sandimmun™.

The compositions according to the invention provide lower mean whole blood exposure to cyclosporin A following oral administration compared to oral administration of Neoral™ at the same dose of cyclosporin A. The whole blood exposure to cyclosporin A may be determined by measuring the area under the Curve (AUC) of the whole blood cyclosporin A concentration-time curve following administration of a single dose of a composition containing cyclosporin A. The area under the concentration-time curve (AUC), calculated from the start of dosing (t=0) to the last measured concentration (t) is designated to be "AUC0-t". Accordingly reference to "AUC0-24hr" is the AUC between t= 0 and the last measurement point at 24 hours ing administration. The AUC0-t may be calculated using well known methods for example by linear trapezoidal analysis. The area under the tration-time curve olated to infinity is "AUC0-inf". The AUC0-inf is calculated using known s as: AUC0-t + Where: Ct = the fitted last non-zero concentration for that treatment, AUC0-t is as defined above; and Kel = the ation rate constant. Kel is ated by regression is of the natural log (Ln) of whole blood concentration values- time profile.

The term "Cmax" refers to the maximum concentration of cyclosporin in whole blood following stration of a single dose of a composition containing cyclosporin A.

P213428WO 81 The term "Tmax" refers to the time taken to reach Cmax following oral administration of a composition containing cyclosporin A.

For statistical analysis, the PK data is log-transformed prior to conducting statistical testing.

In general, statistical tests are carried out using an analysis of variance procedure (ANOVA) and calculating a 90% confidence interval for each pharmacokinetic parameter (Cmax and AUC).

The measurement and analysis of AUC, Cmax and Tmax are well known in the art and can be d out using methods and techniques described in further detail in the examples or by reference to standard textbooks such as Remington, The e and Practice of Pharmacy 22nd edition, or Basic Pharmacokinetics and Pharmacodynamics: An integrated Textbook and Computer Simulations, Sara E. Rosenbaum, 2011 John Wiley& Sons. In all cases references to AUC, Cmax and Tmax are the mean values measured following administration of a composition containing cyclosporin A to a human, preferably a healthy male human in a fasted state. Suitably the subjects used in the PK study are adult male humans with ng about 70 kg (for example 70kg ± 12 kg). Suitably the subjects have a body mass index of about 25 kg/m2 (for example 25 kg/m2 ± 2.5 kg/m2).

In some embodiments the composition of the invention provides an AUC and/or a Cmax value as the AUC or Cmax "following oral administration of a single dose of 75 mg cyclosporin A". It is known that cyclosporin A exhibits an approximately linear pharmacokinetic profile EU HMA’s Public Assessment Report on Ciclosporin "Docpharma" soft capsules DK/H/968/1-3/MR, page 4.

Accordingly reference to an "AUC or Cmax of a particular value after oral administration of the ition as a single dose containing 75 mg cyclosporin A to a human in a fasted state, or an AUC or Cmax ly tional thereto for a total dose other than 75mg" is to be understood to mean that the AUC or Cmax value is ly proportional to mass of the cyclosporin A dose administered. By way of e, if a single dose of 150 mg porin A were to be administered the corresponding AUC and Cmax values will be approximately twice that obtained with a single dose of 75 mg cyclosporin A. Similarly stration of a single dose of 37.5mg of cyclosporin A would be expected to provide a AUC and Cmax values approximately half those observed following administration of 75 mg cyclosporin A. The dose proportionality for porin A is applicable over a broad range of s of cyclosporin A for example from 0.1 to 1000mg, suitably between about 1 mg and about 500 mg, more particularly n about 5 mg and about 350 mg.

In one ment the composition provides a mean whole blood AUC0-inf of from about 140 to about 420 ng.hr/ml, for example from about 140 to about 350 ng.hr/ml, about 140 to about 400ng.hr/ml about 150 to about 350 ng.hr/ml, about 150 to about 300 ng.hr/ml about 180 to about 350 ng.hr/ml, about 200 to about 400 ng.hr/ml or about 180 to about 320 ng.hr/ml and a mean whole blood Cmax of from about 25 to about 45 ng/ml after oral administration of the composition as a single dose containing 75 mg cyclosporin A to a human in a fasted state, or a AUC0-inf and Cmax directly proportional thereto for a total dose other than 75mg. ly the composition provides a Tmax of from about 4 hours to about 8 hours, suitably from about 4 hours to about 6 hours and particularly at about 5 hours; or about 5.5 hours; or about 6 hours.

P213428WO 82 Cyclosporin A concentration in Faecal Samples The cyclosporin composition may release cyclosporin A (preferably in a lised form for example as a solution in an oil droplet or as micelles containing cyclosporin A ) in the lower GI tract and ularly the colon. Accordingly, the composition provides high local cyclosporin A concentrations in the luminal contents and further results in absorption of cyclosporin A into the tissue of the GI tract. The luminal and tissue concentration of cyclosporin A ing oral administration of a composition of the invention is higher ve to that resulting from oral administration of Neoral™ or IV stration as Sandimmun™. However, as sed above, the composition according to the invention results in a relatively low systemic blood exposure to the cyclosporin A. The cyclosporin compositions described herein comprising the surfactant may also reduce the cyclosporin A metabolism following release of the cyclosporin from the composition into the GI tract.

The cyclosporin ition may provide a ratio of the mean concentration of cyclosporin A : the concentration of cyclosporin A metabolites (for example the sum of the mean AM4N and AM9 metabolite concentrations, or the sum of the mean AM1, AM9, and AM4N metabolite concentrations of greater than 12:1. Suitably the metabolite concentration is measured as the sum of the mean concentration of each metabolite present in the faecal sample. In one ment "the concentration of cyclosporin A metabolites" refers to the sum of the mean trations of the AM4N + AM9 metabolites present in the sample. In another embodiment "the tration of cyclosporin A metabolites" refers to the sum of the mean concentrations of the AM4N + AM9 + AM1 metabolites present in the sample. Accordingly In one embodiment ratio of the mean concentration of cyclosporin A : the concentration of AM1, AM9, and AM4N lites is greater than 12:1, for example, from :1 to 40:1, from 20:1 to 35:1, Suitably the ratio of cyclosporin A: metabolite concentration in the faecal sample is determined after orally administering a single dose of 75 mg cyclosporin A. However, other doses and dose regimens such as twice daily dosing may also be used. As described above the concentrations may be determined in a faecal sample collected 12 to 28 hours after dosing the composition. r, the concentrations may be determined in faeces collected at other time points following oral administration of the composition ed sufficient time has elapsed after oral administration of the composition for transit through the gut such that cyclosporin and its metabolites to be present in the collected faecal sample. It is ed that the ratios of cyclosporin to metabolites ed in a collected faecal samples will be approximately the same irrespective of the specific time point at which the faeces is collected. Accordingly, reference herein to collection of a faecal sample at 12 to 28 hours is not intended to be limiting. Suitably, the ratios of cyclosporin to metabolites are ed in s of faeces taken from subjects that have been exposed to a regular daily dose of the composition. After a ged period of daily dosing it is expected that steady-state concentrations of cyclosporin and metabolites will be achieved and as such there may be less variability in the measured concentrations of cyclosporin and metabolites in the faeces.

Accordingly, the concentration of cyclosporin: metabolites may, for example, be measured in a faecal sample collected 4 to 6 hours after oral administration of the last dose of a once daily oral dosing regimen of the composition, the dosing regimen sing once daily oral administration of the composition (for example containing 75 mg cyclosporin A) for seven days.

P213428WO 83 By way of comparison to the compositions according to the invention, the examples herein show that oral administration of Neoral™ results in a ratio of cyclosporin A : Cyclosporin metabolites (AM4N + AM9) of approximately 0.6:1, reflecting the relatively high systemic exposure and relatively low local tissue exposure in the lower GI tract, particularly in the colon. Similarly IV stration of 2 mg/Kg of cyclosporin resulted in a ratio of about 0.3:1 to about 0.45:1.

Cyclosporin A in Luminal Contents and GI Tissue ] The high tration of cyclosporin A in the l contents of the lower GI tract and the concentration of cyclosporin A in the tissue of the GI tract may be determined by measuring cyclosporin A tration in luminal content and tissue s taken at specific points along the GI tract. Cyclosporin A concentrations in olonic faeces and colonic tissue may be measured in human patients as described in the ols described in the Examples. The composition comprising cyclosporin provides high concentrations of cyclosporin A in the mucosa and sub-mucosa (i.e. the inner tissues) of for e the colon. The porin A concentration in the c tissues may be measured by taking a section of the colonic tissue, separating the layers of tissue (for example the mucosa, sub-mucosa and muscularis externa), and measuring the cyclosporin concentration in each of the respective tissue layers.

The presence of a high c luminal cyclosporin A concentration provided by the composition of the invention is expected to provide a concentration gradient which acts to promote absorption of the cyclosporin A (preferably in a solubilised form) into the lamina propria of the colon, where the main target dysregulated immune cells associated with many inflammatory es of the colon predominate. The compositions of the invention therefore provide a local topical treatment of diseased colonic tissue and are expected to be useful in the treatment of conditions such as tive colitis and other inflammatory diseases affecting the at least the colon. In contrast oral administration of Neoral™ provides relatively low luminal concentration of cyclosporin A to the inner colonic tissues. Intravenous administration of cyclosporin A as Sandimmun™ reduces the metabolism in the intestine and may provide r faecal metabolite concentrations as an orally administered composition according to the invention. However, the IV administration of porin results in significantly higher systemic exposure and moreover, relatively high doses of IV cyclosporin may be required to provide therapeutic concentrations of cyclosporin in the colonic tissue compared to oral administration of a composition according to the invention.

The composition comprising cyclosporin may therefore be ed to provide a therapeutic benefit at lower doses than Neoral and /or Sandimmun™, thus further minimising side effects ated with systemic exposure to cyclosporin A. Some release or cyclosporin A from the composition may occur as the composition passes through the GI tract and release of cyclosporin may not be exclusive to the colon. As such the ition of the invention may provide locally acting cyclosporin A in at least the colon and in other parts of the GI tract, for example the rectum and ileum, the composition may therefore provide therapeutic benefit in the treatment or prevention of conditions affecting not just the colon, but also other parts of the GI tract as described herein.

P213428WO 84 Measurement of cyclosporin concentration in the colonic tissue and intracolonic faeces in humans may be performed as bed in the Examples. Suitably samples of colonic tissue are obtained from a patient who has been orally treated with a composition containing cyclosporin by sigmoidoscopy using, for example pinch biopsy s, to obtain samples of colonic tissue. Suitably sigmoidoscopy is a flexible sigmoidoscopy. The sigmoidoscopy is preferably carried out in the unprepared bowel (except for air and water) such that the tissue samples obtained replicate as closely as possible the in-vivo tissue status, which might ise be disturbed by extensive bowel preparation. es are suitably about 5mm in size and ideally at least 5 biopsies are taken approximately 1 cm apart from the subject. Preferably the biopsies are ed as close to the c flexure as possible. Alternatively, biopsies may also be obtained from within the sigmoid colon. Each biopsy should be rinsed with saline, blot dried and then stored at low temperature, suitably at about -70oC, prior to analysis. The tissue samples may be analysed directly for the concentration of cyclosporin A present in the . However, preferably the mucous layer t on the tissue surface is first removed from the sample such that the cyclosporin tration measured is the concentration of cyclosporin t in the epithelial and musosal tissue. The mucous layer may be removed by washing with a le solvent such as N-acetyl cysteine in water. Removal of the mucose layer ensures that the concentration of cyclosporin ed is representative of the concentration which has been absorbed into the tissue, rather than the mucosal layer. The cyclosporin t in the mucosal layer can be determined by ing the l washings.

Suitable methods for the preparation and analysis of the colonic tissue are set out in the examples section.

Samples of olonic faeces are suitably collected from approximately the same location within the colon as the tissue biopsies such that the measurement of cyclosporin concentration in the tissue and intra-colonic faeces represents the concentrations present at imately the same position within the colon.

The tissue biopsies and intra-colonic faecal samples should be obtained after a sufficient duration of cyclosporin dosing to reach steady-state concentrations in the colon. For example, the biopsies and faecal samples are suitably may be carried out after 7 days of daily oral dosing with the composition. The biopsies and intracolonic faecal samples are suitably obtained simultaneously within 4 to 6 hours after the last dose in the 7 day dosage regimen.

The ratio of the mean concentration of cyclosporin A present in intracolonic faeces : the mean concentration of cyclosporin A present in colonic tissue in an adult human patient after oral administration of the composition is greater than 30:1, for example, greater than about 40:1 or greater than about 50:1. The mean concentration of cyclosporin A present in intracolonic faeces : the mean concentration of cyclosporin A present in colonic tissue may be about 30:1 to about 500:1, about 50:1 to about 500:1, optionally from about 80:1 to about 300:1, or optionally about 100:1 to about 250:1. In contrast the Examples show that IV administration of Sandimmun results in an olonic faecal : tissue ratio of cyclosporin A of about 3:1. 8WO 85 References to a "mean" value in relation ot the PK, tissue and faecal analysis described herein is unless ied otherwise a reference to the arithmetic mean value of the measured values. ution Profile The compositions comprising cyclosporin provide compositions with a specific in-vitro dissolution profile for the release cyclosporin A from the composition. The compositions show minimal release of porin A in the stomach and upper GI tract such as the um and jejunum and higher release in at least the colon. The in-vivo release may be modelled using a two stage ro dissolution test in which a composition is exposed to 0.1 N HCl for two hours to simulate pH of the gastric environment and is then exposed to pH 6.8 for twenty two hours (by adding a sufficient quantity of 0.2M tribasic sodium ate solution containing 2% sodium dodecyl sulfate (SDS)) to simulate pH in the small intestine and lower GI tract.

Reference to "a two stage dissolution test using a USP Apparatus II with a paddle speed of 75 rpm and a dissolution medium temperature of 37oC; wherein for the first 2 hours of the dissolution test the dissolution medium is 750 ml of 0.1 N HCl, and at 2 hours 250 ml of 0.2M tribasic sodium phosphate containing 2% SDS is added to the dissolution medium and the pH is adjusted to pH 6.8" is an in- vitro test carried out in accordance with the USP <711> ution test using Apparatus II (paddle apparatus) operated with a paddle speed of 75 rpm and with the dissolution medium at a temperature of 37oC ± 0.5oC. At the start of the test (t=0) the sample is placed in the acidic dissolution medium. After 2 hours an aliquot of the medium is taken for subsequent analysis and immediately (suitably within 5 minutes) the second stage of the dissolution test is initiated. In the second stage of the ution test 250 ml of 0.2M tribasic sodium phosphate containing 2% sodium dodecyl sulfate (SDS) is added to the dissolution medium and the pH is adjusted to 6.8 ± 0.05 using 2N NaOH or 2N HCl as required. Samples of the dissolution medium are taken at time points during the second stage of the test, for example at 4, 6, 12 and 24 hours from the start of the test (i.e. from t=0 at the start of the first stage). The s are analysed for cyclosporin A dissolved in the medium. The "% released" is the amount of cyclosporin A in solution in the tive dissolution medium at a particular time point relative to the amount of cyclosporin in the composition at the start of the test. The cyclosporin A concentrations in a sample may be measured using standard techniques, such as e Phase HPLC as illustrated in the Examples. References to "two stage dissolution test" herein also refer to this test method.

The in-vitro dissolution e of the composition ing to the invention is described above under the Brief Summary.

Manufacturing Processes Various methods may be used to prepare the formulations of the invention.

In those embodiments where the formulation comprises an active ingredient in a waterinsoluble polymer matrix, a basic method for making the core is to mix a fluid form of the matrix material, for example a hydrogel forming polymer matrix al (e.g. poly(amides), poly(aminoacids ), hyaluronic acid; lipoproteins; poly(esters), poly(orthoesters), poly(urethanes) or P213428WO 86 poly(acrylamides), poly(glycolic acid), poly(lactic acid) and corresponding co-polymers (poly(lactideco-glycolide acid; PLGA); siloxane, polysiloxane; dimethylsiloxane/methylvinylsiloxane copolymer; poly(dimethylsiloxane/methylvinylsiloxane/methylhydrogensiloxane) dimethylvinyl or trimethyl copolymer; silicone polymers; alkyl silicone; silica, ium silicate, calcium te, aluminium ium silicate, magnesium silicate, aceous silica etc as described more generally elsewhere ), with an active ingredient to form a mixture that may take the form of a suspension, solution or a colloid. The mixture is processed to form a composition or a core. For example the composition may be shaped into the desired form using a molding or hot-melt extrusion process to form beads.

Methods for preparing a composition and a core comprising the surfactant, cyclosporin, an oil phase and a water-soluble polymer matrix are described below. Generally these cores are coated.

Generally, the manufacturing processes described herein se mixing of liquid(s). Such mixing processes must be performed at temperatures at which the substances to be mixed in the liquid state are in liquid form. For e, thermoreversible g agents must be mixed at a temperature where they are in the liquid state, for e at a temperature of 50 to 75oC, for example 50 to 70oC, or 55-75°C, e.g. 60-70°C and in particular embodiments about 55°C or 65°C in the case of mixing formulations comprising aqueous gelatin. Similarly other components of the formulation may need to be heated to melt the component for example waxes or surfactants which may be used in the disperse phase.

The liquid composition, composition or the core comprising a surfactant, oil phase, hydrogelforming r and cyclosporin as disclosed herein may be made by mixing materials comprising for example water, a hydrogel-forming polymer and a second surfactant to form an aqueous continuous phase, and mixing a disperse phase. At least one of the aqueous phase and the disperse phase comprises cyclosporin, the cyclosporin may be dissolved in the phase which contains it, for example both phases may be a clear liquid before they are mixed together. Preferably, the disperse phase (the oil phase) may comprise the porin, (for example a disperse phase comprising an oil, an optional solvent, cyclosporin and a first surfactant) with the aqueous phase to form a colloid. The colloid may have the form of an on or microemulsion wherein the disperse phase is dispersed in the aqueous continuous phase. This colloid may optionally represent the liquid composition of the invention. In order to prepare the composition of the invention or the core, the hydrogel-forming polymer is then caused or allowed to gel to form a hydrogel forming polymer matrix. Suitably, the process includes formulating or sing the composition into a desired form, e.g. a bead (also termed a minibead), which forming process may comprise moulding but ably comprises ejecting the s colloid h a single orifice nozzle to form droplets which are caused or allowed to pass into a cooling medium, e.g. a water-immiscible cooling liquid, in which the ts cool to form for e.g. beads.

The mixing of the materials may comprise mixing an aqueous premix (or aqueous phase or continuous phase) and a disperse phase premix (e.g. oil phase premix), wherein the aqueous premix ses water and water-soluble substances whilst the disperse phase premix may comprise a P213428WO 87 vehicle containing cyclosporin and the surfactant. The vehicle may be a hydrophobic , for example a liquid lipid, or it may be or comprise a material, for example a surfactant, for forming selfassembly structures. In particular, a disperse phase premix may comprise cyclosporin A, the first tant, an oil and other oil soluble components for example an al solvent. The premixes may contain one or more surfactants suitable for the phase they are to form, as previously mentioned, for e the aqueous premix may comprise a second surfactant.

The aqueous premix comprises, or usually consists of, a solution in water of water-soluble constituents, namely the el-forming polymer and water-soluble ent(s). The aqueous premix may include a plasticiser for the hydrogel-forming r, as bed elsewhere in this specification. The aqueous premix may include a second surfactant, e.g. to increase r viscosity and improve fication and thereby help prevent precipitation of active agent during processing. SDS is an example of such a surfactant. In any event, the constituents of the aqueous premix may be agitated for a period sufficient to dissolve/melt the components, for example, from 1 hour to 12 hours to form the completed aqueous .

The se phase x may comprise the first surfactant and cyclosporin as a dispersion or preferably a solution in a vehicle (for example an oil phase) as described above, for example in a liquid comprising an oil or in a liquid comprising component(s) of self-assembly structures. For example an oil phase pre -mix may therefore be a liquid lipid, for example a medium chain triglyceride (MCT) formulation, the medium chain triglyceride(s) being one or more triglycerides of at least one fatty acid selected from C6-C12 fatty acids, and cyclosporin A and the surfactant comprising or being a medium or long chain fatty acid mono- or di-glyceride. Suitably an oil phase pre-mix is stirred at ambient temperature to form a solution of the cyclosporin in the oil and surfactant.

In some embodiments, the components of the oil phase premix are mixed (or otherwise agitated) for a period of, for example, 10 minutes to 3 hours to form the premix.

The two premixes may be combined and agitated, for example for a period of a few seconds to an hour, for example from 30 seconds to 1 hour, suitably 5 mins to an hour, to form a dispersion of the disperse phase in an aqueous hydrogel-forming polymer to form the liquid ition of the invention. The dispersion may then be further processed to form the composition or a core. The two premixes may be combined into the dispersion by agitation in a mixing ; they may additionally or alternatively be combined in a continuous flow mixer.

The basic method for making a composition or core comprising cyclosporin and hydrogelforming r matrix, therefore, is to mix a liquid form (preferably a solution) of the hydrogel-forming polymer (or mixture of polymers) with the cyclosporin, the surfactant (to avoid any ambiguity the first surfactant) and the oil phase (and any other disperse phase components) to form a dispersion in the polymer, which later in the process forms a hydrogel. The method ly comprises mixing together an aqueous polymer phase premix and a disperse phase premix. Taking t of the final composition required (as described elsewhere ), the disperse phase x and the liquid hydrogel-forming polymer (i.e. the solution or suspension of hydrogel-forming polymer, the aqueous phase) may be mixed in a weight ratio of from 1:1 to 1:10, particularly 1:4 to 1:9, e.g. 1:5 to 1:7. In P213428WO 88 general, only gentle stirring of the components is ed using a magnetic or mechanical system, e.g. overhead stirrer, as would be familiar to a person skilled in the art to achieve a dispersion of the disperse phase in the aqueous phase to form a d (which may be in the form of for example an emulsion or micro emulsion in which the aqueous hydrogel is the continuous phase). Continuous stirring is preferred. Mixing may also be achieved using an in-line mixing system. Any appropriate laboratory stirring apparatus or industrial scale mixer may be utilized for this purpose for example the ic Stirrer actured by Stuart) or ad Stirrer (by KNF or Fisher). It is preferred to set up the equipment in such a way as to minimise evaporation of contents such as, for example, water. In one embodiment of the process of the invention, it is preferred to utilise a closed system for stirring in order to achieve this aim. In-line mixing may be particularly suitable for closed system processing.

Suitably mixing of the two components takes place at a temperature of 50 to 70oC, or 55-75°C, e.g.

The mixing of the two phases results in a colloid wherein the aqueous hydrogel-forming polymer is an aqueous continuous phase and the component(s) not soluble in the aqueous phase are a disperse phase. The colloid may have the form of an emulsion or microemulsion.

] In ments where the disperse phase is or comprises a second surfactant, the amount of the second surfactant may be selected such that, upon combination of the disperse phase premix with the aqueous pre-mix, the second surfactant concentration in the ed mixture s the CMC for the second surfactant used such that micelles are formed in the aqueous phase comprising the hydrogel-forming polymer. Depending on the concentration of second surfactant used, self - assembly structures other than micelles may also form. The CMC for a particular surfactant may be determined using well known methods, for e as described in Surfactants and Polymers in Aqueous Solutions Second Edition, Chapter 2, Holmberg et al. In embodiments mixing of the aqueous phase and a disperse phase which is or comprises a surfactant may result in the ion of a clear liquid, for example a microemulsion, in which the aqueous phase comprising the hydrogelforming polymer is the continuous phase. Microemulsions are a thermodynamically stable dispersion of ssembly structures in the aqueous phase, the size of the self-assembly structures being sufficiently small to give a transparent appearance. The size of the self-assembly structures present as the disperse phase resulting from the mixing of the aqueous and surfactant phases may be from about 0.5 nm to 200 nm, for example about 1 nm to 50 nm, or about 5 nm to 25 nm. The size of the self-assembly structures formed and other characteristics such as the optical isotropicity of the formulation (for example a microemulsion) may be determined using well known techniques such as dynamic light scattering.

Where the polymer matrix substantially consists of gelatin with the addition of sorbitol, the s phase of r matrix is prepared by adding the appropriate quantities of sorbitol (and surfactant if desired) to water, g to imately 50 to 75°C, for example 60-75°C until in solution and then adding gelatin, although the precise order and timing of addition is not critical. A typical "gelatin solution" comprises 8 to 35%, (for example 15-25%, preferably 17-18%) gelatin; 65%- 85% (preferably 77-82%) of water plus from 1-5% rably 1.5 to 3%) sorbitol. When present, a P213428WO 89 second surfactant (e.g. anionic surfactant) in the aqueous phase premix may be present in an amount of 0.1 to 5% (preferably 0.5 to 4%) n all parts are by weight of the aqueous phase.

Optionally the processing temperature required for a standard n can be reduced to a desirable target temperature e.g. 37°C by use of lower melting-point gelatin (or gelatin derivatives or mixtures of gelatins with melting point reducers) or other polymer matrix material such as, for e, sodium alginate. If n droplets are being formed by machine extrusion and immediately cooled, e.g. in a cooling bath, additional appropriate inlet tubing can be used to introduce an oil phase containing cyclosporin A at ambient temperature into the hotter fluid gelatin solution (and the mixture can be immediately homogenized) very shortly before on from a beading nozzle or other dropletting process such that the duration of exposure of the cyclosporin A to the higher temperature gelatin is limited so reducing the degree of any heat-dependent degradation of the active ingredient.

This process may use any appropriate device such as, for example, a nizer, e.g. a screw homogenizer, in conjunction with an extrusion-type tus as described for example in WO 2008/132707 (Sigmoid Pharma) the entirety of which is incorporated herein by reference.

Alternatively, the aqueous- and oil-based solutions can be pumped at appropriate rates and passed through a static mixer to form an emulsion prior to dropping.

The colloid is formed by combining of the disperse phase premix with the liquid s phase with stirring as described above. The resultant colloidal dispersion then has the formulation of a solidified core described above but with liquid water still present in the core formulation.

Optionally the cyclosporin may be added after mixing the aqueous phase and other components of a disperse phase of the type sing a liquid lipid in addition to the cyclosporin, however, it is preferred that the cyclosporin is added together with the other components of the disperse phase as a premix.

The resulting colloid is then poured or introduced into a mould or other vessel or poured onto sheets or between sheets or delivered dropwise (or extruded) into r fluid such that the polymer matrix-containing aqueous phase, on solidification, takes the form of the mould, vessel, sheet or droplet/bead intended. It is preferred to progress to mould-forming e.g. beading without delay.

] Solidification (gelling) can occur in a variety of ways ing on the polymer of the matrix, for example by changing the temperature around the mould, vessel, sheet, droplet/bead etc or by applying a solidification fluid or hardening solution so that the moulded shape is gelled or solidified. In certain ments both temperature change and application of a solidifying fluid or hardening solution are employed together or aneously. For example, when using a dropping method solidification can occur by ng into cooling oil, air or a combination thereof.

In the preferred embodiment in which the composition or core takes the form of beads, the beads may be formed for example by dropping the colloid se into a fluid which effects solidification. Where the viscosity of the composition to be beaded reaches a certain point, drop ion becomes more difficult and specialised apparatus is then preferred.

P213428WO 90 By use of the term "dry", it is not sought to imply that a drying step is necessary to produce the dry core (although this is not ed) rather that the solid or solidified aqueous external phase is substantially free of water or free of available water. Solidification of the aqueous phase (external phase) may have arisen through s means including ally (e.g. by cross-linking) or physically (e.g. by cooling or heating). In this respect, the term "aqueous phase" is nevertheless employed in this document to denote the external (continuous) phase of the core even though water, in certain embodiments, is largely absent from (or trapped within the cross-linked matrix of) the core.

The external phase of the core is however water-soluble and dissolves in s media.

In the case where solidification can be achieved by raising or ng temperature, the temperature of the solidification fluid can be d to achieve fication of the core at a desired rate. For example, when gelatin is used as the hydrogel-forming polymer, the solidification fluid is at a lower temperature than the temperature of the emulsion thus causing solidification, i.e. gelling, of the polymer matrix. In this case, the solidification fluid is termed a cooling fluid.

In the case where solidification can be achieved chemically, e.g. by induction of cross-linking on exposure to a component of the solidification fluid, the concentration of such component in the solidification fluid and/or its temperature (or other characteristic or content) can be adjusted to achieve the desired rate and degree of solidification. For example, if alginate is chosen as the polymer matrix, one component of the solidification fluid may be a calcium-containing entity (such as, for example, calcium chloride) able to induce cross-linking of the alginate and consequent solidification.

Alternatively, the same or similar calcium-containing entity may be included (e.g. disperse) in the aqueous phase of the fluid emulsion prior to beading and red to induce cross-linking e.g. by applying a higher or lower pH to a solidification fluid into which droplets of emulsion fall dropwise or are introduced. Such electrostatic cross-linking can be varied as to the resulting characteristics of the bead by l of calcium ion availability ntration) and other physical conditions (notably temperature).The solidification fluid may be a gas (for example air) or a liquid or both. For example, when gelatin is used as the hydrogel-forming r matrix, the solidification fluid can be initially gaseous (e.g. droplets passing h g air) and then uently liquid (e.g. droplets passing into a cooling liquid). The reverse sequence may also be applied while gaseous or liquid cooling fluids alone may also be used. Alternatively, the fluid may be spray-cooled in which the emulsion is sprayed into a cooling gas to effect solidification.

In the case of gelatin or other water-soluble polymer (or polymer mixture) destined to form an immobilization matrix, it is preferred that the solidification fluid be a non-aqueous liquid (such as, for example, medium chain triglycerides, mineral oil or similar preferably with low HLB to ensure minimal wetting) which can conveniently be placed in a bath ng bath) to receive the droplets of the colloid as they solidify to form the beads of the core. Use of a ueous liquid allows greater flexibility in choice of the temperature at which cooling is conducted.

Where a liquid cooling bath is employed, it is lly maintained at less than 20°C, ably ined in the range 5-15°C, more preferably 8-12°C when standard gelatin is used as P213428WO 91 the hydrogel-forming polymer. If a triglyceride is chosen as the cooling fluid in the cooling bath, a preferred example is Miglyol 840 from Sasol.

If te is selected as the polymer matrix, a typical method of making beads involves se on of a 3% sodium alginate solution in which oil ts are se as described above into a 4°C crosslinking bath containing 0.1 M calcium chloride to produce calcium te (this method can be ed to as "diffusion setting" because the calcium is believed to diffuse into the beads to effect cross-linking or setting). Using a syringe pump, or h machine, droplets can be generated or extruded (egg at 5 mL/h if a pump is used) through a sterile needle or other nozzle (described elsewhere herein) which can be vibrating as discussed elsewhere herein. Airflow of between 15 and 20 L/min through 4.5 mm tubing can be applied rds over the needle to reduce droplet size if desired. Newly formed beads can then be stirred in the calcium chloride bath for up to an hour. If carrageenan is used as the polymer matrix both salt and reduction in temperature e.g. by dropping into cooling oil may be used to obtain solidification.

] An alternative approach when using alginate is internal gelation in which the calcium ions are disperse in the aqueous phase prior to their activation in order to cause gelation of hydrocolloid particles. For example, this can be achieved by the addition of an inactive form of the ion that will cause crosslinking of the alginate, which is then activated by a change in e.g. pH after ient dispersion of the ion is te (see Glicksman, 1983a; Hoefler, 2004 which are both incorporated herein by reference). This approach is particularly useful where rapid gelation is desired and/or where the diffusion approach may lead to loss of API by diffusion thereof into the crosslinking bath.

Where another ionotropic polymer is used than alginate, suitable analogous processes may be used to those described herein in relation to alginate.

Following shape-forming, moulding or beading, the resultant shapes or forms may be washed then dried if appropriate. In the case of beads solidified in a solidification fluid, an optional final step in the method of production described above therefore comprises removal of the solidified beads from the solidification fluid. This may be achieved e.g. by collection in a mesh basket through which the solidification fluid (e.g. medium chain triglycerides) is drained and the beads retained and is preferably ted without delay e.g. as soon as the beads have formed or within 5, 10, 15, 20, 25 or 30 minutes of their formation. Excess solidification fluid may then be removed using a centrifuge (or other apparatus or e adapted to remove excess fluid) followed by drying of the beads to remove water or free water and/or removal of some or all of any onal solvent e.g. ethanol or isopropyl alcohol used to dissolve or facilitate dissolution of the active principle in preceding steps optionally followed by g (e.g. using ethyl acetate) and a subsequent g" step to remove excess solvent (e.g. ethyl e). Isopropyl alcohol is an example of a solvent which is preferably removed later in processing to reduce residues in the oil or aqueous phase. Drying can be achieved by any suitable process known in the art such as use of a drum drier (e.g. Freund Drum dryer which may be part of the Spherex ent train if used) with warm air at between 15°C and 25°C, preferably around 20°C leading to evaporation or entrainment of the water by the air. Alternatively, drying may be carried out using of a fluid bed drier (e.g. Glatt GPCG 1.1) with warm air between 40°C P213428WO 92 and 60°C; or using a vibrational fluid bed drier. Use of gelatin as the r matrix (e.g. as principal constituent of the s immobilisation phase) in most cases requires a drying step and for beads this is preferably achieved by drying in air as above described. Beads can be dried using other techniques, as understood by the person skilled in the art, for example vibrating fluid bed, tray drying or vacuum drying. The resultant formulation (the ation of the invention) is essentially dry as described in more detail above.

] In general, the beads may be generated by the application of surface tension between the liquid dispersion (the mixture of the aqueous and surfactant phases) and an appropriate solidification fluid such as, for example, gas or liquid in order to create the spherical or substantially spherical shape of the ultimate beads.

Alternatively, the beads may be produced through ejection or ion of the liquid dispersion (the aqueous phase pre-mix and the disperse phase premix mixture) through an orifice or nozzle with a certain diameter and optionally subject to vibration (using selected vibrational frequencies) and/or gravitational flow. es of machines which may be used are encapsulation prilling, drop pelletising, spray cooling or spray congealing machines for example the Freund Spherex, ITAS/Lambo, , Inotech, GEA Niro, , Buchi, Gelpell, Brace processing equipment processing equipment. Operation of the Spherex machine manufactured by Freund as may be desired to manufacture beads according to the present invention is described in US patent 5,882,680 (Freund), the entire contents of which are orated herein by reference. It is preferred to select a vibrational frequency in the region of 2-200 Hz, suitably 10-50 Hz, although the ultimate choice (and separately the amplitude of vibration selected) depends on the viscosity of the dispersion to be beaded. If the polymer matrix is chosen to solidify at lower temperature, it may be appropriate to maintain the lines to the orifice/nozzle at a n temperature to maintain the fluidity of the solution.

Suitably the colloid is ejected through a single-orifice nozzle, e.g. having a diameter of from 0.1 mm to mm (for example 0.5-5 mm), to form drops which are then caused or allowed to fall into a g oil or other hardening medium and allowed to harden to form seeds, after which the seeds are recovered from the cooling oil and dried.

It will be iated, therefore, that the invention includes a process for manufacturing a composition of the invention or a core comprising cyclosporin, a first surfactant being or comprising a medium chain or long chain fatty acid mono- or di-glyceride or a combination f, wherein the surfactant does not comprise or is not a polyethyleneglycol ether or ester, and an oil phase in a hydrogel forming polymer matrix, which s comprises: g an aqueous premix which comprises water and water soluble/dispersible materials (including ore a el-forming polymer) and a disperse phase premix (e.g. an oil phase premix) which ses the oil phase, the cyclosporin and the first surfactant optionally other excipients (e.g. oil(s) and oil soluble/dispersible materials), and combining the two premixes to form a colloid (disperse phase) within an aqueous phase comprising the hydrogel-forming polymer. The colloid may then be formed into a shaped unit, for example a bead to provide the core comprising the active ingredient. More particularly the manufacture of a composition or core as defined above may comprise: (i) g an aqueous phase pre-mix comprising a solution in water of water-soluble P213428WO 93 constituents (e.g. of a hydrogel-forming polymer, any water-soluble excipient(s), as described elsewhere herein); (ii) forming a disperse phase pre-mix typically comprising a dispersion or preferably a solution of pori, in a liquid lipid, and the first surfactant, optionally together with other disperse phase constituents (e.g. surfactant, solvents etc as described elsewhere herein); (iii) mixing the aqueous phase pre-mix (i) and the disperse phase x (ii) to form a colloid; (iv) ejecting the colloid h a nozzle to form droplets; (v) causing or allowing the a el-forming polymer to gel or solidify to form a water soluble polymer matrix; and (vi) drying the solid.

The manufacture of a liquid composition of the invention may comprise: (i) forming an aqueous phase pre-mix comprising a solution in water of water-soluble constituents (e.g. of a hydrogel-forming polymer, any water-soluble excipient(s), as described elsewhere ); (ii) forming a disperse phase pre-mix typically comprising a sion or preferably a solution of cyclospori, in a liquid lipid, and the first surfactant, optionally together with other se phase constituents (e.g. surfactant, solvents etc as described elsewhere herein); and (iii) mixing the aqueous phase pre-mix (i) and the disperse phase x (ii) to form a colloid.

Some manufacturing processes comprise steps (A) to (D) below or, alternatively, a manufacturing process may comprise a single one or any combination of steps (A) to (D).

(A) Exemplary Preparation of Aqueous Phase: Aqueous phase ents are added to water, e.g. ed water, under agitation e.g. sonication or stirring. The temperature is gradually increased, for example to 60-70° C and in particular 65°C, to achieve complete dissolution of the . The aqueous phase ents include a hydrogelforming polymer, e.g. gelatin or agar and optionally one or more other excipients, for example D- sorbitol (a plasticiser) and surfactant (for example SDS). Possible aqueous phase components are described elsewhere .

] The gelatin may be Type A n. In some less preferred implementations, the gelatin is Type B. The gelatin may have a Bloom strength of 125-300, optionally of 200-300, for example of 225- 300, and in particular 275. The components of the aqueous phase may be agitated for a period of, for e, from 1 hour to 12 hours to complete preparation of the aqueous phase (aqueous premix).

(B) Exemplary Preparation of Disperse Phase: Cyclosporin is mixed with the first surfactant, an oil and other disperse phase components (for example a co-solvent) under agitation e.g. sonication or stirring, suitably at t temperature to disperse or preferably dissolve the active ingredient.

(C) Exemplary Mixing of the two phases P213428WO 94 The aqueous phase and the disperse phase are mixed. The two phases may be mixed in a desired weight; for example, the weight ratio of disperse phase to aqueous phase may be from 1:1 to 1:10, e.g. from 1:4 to 1:9 and ally from 1:5 to 1:8 such as about 1:5 or about 1:7. The resulting colloid is agitated, e.g. sonicated or stirred, at a temperature of 60-70°C and in particular 65°C, to achieve a homogeneous dispersion, then the nous dispersion is formed into beads. In particular, the homogenous sion is d through a single orifice nozzle to form droplets which fall into a cooling medium. The nozzle is suitably vibrated to facilitate droplet formation. The nozzle may be vibrated at a frequency of 2-200 Hz and optionally 15-50 Hz.

The cooling medium may for example be air or an oil; the oil is suitably physiologically acceptable as, for example, in the case of medium chain triglycerides e.g. Miglyol 810N. The cooling medium may be at a cooling temperature often of less than 15°C, for example of less than 10°C but above 0°C. In some ments the cooling temperature is 8-10°C. The nozzle size (diameter) is lly from 0.5 to 7.5mm, e.g. from 0.5 to 5mm and optionally from 0.5 to 4mm. In some embodiments, the nozzle diameter is from 1 to 5mm for example from 2 to 5mm, and optionally from 3 to 4mm, and in particular may be 3.4mm. The nozzle er may be from 1 to 2mm.

The flow rate through a 3.4mm nozzle or through a 1.5mm nozzle is 5 to 35 g/min and optionally 10 to 20 g/min and for nozzles of different sizes may be adjusted suitably for the nozzle area.

(D) Exemplary Processing of beads Cooled beads are recovered, for example they may be recovered from g oil after a residence time of 15-60 minutes, for example after approximately 30 minutes. Beads red from a cooling liquid (e.g. oil) may be centrifuged to eliminate excess cooling liquid, and then dried. Suitably, drying is carried out at room temperature, for example from 15-40°C and optionally from 20-35°C. The drying may be performed in a drum drier, for example for a period from 6 to 24 hours, e.g. of about 12 hours in the case of beads dried at room temperature. The dried beads may be washed, suitably with a volatile non-aqueous liquid at least lly miscible with water, e.g. they may be washed with ethyl acetate. The washed beads may be dried at room temperature, for example from C and optionally from 20-25°C. The drying may be performed in a drum drier, for example for a period from 6 to 48 hours, e.g. of about 24 hours in the case of beads dried at room temperature. Drying may be achieved by any suitable means, for example using a drum dryer, suitably under vacuum; or by simply passing warm air h the batch of beads, or by fluidising the beads in a suitable equipment with warm air, for example if a fluid bed dryer. Following drying, the beads are passed through a 1 to 10 mm, optionally 2 to 5 mm to remove oversized beads and then through a sieve with a pore size of 0.5 to 9 mm optionally 1 to 4 mm to remove undersized beads.

It can be iated that it is possible to recycle the beads that are rejected by the sieving process.

As a further aspect of the invention there is provided a formulation obtainable by g the characteristic of) any of the processes described herein. It is to be understood that the processes P213428WO 95 described herein may therefore be used to provide any of the specific cores described in embodiments herein by dispersing the riate components which form the se phase of the core in the appropriate components which form the aqueous continuous matrix phase of the core.

The preceding paragraphs describe the formation of uncoated compositions or cores. The ition may se a coating. Cores may be coated. The composition or the core may be coated with a subcoat and/or coated with a second coating (also referred to as a modified release coating or outer coat). Suitable sub coats and modified release coatings (second coating or outer coat) are any of those described herein and any of the first coating (for the subcoat) or the second coating (for the modified release coating). The coating(s) may be applied using well known methods, for example spray coating as described below to give the desired sub coat and modified e coating weight gains.

With regard to one of the methods described above (ejection of emulsion through an optionally vibrating nozzle) with two tric orifices (centre and outer), the outer fluid may form a coating (outside the bead) as described herein. The Spherex machine manufactured by Freund (see US patent 5,882,680 to Freund) is preferably used (the entire contents of this patent is incorporated herein by reference). Other similar ejection or ion apparatus may also be used, for example the ejection apparatus described hereinbefore.

] Use of the Spherex machine achieves very high monodispersity. For example, in a typical 100g, batch 97g of beads were between 1.4 to 2 mm er or between 1 and 2 mm. d size ranges can be achieved by methods known in the art for rejecting/screening different sized particles.

For example, it is possible to /screen out the larger/smaller beads by passing a batch first through e.g. a 2 mm mesh and subsequently through a 1.4 mm mesh.

The 1.4 to 2mm diameter range is a good size if it is desired to spray coat the beads (if smaller, the spray of the coating machine may bypass the bead; if too large, the beads may be harder to fluidise, which is necessary to achieve consistent coating).

Coating Process The coating process can be d out by any suitable means such as, for e, by use of a coating machine which applies a solution of a polymer coat (as bed above in particular) to the formulation. Polymers for coating are either provided by the manufacturer in ready-made solutions for direct use or can be made up before use following manufacturers’ instructions.

Coating is ly carried out using a fluid bed coating system such as a Wurster column to apply the coating(s) to the composition or the core. Appropriate coating machines are known to persons skilled in the art and include, for example, a perforated pan or fluidized-based system for e the GLATT, Vector (e.g. CF 360 EX), ACCELACOTA, Diosna, O’Hara and/or HICOATER processing equipment. To be mentioned is the MFL/01 Fluid Bed Coater (Freund) used in the "Bottom Spray" uration.

Typical coating conditions are as follows: P213428WO 96 Process Parameter Values sing airflow (m3/h) 20-60 (preferably 30-60) Inlet air temperature (°C) 20 – 65 Exhaust air temperature (°C) 20 – 42 Product ature (°C) 20 – 45 (preferably 40 to 42) Atomizing air pressure (bar) Up to 1.4 e.g. 0.8-1.2 Spray rate (g/min) 2-10 and 3-25 RPM Suitably the coating is applied as a solution or dispersion of the polymers (and other components) of the coating. Generally the gs are applied as an aqueous, solution or dispersion, although other solvent systems may be used if required. The coating dispersion is applied to the composition or the core as a spray in the fluid bed coater to give the required coating weight gain.

Generally the g process is carried out at a temperature which maintains the cores at a ature of from 35 to 45oC, preferably 40 to 42oC.

After applying the coating, the ition may be dried, for example by drying at 40 to 45oC.

The ion further provides a product having the characteristics of a composition obtained as described herein, a product defined in terms of its teristics being defined by the characteristics of the composition to the exclusion of the method by which it was made.

As ned herein the processes described may be used to provide any of the composition described in the various ments herein. By way of example there is provided a composition of the invention comprising a core and a first g sing a water-soluble cellulose ether or a water soluble derivative of a cellulose ether and/or a second coating comprising a delayed release polymer wherein the core comprises a hydrogel-forming polymer matrix comprising gelatin, cyclosporin A, medium chain mono-di- and/or tri-glycerides, a first surfactant being or comprising a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof that does not comprise or is not a polyethyleneglycol ether or ester, a co-solvent and optionally a second surfactant, the core having the characteristics of a core obtained by the process sing steps (i) to (vi) bed above for forming the core, wherein the aqueous phase pre-mix in step (i) of the process comprises gelatin and optionally a second surfactant (suitably an anionic surfactant), and the oil phase pre-mix in step (ii) of the process comprises medium chain mono-di- or tri-glycerides, hydrophobic active ingredient, surfactant (suitably a non-ionic surfactant) and cosolvent; and the wherein the core is optionally coated with a first coating comprising a soluble cellulose ether or a water soluble derivative of a cellulose ether and/or a second coating comprising a delayed release polymer; wherein the coatings are any of those described herein. Accordingly, the process may produce a composition as described above comprising a first coating and/or a second coating. The process may additionally produce a ition comprising a first coating and a second coating being e the first coating.

In the cores described herein to which the following characteristics are applicable, e.g. in the immediately preceding paragraph, the following characteristics may be present: P213428WO 97 gelatin may be present in an amount of 300 to 700 mg/g; the medium chain mono-, di- or tri-glycerides (for example caprylic/capric triglyceride) may be present in an amount of 20 to 200 mg/g; co-solvent (for example 2-(ethoxyethoxy)ethanol) may be t in an amount of 150 to 250 mg/g; non-ionic surfactant (for example sorbitan-based surfactants, PEG-fatty acids, or glyceryl fatty acids or poloxamers or particularly a polyethoxylated castor oil for example Kolliphor EL) may be present in an amount of 80 to 200 mg/g; c surfactant (for example, alkyl sulphates, ylates or phospholipids cularly SDS)) may be present in an amount of 15 to 50 mg/g; and cyclosporin A, may be present in an amount of from 60 to 180 mg/g, suitably 60 to 150 mg/g, 90 to 150mg/g, or 80 to 100mg/g, for example 81 to 98 mg/g; wherein all weights are based upon the dry weight of the core before coating.

The composition may comprise or the core may be coated with a first coating oating) which is or comprises a water-soluble compound selected from ose ethers and their derivatives, particularly hydroxypropylmethyl cellulose; the first coating being present in an amount corresponding to a weight gain due to the first coating in a range selected from: (i) from 8% to 12%, for example about 10%; or (ii) from 4% to 6%, for example about 5% by weight based upon the weight of the core prior to applying the first coating. The first g may have a modified release coating (or second coating) applied to it.

Preferably, any modified release coating (second coating), ally in the embodiments of the immediately ing paragraphs, is or ses a pH independent modified release coating, more especially the second coating may be a modified release coating comprising ethyl cellulose (eg Surelease) still more particularly a modified release coating comprising ethyl cellulose and a watersoluble polysaccharide, pectin (e.g. a ase-pectin coating as described herein); and wherein the modified release coating is present in an amount corresponding to a weight gain of the formulation due to the second g selected from (a) from 10% to 12%, for example about 11% or about 11.5%; or (b) from 16% to 18%, for example about 17% by weight based upon the weight of the formulation prior to applying the second coating.

In addition the process to form a composition of the invention may comprise the steps of mixing a first population and a second population, n the first population has a coating that is or comprises a water-soluble cellulose ether but having no outer coating, e.g. as bed herein; and the second population has a first coating that is or comprises a water-soluble cellulose ether and a second coating that is or comprises a delayed release coating, for example as described herein e.g. a coating that is or comprises a delayed release polymer.

Applications The composition of the invention may advantageously be used for oral delivery pharmaceutically active ingredients by virtue of the enhanced ution profiles achieved.

P213428WO 98 The compositions of the invention include modified release compositions which comprise cyclosporin A and a modified release coating, for example comprising a pH independent polymer, to target cyclosporin release to the lower intestine. Such compositions result in low systemic exposure to cyclosporin A, whilst providing high levels of cyclosporin A in the lower GI tract, particularly in the colon. Such compositions release the cyclosporin A in an active form for example as a solution, which provides enhanced absorption of cyclosporin A in the local tissue of the lower GI tract. When the composition is used in the form of minibeads, the minibeads are ageously dispersed along large sections of the GI tract following oral administration and are therefore expected e a more uniform exposure to cyclosporin to large sections of for example the colon.

Accordingly the modified release compositions ing to the invention comprising cyclosporin for local treatment of the lower GI tract are expected to be useful in the treatment or prevention of a ion of the GIT. In particular the composition of the invention may comprise cyclosporin A and/or another immunosuppressant and be useful in the prevention or ent of inflammatory conditions affecting the lower GI tract, particularly conditions affecting the colon.

The ition of the invention is administered orally. The dose required will vary ing upon the specific condition being treated and the stage of the condition. In the case of compositions containing cyclosporin A, the composition will generally be administered to provide a dose of cyclosporin A of from 0.1 to 100mg, for example a dose of 1 to 500 mg or particularly a dose of 25 to 250mg cyclosporin A. The composition is suitably administered as a single daily dose.

In one aspect of the invention there is ed a composition of the invention for use in the treatment or prophylaxis of an matory bowel disease, Crohn’s disease, ulcerative colitis, graftversus-host disease, gastrointestinal graft-versus-host disease, myasthenia gravis , irritable bowel syndrome (e.g. with constipation, diarrhea and/or pain symptoms), celiac disease, stomach ulcers, diverticulitis, pouchitis, proctitis, mucositis, chemotherapy-associated enteritis, radiation-associated enteritis, short bowel disease, or chronic ea, gastroenteritis, duodenitis, tis, peptic ulcer, Curling’s ulcer, appendicitis, colitis, iculosis, endometriosis, colorectal oma, arcinoma, inflammatory disorders such as diversion colitis, ischemic colitis, infectious colitis, chemical colitis, microscopic colitis ding collagenous colitis and lymphocytic colitis), atypical colitis, pseudomembraneous colitis, fulminant colitis, autistic enterocolitis, interdeminate colitis, iletis, ileitis, ileocolitis or granulomatous colitis, the prevention of ion following bone marrow transplantation, psoriasis, atopic dermatitis, rheumatoid arthritis, nephrotic syndrome primary sclerosing cholangitis, familial adenomatous sis, or perinanal Crohn’s, including perianal fistulae.

In one embodiment the composition of the invention is for use in the treatment of an matory bowel disease. The main forms of inflammatory bowel disease are Crohn’s disease and ulcerative colitis. Accordingly the composition of the invention may be useful in the treatment of both of these conditions.

The composition of the invention may be for use in the treatment or prevention of irritable bowel syndrome (e.g. with constipation, diarrhea and/or pain symptoms), celiac disease, stomach P213428WO 99 ulcers, diverticulitis, pouchitis, tis, mucositis, radiation-associated enteritis, short bowel disease, or chronic diarrhea, gastroenteritis, duodenitis, jejunitis, peptic ulcer, Curling’s ulcer, appendicitis, colitis, diverticulosis, endometriosis, colorectal carcinoma, adenocarcinoma, inflammatory disorders such as diversion colitis, ischemic colitis, infectious colitis, chemical colitis, microscopic colitis (including collagenous colitis and lymphocytic colitis), atypical colitis, pseudomembraneous colitis, fulminant colitis, autistic enterocolitis, interdeminate colitis, jejunoiletis, s, ileocolitis, granulomatous colitis, fibrosis, graft-versus-host disease, gastrointestinal versus-host disease, HIV prophylaxis and treatment (for e HIV enteropathy) or gastrointestinal enteropathies. The composition may also be for use in the treatment or prevention of idium difficile colitis.

Crohn’s disease may affect the entire GI tract including the colon. However, ulcerative colitis is a condition which affects only the colon and the rectum. Accordingly, the release profile provided by the colon-targeted, immunosuppressant-containing (e.g. cyclosporin A-containing), ition according to the invention is expected to be especially beneficial in the treatment of ulcerative colitis.

The colon-targeted, composition of the ion primarily releases cyclosporin A, in the colon. r, cycosporin may also be released higher in the GI tract and accordingly the composition may also e therapeutic benefit in conditions which affect other parts of the lower GI tract, for example s disease, irritable bowel syndrome (e.g. with constipation, diarrhea and/or pain symptoms), celiac disease, stomach ulcers, diverticulitis, collagenous colitis, pouchitis, proctitis, mucositis, radiation-associated enteritis, short bowel e, chronic diarrhea, gastroenteritis, duodenitis, jejunitis, peptic ulcer, Curling’s ulcer, icitis, diverticulosis, endometriosis, colorectal carcinoma, adenocarcinoma, inflammatory disorders such as, jejunoiletis, ileitis, litis, celiac disease, fibrosis, graft-versus-host disease, intestinal graft-versus-host e, HIV prophylaxis and treatment (for example HIV enteropathy) or enteropathies.

Gastrointestinal Graft-Versus-Host-Disease (GI-GVHD) is a life-threatening condition and one of the most common causes for bone marrow and stem cell transplant failure. In patients with GIGVHD it is the donor cells that begin to attack the t’s body – most frequently the gut, liver and skin. Patients with mild-to-moderate GI-GVHD typically develop symptoms of anorexia, nausea, ng and diarrhoea. If left untreated, GI-GVHD can progress to ulcerations in the lining of the GI tract, and in its most severe form, can be fatal. Accordingly, in one embodiment the composition is for use in the ent or prophylaxis of Gastrointestinal Graft-Versus-Host-Disease (GI-GVHD).

In a further embodiment there is ed a composition of the invention for use in the treatment of celiac disease.

In a r embodiment there is provided a composition of the invention for use in the treatment or prophylaxis of tive colitis.

Also provided is a composition of the invention for use in the treatment of neurodegenerative es (for example Parkinson’s disease, Alzheimer’s disease or ar dementia) or paediatric diseases, including, but not limited to ulcerative colitis, Crohn’s disease and GvHD.

P213428WO 100 ] Chronic inflammation of the GI tract may result in cellular transformation and the onset of cancer through tumourigenesis. Cyclosporin has been shown to be effective in inhibiting cell growth in a number of colorectal cancer cell lines eck et al Cell Cycle 11:21; 2012; 3997-4008). It has also been shown that cyclosporin may be an effective inhibitor of tumourigenesis (Kawahara et al, Cyclosporine A and Tacrolimus Inhibit Urothelial Tumorigenesis; Molecular Carcinogenesis, 2015).

Accordingly, cyclosporin may be beneficial in providing a cytostatic ancer effect thereby inhibiting the growth of a tumour. Cyclosporin may be beneficial in preventing or delaying the onset of colorectal cancer in patients with chronic inflammatory conditions ing the GI tract, particularly the colon, for example the inflammatory ions described herein such as ulcerative colitis or Crohn’s disease. As discussed above the compositions sing cyclosporin e high levels of cyclosporin in a solubilised form into the colon and may therefore by particularly cial in the treatment of colorectal cancer.

A further aspect of the invention provides a composition comprising cyclosporin as defined herein for use in the treatment of a cancer affecting the GI tract, particularly the lower GI tract and especially the colon. Accordingly the composition comprising cyclosporin may be for use in the treatment of colorectal cancer. The composition comprising cyclosporin may be for use in providing a cytostatic effect in a cancer affecting the GI tract, ularly a colorectal cancer.

Also provided is a composition comprising cyclosporin for use in the preventing or delaying the onset of a cancer of the GI tract in a t with c inflammatory condition affecting the GI tract, ularly the lower GI tract and especially the colon. Accordingly the composition comprising cyclosporin may be for use in inhibiting tumourigenesis in the GI tract, particularly the colon.

The composition comprising cyclosporin may be used alone or together with another anticancer agent, for example the composition comprising cyclosporin may be used together with an antineoplastic agent to treat or delay the onset of a cancer ing the GI tract. The composition comprising cyclosporin may be stered to a subject as a fixed dose combination with one or more additional anticancer agents. The composition comprising cyclosporin may be administered separately, tially or substantially simultaneously with another ncer agent.

Anti-cancer agents which may be suitable for use with the composition comprising cyclosporin include, but are not limited to one or more agents selected from (i) antiproliferative/antineoplastic drugs and combinations thereof, such as alkylating agents (for example cis-platin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen d, uracil mustard, bendamustin, melphalan, chlorambucil, chlormethine, busulphan, temozolamide, nitrosoureas, ifosamide, melphalan, pipobroman, triethylene-melamine, triethylenethiophoporamine, carmustine, lomustine, stroptozocin and dacarbazine); antimetabolites (for example gemcitabine and antifolates such as fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed, methotrexate, pemetrexed, ne arabinoside, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, abine phosphate, pentostatine, and gemcitabine and hydroxyurea); antibiotics (for example cyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine P213428WO 101 and vinorelbine and taxoids like taxol and taxotere and polokinase inhibitors); proteasome inhibitors, for example carfilzomib and bortezomib; interferon y; and topoisomerase inhibitors (for example epipodophyllotoxins like ide and teniposide, amsacrine, topotecan, irinotecan, ntrone and camptothecin); bleomcin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, axel (Taxol™), nabpaclitaxel, docetaxel, mithramycin, deoxyco-formycin, mitomycin-C, L- asparaginase, interferons (especially IFN-alpha), etoposide, teniposide, DNA-demethylating agents, (for example, azacitidine or decitabine); and histone de-acetylase (HDAC) inhibitors (for example vorinostat, , panobinostat, romidepsin, valproic acid, mocetinostat (MGCD0103) and pracinostat SB939); (ii) cytostatic agents such as antiestrogens (for example tamoxifen, fulvestrant, fene, raloxifene, droloxifene and fene), antiandrogens (for example bicalutamide, ide, mide and cyproterone e), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for e as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5-reductase such as finasteride; and navelbene, CPT-ll, anastrazole, letrazole, capecitabine, reloxafme, cyclophosphamide, ifosamide, and afine; (iii) anti-invasion agents, for e dasatinib and bosutinib (SKI-606), and metalloproteinase inhibitors, inhibitors of urokinase plasminogen activator receptor function or antibodies to Heparanase; (iv) inhibitors of growth factor function: for e such inhibitors include growth factor antibodies and growth factor receptor antibodies, for example the anti-erbB2 antibody trastuzumab [Herceptin™], the anti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab, tyrosine kinase inhibitors, for example inhibitors of the mal growth factor family (for example EGFR family tyrosine kinase inhibitors such as gefitinib, erlotinib, 6-acrylamido-N-(3-chlorofluorophenyl)(3- morpholinopropoxy)-quinazolinamine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib) and antibodies to costimulatory molecules such as CTLA-4, 4-lBB and PD-l, or antibodies to cytokines (IL-I0, TGF-beta); inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor ; modulators of protein regulators of cell apoptosis (for example Bcl-2 inhibitors); inhibitors of the platelet-derived growth factor family such as imatinib and/or nib (AMN107); inhibitors of serine/threonine kinases (for example Ras/Raf signalling inhibitors such as farnesyl transferase inhibitors, for example sorafenib , tipifarnib and rnib), inhibitors of cell signalling h MEK and/or AKT kinases, c-kit inhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1R kinase inhibitors, IGF receptor, kinase tors; aurora kinase inhibitors and cyclin dependent kinase tors such as CDK2 and/or CDK4 inhibitors; and CCR2, CCR4 or CCR6 antagonists; (v) giogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti-vascular endothelial cell growth factor antibody bevacizumab (Avastin™)]; thalidomide; domide; and for example, a VEGF receptor tyrosine kinase inhibitor such as vandetanib, vatalanib, sunitinib, axitinib and pazopanib; P213428WO 102 (vi) gene therapy approaches, including for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2; (vii) immunotherapy approaches, including for example antibody therapy such as alemtuzumab, rituximab, ibritumomab tiuxetan (Zevalin®) and umab; interferons such as interferon α; interleukins such as IL-2 (aldesleukin); interleukin inhibitors for example IRAK4 inhibitors; cancer vaccines including prophylactic and ent vaccines such as HPV vaccines, for example Gardasil, ix, Oncophage and Sipuleucel-T (Provenge); gp100;dendritic cell-based vaccines (such as Ad.p53 DC); toll-like receptor modulators for example TLR-7 or TLR-9 agonists; PD-1, PD-L1, PD-L2 and CTL4-A modulators (for example Nivolumab), dies and vaccines; other IDO inhibitors (such as indoximod); anti-PD-1 monoclonal dies (such as MK-3475 and mab); anti-PDL1 monoclonal dies (such as MEDI-4736 and RG-7446); anti-PDL2 monoclonal antibodies; and anti-CTLA-4 antibodies (such as umab; and (viii) cytotoxic agents for example ibine (fludara), cladribine, tatin (NipentTM); The coating containing the water-soluble cellulose ether of the present invention may be useful in reducing the variability between release es of different batches of minibeads.

A "batch" is a specific quantity of a drug or other material that is intended to have uniform character and y, within specified limits, and is produced according to a single cturing order during the same cycle of manufacture. A "lot" means a batch, or a specific identified portion of a batch, having uniform character and quality within specified limits; or, in the case of a drug product ed by continuous process, it is a specific identified amount produced in a unit of time or quantity in a manner that assures its having uniform character and quality within specified limits. "Lot number", "control number", or "batch number" means any distinctive combination of letters, numbers, or symbols, or any combination of them, from which the complete history of the manufacture, processing, packing, holding, and distribution of a batch or lot of drug t or other material can be determined." EXAMPLES e 1a: Preparation of a Liquid Composition of the Invention An aqueous phase was prepared by mixing sodium dodecyl sulphate (SDS) and D-sorbitol with purified water under constant stirring. Gelatin was then added to this solution and gentle heat was applied to approximately 60-70° C to achieve complete melting of gelatin. The composition of the aqueous phase is shown in Table 1 below.

Table 1 Component w/w% water 79.6 SDS 1.3 Sorbitol 2.0 P213428WO 103 Gelatin 17.1 An oil phase was prepared by mixing together thoxyethoxy)ethanol (Transcutol HP), glyceryl monooleate/dioleate l GMO-50) and capric/caprylic ceride (Miglyol 810) with stirring at room temperature to form a solution. Ciclosporin A was added and mixed until a clear solution was obtained. The composition of the oil phase is shown below in Table 2.

Table 2 Component w/w% Cyclosporin A 24.5 Miglyol 810 N 12.5 Transcutol HP 37.0 Capmul GMO-50 26 The oil phase was mixed with the heated aqueous phase in a ratio of approximately 1:5 (oil phase:aqueous phase). The resulting mixture was stirred at 60-70°C, 250-350rpm using a magnetic stirrer to achieve homogeneity.

Example 1b: Preparation of a r Liquid Composition Following the procedure of Example 1a a further liquid composition with glyceryl ate/caprate (Capmul MCM) as the surfactant in the oil phase in place of the glyceryl monooleate/dioleate (Capmul GMO-50) was prepared. The aqueous phase of the ition is shown in Table 3 and the oil phase of the composition is shown in Table 4. The oil phase was mixed with the heated aqueous phase in a ratio of approximately 1:5 (oil phase:aqueous phase).

Table 3 Table 4 Component w/w% Component w/w% water 79.6 Cyclosporin A 24.5 SDS 1.3 Miglyol 810 N 12.5 Sorbitol 2.0 Transcutol HP 37.0 Gelatin 17.1 Capmul MCM 26 Example 1c: Preparation of a Further Liquid Composition Following the procedure of Example 1a a further liquid composition with glycerol linoleate (Maisine 35-1) as the surfactant in the oil phase in place of the yl monooleate/dioleate (Capmul GMO-50) was prepared. The aqueous phase of the composition is shown in Table 5 and the oil phase P213428WO 104 of the composition is shown in Table 6. The oil phase was mixed with the heated aqueous phase in a ratio of approximately 1:5 (oil phase:aqueous phase).

Table 5 Table 6 Component w/w% Component w/w% water 79.6 Cyclosporin A 24.5 SDS 1.3 Miglyol 810 N 12.5 ol 2.0 Transcutol HP 37.0 Gelatin 17.1 Maisine 35-1 26 Example 2: Preparation of a Minibead.

A minibead as described herein may be a composition of the invention. Alternatively the minibead may be a core. The minibead was generally prepared by forming a minibead according to the following procedure The composition or core in the form of seamless minibeads were prepared using Spherex process as follows.

] An aqueous phase and oil phase mixture was prepared following the procedure described in Example 1a.

The mixture was then fed (via temperature controlled tubing) through a vibrating nozzle, with a single nozzle outlet with a diameter of 3mm. Seamless minibeads were formed as the solution flowed h the vibrating nozzle into a cooling chamber of constantly flowing medium chain ceride ol 810) cooling oil at a temperature of 10°C.

The minibeads were removed from the cooling oil and placed in a centrifuge to remove the excess oil. Following centrifugation, a first drying step was initiated with a set refrigerator temperature of 10°C and the heater temperature of 20°C. The dryer was rotated at 15 RPM. When the beads were ed to be freely rotating in the drying drum, they were considered to be dry.

The ads were washed with ethyl e and then dried for a further 24h under the same drying conditions as those mentioned above in the first drying step . The dried minibeads were then sieved to remove ze and undersize beads resulting in cores m in diameter. This procedure provided cores with the composition shown in Table 7, the values being the weight percent of the total weight for each component.

Table 7 Component w/w% Cyclosporin A 12.1 P213428WO 105 l 810 N 6.2 Transcutol HP 18.3 Capmul GMO-50 12.9 SDS 3.2 Sorbitol 4.9 Gelatin 42.4 Example 3: Preparation of a Minibead with a First Coating (Sub-Coat).

A coated minibead can be produced by coating a minibead ed in Example 2 with a dispersion of Opadry White 20A28380 (supplied by Colorcon). The minibeads were loaded into a fluid bed coater (Wurster column) and coated with Opadry White 20A28380 (supplied by Colorcon Limited) as a dispersion. The processing parameters, such as inlet air temperature and inlet air volume, were adjusted to keep the minibead temperature between 40°C and 42°C until the required coating weight gain was reached. The resulting subcoated ads were dried for 5 minutes at 40oC in the coater.

Composition of the Coated ad A minibead with the composition shown in Table 8 below was produced by the above ure. A minibead with an Opadry weight gain of 7.5% relative to the weight of the core is shown in Table 8. Table 9 shows the composition of a minibead coated with an Opadry weight gain of 5% relative to the weight of the core. Table 10 shows the composition of a minibead coated with an Opadry weight gain of 10% ve to the weight of the core.

Table 8 Component w/w% Cyclosporin A 11.3 Miglyol 810 N 5.8 Transcutol HP 17.0 Capmul GMO-50 12.0 SDS 2.9 Sorbitol 4.6 Gelatin 39.4 Opadry 7.0 Table 9 P213428WO 106 Component w/w% Cyclosporin A 11.5 Miglyol 810 N 5.9 Transcutol HP 17.4 Capmul GMO-50 12.3 SDS 3.1 Sorbitol 4.7 Gelatin 40.3 Opadry 4.8 Table 10 Component w/w% Cyclosporin A 11.0 Miglyol 810 N 5.6 Transcutol HP 16.7 Capmul GMO-50 11.7 SDS 2.9 Sorbitol 4.5 Gelatin 38.5 Opadry 9.1 Example 4a: ation of a ad with a Second Coating of Ethylcellulose.

A minibead coated with Opadry, the first coating (also referred to as a subcoat), was produced ing the procedure in Example 3. The minibead produced by the procedure of Example 3 was then further coated with a second coating (also referred to as an overcoat) of Surelease® (an ethylcellulose dispersion).

The Surelease® overcoat was applied by the following procedure. ase® was slowly added to a stainless steel vessel and mixed to provide the required coating suspension of Surelease® for the overcoat. The resulting g suspension was then applied onto the e of the subcoated minibeads using an analogous coating method to that described for the Opadry coating in P213428WO 107 Example 3 until the desired weight gain of Surelease® was reached. The over-coated minibeads were then dried in the coater for an hour at 40-45°C.

The minibead was coated with a 9.5% weight gain of Surelease®.

Minibeads with no Opadry coating may be produced by coating a minibead described in Example 2 with Surelease® as described above.

The minibead with a first and second coating has the composition shown in Table 11.

Table 11 ent w/w% porin A 10.3 Miglyol 810 N 5.3 Transcutol HP 15.5 Capmul GMO-50 10.9 SDS 2.7 Sorbitol 4.2 Gelatin 36.0 Opadry 6.4 Surelease 8.7 Similarly, the ition of ads coated with 5% Surelease and 7.5% Opadry are shown in Table 12 and the composition of minibeads coated with 20% Surelease and 7.5% Opadry are shown in Table 13.

Table 12 Component w/w% Cyclosporin A 10.7 Miglyol 810 N 5.5 Transcutol HP 16.2 Capmul GMO-50 11.4 SDS 2.9 Sorbitol 4.4 Gelatin 37.5 P213428WO 108 Opadry 6.6 Surelease 4.8 Table 13 Component w/w% Cyclosporin A 9.4 Miglyol 810 N 4.8 Transcutol HP 14.2 Capmul GMO-50 10.0 SDS 2.5 Sorbitol 3.8 Gelatin 32.8 Opadry 5.8 Surelease 16.7 e 4b: Preparation of a Minibead with a Second Coating of Ethylcellulose/Pectin.

A ad coated with , the first coating (also referred to as a subcoat), was produced following the procedure in Example 3. The minibead produced by the procedure of Example 3 was then further coated with a second coating (also referred to as an overcoat) of a mixture of Surelease® (an ellulose dispersion) and Pectin.

The Surelease®/pectin overcoat was applied by an analogous method to the Surelease g of Example 4a. Pectin was added to purified water in a stainless steel vessel and mixed to obtain a solution. Surelease® was slowly added to the vessel whilst ining mixing to provide the required Pectin concentration in the Surelease® for the overcoat. The ing coating suspension was then applied onto the surface of the sub-coated minibeads using an analogous coating method to that described for the Opadry coating in Example 3 until the d weight gain of Surelease®/Pectin was reached. The over-coated minibeads were then dried in the coater for an hour at 40-45°C.

The minibead was coated with a 9.5% weight gain of Surelease®/Pectin.

The minibead with a first and second coating has the composition shown in Table 14.

Table 14 P213428WO 109 Component w/w% porin A 10.3 Miglyol 810 N 5.3 Transcutol HP 15.5 Capmul GMO-50 10.9 SDS 2.7 Sorbitol 4.2 Gelatin 36.0 Opadry 6.4 Surelease 8.5 Pectin 0.2 Example 5: Crystallisation Experiments Comparative Example 1 ] Three liquid compositions comprising Cremophore EL (also known as Kremophore EL), a hoxylated castor oil surfactant with an HLB value of greater than 10 were prepared. The three liquid compositions are different s from the same batch. Each of the liquid compositions has an oil phase comprising: cyclosporin A 26.3%, Transcutol HP 40%, Cremophor EL 22.5%, and Miglyol 810 11.2% (% amounts are of the oil phase); and an s phase comprising: gelatin 17.1%, Sorbitol 2.0%, SDS 1.4%, and water 79.5% (% amounts are of the aqueous phase. The liquid composition was prepared by mixing the oil phase and aqueous phases in an oil phase to aqueous phase ratio of 1:7.

Example 5a Three liquid compositions of Example 1a comprising Capmul GMO-50, replacing the Cremophore EL were prepared. The three liquid compositions are 3 sublots of the same batch.

Capmul GMO-50 has an HLB value of 3.

] Example 5b A liquid composition of Example 1b comprising Capmul MCM was prepared Capmul MCM has an HLB value of 6-7.

Example 5c P213428WO 110 ] A liquid composition of Example 1c comprising Maisine 35-1 was prepared. Maisine 35-1 has an HLB value of The crystallisation rate of the liquid compositions of Comparative Example 1, Example 5a, Example 5b and Example 5c were tested to determine the length of time for cyclosporin crystallisation to occur. Each of the liquid compositions was stirred at 250-350rpm to form an emulsion. Samples of the three emulsions were taken at 30 minute intervals and viewed under a microscope at 50x or 100x magnification. The time when crystals appeared in the sample is shown in Table 15.

Table 15 Beads Example Surfactant HLB llization time (h) formation 5a Capmul GMO-50 3 2 Yes 5b Capmul MCM 5-6 >7 No 5c Maisine 35-1 4 3 Yes Comparative Cremophor EL 14 0.5 Yes Ex. 1 Figure 1 shows images of the three liquid compositions of Comparative Example 1 comprising hore EL (also known as Kremophore EL), a polyethoxylated castor oil surfactant with an HLB value of greater than 10.

Figure 2 shows images of three liquid compositions of e 5a sing Capmul GMO-50 which is replacing the Cremophore EL. Capmul GMO-50 has an HLB value of 3.The three compositions comprising Capmul GMO-50 are Figure 3 shows images of a liquid composition of e 5b, comprising Capmul MCM.

Figure 4 shows images of a liquid composition of Example 5c comprising Maisine 35-1.

As is t from the images the liquid compositions of Example 5a, 5b and 5c had a much longer period before crystals appeared. The Capmul GMO-50 and Capmul MCM compositions were essentially free of l formation throughout the test period. The Capmul GMO-50 compositions were essentially free of crystal up to 240 min, whereas the Cremophore EL compositions had noticeable crystal formation after 120 min. The Capmul MCM itions were crystal free for 7 hours. The Maisine 35-1 composition had crystal formation at 3 hours.

Example 6: In-Vitro ution Profile of Minibeads of Example 2.

The ro dissolution profiles of a sample of the minibeads produced in Example 2 were measured using the following dissolution test. The dissolution testing was carried out in accordance with USP <711> Dissolution using Apparatus II (paddle apparatus) operated with a paddle speed of 75 P213428WO 111 rpm and with the ution medium at a temperature of 37oC ± 0.5oC. The dissolution medium was deionised water. The sample of ads were placed in the dissolution medium and at the start of the test (t=0). The dissolution medium was sampled at regular intervals. The obtained dissolution data is shown in the Table 16 and Table 17 below. The dissolution profile for two batches of the minibead of Example 2 is shown in Figure 5.

Table 16 Timepoint ESN-740 ESN-740 ESN-740 ESN-740 ESN-740 ESN-740 (Hrs) Pot 1 Pot 2 Pot 3 Pot 4 Pot 5 Pot 6 %RSD 0.08 18.7 21.7 18.3 19.7 16.5 15.4 12.2 0.17 49.3 51.6 49.1 49.9 45.2 36.6 11.7 0.25 67.0 67.0 72.6 67.4 64.2 54.0 9.5 0.5 76.4 77.1 79.0 76.2 89.5 63.7 10.7 1 81.5 82.6 84.0 85.5 77.7 77.2 4.1 2 92.1 90.5 87.2 95.4 82.4 87.9 5.0 4 101.5 95.7 85.5 97.4 82.6 93.2 7.8 6 99.5 97.1 91.2 98.7 96.2 94.8 3.1 12 98.7 94.8 94.8 90.2 91.9 95.5 3.1 24 100.5 99.1 52.3 85.3 93.1 88.0 20.5 P213428WO 112 Table 17 Timepoint 0 ESN-760 ESN-760 ESN-760 ESN-760 ESN-760 (Hrs) Pot 1 Pot 2 Pot 3 Pot 4 Pot 5 Pot 6 %RSD 0.08 16.1 15.5 18.2 20.2 17.5 6.4 30.8 0.17 47.6 44.2 52.3 51.4 44.2 24.2 23.4 0.25 65.6 61.7 68.7 69.6 57.3 34.6 21.9 0.5 75.7 71.2 75.2 75.9 68.0 45.7 17.0 1 78.9 78.0 79.0 84.8 76.4 61.6 10.2 2 94.6 90.7 97.9 97.9 90.7 68.9 12.1 4 102.7 97.5 98.2 101.0 97.9 82.3 7.6 6 102.2 99.1 97.5 102.1 101.0 84.0 7.1 12 104.5 99.1 100.0 94.9 99.5 94.9 3.6 24 102.0 100.2 101.0 100.0 102.7 101.2 1.0 Comparative Example 2 Minibeads corresponding to those of Example 2 were prepared with Cremophore EL as the surfactant in place of Capmul .These minibeads had the composition shown in Table 18.

These minibeads were submitted to the same ution test in deionised water. The dissolution profile of 5 batches of these minibeads is shown in Figure 6.

Table 18 Component w/w% porin A 10.8 Miglyol 810 N 4.6 Transcutol HP 16.4 Cremophor EL 9.2 SDS 4.0 Sorbitol 5.8 n 49.2 Example 6a As a further comparison with Comparative Example 2, minibeads were prepared using almost identical quantities of each excipient to those of Comparative Example 2, except that the 9.2 w/w% Cremophor was replaced with 9.3% Capmul GMO-50 to give the minibead ition shown in Table 19. The minibeads of Table 19 were prepared in an analogous manner to those of Example 2. The w/w% in Tables 18 and 19 refer to the dry weight of the composition.

P213428WO 113 Table 19 Component w/w% Cyclosporin A 10.9 Miglyol 810 N 4.6 Transcutol HP 16.6 Capmul GMO-50 9.3 SDS 4.0 Sorbitol 5.7 Gelatin 49.0 Cyclosporin A 10.8 These minibeads were submitted to the same dissolution test in deionised water. The dissolution profile of 3 batches of these minibeads is shown in Figure 7.

It is apparent from the dissolution profiles shown in Figures 5, 6 and 7 that compositions sing Capmul GMO-50 (Figures 5 and 7) have superior dissolution profiles compared to compositions comprising Cremophor EL (Figure 6). The dissolution profile of the Capmul GMO-50 compositions have a higher maximum release and this maximum release of cyclosporin is generally maintained in solution for longer than that observes with Comparative Example 2, which contained Cremophor. Figure 6 shows that the % release of porin from the Cremophor EL compositions is lower and that the cyclosporin tration reduces over time from a maximum compared to the compositions containing Capmul GMO-50. e 7: In-Vitro Dissolution Profile of Minibeads of Example 4a Minibeads corresponding to those of Example 4a, ically minibeads with the composition shown in Table 11 (7.5% Opadry subcoat, first coating and 9.5% Surelease overcoat, second coating) were prepared, having Capmul GMO-50 as the surfactant. These minibeads were submitted to a 2 stage ution test.

In the first stage of the test the dissolution medium was 750ml of 0.1N HCl simulating the pH of the gastric environment. At the start of the test (t=0) the sample was placed in the dissolution medium. After 2 hours an aliquot of the medium is taken for subsequent analysis and immediately (suitably within 5 s) the second stage of the dissolution test is ted. In the second stage 250 ml of 0.2M tribasic sodium phosphate containing 2% sodium dodecyl sulphate (SDS) is added to the dissolution medium and the pH adjusted to 6.8 ± 0.05 using 2N NaOH or 2N HCl as required.

Samples of the dissolution medium were taken at the following time points during the second stage of the test: 4 hours; 6 hours; 12 hours; and 24 hours from the start of the test (i.e. from t=0 at the start of the first stage).

P213428WO 114 The sample taken at the end of the first stage (2 hours) and the samples from the second stage were analysed for cyclosporin A using Reverse Phase HPLC with UV detection at 210nm.

The amount of dissolved cyclosporin A in the dissolution medium is expressed as a percentage based upon the original cyclosporin content in the test formulation (the % released). The percentage release is given in Table 21 and the dissolution profile of minibeads with the ition shown in Table 11 is shown in Figure 8.

Table 21 Timepoint % Drug release (hours) Batch 1 Batch 2 Batch 3 Batch 4 0 0 0 0 0 2 0 0 0 0 4 15 15 15 17 6 39 38 36 40 12 70 69 64 71 24 99 99 95 100 Example 8: Droplet Size - c Light Scattering.

The size of the droplets was measured using dynamic light scattering. Coated minibeads of Example 4a, Table 11 (minibeads coated with 7.5% wt gain Opadry and 9.5% wt gain ase) (0.5 g) comprising Capmul GMO-50 as the first surfactant were added to a beaker containing 50 g of deionised water. The beaker contents were mixed at 250 rpm throughout the study. Samples of the beaker contents were taken at 0, 1, 2, 3, 4, 5, 6 and 24 hours.

Samples of the beaker contents were filtered using 0.65µm pore size filters (Merck Millipore Ultrafree-CL Centrifugal Filter). The particle size and zeta potential of each sample was measured and analysed using a Malvern Nano-Zetasiser. The resulting data is shown in Figure 9.

The coated minibeads tested in this example t very stable droplet e for up to 4 hours with droplet size ranging from 120 to 240 nm in water media. As time passes and further droplets are released from the coated minibeads after 4 hours the droplet size range is broader.

Without wishing to be bound by theory, it is le that as time passes and the dissolution media becomes more ted with ed droplets a re-equilibration between the droplets already present in the media and the ones freshly released may occur. The variability in droplet size after 4 hours is potentially caused by this re-equilibration s rather than a representation of the size of the droplets being released from the minibeads.

Example 9: In-Vivo Study in y Male Volunteers In the study described below "CyCol®" is a reference to the minibeads of Example 4a, and bed in Table 11 (comprising a core with Capmul GMO-50 as the surfactant, a 7.5% weight gain ® first coating on the core, and a 9.5% weight gain of a Surelease® second coating). The P213428WO 115 minibeads were loaded into HPMC capsules to provide a unit dose of 37.5 mg cyclosporin A per capsule.

LIST OF ABBREVIATIONS AND DEFINITION OF TERMS Abbreviation Definition AUC%extrap Residual area/Percentage of AUC0-inf extrapolated nf Area under the concentration-time curve from time zero to infinity polated) AUC0-t Area under the tration-time curve from time zero to the last non-zero concentration t Area under the concentration-time curve from time zero to last quantifiable concentration Cav Average steady state concentration CYP Cytochrome P-450 kel Elimination rate constant LLQ Lower limit of quantification LR Linearity ratio SD Standard deviation t½ Terminal half-life UC Ulcerative colitis STUDY OBJECTIVES Primary Objectives:  To characterise whole blood cokinetics of CyCol® following single and multiple oral doses, and compare to a single Sandimmun® IV administration pharmacokinetic profile in healthy male subjects.

 To evaluate the colonic mucosa (epithelial, mucosal and sub-mucosal tissue) concentrations of cyclosporin and its metabolites ing multiple oral doses of CyCol® and compare to concentrations following a single Sandimmun® IV administration. 8WO 116 Secondary Objectives:  To obtain safety and tolerability information following multiple oral doses of CyCol® at the selected doses in y male subjects.

Exploratory Objectives:  To evaluate the amount of unchanged porin and its metabolites excreted in the faeces after administration of multiple doses of CyCol® and compare to amounts ing a single Sandimmun® IV administration.

INVESTIGATIONAL PLAN Overall Study Design and Plan – Description This was a Phase I, single , multi-stage open study designed to te the safety, tolerability, pharmacokinetics and ve colonic mucosal concentrations of cyclosporin capsules (CyCol®) compared to IV cyclosporin in healthy male eers. This study also investigated the amount of unchanged cyclosporin recovered from faecal samples and relative concentrations of its metabolites AM9, AM4N and AM1. The concentrations of cyclosporin metabolites were also examined in faecal and colonic mucosa samples.

In Stage 1 of the study, a total of 24 eligible subjects received either Sandimmun® IV over 24 hours (two consecutive 12 hour infusions) at a dose of 2 mg/kg (2 mg/kg/day), a once daily oral dose of CyCol® 75 mg for 7 days or a twice daily (BID) oral dose of CyCol® 75 mg for 7 days. Subjects who received CyCol® 75 mg BID only received a single dose on Day 7 (morning dose).

At the end of Stage 1 the data were reviewed and based on the evidence available, the protocol allowed for further investigations to be conducted with alternative CyCol® doses and dose frequencies in uent . Following this review, 8 subjects were recruited to Stage 2 of the study and received a once daily oral dose of CyCol® 37.5 mg for 7 days. A further 8 subjects were recruited to Stage 3 and received a BID oral dose of CyCol® 150 mg for 7 days.

Details of the dosing sequence are presented in Figure 10.

Study Design, Including Choice of Control Groups A multi stage design was used for this study to reduce the number of dose levels investigated and the number of subjects exposed to cyclosporin and study procedures. The dosing regimens chosen for Stages 2 and 3 were chosen following review of data (safety and tolerability, ic re and colonic mucosa tissue concentrations) ed at other doses.

The study was open-label because of the different mode of administration of the comparator product (IV versus oral for investigational medicinal product), and the objective endpoints of the study (cyclosporin concentrations). The single dose pharmacokinetic profile was examined over a 24 hour period, based on previous experience that demonstrated this duration adequately characterised the concentration-time e.

The study recruited healthy male volunteers. Females were not included in this study as the Food and Drug Administration pregnancy category for cyclosporin is C.

P213428WO 117 The comparator chosen for this study was mun®. The current en t regimen for UC can often involve the use of several agents administered ly, orally or intravenously depending on the severity of the disease. Cyclosporin is unlicensed for UC. However, ing to the 2013 National Institute for health and al Excellence guidelines for UC, it is ended that treatment with IV porin should be considered for subjects with acute severe colitis and not responding to or unsuitable for first line therapy with osteroids. An IV dose of 2 to 4 mg/kg/day or an oral dose of to 8 mg/kg/day is recommended for severe ulcerative colitis treatment. The rationale for choosing IV administration of cyclosporin over the oral formulation is based on studies documenting variable absorption and the extensive first pass metabolism following oral ingestion. In addition, an IV dose of 2 to 4 mg/kg/day is known to be efficacious in the treatment of patients with UC and to attain the same trations using a currently available oral dose form, approximately 3 times the IV dose would be ed. Furthermore, when cyclosporin is administered by IV infusion blood concentrations are constant and lites are t in lower concentrations compared to oral administration. CyCol® has been developed to delay the release of active cyclosporin until it reaches the colon, it thereby bypasses the site of absorption in the jejunum and thus limits the amount of metabolism (both presystemic metabolism in the gastrointestinal tract and systemic metabolism by the hepatic system) by cytochrome P450 enzymes, including the CYP3A4 . Therefore, the concentration of metabolites following CyCol® administration is expected to be similar to IV cyclosporin. Furthermore, colon tissue concentrations of cyclosporin are 10 times higher in healthy volunteers given cyclosporin parentally compared to those of healthy volunteers given the drug orally.

Treatments Treatments Administered Subjects who met all the eligibility criteria for Stage 1 of the protocol were assigned to one of the following dose groups on Day 1:  Sandimmun® 2 mg/kg IV infusion over 24 hours (2 mg/kg/day).

 CyCol® 75 mg (2 x 37.5 mg CyCol® capsules) once daily orally for 7 days.

 CyCol® 75 mg (2 x 37.5 mg CyCol® capsules) BID orally for 7 days (only a single [morning] dose was administered on Day 7).

Following review of the Stage 1 data, the following CyCol® dosing regimen was explored in Stage 2:  CyCol® 37.5 mg (1 x 37.5 mg CyCol® capsule) once daily orally for 7 days.

Following review of the data from Stages 1 and 2, the following CyCol® dosing regimen was explored in Stage 3:  CyCol® 150 mg (4 x 37.5 mg CyCol® capsule) BID orally for 7 days (only a single [morning] dose was administered on Day 7).

The dosing regimens chosen for Stages 2 and 3 were based upon safety and bility, systemic exposure and c tissue concentrations observed at other doses. To investigate higher doses the lower doses had to have been well tolerated and had to have maximum systemic exposure P213428WO 118 ≤250 ng/ml. Providing these ia were met, doses could have been escalated higher in search of a regimen that yielded colonic tissue trations >300 ng/ml.

Identity of Investigational Products Sandimmun® was provided in commercial packaging of 1 mL ampoules ning 50 mg of cyclosporin. The 1 mL es of Sandimmun® were diluted with normal saline in accordance with the Summary of Product teristics (SmPC) and prepared suitably for the two consecutive 12 hour infusions. Each infusion was prepared such that the total dose administered was 2 mg/kg/day.

Methods of Assigning Subjects to Treatment Groups In Stage 1, subjects were ed sequentially to study treatment (Sandimmun® IV, CyCol® 75 mg once daily or CyCol® 75 mg BID).

All subjects ted to Stage 2 received CyCol® 37.5 mg once daily orally for 7 days and all subjects recruited to Stage 3 received CyCol® 150 mg BID orally for 7 days.

Selection and Timing of Dose for each Subject Sandimmun® On the morning of Day 1, following an overnight fast, subjects started their IV infusion via an IV catheter. Sandimmun® IV was administered over 24 hours, as two consecutive 12 hour infusions, at a dose of 2 mg/kg/day ing to preparation and administration detailed in the Sandimmun® SmPC.

CyCol® On the morning of Day 1, following an overnight fast, subjects assigned to the CyCol® groups were administered the morning dose with imately 240 mL of water. A hand and mouth check was performed to ensure ingestion of the study drug. Time of dosing was set to the time the first e was administered.

On Day 2, prior to rge from the study centre, subjects received the morning dose with approximately 240 mL of water. For those subjects assigned to the BID dosing regimen, the pharmacy dispensed the evening dose to the subject with instructions to take the study drug as prescribed and within 10 to 12 hours following the morning dose.

Subjects returned to the study centre daily to receive the morning dose of CyCol® until Day 6 when the subjects were readmitted for an overnight stay. For those subjects ed to the BID dosing regimen, at each daily visit to the study centre, the pharmacy dispensed the evening dose to subjects with instructions for the evening dose administration.

If the subject was expected to return to the study centre on Day 6 after the scheduled time of the Day 6 morning dose, the pharmacy could have dispensed the Day 6 morning dose to the subject upon discharge from the study centre on Day 5, with instructions for the morning dose administration.

The last dose of CyCol® administration was on the morning of Day 7 for all subjects.

P213428WO 119 Blinding: This was an open label study.

Pharmacokinetic assessments – whole blood Blood samples for pharmacokinetic analysis were collected from all subjects on Day 1 at 0 (pre dose), 2, 3, 4, 5, 6, 8, 10, 12, 16, 20 and 24 hours post-dose (post start of infusion for the Sandimmun® IV group).

For subjects in the Sandimmun® IV group blood samples were also collected at 2, 4, 6 and 8 hours after completion of the on on Day 2.

For subjects in the CyCol® groups trough pharmacokinetic samples were obtained pre morning dose on Day 4. For subjects who received CyCol® once daily additional pharmacokinetic samples were obtained at 6, 12 and 16 hours post morning dose on Day 6. For subjects who received CyCol® BID additional pharmacokinetic s were obtained at 6 and 12 hours post morning dose (prior to evening dose) on Day 6, and 4 hours post g dose. On Day 7, all subjects in the CyCol® groups had blood samples for pharmacokinetics obtained at 0 (pre dose), 2, 4, 6 8, and 12 hours post-dose.

Actual sampling times were used for statistical analyses and so each time was ed accurately.

Flexible doscopy and colon biopsies All sigmoidoscopies were performed in an unprepared bowel (except for air and water), unless the subject had not defaecated within 24 hours of sigmoidoscopy.

Subjects who received Sandimmun® IV were required to have the sigmoidoscopy performed within the last hour of their infusion ion had to be on-going).

Subjects who received CyCol® were required to have the sigmoidoscopy performed within 4 to 6 hours of the Day 7 morning/last dose.

The sigmoidoscopy was conducted by an appropriately trained endoscopist who was familiar with the study protocol and obtaining biopsies. Per protocol, whenever possible, the same endoscopist was to perform all the sigmoidoscopies. In this study two opists med the sigmoidoscopies.

For ts who had not defaecated within 24 hours of the sigmoidoscopy, a rectal enema (Fleet saline, or similar) was administered 15 minutes before the scheduled biopsy time and the time and volume of enema stration was recorded. The time and total weight of the voided gut contents post enema were recorded. A representative sample (approximately 5 g) was obtained from the voided gut for processing as described herein.

The colon biopsies were collected (as described below) as soon as possible and ideally within minutes of the enema being voided.

Standard pinch biopsy forceps were used to obtain the colonic mucosa biopsies. Each biopsy was imately 5 mm in size (and included mucosa and submucosa layers). A total of 5 biopsies, imately 1 cm apart were obtained from as close to the sigmoid colon as possible. The time of the biopsy collection and distance from the anal verge to the region where es were obtained were recorded in the CRF.

P213428WO 120 In the event that access to the sigmoid colon was limited, the 5 biopsies could have been obtained from the rectum.

Each biopsy was rinsed with saline, blot dried and then transferred to a pre-weighed collection tube.

The tube was weighed to enable determination of the biopsy . The biopsy, without any further preparation or processing was transferred to a cryovial and stored at -700C and transported to the bioanalytical laboratory on dry ice.

Concentrations of cyclosporin and its metabolites in the l tissue were assessed by a lytical tory. Analysis of the tissue samples was carried out as described below under "Colonic Tissue Analysis" Faecal Sample Collection Subjects were recommended to defaecate upon admission to the study centre on Day 0. This sample was collected and one aliquot was collected as a blank matrix.

From Day 0 through to completion of the study, subjects were requested to collect their faeces. For subjects who received Sandimmun® IV, samples collected during the infusion were kept separate from those collected after completion of the infusion.

The time and date of each sample were recorded. Each sample was collected and weighed. Faecal samples were homogenised as soon as possible following collection.

Subjects who produced faecal s as an outpatient were instructed to collect the samples in the riate containers and store the samples at room temperature until return to the study centre.

After homogenisation one aliquot of imately 5 g was ted from each sample and frozen at- 70°C (with no additives).

The three intracolonic samples ximately 500 mg to 1 g), if available, were collected and stored in individual containers without any additives at -70OC.

For subjects who were administered a rectal enema, the time and total weight of the voided gut contents post enema were recorded. A representative sample (approximately 5 g) was obtained from the voided gut and transferred to a container t any additives or additional processing.

All samples were stored at -700C and transported to a bioanalytical laboratory on dry ice.

Drug Concentration Measurements Blood samples were collected as described herein for determination of cyclosporin trations.

The amount of unchanged porin and relative concentrations of its metabolites (AM9, AM4N and AM1) in faeces were also assessed using faecal samples collected as described herein.

P213428WO 121 Primary Endpoints Whole blood pharmacokinetics The following single dose pharmacokinetic ters were derived after dosing on Day 1 for both the Sandimmun® IV and CyCol® groups using standard non-compartmental methods:  Cmax: maximum observed concentration  Tmax: time of ed Cmax  AUC0-t: area under the concentration-time curve from time zero to the last non-zero concentration, ined using the linear/log trapezoidal method  AUC0-inf: area under the tration-time curve from time zero to infinity (extrapolated) determined by AUC0-t + (Clast/kel),where Clast is the predicted concentration at the last quantifiable time point estimated from the log-linear sion analysis.  t½: elimination half-life calculated as loge(2)/kel  kel: elimination rate constant (derived to calculate AUC0-inf), determined by a linear regression of the log-linear concentration-time curve. Only those data points judged to describe the terminal log-linear e were used in the regression  Residual area (% extrapolated; AUC%extrap): calculated as100*(1-AUC0-t/ AUC0-inf) and derived to quantify the area extrapolated to infinity  AUC0-τ: area under the concentration-time curve over the dosing interval (derived to calculate the accumulation ratio discussed below).

The following steady state ters were derived for the CyCol® groups: - Cmax, Tmax, AUC0-τ and average steady state concentration (Cav) onally the accumulation ratio was derived to estimate the accumulation over the dosing interval from single dose to steady state.

AC = AUC0-τ (steady state) / AUC0-τ (single dose) The linearity ratio (LR) was derived to te the relative exposure per dose with steady state compared to a single dose.

LR = AUC0-τ (steady state) / AUCinf (single dose) For the mun® IV group simulations to steady state were conducted by the clinical pharmacokineticist to predict the concentration time profile and estimate Cmax, Tmax and AUC0-τ for the group (i.e., group values, not individual values).

To determine the steady state CyCol® parameters both the Day 6 and Day 7 concentrations were used as summarised below.

P213428WO 122 Time Days used for Days used for post-dose once daily BID 0 7 7 2 7 7 4 7 6, 7 6 6, 7 6, 7 8 7 7 12 6, 7 6, 7 16 6 NA 24 Day 7 pre-dose NA Actual sampling times were used in the derivation of parameters.

Cyclosporin concentrations were listed and summarised by group, day and nominal time post-dose.

Individual subject and median profiles of the concentration-time data (by day and by group) were plotted by dose group using actual and nominal times respectively. Median profiles were plotted on both -linear and log-linear.

Each pharmacokinetic parameter was ised by group and by day. For the single dose, pharmacokinetic parameters, the Day 1 parameters were summarised combining the once daily and BID groups using the same dose.

To assess the relationship n the pharmacokinetic parameters and dose, dose normalised AUC0-inf, AUClast and Cmax were plotted against dose for Day 1 and dose normalised Cav and Cmax were plotted against dose for Day 7 (using a logarithmic scale). These plots ed individual subject values and the geometric means for each dose. The values were dose ised (to a 1 mg dose) by dividing the individual values and raw ric means by dose.

Colonic mucosal tissue and mucous layer pharmacokinetics Concentrations of cyclosporin and its lites (AM9, AM4N and AM1) in both the colonic mucosal tissue and mucous layer were determined. For the Sandimmun® IV group simulations were conducted by the clinical pharmacokineticist to estimate the steady state concentrations.

Concentrations of cyclosporin and its metabolites (AM9, AM4N and AM1) were listed. Concentrations were also plotted against distance from the anal verge.

The concentration of cyclosporin in the colonic tissue was measured as follows.

Colonic Tissue Analysis Concentrations of cyclosporin and its lites (AM1, AM9 and AM4N) in colonic tissue was determined using the following protocol: P213428WO 123 Principle -liquid extraction with internal standardisation and HPLC tion using a C18-column, followed by MS/MS detection.

Internal Standard - D12 – Cyclosporin A Sample Matrix - Human Tissue Calibration standards and quality control samples are prepared in 50% EtOH.

Solutions The IS stock on and respective dilutions are ed by using DMSO/MeOH (1/1). The internal standard (IS) working solution is prepared by dilution of the IS stock solution or one of its dilutions with DMSO/MeOH (1/1), and should have a concentration of ~50 ng/mL Storing of Samples and Solutions s / solutions should be stored at -20°C to -80°C Sample Handling and Sample Preparation for Analysis Step Thawing / transfer procedure (step by step) The following thawing procedures are possible: • Thawing at approximately 20 to 25°C in a water bath for approx. 10 minutes • Thawing air exposed at approximately 20 to 25°C for at least 30 minutes (depends on sample volume) 2 If able: Vortexing for 30 seconds 3a Cal. Stds. & QCs: Transfer of 1000 µL of each sample into a sample vial 3b Study : weight: approx.. 2 – 20 mg Re-freezing of original samples between -20°C and -80°C Unless used for immediate preparation -> freezing of transferred samples between -20°C and -80°C Chromatographic and ampler Parameters Parameter Scheduled range / description Mobile phase solvent A 10 mM Ammonium acetate in water Mobile phase solvent B ACN/THF (8/2) Mobile phase solvent loading mM Ammonium acetate in water tographic run 0.0 – 4.5 min linear gradient: 40 % B → 52 % B 4.5 – 6.0 min linear nt: 52 % B → 85 % B 6.0 – 6.01 min linear gradient: 85 % B → 0 % B 6.01 – 7.0 min isocratic: 0 % B Flow 0.8 mL/min Injection volume 10 µL Pre-column / Column Luna C18, 4 x 2 mm / ACE3AQ; 100 x 2.1 mm, 3 µm (ACT, UK) P213428WO 124 Column temperature 80°C Parameter led range / description Cooling set point (T) 25°C Detection Parameter Scheduled range / ption MS Ionisation mode ESI MS polarity Positive MS detection mode MRM Vaporizer temperature 600°C Ionisation e 5.5 kV Gas 1 Pressure = 75 psi Gas 2 Pressure = 75 psi Curtain gas pressure = 40 psi Lateral position 5 units ± 2 units (default) Vertical position 4 units ± 2 units (ESI default) Quadrupole resolution low → low 1203.0 ± 0.3 → 99.9 ± 0.3 m/z: Cyclosporin A (CE: 125 eV, CXP: 16V) 1215.0 ± 0.3 → 99.9 ± 0.3 m/z: D12-Cyclosporin A Transitions (CE: 125 eV, CXP: 16V) 1219.0 ± 0.3 → 224.0 ± 0.3 m/z: AM1 (CE: 65 eV, CXP: 15V) 1219.0 ± 0.3 → 99.9 ± 0.3 m/z: AM9 (CE: 125 eV, CXP: 16V) 1189.0 ± 0.3 → 224.0 ± 0.3 m/z: AM4N (CE: 65 eV, CXP: 15V) DP (declustering potential) 130 V ± 20 V Acceptance Criteria for Chromatograms ter Scheduled range / acceptance criteria / description AM1 Retention time for SST 4.2 min ± 0.5 min AM4N Retention time for SST 5.4 min ± 0.5 min AM9 Retention time for SST 4.4 min ± 0.5 min Calibration rds and Quality Conrol Samples: P213428WO 125 ation of blank samples and processed : • Preparation as described below, but taking DMSO / MeOH (1/1) instead of IS working solution.

Step Preparation procedure (step by step) I [if not stored /available as 1000 µL aliquots already -> see transfer above] II [if frozen -> thawing at 20°C to 25°C in a water bath for approx.. 5 min] 1 Addition of 25 µL of internal standard working solution 2 Addition of 4 mL of DIPE Conversion Point : 3 Extraction by shaking the test tubes vigorously for approx. 5 minutes using a DVX- 2500 Multi-tube Vortexer (1700 rpm; cycle: 5 seconds run, 1 second pause time) 4 Centrifugation (phase separation) at 4000 rpm for 2 minutes Storage at -75°C for about 10 minutes 6 Decanting of the organic, liquid phase into a centrifuge vial 7 Evaporation of the organic phase using compressed air (Turbovap) at about 40°C for 14 minutes 8 Addition of 50 µL of 50 % EtOH Vortexing for approx. 2 minutes using a DVX-2500 tube Vortexer (2500 rpm; cycle: 5 seconds run, 1 second pause time) Centrifugation at 4000 rpm for 1 minute Carry-over samples: • Transfer of approx. 100 µL 50% EtOH into appropriate auto-sampler vials Matrix samples (human tissue) – PART A: Step Preparation procedure (step by step) I [if not stored /available as approx.. 2 - 20 mg ts already -> see transfer above] II [if frozen -> thawing at 20°C to 25°C in a water bath for approx.. 5 min] 1 Addition of 500 µL 2 % N-Acetyl-L-Cysteine in water 2 ing for approx. 10 min using a DVX-2500 Multi-tube er (1000 rpm) 3 Centrifugation (phase separation) at 13000 rpm for 2 minutes using biofuge pico 4 Decanting of the liquid phase into a sample vial (volume: . 10 mL) 4a Caution: The remaining residue will be prepared separately (described in part B) Addition of 500 µL EtOH to the liquid phase 9 Addition of 25 µL of internal standard working solution Addition of 4 mL of DIPE P213428WO 126 Conversion Point : 11 Extraction by shaking the test tubes vigorously for approx. 5 minutes using a DVX- 2500 Multi-tube Vortexer (1700 rpm; cycle: 5 seconds run, 1 second pause time) 12 Centrifugation (phase tion) at 4000 rpm for 2 s 13 Storage at -75°C for about 10 minutes 14 Decanting of the organic, liquid phase into a centrifuge vial Evaporation of the organic phase using compressed air (Turbovap) at about 40°C for 14 minutes 16 Addition of 50 µL of 50 % EtOH Vortexing for approx. 2 minutes using a DVX-2500 Multi-tube Vortexer (2500 rpm; cycle: 5 seconds run, 1 second pause time) 18 Centrifugation at 4000 rpm for 1 minute • Matrix s (human tissue) – PART B (Samples are taken from part A, step 4a): Step Preparation procedure (step by step) 1 Addition of 500 µL 50 % EtOH to the remaining residue 2 Addition of 25 µL of al standard working solution 3 Destroying of the tissue by using an ultrasonic processor : 0.5 s, max. amplitude) for 30 s 4 Decanting of the liquid phase into a sample vial (volume: approx. 10 mL) Addition of 500 µL 50 % EtOH to the remaining residue to the remaining residue ing for approx. 1 min using a DVX-2500 Multi-tube Vortexer (2500 rpm; cycle: 5 seconds run, 1 second pause time) 7 Decanting of the liquid phase including all tissue into the same sample vial as used in step 4 8 Addition of 4 mL of Diisopropylether (DIPE) sion Point : 9 tion by shaking the test tubes vigorously for approx. 5 minutes using a DVX- 2500 Multi-tube Vortexer (1700 rpm; cycle: 5 seconds run, 1 second pause time) Centrifugation (phase separation) at 4000 rpm for 2 minutes 11 Storage at -75°C for about 10 minutes 12 Decanting of the organic, liquid phase into a centrifuge vial 13 Evaporation of the organic phase using compressed air (Turbovap) at about 40°C for 14 minutes 14 Addition of 50 µL of 50 % EtOH Vortexing for approx. 2 minutes using a DVX-2500 Multi-tube er (2500 rpm; cycle: 5 seconds run, 1 second pause time) 16 Centrifugation at 4000 rpm for 1 minute P213428WO 127 Faceal Analysis The concentration of unchanged cyclosporin and the relative concentrations of the lites (AM9, AM4N and AM1) were reported for each intracolonic faecal sample and the faeces collected over the duration of the study.

If an enema was used prior to sigmoidoscopy then the appropriate faecal sample had the reported concentrations adjusted for the extra weight of enema used (multiply by [total faecal weight/totalenema ing collection of sigmoidoscopy biopsy samples, three intracolonic faecal s were taken from the region of the biopsy collection site to test for cyclosporin concentrations.

Amounts of unchanged cyclosporin and the metabolites were plotted for each subject against the collection time. The amount per hour was calculated and d for the collection interval. Hence the plot consists of stepped lines where the area under each step equates to the amount of drug measured for the collection interval.

Times since doses taken since the start of the collection interval were listed.

The concentrations from the intracolonic faecal samples taken after the sigmoidoscopy were listed with the colonic mucosal tissue and mucous layer concentrations.

Determination of cyclosporin-A and its metabolites, AM1, AM9 and AM4N, in faecal samples The faecal samples were analysed by the RP-LC-MS/MS method using the ol below.

Method: Preparation of Solutions and tion Samples Concentrated Solutions and Dilutions To spike calibration standards, quality control samples and other control samples, concentrates and dilutions were prepared as shown in the following table using the reference items and the internal standard with purities as described above. ons Used for Preparation Name / Preparation Conc. [µg/mL] date Cyclospo AM1 AM4N AM9 rin A 1640 5.27 mg of Cyclosporin A were dissolved 500.00 - - - in 10.3819 mL of DMSO / MeOH (1/1) K2733-1634 2.57 mg of AM1 were dissolved in 10 mL - 251.86 - - of DMSO / MeOH (1/1) K2731-1649 1.76 mg of AM4N were dissolved in 10 - - 149.60 - mL of DMSO / MeOH (1/1) 1648 5.03 mg of AM9 were dissolved in 10 mL - - - 367.19 of DMSO / MeOH (1/1) 420 µL of K2718-1640, 417 µL of K2733- V1-B-812 1634, 702 µL of K2731-1649, and 286 µL 21.000 10.503 10.502 10.502 of K2732-1648 were put together and filled up to 10 mL with DMSO / MeOH P213428WO 128 (1/1) Conc. [µg/mL] 1.9 mg of D12-Cyclosporin A were K2711-1577 dissolved in 10 mL of DMSO / MeOH D12-Cyclosporin A (1/1) V1-IS811 500 µL of K2711-1577 were filled up to 10 9.2720 mL of DMSO / MeOH 1/1 IS-WS814- 865 µL of V1-IS811 were mixed with Faeces 0.16013 50 mL of DMSO / MeOH 1/1 Calibration Standards and Quality Control s For analytical calibration purposes calibration standards and quality control samples were spiked with either a defined volume of a concentrated solution described above or a higher concentrated ation rd into 50 % Ethanol at eight concentration levels / three concentration levels respectively: Preparation of Calibration Standards Std Volume Added Matrix Matrix Conc. ] of of added solution volume Cyclospo AM1 AM4N AM9 solution [mL] rin A Std0B-812 --- ---- --- --- --- Std1B-812 102.6 Std4B-812 4 2.00 1.00 1.00 1.00 Std2B-812 65.2 Std5B-812 4 4.00 2.00 2.00 2.00 812 48.6 Std8B-812 4 12.0 6.00 6.00 6.00 Std4B-812 348 Std8B-812 4 80.0 40.0 40.0 40.0 Ethanol Std5B-812 48.1 V1-B-812 4 250 125 125 125 Std6B-812 97.6 V1-B-812 4 500 250 250 250 Std7B-812 148.2 V1-B-812 4 750 375 375 375 Std8B-812 200 V1-B-812 4 1000 500 500 500 QC-A2-812 187.4 QC-B2-812 6 5.00 2.50 2.50 2.50 QC-B2-812 63.4 V1-B-812 8 165 82.6 82.6 82.6 Ethanol 812 338 V1-B-812 8 851 426 426 426 Other Control Samples Preparation of Other Control Samples (about 1 mg Blank Faeces added per aliquot) Std Volume Added Matrix Matrix Conc. [ng/mL] of of added solution volume Cyclospo AM1 AM4N AM9 solution [mL] rin A QC-A2-812 187.4 QC-B2-812 6 5.00 2.50 2.50 2.50 QC-B2-812 63.4 V1-B-812 8 165 82.6 82.6 82.6 Ethanol 812 338 V1-B-812 8 851 426 426 426 Sample Preparation for Analysis (Processing) P213428WO 129 Calibration Standards and Quality Control Samples: Preparation of blank samples and (if applicable) sed :  Preparation as described below, but taking DMSO / MeOH (1/1) instead of IS working on.

Step Preparation procedure (step by step) 1 Addition of 0.4 mL of water 2 Addition of 25 µL of internal standard working solution 3 on of 4 mL of DIPE Extraction by shaking the test tubes vigorously for approx. 5 minutes using a DVX-2500 Multi-tube er (1700 rpm; cycle: 5 s run, 1 second pause time) Centrifugation (phase separation) at 4000 rpm for 2 minutes 6 Storage at -75°C for about 10 minutes 7 Decanting of the organic, liquid phase into a centrifuge vial Evaporation of the organic phase using compressed air (Turbovap) at about 40°C for 14 minutes 9 on of 750 µL of 50 % EtOH Vortexing for approx. 2 minutes using a DVX-2500 Multi-tube Vortexer (2500 rpm; cycle: 5 seconds run, 1 second pause time) 11 Centrifugation at 4000 rpm for 1 minute 12 Transfer of an volume adequate to injection purposes into appropriate auto-sampler vials 13 Crimping the vials with appropriate vial caps Matrix s (human faeces): Step Preparation procedure (step by step) I [if not stored /available as 100 mg aliquots already] II [if frozen -> g at 20°C to 25°C in a water bath for approx.. 5 min] 1 Fill up the volumetric flask (5 mL) with 50% EtOH 2 Vortexing for about 1 min using a vortex mixer 3 Wait e down) for 3 min 4 Transfer of 50 µL into a sample vial Addition of 950 µL 50 % EtOH 6 Vortexing for approx. 30 s using a 00 Multi-tube Vortexer (2500 rpm) 7 Transfer of 50 µL into a sample vial 8 Addition of 0.4 mL of water 9 Addition of 25 µL of internal standard working solution Addition of 4 mL of DIPE Extraction by g the test tubes vigorously for approx. 5 minutes using a DVX-2500 Multi- tube Vortexer (1700 rpm; cycle: 5 seconds run, 1 second pause time) 12 Centrifugation (phase separation) at 4000 rpm for 2 minutes 13 Storage at -75°C for about 10 minutes 14 Decanting of the organic, liquid phase into a centrifuge vial Evaporation of the organic phase using compressed air (Turbovap) at about 40°C for 14 minutes 16 Addition of 750 µL of 50 % EtOH Vortexing for approx. 2 minutes using a DVX-2500 Multi-tube Vortexer (2500 rpm; cycle: 5 seconds run, 1 second pause time) 18 Centrifugation at 4000 rpm for 1 minute 19 Transfer of an volume adequate to injection purposes into appropriate auto-sampler vials Crimping the vials with appropriate vial caps 8WO 130 Apparatus Instruments and Materials Instrument / material Code Manufacturer Work station API 6500 Mass Spectrometer 6500 Q-Trap AB SCIEX, USA/Canada Software Data acquisition Analyst 1.6.2 (AB SCIEX, USA/Canada) Data processing Analyst 1.6.2 (AB SCIEX, USA/Canada) Statistics and calculations Analyst 1.6.2 (AB SCIEX, USA/Canada) Lotus 123 (Lotus Corp, USA) Chromatographic Conditions and Detection ters Chromatographic Conditions Parameter Scheduled range / description Mobile phase solvent A 10 mM Ammonium acetat in water Mobile phase solvent B ACN/THF (8/2) Chromatographic run 0.0 – 4.5 min linear nt: 40 % B → 52 % B 4.5 – 6.0 min linear nt: 52 % B → 85 % B 6.0 – 6.01 min linear gradient: 85 % B → 0 % B 6.01 – 7.0 min isocratic: 0 % B Flow 0.8 mL/min Injection volume 10 μL Injector flush DMSO / MeOH / Water (1/1/1) Pre-column / Column Luna C18, 4 x 2 mm / ACE3AQ; 100 x 2.1 mm, 3 µm (ACT, UK) Column temperature 80°C Cooling set point (T) 25°C Detection Parameters Parameter Scheduled range / description MS tion mode ESI MS polarity Positive MS detection mode MRM Vaporizer temperature 600°C Ionisation voltage 5.5 kV Gas 1 Pressure = 75 psi Gas 2 Pressure = 75 psi Curtain gas pressure = 40 psi Quadrupole resolution low  low 1203.0  99.9 m/z: porin A Transitions (CE: 125 eV, CXP: 16V) 1215.0  99.9 m/z: D12-Cyclosporin A (CE: 125 eV, CXP: 16V) 1219.0  224.0 m/z: AM1 (CE: 65 eV, CXP: 15V) 1219.0  99.9 m/z: AM9 (CE: 125 eV, CXP: 16V) P213428WO 131 Parameter Scheduled range / description 1189.0  224.0 m/z: AM4N (CE: 65 eV, CXP: 15V) DP (declustering potential) 130 V Data Evaluation trations were evaluated using an internal standard method.

The concentrations of each analyte were determined using the following regression model, weighting factor and formula: Weighting Analyte Regression model a for concentration factor -b  b2  4a(c peakarearatio) all 4 y = ax2 + bx + c 1/conc. concentration Based thereon (arithmetic) mean values and relative standard deviations (CV) (formulas shown below) were calculated using the program "Lotus 123". xi calculated concentration 1 x mean calculated concentration standard deviation = ( − ̅) N number of values i index of value standard ion relative standard deviation (%) = ∗ 100 mean calculated concentration The concentration of porin A and the metabolites AM4N, AM9 in faecal samples collected on day 2 of the study is shown in Table 22.

Table 22: Total 2 Total 1 Mean Mean Mean CyA + Total AM4N+ Ratio Group CyA AM4N AM9 AM4N l AM9 CyA:AM4:AM9 (ng/g) (ng/g) (ng/g) +AM9 2 % (ng/g) (ng/g) IV Group 8 2194.52 3343.21 5537.73 8017.31 69.1% 0.45 IV Group 1215.75 1110.19 2942.91 4053.1 5268.85 77.9% 0.30 37.5mg 1918.8 3687.9 5606.7 356865.20 1.6% 62.65 OD 351258.5 .4 1419.76 2384.75 3804.51 126744.91 3.0% 32.31 159430.6 1069.01 2431.42 3500.43 162931.00 2.2% 45.55 150mg 14357.91 1082493.61 1.3% 74.39 BID 1068136 4388.83 9969.08 P213428WO 132 Pharmacokinetic Evaluation Demographic and Other Baseline Characteristics aphic teristics are summarised in Table 23. All subjects were male and aged between 19 and 54 years. Demographic characteristics were similar across the treatment groups; any differences were not considered to affect the results of the study.

Table 23 Demographics Sandimmun® CyCol® CyCol® CyCol® Sandimmun® CyCol® IV 75 mg 75 mg 150 mg IV 37.5 mg l Group 1 once BID BID Group 2 once daily N=48 N=8 daily N=8 N=8 N=8 N=8 Age, years Mean 30.4 29.4 37.4 38.8 32.1 31.9 33.3 SD 9.21 7.95 10.81 12.44 6.22 6.92 9.38 Median 27.0 28.5 36.5 36.5 32.5 30.0 32.0 Min, Max 23, 50 19, 39 21, 54 20, 54 22, 44 24, 45 19, 54 Age categories, n 18-30 5 (62.5) 4 (50.0) 2 (25.0) 2 (25.0) 3 (37.5) 5 (62.5) 21 (43.8) 31-55 3 (37.5) 4 (50.0) 6 (75.0) 6 (75.0) 5 (62.5) 3 (37.5) 27 (56.3) Mean 75.0 77.3 78.4 84.5 79.4 73.9 78.1 SD 9.28 12.93 15.61 13.91 11.14 11.79 12.41 Median 70.8 72.3 77.4 87.1 83.7 73.7 77.4 Min, Max 65, 89 66, 103 58, 100 64, 100 59, 91 54, 91 54, 103 Height, cm Mean 176.0 175.9 177.3 181.4 180.4 175.4 177.7 SD 5.76 6.75 9.33 9.47 5.07 6.05 7.27 Median 176.0 173.0 179.0 180.5 179.0 176.5 177.0 Min, Max 166, 185 169, 188 160, 191 171, 200 174, 188 163, 183 160, 200 Race, n (%) Black 0 0 1 (12.5) 1 (12.5) 2 (25.0) 1 (12.5) 5 (10.4) Caucasian 4 (50.0) 6 (75.0) 4 (50.0) 5 (62.5) 6 (75.0) 6 (75.0) 31 (64.6) Asian/Pacific 3 (37.5) 1 (12.5) 0 1 (12.5) 3 (37.5) 2 (25.0) 10 (20.8) Islander Mixed 1 (12.5) 0 0 1 (12.5) 0 0 2 (4.2) Ethnicity, n (%) Not Hispanic 8 (100.0) 7 (87.5) 8 (100.0) 8 (100.0) 8 (100.0) 8 (100.0) 47 (97.9) Hispanic 0 1 (12.5) 0 0 0 0 1 (2.1) Body mass index, kg/m2 Mean 24.2 24.6 24.9 25.6 24.4 23.9 24.6 SD 2.89 2.71 3.84 2.80 3.11 3.21 2.99 Median 23.5 23.5 25.8 26.1 24.1 23.2 24.1 Min, Max 21, 29 22, 29 19, 29 21, 30 20, 29 20, 29 19, 30 BID=twice daily; IV=intravenous; Max=maximum; Min=minimum; n=number of subjects in assigned category; N=number of subjects in Safety Population; SD=standard ion.

Percentages are based on the number of subjects in the Safety Population. 8WO 133 Pharmacokinetic Results and Tabulations of Individual t Data Analysis of cokinetics Cyclosporin Pharmacokinetics Median whole blood cyclosporin concentration-time profiles (linear-linear) for the CyCol® groups on Day 1 and Day 7 are provided in Figures 11 and 12 respectively. Cyclosporin concentrations generally increased with increasing dose of Cycol®.

Single dose pharmacokinetics Whole blood cyclosporin pharmacokinetic parameters following a single dose are summarised in Table 24. Exposure (AUC ® inf) was considerably lower following treatment with all doses of CyCol compared with Sandimmun® IV. Cyclosporin concentrations (AUClast and AUCinf) increased with increasing dose of CyCol®. Median T ® (between 5.0 and max was similar at all doses of CyCol .5 .

Median percent extrapolation was low in the two Sandimmun® IV groups (<5%). Median percent extrapolation was highest in the CyCol® 75 mg BID and 150 mg BID groups (24.7% and 28.9%, respectively.

Table 24 y of Single Dose Whole Blood porin Pharmacokinetic ters – PK Population Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 AUClast (ng.h/mL) n 8 8 8 8 Arithmetic mean 118.1 241.2 250.5 554.3 SD 62.10 158.71 162.89 355.15 Geometric mean 104.7 191.7 199.1 463.7 CV% 52.6 65.8 65.0 64.1 Median 96.9 208.5 228.0 437.5 55.6, 470 64.8, 146, Minimum, Maximum 46.4, 228 480 1231 AUCinf (ng.h/mL) n 8 8 8 8 8 8 Arithmetic mean 8836.0 8397.3 132.8 265.2 329.5 821.8 SD 1180.58 1609.76 70.81 163.83 208.07 460.76 Geometric mean 8769.5 8274.5 117.9 218.3 265.9 700.4 CV% 13.4 19.2 53.3 61.8 63.1 56.1 Median 8649.5 8110.5 105.9 228.0 314.5 754.0 55.0, 263 67.9, 502 100, 637 220, Minimum, Maximum 7457, 11088 6316, 11845 Cmax (ng/mL) n 8 8 8 8 Arithmetic mean 18.38 41.89 28.84 59.44 SD 9.810 32.574 33.206 54.306 Geometric mean 15.70 29.86 17.66 40.93 CV% 53.4 77.8 115.2 91.4 Median 8649.5 8110.5 105.9 228.0 314.5 754.0 P213428WO 134 Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Minimum, Maximum 7457, 11088 6316, 11845 55.0, 263 67.9, 502 100, 637 Cmax (ng/mL) n 8 8 8 8 Arithmetic mean 18.38 41.89 28.84 59.44 SD 9.810 32.574 33.206 54.306 Geometric mean 15.70 29.86 17.66 40.93 CV% 53.4 77.8 115.2 91.4 Median 17.70 30.40 14.40 36.60 6.03, 30.8 7.20, 86.9 3.73, 10.5, Minimum, m 101 156 t½ (h) n 8 8 Arithmetic mean 5.04 3.83 SD 0.753 0.556 CV% 14.9 14.5 Median 4.87 3.88 m, Maximum 4.25, 6.28 2.70, 4.42 Tmax (h) n 8 8 8 8 Median 5.0 5.5 5.0 5.5 Minimum, Maximum 4, 8 4, 12 4, 12 3, 8 AUC0-τ (ng.h/mL) n 8 8 Arithmetic mean 8448 8122 SD 1081.5 1562.3 Geometric mean 8389 8002 CV% 12.8 19.2 Median 8422 7867 Minimum, Maximum 7119, 10472 6054, 11465 F (%) n 8 8 8 8 Arithmetic mean 6.006 6.000 3.729 4.650 SD 3.2020 3.7087 2.3575 2.6082 ric mean 5.336 4.938 3.007 3.962 CV% 53.3 61.8 63.2 56.1 Median 4.790 5.150 3.560 4.265 2.49, 11.9 1.54, 11.4 1.13, 1.24, Minimum, Maximum 7.21 9.40 Cmin (ng/mL) n 8 8 8 8 Arithmetic mean 1.21 2.67 4.23 11.32 SD 0.691 0.886 1.979 7.461 Geometric mean 1.04 2.53 3.73 9.69 CV% 57.2 33.2 46.9 65.9 Median 1.12 2.44 4.26 8.74 0.333, 1.37, 4.00 1.17, 5.06, Minimum, Maximum 2.58 7.40 27.2 Percent extrapolated (%) n 8 8 8 8 8 8 Median 4.5 3.3 11.6 9.0 24.7 29.8 7.15, 15.6 5.75, 23.7 10.6, 17.2, Minimum, Maximum 1.64, 6.03 2.27, 4.14 .7 56.6 BID=twice daily; IV=intravenous; N=number of subjects in Safety Population.

P213428WO 135 Steady state cokinetics Whole blood cyclosporin pharmacokinetic parameters at steady state are summarised in Table 25.

Cyclosporin concentrations (AUC0-τ) sed with increasing dose of CyCol®. Median T max was similar at all doses of CyCol® (between 5.5 and 6.0 hours).

Table 25 y of Steady State Whole Blood Cyclosporin Pharmacokinetic Parameters – PK Population CyCol® CyCol® CyCol® CyCol® 37.5 mg 75 mg 75 mg 150 mg once daily once daily BID BID N=8 N=8 N=8 N=8 AUC0-τ mL) n 8 8 7 7 Arithmetic mean 128 225 298 585 SD 53.6 113.5 121.4 228.1 Geometric mean 117 190 266 539 CV% 41.9 50.4 40.7 39.0 Median 130 257 343 608 Minimum, Maximum 49.3, 223 54.2, 354 84.2, 410 228, 886 Cmax (ng/mL) n 8 8 7 7 Arithmetic mean 19.47 38.57 36.67 45.30 SD 10.155 24.028 13.866 25.682 Geometric mean 16.63 28.89 33.72 37.95 CV% 52.2 62.3 37.8 56.7 Median 21.25 48.50 41.10 48.40 Minimum, Maximum 6.23, 35.9 4.94, 75.4 14.1, 49.7 12.9, 82.1 Tmax (h) n 8 8 7 7 Median 5.5 6.0 6.0 6.0 Minimum, Maximum 5, 8 2, 6 1, 6 4, 8 Cmin (ng/mL) n 8 8 7 7 Arithmetic mean 1.11 2.68 5.69 10.06 SD 0.509 1.279 3.355 1.859 Geometric mean 1.01 2.39 4.84 9.90 CV% 46.0 47.8 58.9 18.5 Median 1.02 2.58 4.63 9.70 Minimum, Maximum 0.56, 1.99 1.17, 4.50 2.28, 10.4 6.90, 12.2 F (%) n 8 8 7 7 Arithmetic mean 5.789 5.099 3.374 3.313 SD 2.4267 2.5700 1.3748 1.2895 Geometric mean 5.313 4.300 3.012 3.049 CV% 41.9 50.4 40.7 38.9 Median 5.840 5.810 3.880 3.440 Minimum, Maximum 2.23, 10.1 1.23, 8.01 0.95, 4.64 1.29, 5.01 Cav (ng/mL) n 8 8 7 7 Arithmetic mean 5.33 9.38 24.86 48.77 SD 2.234 4.728 10.116 19.006 Geometric mean 4.89 7.91 22.20 44.89 CV% 41.9 50.4 40.7 39.0 Median 5.40 10.69 28.58 50.67 Minimum, Maximum 2.05, 9.29 2.26, 14.75 7.02, 34.17 19.00, 73.83 P213428WO 136 CyCol® CyCol® CyCol® CyCol® 37.5 mg 75 mg 75 mg 150 mg once daily once daily BID BID N=8 N=8 N=8 N=8 Linearity Ratio n 8 8 7 7 Arithmetic mean 1.295 1.403 0.968 0.883 SD 0.7862 1.4995 0.4929 0.3378 Geometric mean 0.996 0.870 0.871 0.823 CV% 60.7 106.9 50.9 38.3 Median 1.5 0.9 0.7 0.8 Minimum, Maximum 0.238, 2.309 0.154, 4.786 0.492, 1.762 0.422, 1.300 BID=twice daily; IV=intravenous; er of subjects in Safety tion.

Colon Tissue Distribution Cyclosporin distribution Tissue, mucous and intracolonic cyclosporin trations are summarised in Table 26.

Tissue cyclosporin concentrations generally increased with increasing dose of CyCol® and concentrations were higher in the CyCol® 75 mg BID and 150 mg BID groups than in the Sandimmun® IV groups. There was no relationship between tissue cyclosporin concentrations and distance from anal verge.

Mucous cyclosporin concentrations were higher in the CyCol® 150 mg group compared with the other CyCol® groups and the Sandimmun® IV groups. There was no relationship between mucous cyclosporin concentrations and ce from anal verge.

Intracolonic faecal cyclosporin concentrations generally increased with sing dose of CyCol® and were considerably higher in all of the CyCol® groups ed with the Sandimmun® IV groups.

Table 26 Summary of Tissue, Mucous and Intracolonic Faecal Cyclosporin Concentrations – PK Population Sandimmun CyCol® CyCol® CyCol® CyCol® Sandimmun ® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Tissue cyclosporin concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 802 1100 1045 1094 1579 5210 SD 397.0 276.1 712.5 881.3 502.3 4417.5 Geometric mean 717 1071 863 752 1493 4043 CV% 49.5 25.1 68.2 80.6 31.8 84.8 Median 789 974 727 906 1576 3114 Minimum, 326, 836, 352, 139, 669, 1945, Maximum 1487 1541 2431 2497 2279 14269 Mucous cyclosporin concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 73.68 126.93 103.56 101.09 78.27 596.38 SD 47.311 60.086 58.416 63.478 40.067 458.366 Geometric mean 60.46 116.36 90.72 70.42 69.93 457.16 CV% 56.0 43.6 49.0 62.2 43.1 98.2 P213428WO 137 Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Median 1535 3230 96957 277626 325550 294153 Minimum, 955, 1885, 66664, 8212, 142242, 88100, Maximum 3748 6529 234285 433770 557300 1569913 Table 26 shows that oral administration of the CyCol™ compositions comprising the surfactant provided similar or higher cyclosporin A concentrations in c tissue compared to IV administration of 2mg/kg cyclosporin. However, the CyCol™ compositions resulted in significantly lower systemic re compared to IV stration of cyclosporin (see the AUC and Cmax values in Tables 24 and ). Reference to Table 22 also shows that the CyCol™ compositions resulted in much lower cyclosporin lism as evidenced by the high ratio of cyclosporin to the AM4+AM9 metabolites in collected faecal samples.

AM1 concentrations Tissue, mucous and olonic faecal AM1 concentrations are summarised in Table 27.

Tissue AM1 concentrations increased with increasing dose of CyCol® but were slightly lower in the CyCol® 150 mg BID compared with the Sandimmun® IV groups. There was no relationship between tissue AM1 concentrations and distance from anal verge.

Mucous AM1 concentrations were similar in the CyCol® 150 mg BID and mun® IV groups and lower in the other CyCol® groups. There was no relationship between mucous AM1 concentrations and distance from anal verge.

Intracolonic faecal AM1 concentrations were r in all treatment groups.

Table 27 y of Tissue, Mucous and Intracolonic Faecal AM1 Concentrations – PK Population Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Tissue AM1 concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 517.6 345.2 70.0 101.7 232.7 303.7 SD 294.49 89.95 42.48 74.12 198.53 156.50 Geometric mean 453.4 334.5 61.8 82.5 169.3 272.5 CV% 56.9 26.1 60.7 72.9 85.3 51.5 Median 534.9 320.3 63.0 75.9 155.5 330.7 Minimum, 217, 205.9, 34.3, 27.8, 38.0, 132.6, Maximum 1143.0 467.8 166.3 258.2 624.6 605.0 P213428WO 138 Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Mucous AM1 concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 304.3 307.3 60.3 67.0 56.9 233.8 SD 146.10 163.66 35.63 33.47 43.74 102.73 Geometric mean 275.6 276.5 53.7 61.0 46.1 215.7 CV% 48.0 53.3 59.1 50.0 76.8 43.9 Median 268.1 245.1 48.6 61.2 44.3 203.3 Minimum, 131.2, 169.4, 31.6, 32.9, 21.2, 122.0, Maximum 566.1 568.8 141.1 138.9 145.4 378.6 Intracolonic faecal AM1 concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 2472 4944 2939 4719 5386 5870 SD 1477.2 3375.2 645.8 2618.3 5528.3 3249.3 Geometric mean 2094 4029 2876 3921 3564 5236 CV% 59.8 68.3 22.0 55.5 102.6 55.4 Median 2160 3322 2938 4424 4441 4933 Minimum, 1027, 1885, 2158, 1002, 948, 3041, Maximum 4712 10456 3749 8838 16997 12251 AM4N concentrations Tissue, mucous and intracolonic faecal AM4N concentrations are summarised in Table 28.

Tissue AM4N concentrations generally increased with increasing dose of CyCol®. Concentrations in the CyCol® 150 mg BID were lower compared with the mun® IV groups. There was no relationship between tissue AM4N concentrations and distance from anal verge.

Mucous AM4N concentrations were higher in the CyCol® 150 mg BID group compared with the other CyCol® groups but concentrations were highest in the Sandimmun® IV groups. There was no relationship between mucous AM4N concentrations and distance from anal verge. Intracolonic faecal AM4N concentrations were r in all treatment groups.

Table 28 Summary of Tissue, Mucous and Intracolonic Faecal AM4N Concentrations – PK Population Sandimmun® CyCol® CyCol® CyCol® CyCol® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Tissue AM4N trations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 19.816 24.272 3.013 3.862 3.657 9.686 SD 23.4043 28.1267 2.8647 3.4242 2.8473 5 Geometric mean 11.746 11.188 2.224 2.247 2.809 6.146 CV% 118.1 115.9 95.1 88.7 77.9 113.4 Median 12.011 15.464 1.581 3.014 2.246 4.523 Minimum, 3.49, 1.36, 1.12, 0.40, 0.78, 2.30, Maximum 71.45 81.84 8.99 8.41 9.11 31.58 P213428WO 139 Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Mucous AM4N concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 3.787 10.458 1.194 1.338 0.632 2.931 SD 1.7635 5 0.9826 0.8601 0.4757 1.8756 Geometric mean 3.440 4.332 0.913 0.974 0.513 2.510 CV% 46.6 138.0 82.3 64.3 75.2 64.0 Median 3.291 5.455 0.797 1.275 0.385 2.179 Minimum, 1.64, 0.40, 0.34, 0.19, 0.28, 1.41, Maximum 6.95 43.78 2.85 2.31 1.44 5.90 Intracolonic faecal AM4N concentrations (ng/g) n 8 8 8 8 7 7 Arithmetic mean 1390 2739 2670 2143 2134 2788 SD 665.9 983.7 453.1 819.3 1554.1 1123.3 Geometric mean 1285 2599 2639 1987 1768 2627 CV% 47.9 35.9 17.0 38.2 72.8 40.3 Median 1119 2536 2599 2282 2020 2685 Minimum, 875, 1785, 2158, 895, 815, 1842, Maximum 2863 4554 3425 3171 5361 5060 AM9 concentrations Tissue, mucous and intracolonic faecal AM9 trations are summarised in Table 29.

Tissue AM9 concentrations generally increased with increasing dose of CyCol®. Concentrations in the CyCol® 150 mg BID were lower compared with the Sandimmun® IV groups. There was no relationship between tissue AM4N concentrations and distance from anal verge.

Mucous AM9 concentrations were higher in the CyCol® 150 mg BID group compared with the other CyCol® groups but concentrations were t in the Sandimmun® IV groups. There was no relationship between mucous AM9 concentrations and distance from anal verge. olonic faecal AM9 concentrations were r in all treatment groups.

Table 29 Summary of Tissue, Mucous and Intracolonic Faecal AM9 Concentrations – PK Population Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Tissue AM9 concentrations (ng/g) N 8 8 8 8 7 7 Arithmetic mean 64.41 80.81 11.81 15.80 28.53 50.28 SD 57.022 74.901 8.842 12.491 36.438 44.724 Geometric mean 45.98 53.22 9.57 11.38 18.15 36.42 CV% 88.5 92.7 74.9 79.1 127.7 88.9 Median 38.90 49.76 8.37 13.10 13.52 23.97 Minimum, 15.21, 15.18, 4.49, 3.17, 5.38, 15.92, Maximum 159.11 208.30 26.72 37.45 109.85 120.30 P213428WO 140 Sandimmun® CyCol® CyCol® CyCol® CyCol® Sandimmun® IV 37.5 mg 75 mg 75 mg 150 mg Group 1 once once BID BID Group 2 N=8 daily daily N=8 N=8 N=8 N=8 Mucous AM9 concentrations (ng/g) N 8 8 8 8 7 7 Arithmetic mean 31.48 193.81 9.20 11.31 9.62 29.57 SD 16.406 180.450 6.570 7.819 9.460 18.643 ric mean 28.40 134.27 7.70 8.88 7.37 25.14 CV% 52.1 93.1 71.4 69.1 98.3 63.0 Median 27.82 100.41 6.45 9.21 6.75 20.38 Minimum, 15.76, 30.83, 4.23, 2.82, 3.77, 10.73, Maximum 66.11 558.92 21.96 25.20 30.64 61.55 olonic faecal AM9 concentrations (ng/g) N 8 8 8 8 7 7 Arithmetic mean 2254 5646 3637 5479 5862 6351 SD 1121.2 4390.0 1215.2 3550.3 6721.1 4584.7 Geometric mean 2002 4279 3464 4292 3820 5296 CV% 49.7 77.8 33.4 64.8 114.7 72.2 Median 2116 3226 3352 6123 4023 5516 Minimum, 1032, 1878, 2158, 912, 953, 2438, Maximum 3916 11714 5396 12100 20598 15941 Pharmacokinetic Conclusions  Systemic exposure to cyclosporin A was lower ing treatment with CyCol® at doses up to 150 mg BID once daily for 7 days compared with a single IV on of Sandimmun® 2 mg/kg over 24 hours (2 mg/kg/day).

 Tissue and mucous concentrations of cyclosporin A were higher in the CyCol® 75 mg BID and 150 mg BID groups ed with the Sandimmun® IV groups suggesting that these doses should be efficacious. Tissue and mucous concentrations of AM1, AM4N and AM9 were generally higher following treatment with Sandimmun® IV compared with CyCol®.  olonic faecal trations of cyclosporin A were considerably higher in the CyCol® groups compared with the Sandimmun® IV groups indicating that CyCol® is predominantly excreted in faeces.

SION AND OVERALL CONCLUSIONS Systemic exposure to cyclosporin was lower following treatment with CyCol® at doses up to 150 mg BID once daily for 7 days compared with a single IV infusion of Sandimmun® 2 mg/kg over 24 hours (2 mg/kg/day). This may result in a lower incidence of side effects related to cyclosporin following treatment with CyCol® compared with Sandimmun®.

Tissue and mucous concentrations of cyclosporin were higher in the CyCol® 75 mg BID and 150 mg BID groups compared with the Sandimmun® IV groups suggesting that these doses should be efficacious. Tissue and mucous concentrations of AM1, AM4N and AM9 were generally higher following treatment with mun® IV compared with CyCol®.

Intracolonic faecal concentrations of cyclosporin were considerably higher in the CyCol® groups compared with the Sandimmun® IV groups indicating that CyCol® is predominantly excreted in faeces.

P213428WO 141 Administration of CyCol® at doses up to 150 mg BID was generally well tolerated; the majority of AEs were mild and no severe or serious AEs were reported. There was a higher incidence of gastrointestinal disorders at the highest dose of CyCol® but none led to discontinuation.

Comparative Example 10: In -Vitro Study Using Minibead Composition Comprising Cremophor Preparation of ad Modified e Compositions Minibead Formulations I and II and a Fast Release Formulation were prepared using an analogous process to that described above.

Formulation I: "Medium" coating level (10% weight gain Opadry subcoat; 11% weight gain Surelease™/Pectin overcoat) Component % Core Cyclosporin A 8.8 Miglyol 810 N 3.8 Transcutol HP 13.5 Kolliphor™ EL 7.6 SDS 3.3 Sorbitol 4.7 Gelatin 40.3 at Opadry 8.2 Overcoat Surelease™ (solid contents) 9.7 Pectin 0.2 Formulation II: "High" coating level (10% weight gain Opadry subcoat; 17% weight-gain ase™/Pectin overcoat) Component % Core Cyclosporin A 8.4 l 810 N 3.6 utol HP 12.8 Kolliphor™ EL 7.2 SDS 3.1 Sorbitol 4.4 Gelatin 38.3 Sub-coat Opadry 7.8 Overcoat Surelease™ (solid contents) 14.2 Pectin 0.3 P213428WO 142 Fast Release Formulation (No Surelease™ Pectin overcoat) Component % Core Cyclosporin A 9.8 Miglyol 810 N 4.2 Transcutol HP 14.9 Kolliphor™ EL 8.4 SDS 3.6 Sorbitol 5.2 Gelatin 44.8 Sub-coat Opadry 9.1 Human Pharmacokinetic Study Study Objectives: Objective 1: To compare the rate and extent of absorption of cyclosporin-A following administration of the Fast Release Formulation (fast-release capsule; Test 1), Formulation I (mediumrelease capsule; Test 2), and ation II (slow-release capsule; Test 3) with Neoral™ immediaterelease capsule ence), administered as a single 75 mg dose under fasting conditions.

] Objective 2: To evaluate the amount of unchanged cyclosporin-A excreted in the faeces after administration of the Comparative Formulation (fast-release capsule; test 1), Formulation I (mediumrelease capsule; Test 2), Formulation II (slow-release capsule; Test 3) versus Neoral, administered as a single 75 mg dose under fasting ions.

Study Design: A single centre, ised, single-dose, open-label, od, 4-sequence crossover comparative BA study, performed under fasting conditions.

Subjects: ed and randomised: 18 (12 females and 6 males) Withdrew consent: 0 Withdrawal: 1 (was withdrawn) Completed all 4 periods: 16 Safety population: 18 Pharmacokinetic (PK) population: 18 Pharmacokinetic Analysis The mean pharmacokinetic values obtained in the study are summarized in the Table 30.

Table 30: Summary of pharmacokinetic parameters for cyclosporin-A for each treatment P213428WO 143 Mean ± SD Whole Blood Cyclosporin-A (CV%) Cyclosporin-A Cyclosporin-A Cyclosporin-A Neoral (Test 1) (Test 2) (Test 3) Fast e Formulation I Formulation II Formulation N 17 17 18 17 1582.20 ± AUC0-t 2 ± 297.62 609.89 ± 280.15 408.49 ± 231.01 358.09 (ng•hr/mL) (24.55) (45.93) (56.55) (22.63) 1639.78 ± AUC0-inf 1257.83 ± 312.14 672.07 ± 296.71 474.37 ± 247.93 371.52 (ng•hr/mL) (24.82) ) (52.27) (22.66) 594.66 ± Cmax 321.33 ± 87.61 138.28 ± 63.54 82.81 ± 48.01 117.01 (ng/mL) (27.27) (45.95) (57.98) (19.68) Residual 3.55 ± 0.71 10.72 ± 8.10 15.38 ± 12.69 3.52 ± 0.77 (%) (20.12) (75.50) (82.52) (21.87) Tmaxa 2.00 5.00 5.00 1.25 (hr) (1.25 - 3.00) (5.00 - 8.00) (5.00 - 10.0) (1.00 - 1.75) 0.1037 ± Kel 0.1105 ± 0.0113 0.0863 ± 0.0259 0.0822 ± 0.0232 0.0103 (1/hr) (10.25) (30.01) (28.20) (9.97) T½ el 6.33 ± 0.61 8.72 ± 2.76 9.49 ± 4.55 6.75 ± 0.77 (hr) (9.70) (31.66) (47.96) (11.43) a Median (Min - Max) In Table 30 the AUC and Cmax values are the mean value ± standard deviation (SD) The whole blood concentration of cyclosporin A for each of these composition is shown in Figure 13.

A comparison of Figures 11 and 12 using the minibeads of the present invention containing Capmul Capmul GMO-50 (glyceryl monooleate/dioleate) as the surfactant with Figure 13 (and the corresponding data tables) shows that the Capmul containing composition according to the ion provided lower Cmax and AUC values indicating lower ic exposure to cyclosporin A.

Determination of cyclosporin-A and its metabolites, AM9 and AM4N, in faecal samples P213428WO 144 Faecal samples collected during the PK trial were analysed by RP-LC-MS/MS as described previously. The results are shown in Table 31.

Table 31: Total 2 Mean Total 1 Total Mean Mean CyA + Ratio CSA% AM4N+ 1/Total AM4N% AM9% AM4N CyA:AM4:AM9 AM9% 2 % +AM9% 73.8 12 14.2 26.2 100 26.20% 2.82 Release Formulation 86.9 5.5 7.6 13.1 100 13.10% 6.63 ation 91.5 3.4 5.1 8.5 100 8.50% 10.76 Neoral 37.1 26.2 36.7 62.9 100 62.90% 0.59 A comparison of the faecal data in Table 31 with the faecal data obtained with the Capmul containing compositions of Example 9 (Table 22) shows that the Capmul itions of Example 9 resulted in much lower cyclosporin metabolism than the comparative compositions containing Cremophor. This is illustrated by the lower relative % of the (AM9 + AM4N) metabolites in the collected faeces for the Capmul compositions in Table 22 compared to the Cremophor itions in Table 31. This difference is clearly illustrated in Figure 14 which shows the ratio of cyclosporin to (AM4N + AM9) measured in the faecal samples for each of the tested formulations.

The compositions of the ion comprising Capmul GM0-50 resulted in significantly less cyclosporin A metabolism compared to the compositions containing Cremophor. The compositions of the invention therefore provide higher local levels of cyclosporin in the colon as a result of the reduced systemic and non-systemic metabolism of the cyclosporin released from the composition. The compositions of the invention may enable a lower dose of cyclosporin to be administered whilst maintaining a therapeutic , thereby further increasing the therapeutic window.

Figure 15 es the in-vitro dissolution profile of the Capmul formulation used in Example 9 with the hor compositions used in Comparative Example 10 using the two-stage dissolution test bed herein. Figure 15 shows that the release profiles for the Capmul composition, the Slow Release, and the Medium Release are all very similar in the 2 to 5 hour period. During this time t he compositions are expected to release cyclosporin in the small intestine and would be prone to both systemic and enteric P450 metabolism of the cyclosporin. Despite the similarities in the in-vitro release profiles, Figure 14 shows that the Capmul composition of the present invention icantly reduced the metabolism of cyclosporin A compared to the Fast, Medium and Slow compositions containing Cremophor as a tant. Additionally the Capmul composition exhibited a lower AUC and Cmax illustrating a lower ic exposure to cyclosporin than the Cremophor compositions following oral administration (see Tables 24 and 30).

P213428WO 145 Example 11: Emulsion Stability and Bead Formation: Effect of Surfactant and Surfactant Concentration itions with differing aqueous phase tants were investigated. The different s phase surfactants were compared to SDS. Three different families of surfactant were chosen to test: Sucrose Fatty Acid Esters, Sodium n-Alkyl Sulfates and Fatty Alcohol Ethoxylates (Brij series). ons were prepared by mixing an oil phase and an aqueous phase, as described in Example 1.

The oil phase was consistent for all emulsions of this example. The emulsion aqueous phase mixed was one of three s mixtures. The three aqueous phases differed in the amount of surfactant, 0.7%, 1.3 % and 2.5%, and water. The oil and aqueous phase mixtures are shown in tables 32 to 34.

Table 32 Aqueous Phase Oil Phase Component % Component % H2O 79.56% Transcutol HP 37.00% Gelatin 17.14% Capmul GMO 50 26.00% D-Sorbitol 2.00% CyA 24.50% Surfactant 1.30% Miglyol 12.50% Table 33 Aqueous Phase Oil Phase Component % Component % H2O 79.62% Transcutol HP 37.00% Gelatin 17.14% Capmul GMO 50 26.00% D-Sorbitol 2.00% CyA 24.50% Surfactant 0.70% Miglyol 12.50% Table 34 Aqueous Phase Oil Phase Component % Component % H2O 78.36% Transcutol HP 37.00% Gelatin 17.14% Capmul GMO 50 26.00% D-Sorbitol 2.00% CyA 24.50% Surfactant 2.50% Miglyol 12.50% P213428WO 146 The on was prepared by mixing the oil phase and aqueous phase in a ueous phase ratio of 1:5. The experiments were performed in triplicate, N = 3. In order to ate current cturing ions as much as possible the following procedure was carried out: - Identical size beakers were used to ensure minimal variation between experiments - cal submersible magnetic stirrers were placed in each beaker to keep the emulsions under constant ic stirring, and the same rpm stir rate used for each - Hot plates were used to maintain a constant emulsion temperature of 65 o C - Emulsions were kept covered with aluminium foil during the process Sampling of the emulsion was carried out at various time points using disposable pipettes to avoid cross contamination. on Analysis Samples were withdrawn from each emulsion at half hour time intervals starting at time zero (t0) using a disposable pipette. Drops of emulsion were placed on a glass slide, pre heated on a hot plate. The sample was covered with a cover slip and a small amount of pressure applied to form a thin film of emulsion. The samples were allowed to solidify to room temperature before being viewed at x50 and x100 objective lens under polarised light to check for the presence of crystals. Results were recorded as optical photomicrographs with three images taken of each emulsion at each time point. Images documenting crystal growth and size with respect to time within the sample emulsion as well as background oil droplet size were collected. Table 35, shown below, gives the time point when crystallisation was observed.

Photomicrographs of the emulsions enabled the investigation of the droplet size as well as crystal formation. Smaller, more uniform droplet size was preferred. These preferred droplet characteristics were observed in some of the tested ons, particularly the emulsions containing Sodium n-Octyl Sulfate and Sodium n-Octadecyl Sulfate.

The icrographs of an emulsion for each surfactant at specified time points are shown in Figures 16 to 22.

Bead ion Bead formation was attempted, to examine how bead formation was affected by the different surfactants. Samples (1ml) were withdrawn from each emulsion using a Gilson pipette when the on stability experiment had been terminated, before the emulsions were discarded. Beads were formed by dropping the on at a steady rate into a cooling bath of medium chain triglyceride oil which was kept refrigerated. Beads were retrieved using a sieve and placed on tissue paper within a container, gently patted with tissue paper to remove excess surface oil and left to dry overnight on the worktop at room temperature. Bead formation was deemed to have occurred if a spherical or nearly spherical bead was formed by dropping an emulsion into a cooling oil or expelling the emulsion when P213428WO 147 under the surface of the g oil. Formation of oval or elongated beads was not deemed as bead ion. It has to be acknowledged that due to the manual nature of the method of producing beads employed here compared with bead production using the Spherex equipment at a manufacturing scale, formulations which may not form beads at this lab scale study may in fact form beads when processed using the Spherex equipment.

Table 35 Concentration Emulsion Beads tant in aqueous Onset Crystallisation images in Formation phase % w/w Figure sodium dodecyl 1.4 (t = 2 hours Y sulphate 0.7 (t = 2 hours) Y 16A Sucrose Laurate 1.3 (t = 1 hour 30 mins) N 16B 2.5 (t = 1 hour ) N 16C 0.7 (t = 2 hours 30 mins) Y 17A Sucrose Palmitate 1.3 (t = 1 hour 30 mins ) Y 17B 2.5 (t = 30 mins) N 17C Sodium n- 0.7 (t = 2 hours 30 mins) Y 18A Octyl 1.3 (t = 4 hours) Y 18B Sulphate 2.5 (t = 1 hour 30 mins) Y 18C Sodium n- 0.7 (t = 3 hours) Y 19A Octadecyl 1.3 (t = 2 hours) Y 19B Sulfate 2.5 (t = 2 hours) Y 19C Brij L4 0.7 (t = 1 hour 30 mins) Y 20A (Polyethylene 1.3 (t = 2 hours 30 mins) N 20B Glycol Dodecyl Ether) 20C 2.5 (t = 1 hour 30 mins) N Brij C10 (PEG 0.7 (t = 1 hour 30 mins) N 21A Hexadecyl 1.3 (t = 1 hour) N 21B Ether) 2.5 (t = 1 hour) N 21C Brij S10 (PEG 0.7 (t = 1 hour) Y 22A Octadecyl 1.3 (t = 1 hour) N 22B Ether) 2.5 (t = 1 hour) N 22C P213428WO 148 The anionic surfactants Sodium n-Octyl Sulfate and Sodium n-Octadecyl Sulfate in particular ed stability that was comparable to that of sodium dodecyl sulfate. Sodium n-Octadecyl Sulfate provided the best stability for each concentration. Non-ionic surfactants provided on stability but with more rapid onset of crystallization compared to SDS, except for Brij L4 at 1.3% aqueous concentration which gave an increase in on stability.

Claims

1. An oral composition sing porin, a hydrogel forming polymer matrix, a surfactant and an oil phase being dispersed in the hydrogel forming polymer matrix, n the surfactant comprises a medium chain or long chain fatty acid mono- or di-glyceride or a combination 5 thereof and does not comprise a polyethyleneglycol ether or ester; wherein the hydrogelforming polymer matrix is in an amount of at least 25% by weight based upon the dry weight of the composition; and wherein the hydrogel forming r matrix comprises one or more of gelatin, agar, , casein, chitosan, soya bean protein, safflower protein, alginates, gellan gum, carrageenan, xanthan gum, phthalated gelatin, succinated gelatin, cellulosephthalate- 10 acetate, oleoresin, polyvinylacetate, polymerisates of acrylic or methacrylic esters and polyvinylacetate-phthalate.

2. The composition of claim 1 wherein the oil phase comprises a solution of the cyclosporin.

3. The composition of claim 1 or claim 2, n the composition comprises the surfactant comprising a medium chain or long chain fatty acid mono- or di-glyceride or combination thereof 15 that does not comprise a hyleneglycol ether or ester present in an amount of more than 6 wt% of the dry weight of the composition.

4. The composition of any one of claims 1 to 3, wherein the surfactant comprises a surfactant selected from: glyceryl prate, glyceryl dicaprate, glyceryl prylate, glyceryl dicaprylate, 20 yl monocaprylate/caprate, glyceryl dicaprylate/caprate, glyceryl monooleate/dioleate, glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitostearate, glyceryl dipalmitostearate, glyceryl monobehenate, yl dibehenate, glyceryl monolinoleate, glyceryl dilinoleate, polyglyceryl dioleate, and a combination thereof. 25

5. A composition of claim 4, wherein the surfactant is selected from: glyceryl monocaprylate/caprate, glyceryl dicaprylate/caprate, glyceryl monooleate, yl dioleate, glyceryl monostearate, glyceryl rate, glyceryl monopalmitostearate, glyceryl dipalmitostearate, glyceryl monobehenate, glyceryl dibehenate, yl monolinoleate, glyceryl dilinoleate, polyglyceryl dioleate and a combination thereof. 30

6. The ition of any one of claims 1 to 3, wherein the surfactant comprises a surfactant selected from: yl caprylate/caprate, glyceryl monooleate/dioleate, glyceryl monolinoleate, and a combination f; or wherein the surfactant ses glyceryl caprylate, glyceryl caprate, glyceryl monooleate, glyceryl dioleate, glyceryl monolinoleate or a combination thereof. 35

7. The composition of any one of the preceding claims, wherein the oil phase comprises a liquid lipid.

8. The composition of claim 7, wherein the oil phase comprises a solvent miscible with the liquid lipid.

9. The composition of claim 7 or claim 8, wherein the liquid lipid comprises a glyceride composition. 5

10. The ition of claim 9, wherein the liquid lipid comprises a medium chain fatty acid ceride or a combination thereof, or the liquid lipid ses a caprylic/capric triglyceride composition.

11. The composition of any one of claims 1 to 6, n the oil phase comprises an oil selected from a vegetable oil and a petrochemical oil. 10

12. The composition of any one of claims 1 to 6, wherein the oil phase comprises a polyunsaturated fatty acid.

13. The composition of claim 12, wherein the poly-unsaturaturated fatty acid is ed from omega-3 oils.

14. The composition of claim 13, wherein the omegaoil is selected from eicosapentanoic acid, 15 hexaenoic acid, alpha-linoleic acid and conjugated linoleic acid.

15. The composition of any of claims 1 to 6, wherein the oil phase comprises an oil selected from olive oil, sesame oil, coconut oil, palm kernel oil and neem oil.

16. The composition of any of claims 1 to 6, wherein the oil phase ses an oil selected from caprylic/capric triglyceride; caprylic/capric/linoleic triglyceride; caprylic/capric/succinic 20 triglyceride; and propylene glycol dicaprylate/dicaprate.

17. The composition of any of claims 1 to 6, wherein the oil phase comprises an oil with an HLB of from 0 to 10.

18. The composition of claim 17, wherein the oil phase comprises an oil with an HLB of from 0 to 5, 0 to 3, or 0 to 2. 25

19. The composition of any of claims 1 to 6, wherein the oil phase comprises an oil comprising a medium chain triglyceride and 2-(ethoxyethoxy)ethanol.

20. The composition of any one of claims 1 to 19, wherein the composition further comprises a solvent, wherein the solvent is miscible with the oil phase and water.

21. The composition of claim 20, wherein the solvent is selected from 2-(2-ethoxyethoxy)ethanol 30 and a poly(ethylene glycol).

22. The composition of claim 21, wherein the solvent is thoxyethoxy)ethanol.

23. The composition of any one of claims 1 to 22, wherein the composition r comprises a high HLB surfactant having an HLB value of at least 10.

24. The composition of claim 23, n the surfactant is an c surfactant.

25. The composition of claim 22 or claim 23, wherein the high HLB surfactant is selected from fatty acid salts and bile salts.

26. The ition of claim 25, wherein the high HLB surfactant is an alkyl sulphate. 5

27. The composition of claim 26, wherein the alkyl sulphate issodium dodecyl sulphate.

28. The composition of any one of claims 1 to 27 wherein the hydrogel forming polymer matrix comprises a hydrocolloid selected from eenan, gelatin, and agar, or a combination thereof.

29. The ition of claim 28, wherein, the polymer of the hydrogel forming r matrix 10 comprises gelatin.

30. The ition of any one of claims 1 to 29, wherein the surfactant is present in an amount of more than 12 wt% of the oil phase.

31. The composition of claim 1 in the form of a solid colloid, the colloid comprising a continuous phase and a disperse phase, 15 wherein the disperse phase comprises: cyclosporin in an amount of 60 – 180 mg/g; caprylic/capric triglyceride in an amount of 40 – 80 mg/g; 2-(2-ethoxyethoxy) l in an amount of 100 – 200 mg/g; and glyceryl monooleate and/or glyceryl dioleate in an amount of 100 – 150 mg/g; 20 and wherein the continuous phase comprises: a hydrogel-forming polymer matrix comprising gelatin in an amount of 300 – 700 mg/g; sodium dodecyl sulphate in an amount of 15 – 50 mg/g; and a plasticiser; 25 wherein weights are based upon the dry weight of the composition.

32. The composition of claim 31, wherein the plasticiser is selected from glycerin, a polyol, polyethylene glycol and triethyl citrate or a mixture thereof.

33. The composition of claim 31, wherein the plasticiser is sorbitol.

34. The composition of claim 1 in the form of a minibead comprising: Cyclosporin A (12.1 % w/w) capric/caprylic triglyceride (Miglyol 810 N) (6.2 % ww) thoxyethoxy)ethanol (Transcutol HP) (18.3 % ww) glyceryl monooleate/dioleate l GMO-50) (12.9 % ww) 5 SDS (3.2 % ww) Sorbitol (4.9 % ww) Gelatin (42.4 % ww) n weights are based upon the dry weight of the composition.

35. The composition of any one of claims 1 to 34, n the composition is a solid composition 10 and further comprises at least one coating.

36. The composition of claim 35, wherein the at least one coating is an outer g (a second coating).

37. The solid composition of claim 35 or claim 36, wherein the at least one coating is adapted to release the cyclosporin in at least the colon. 15

38. The solid composition of claim 37 that is adapted to release the porin in the ileum and colon or that is adapted to release substantially all the cyclosporin in the colon.

39. The solid composition of any one of claims 35 to 38, wherein the at least one coating comprises a pH-independent coating.

40. The solid composition of claim 39, wherein the ependent coating comprises 20 ethylcellulose; and/or wherein the pH-independent coating comprises a polymer susceptible to degradation by enzyme(s) of colonic bacteria.

41. The solid composition of any one of claims 36 to 40, wherein the at least one coating is present in an amount corresponding to a weight gain due to the coating of from 2% to 40%, relative to 25 the weight of the uncoated composition.

42. The solid composition of claim 41, wherein the weight gain due to the second coating is selected from a range of from: 4% to 30%, 4% to 7%, 7% to 40%, 7% to 30%, 8% to 25%, 8% to 20%, 2% to 25%, 2% to 20%, 4% to 25%, 4% to 20%, 4% to 15%, 4% to 13%, 7% to 15%, 7% to 13%, 8% to 12%, 9% to 12% and 20% to 25%, ve to the weight of the uncoated 30 composition.

43. The solid composition of any one of claims 36 to 40, wherein the composition comprises two coatings, a first coating and a second coating.

44. The solid composition of claim 43, n the composition comprises a first coating which comprises a water-soluble cellulose ether, and a second coating as specified in any one of claims 37 to 42.

45. The solid composition of claim 44, wherein the water-soluble cellulose ether is 5 hydroxypropylmethyl cellulose.

46. The solid composition of any one of claims 44 to 45, wherein the first g is present in an amount corresponding to a weight gain due to the coating of from 0.5% to 20% relative to the weight of the uncoated composition.

47. The composition of any one of claims 1 to 33, or 35 to 46, wherein the ition is a solid 10 ition that is in the form of a minibead.

48. The composition of claim 47, wherein the minibead has a size of from 0.5mm to 5mm.

49. The composition according to claim 1, wherein the composition is colloidal, and wherein: the hydrogel forming polymer matrix is a continuous phase and is present in an amount of 300 to 700 mg/g, the hydrogel forming polymer matrix comprising gelatin; the surfactant has an HLB of up to 8 and is t in an amount of 80 to 200 mg/g; wherein the surfactant comprises a medium or long chain fatty acid mono- or di-glyceride or a combination thereof which does not comprise a polyethyleneglycol ether or ester, , and the composition r comprises a disperse phase comprising: 20 the cyclosporin in an amount of 90 to 200 mg/g; and a medium chain yceride in an amount of 20 to 200 mg/g; and the composition further comprises: a t in an amount of 100 to 250 mg/g; and an anionic surfactant in an amount of 15 to 50 mg/g. 25

50. The composition of claim 49, wherein the surfactant is ed from yl caprylate, glyceryl caprate, yl monooleate, glyceryl dioleate, glyceryl monolinoleate or a combination thereof.

51. The composition of any one of claims 1 to 30 or 35 to 50, wherein the surfactant comprises glyceryl monooleate, glyceryl dioleate or a combination thereof.

52. The composition of any one of claims 1 to 30 or 35 to 50, n the hydrogel forming polymer 30 matrix comprises gelatin, the surfactant comprises glyceryl monooleate, glyceryl dioleate or a combination thereof, and the oil phase comprises a medium chain triglyceride.

53. A process for making an oral composition, the s comprising mixing an oil phase with an aqueous phase comprising a hydrogel forming polymer matrix present in an amount of at least 25% by weight of the composition based upon the dry weight of the composition, wherein the 35 hydrogel forming polymer matrix comprises one or more of gelatin, agar, , casein, chitosan, soya bean protein, safflower protein, alginates, gellan gum, carrageenan, xanthan gum, phthalated gelatin, succinated gelatin, cellulosephthalate-acetate, oleoresin, polyvinylacetate, polymerisates of acrylic or methacrylic esters and polyvinylacetate-phthalate, wherein the oil phase has cyclosporin in solution and comprises a surfactant sing a medium chain or long chain fatty acid mono- or di-glyceride or a combination thereof and not 5 comprising a polyethyleneglycol ether or ester.

54. The process of claim 53, wherein the oil phase and the aqueous phase are mixed in an oil phase to aqueous phase ratio of from 1:2 to 1:12; and/or wherein mixing the oil phase with the aqueous phase forms an emulsion; and/or wherein the process further comprises causing the on to solidify. 10

55. Use of the composition of any one of claims 1 to 54 in the manufacture of a medicament for treating a condition.

56. The use of claim 55, wherein the condition is a condition of the gastrointestinal tract.

57. The use of claim 56, wherein the ion is selected from inflammatory bowel disease, irritable bowel syndrome, Crohn’s disease, ulcerative colitis, celiac disease, graft vs host disease, 15 gastrointestinal graft-versus-host e, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, pouchitis, Curling’s ulcer, appendicitis, colitis, pseudomembraneous colitis, diverticulosis, diverticulitis, microscopic colitis, enous colitis, lymphocytic colitis, colorectal carcinoma, arcinoma, myasthenia gravis, stomach , proctitis, mucositis, chemotherapyassociated enteritis, radiation-associated enteritis, short bowel e, or chronic diarrhea, 20 endometriosis, inflammatory disorders such as diversion colitis, ischemic colitis, infectious colitis, chemical colitis, fulminant colitis, autistic enterocolitis, rminate colitis, jejunoiletis, atypical s, , ileocolitis or granulomatous colitis, the prevention of rejection following bone marrow transplantation, psoriasis, atopic dermatitis, rheumatoid arthritis, nephrotic syndrome primary sing cholangitis, familial adenomatous polyposis, or perianal s, including 25 perianal fistulae; or wherein the condition is selected from the prevention of rejection following bone marrow transplantation, sis, atopic dermatitis, and rheumatoid arthritis.

58. The use of any one of claims 55 to 57, wherein the condition is ulcerative colitis. SublotA at 80' Sublot B at 60' Sublot C at 60' *S'UbléfAW' sum 10' “0120' SublotA at 210' Sublot B at 180' Sublot at 180' Cremophor formulation SublotAat60' Sublot B at 60' SublotC at 6' SublotA at 120' Sublot B at 120' Sublot C at 120' A at 240' Sublot B at 240' Sublot C at 240' Capmul formulation WO 71515 1.5h 2h 120.0 100.0 released CyA ..- Capmul GMO batch 1 + Capmul GMO batch 2 20.0 - 0.0 —‘- . 0 5 10 15 20 Timepoint (Hrs) 80 - ----- ...... 70 ML - o -CremaphorBatch1 8 60 2! ~. - I - CremaphorBatchZ """"- (D a.» . “a 'N. ..... g 50 am R. +Cremaphor Batch 3 . ....... § D "x. ....... ——x— CremaphorBatch4 \o 40 . ° . ..... ‘~~., ...... - >K - Cremaphor Batch 5 30 ~. ...... 1o ~ O 2 4 6 81012141618 20 2224 Time (hrs) H N m :38 £38 £38 056 056 056 338+ SEQSIII .258qu mu_: