Pharmaceutical composition comprising budesonide for treating IgA nephropathy

Abstract

The present invention provides a method of treating IgA nephropathy, said method comprising: (i) identifying a pharmaceutically acceptable composition intended for treating IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of said budesonide following administration to the gastrointestinal tract, said composition being in a standard in vitro USPlt; 711gt, 711gt; in a dissolution test in European Pharmacopoeia 2.9. 3, dissolution equipment using a second method (paddle method) according to the test meets the following requirements; (a) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 1.2; (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) the composition satisfying the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8; (ii) wherein said method comprises the step of administering said composition to a patient suffering from IgA nephropathy in need of said treatment.

Description

[0001] This application is a divisional application of the invention application with the application date of January 24, 2023, Chinese application number 202380018644.0, and invention name “Pharmaceutical composition containing budesonide for treating IgA nephropathy”. Technical Field

[0002] The present invention relates to a method for treating IgA nephropathy and a method for determining whether a pharmaceutical composition can safely and effectively treat IgA nephropathy. The present invention also relates to compositions for treating IgA nephropathy and methods for producing those compositions. Background Art

[0003] The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an admission that the document is prior art or part of the common general knowledge.

[0004] IgA nephropathy (IgAN), sometimes called Berger's disease, is a serious, progressive autoimmune disease of the kidneys, with up to 50% of patients at risk of developing end-stage renal disease (ESRD) within ten to twenty years.

[0005] IgAN is a rare disease, affecting an estimated 130,000 to 150,000 people in the United States and approximately 200,000 in Europe. Significantly higher prevalence has been observed in Asia, including Greater China, where IgAN has historically been a leading cause of ESRD. It is estimated that IgAN affects approximately two million people in Greater China.

[0006] Although IgAN is expressed in the kidneys, most scientific research has found that the pathogenesis of IgAN begins in the ileum, the last part of the small intestine before the large intestine. Clumps of lymphoid tissue, called Peyer's patches, are primarily found in the ileum, where they produce secretory IgA antibodies. IgA antibodies play a key role in the immune system by protecting the body from foreign substances, such as food-borne agents, bacteria, and viruses.

[0007] Patients with IgAN have elevated levels of a subclass of IgA antibodies produced in the intestine that lack a galactose unit at their hinge region. The hinge region is a flexible stretch of amino acids in the central portion of the heavy chain of an IgA antibody. In patients with IgAN, it is hypothesized that a combination of genetic predisposition and environmental, bacterial, or dietary factors leads to increased production of these galactose-deficient IgA antibodies, possibly combined with increased intestinal permeability, leading to the appearance of these antibodies in the blood. Galactose-deficient IgA antibodies (also referred to herein as poorly O-galactosylated IgA1) are immunogenic when found in the circulation, triggering autoantibodies or antibodies produced by the body in response to components of its own tissues. This, in turn, leads to the formation of pathogenic immune complexes or antibody clusters that are deposited in the membranes of the kidney's filtration apparatus, the glomeruli. These trapped immune complexes trigger an inflammatory cascade that damages the membranes, causing proteins and blood to leak into the urine. Ultimately, the glomeruli are destroyed, reducing the kidney's ability to remove waste products from the blood. As the disease progresses, waste products that are normally removed from the blood accumulate, leading to potentially life-threatening complications that in many patients will result in the need for dialysis or a kidney transplant.

[0008] The standard of care for ESRD is dialysis or kidney transplantation, which presents a substantial health economic burden and a significant impact on patients' quality of life.

[0009] Despite the need for new therapies, few new drugs have been developed for chronic kidney disease over the past decade, and until recently, there have been no approved therapies for the direct treatment of IgAN itself. Patients with IgAN are generally initially given antihypertensive drugs. This treatment regimen initially attempted to manage the symptoms of IgAN by lowering blood pressure and reducing proteinuria, but has not yet been proven to address the underlying cause of IgAN. Over time, physicians have attempted to control disease progression with a variety of off-label treatments such as statins, ω-3-acids, and diuretics, but a significant proportion of patients experience continued deterioration of renal function, and until recently, no approved treatment options were available.

[0010] For patients with IgAN whose disease progresses, clinicians can treat them with systemic immunosuppressive agents, primarily consisting of high-dose systemic corticosteroids, such as prednisone, prednisolone, and methylprednisolone. Although some published reports suggest that these agents can reduce proteinuria, these high-dose systemic corticosteroids are also associated with a wide range of adverse events, including hypertension, weight gain, diabetes, serious infections, and osteoporosis. Furthermore, any possible effect on the underlying disease in terms of renal function, as measured by estimated glomerular filtration rate (eGFR), has not been demonstrated.

[0011] Thus, in trying to meet the current clinical need for an effective treatment of IgAN, there is a clear need for new and / or improved treatments for IgAN, wherein an effective local treatment with an immunosuppressive agent is obtained without such unwanted side effects.

[0012] Peyer's patches (aggregated lymphatic nodules) are small patches of lymphatic tissue found in the ileal region of the small intestine. As it monitors the intestinal bacterial population and prevents the growth of pathogenic bacteria in the intestinal tract, it is an important part of the immune system.

[0013] As Peyer's patches are responsible for the synthesis of most of the IgA in the body, a dose of a locally acting immunosuppressive agent targeted to the ileum (and especially the terminal / distal ileum) where Peyer's patches are predominantly located can be used to reduce the formation of IgA molecules that ultimately drive the formation of immune complexes in IgAN by reducing the formation of secretory galactose-deficient IgA antibodies and their appearance in the blood. Such targeted release can also limit systemic exposure of locally acting immunosuppressive agents, such as certain corticosteroids, to avoid unwanted side effects.

[0014] Peyer's patches are sites of strong B cell activation in the human body. Thus, it can be considered that monitoring survival factors associated with B cell activation will provide an indication of the efficacy of locally acting immunosuppressive treatment.

[0015] The tumour necrosis factor (TNF) family members, B cell activating factor (BAFF) and its homologue a proliferation-inducing ligand (APRIL) are key survival factors for peripheral B cells and are expressed by cells including monocytes, dendritic cells, neutrophils, basophils, stromal cells, activated T cells, intestinal mucosal cells, activated and malignant B cells and epithelial cells (Mackay and Schneider, 2009. Nat. Rev. Immunol., 9: 491-502; Schneider et al., 1999. J. Exp. Med., 189: 1747-1756; Yu et al., 2000. Nat. Immunol., 1: 252-256).

[0016] BAFF is a ligand for transmembrane activator and CAML interactor (TACI), also known as tumor necrosis factor receptor superfamily member 13B (TNFRSF13B); B-cell maturation antigen (BCMA), also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17); and B-cell activating factor receptor (BAFF-R), also known as tumor necrosis factor receptor superfamily member 13C (TNFRSF13C). BAFF-R is specific for BAFF, while TACI and BCMA also bind to APRIL (Mackay and Schneider, 2009. Nat. Rev. Immunol., 9:491-502).

[0017] BAFF is a potent B cell activating factor and is crucial for B cell homeostasis and regulating B cell selection. Excessive BAFF has been shown to be associated with the development of autoimmune disorders such as IgAN in animal models, and high levels of BAFF have been detected in the serum of patients with various autoimmune conditions. Elevated levels of BAFF have been associated with upregulation of humoral immunity by increased levels of B cells and immunoglobulins (Steri et al., 2017. N. Engl. J. Med., 376: 1615-1626).

[0018] Elevated serum levels of BAFF and APRIL are found in patients suffering from IgAN, and this has led to the development of drugs that attempt to inhibit those molecules. The focus of those drugs is to block the interaction between BAFF and / or APRIL and their receptors. For example, but not limited to, the BAFF inhibitor Blisibimod (Anthera Pharmaceuticals, discontinued) is a fusion protein consisting of four BAFF binding domains fused to the N-terminus of the Fc region of a human antibody, which binds to BAFF and inhibits the interaction with the BAFF receptor. Similarly, the combined BAFF / APRIL antagonist Atacicept (MerckSerono, licensed from Vera Therapeutics) is also a recombinant fusion protein that combines the BAFF and APRIL binding domains with the antibody Fc region and blocks the interaction with TACI. The APRIL antagonist VIS649 (Visterra, a subsidiary of Otsuka) and the BAFF inhibitor Belimumab (GlaxoSmithKline) are monoclonal antibodies that directly bind to APRIL and BAFF, respectively, and block their interaction with their receptors. Therefore, drugs targeting BAFF and APRIL attempt to block the activity of endogenous BAFF / APRIL molecules in order to reduce B cell activation and proliferation and related immune effects.

[0019] The current understanding of the pathogenesis of IgAN and current treatments is summarized in J. Barratt et al., Treatment of IgA Nephropathy: Evolution Over Half a Century, Seminars in Nephrology, 2018, 38(5), 531-540, see also Boyd et al., Kidney International, 2012, 81, 833-843. The current understanding of the pathogenesis of IgAN is also summarized in Seikrit et al., The Immune Landscape of IgA Induction in the Gut, Seminars in Immunopathology, 2021, 43, 627-637.

[0020] Surprisingly, we have found that oral administration of a budesonide formulation with a unique in vitro release profile resulted in a significant reduction in serum levels of BAFF in those subjects relative to levels observed prior to budesonide administration. Furthermore, the observed reduction in serum BAFF levels can coincide with a reduction in the levels of biomarkers associated with B cell activation and proliferation. Thus, the in vitro release profile can indicate successful targeted release in the intestine of a subject (i.e., successful targeted release to the distal ileum).

[0021] Of particular interest is that systemic glucocorticoid treatment of IgAN has been shown to reduce total serum IgA and poorly O-galactosylated IgA1 (Kosztyu P et al., Glucocorticoids Reduce Aberrant O-Glycosylation of IgA1 in IgA Nephropathy Patients. Kidney Blood Press Res 2018; 43: 350-359). However, under the treatment of the present invention with oral administration of the budesonide formulation as defined herein, no difference in the level of total functional IgA antibodies and IgG, including IgA1, was observed under budesonide capsule treatment, but serum levels of galactose-deficient IgA (poorly O-galactosylated IgA) did decrease. This finding leads to the conclusion that local ileal treatment with budesonide capsules has a selective effect on pathogenic antibodies but is ineffective against the total pools of IgA, IgA1, and IgG.

[0022] These results demonstrate that treatment with the budesonide formulation as defined herein supports a direct effect on the underlying pathogenic pathways in IgAN and that the budesonide payload has primarily local rather than systemic effects, resulting in reduced side effects in patients when treated with Nefecon budesonide.

[0023] In support of this notion, computer simulation modeling of a budesonide formulation with a unique in vitro release profile showed that the loaded drug was released primarily into the ileum, especially the distal ileum. In the case of budesonide, which is primarily metabolized through the intestinal wall in the small intestine with a high first-pass rate ( J et al., Presystemic elimination of budesonide in man when administered locally at different levels in the gut, with and without local inhibition by ketoconazole. Eur J Pharm Sci. 2008 Nov 15;35(4):264-70; Raje et al. Evaluation of separate role of intestine and liver in first pass metabolism of budesonide in rat Xenobiotica. 2018 Dec;48(12):1206-1214), these results further indicate that budesonide formulations have local rather than systemic effects.

[0024] Taken together, the results described herein demonstrate that budesonide formulations exhibiting a unique in vitro release profile as defined herein are effective treatments for IgAN. Due to their targeted local release and action of the topical corticosteroid, lower levels of undesirable side effects are achieved.

[0025] Formulations of the corticosteroid budesonide have been previously described in International Patent Application WO 2009 / 138716 A1. Summary of the Invention

[0026] According to a first aspect of the present invention, there is provided a method for treating IgA nephropathy, the method comprising:

[0027] (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements: <711> / European Pharmacopoeia (Ph.Eur.) 2.9.3 dissolution test using a dissolution apparatus according to Method 2 (paddle method) of said test (as described below);

[0028] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0029] (b) the composition meets the requirement of releasing no more than about 10% of the budesonide into a pharmaceutically relevant dissolution medium within about 30 minutes; and

[0030] (c) the composition satisfies the requirement of releasing at least about 70% of the budesonide into a pharmaceutically relevant dissolution medium within about 120 minutes, followed by

[0031] (ii) administering the composition to a patient with IgA nephropathy in need of such treatment,

[0032] Said method is hereinafter referred to as "the method of the invention".

[0033] The term "pharmaceutically relevant dissolution medium" includes media suitable for in vitro dissolution analysis, the results of which are indicative of in vivo release at relevant portions of the intestinal tract. For example, a pharmaceutically relevant dissolution medium may alternatively be referred to as an "enterically pharmaceutically relevant dissolution medium" or a "pharmaceutically relevant enteric dissolution medium" and may be any such medium that simulates dissolution and release in the small intestine or relevant portions thereof.

[0034] The pharmaceutically relevant dissolution medium is preferably aqueous.

[0035] The pH of a pharmaceutically relevant dissolution medium may be from about 6.2 to about 7.5, such as from about 6.5 to about 6.8.

[0036] Pharmaceutically relevant dissolution media can be a phosphate buffered medium having a pH of about 6.2, a Fasted State Simulated Intestinal Fluid (FaSSIF) having a pH of about 6.5 (e.g., the FaSSIF buffer defined below under the heading "Release in Level 1 Fasted State Simulated Intestinal Fluid at a pH of about 6.5"), a phosphate buffered medium having a pH of about 6.8 (e.g., the phosphate buffer defined below under the heading "Release in medium at pH 6.8"), or a phosphate buffered medium having a pH of about 7.2 or about 7.5.

[0037] The method of the present invention may comprise (I) combining budesonide with one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract to produce a pharmaceutically acceptable composition intended for use in treating IgA nephropathy, and then (II) conducting a standard in vitro USP test as described above. <711> / European Pharmacopoeia 2.9.3 dissolution test, and if the composition meets requirements (a) to (c) as described above, administering the composition to a patient with IgA nephropathy in need of such treatment.

[0038] As an alternative embodiment of the present invention, there is provided a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above for use in treating IgA nephropathy.

[0039] As another alternative embodiment of the present invention, there is provided the use of a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above for the manufacture of a medicament for the treatment of IgA nephropathy.

[0040] As referred to herein, the term "treatment" of IgA nephropathy further includes, in addition to therapeutic, symptomatic and / or palliative treatment of the relevant condition, the prevention or diagnosis thereof.

[0041] For the avoidance of doubt, when referring to USP <711> When referring to the USP, this refers to the test published on May 1, 2016, and when referring to the European Pharmacopoeia 2.9.3, this refers to section 2.9.3 of the European Pharmacopoeia 10.0. It should be understood that jurisdictions outside the United States and Europe may have equivalent pharmacopoeias that reflect the same or similar tests as outlined in the USP and the European Pharmacopoeia, such as the Chinese Pharmacopoeia.

[0042] For the avoidance of doubt, step (ii) of administering the composition to the patient will only proceed if the average (average / mean) of the tested composition meets each and all of the criteria (a), (b) and (c) of step (i).

[0043] As used herein, the term "budesonide" refers to a compound according to Formula I:

[0044]

[0045] Budesonide is also commonly referred to by its IUPAC name (16α,17-[(1RS)-butylenebis(oxy)]-11β,21-dihydroxypregna-1,4-diene-3,20-dione).

[0046] Although the compositions of the present invention comprise budesonide, it will be appreciated that the compositions may alternatively comprise different corticosteroids capable of having a local effect in a manner similar to budesonide. Such suitable alternative corticosteroids include, but are not limited to, aclometasone, beclomethasone, betamethasone, clobetasol, hydrocortisone, dexamethasone, flunisolide, methylprednisolone, mometasone furoate, prednisolone, triamcinolone, fluticasone, ciclesonide, fludrocortisone, and mixtures thereof, including mixtures comprising budesonide.

[0047] The paddle method of method 2 can be operated at about 50 revolutions per minute (rpm), about 75 rpm, or about 100 rpm. Preferably, the paddle method of method 2 is operated at about 100 rpm or about 50 rpm.

[0048] The pharmaceutically relevant dissolution medium of criteria b) and c) may comprise a surfactant in an amount of about 0.5 mg / mL (0.05% w / v). The surfactant may be a polysorbate, preferably wherein the surfactant is polysorbate 80 (e.g., Tween 80).

[0049] In criterion a) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 120 minutes.

[0050] In criterion a) of step (i) of the method, the amount of budesonide released may be from about 0% to about 10%, such as from about 0% to about 5%, for example from about 0% to about 2.5% within about 120 minutes.

[0051] In criterion b) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 30 minutes.

[0052] In criterion b) of step (i) of the method, the amount of budesonide released can be about 0% to about 10%, such as about 0% to about 5%, for example about 0% to about 2.5% in about 30 minutes.

[0053] In criterion c) of step (i) of the method, the amount of budesonide released can be at least about 75%, for example about 80%, such as about 84% or about 85% in about 120 minutes.

[0054] In criterion c) of step (i) of the method, the amount of budesonide released can be about 70% to about 100%, such as about 75% to about 100%, for example about 84% to about 100%, such as about 85% to 100% in about 120 minutes.

[0055] In criterion b) of step (i) of the method, the composition can further meet the requirement that no more than about 10% of the budesonide is released into a pharmaceutically relevant dissolution medium in about 37.5 minutes, such as no more than about 5%, for example no more than about 2.5% of the budesonide is released into a pharmaceutically relevant dissolution medium in about 37.5 minutes. For example, the amount of budesonide released into a pharmaceutically relevant dissolution medium in about 37.5 minutes can be about 0% to about 10%, such as about 0% to about 5%, for example about 0% to about 2.5%. Optionally, the release in about 37.5 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 2nd Method paddles at 50 rpm.

[0056] In criterion b) of step (i) of the method, the composition can further meet the requirement that at least about 20% of the budesonide is released into a pharmaceutically relevant dissolution medium in about 75 minutes, such as at least about 21%, for example at least about 22% or 23% of the budesonide is released into a pharmaceutically relevant dissolution medium in about 75 minutes. For example, the amount of budesonide released into a pharmaceutically relevant dissolution medium in about 75 minutes can be about 23% to about 74%. Optionally, the release in about 75 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 2nd Method paddles at 50 rpm.

[0057] In criterion c) of the method, the composition can further meet the following requirement: at least about 75% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 150 minutes, such as at least about 76%, for example, at least about 77% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 150 minutes. For example, the amount of budesonide released into the pharmaceutically relevant dissolution medium within about 150 minutes can be about 77% to about 100%. Optionally, the release within about 150 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 50 rpm in a Phase 2 paddle method.

[0058] In criterion b) of the method, the composition can further meet the following requirement: no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 45 minutes, such as no more than about 5%, such as no more than about 2.5% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 45 minutes. For example, the amount of budesonide released into the pharmaceutically relevant dissolution medium within about 45 minutes can be from about 0% to about 10%, such as from about 0% to about 5%, for example, from about 0% to about 2.5%. Optionally, the release within about 45 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm for the Phase 2 paddle method.

[0059] In criterion b) of the method, the composition can further meet the following requirement: no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 60 minutes, such as no more than about 5%, such as no more than about 2.5% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 60 minutes. For example, the amount of budesonide released into the pharmaceutically relevant dissolution medium within about 60 minutes can be from about 0% to about 10%, such as from about 0% to about 5%, for example, from about 0% to about 2.5%. Optionally, the release within about 60 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm for the Phase 2 paddle method.

[0060] In criterion b) of the method, the composition can further meet the following requirement: 50 to 90% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 90 minutes. Optionally, the release within about 90 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm in the Phase 2 paddle method.

[0061] In criterion c) of the method, the composition can further meet the following requirement: at least about 80% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 180 minutes, such as at least about 85%, for example, about 80% to about 100%, or about 85% to about 100% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 180 minutes. Optionally, the release within about 180 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm for the Phase 2 paddle method.

[0062] In criterion c) of the method, the composition can further meet the following requirement: at least about 85% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 240 minutes, such as at least about 90%, for example, about 90% to about 100% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 240 minutes. Optionally, the release within about 240 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm for the Phase 2 paddle method.

[0063] In criterion c) of the method, the composition can further meet the following requirement: at least about 90% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 360 minutes, such as at least about 95%, for example, about 95% to about 100% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 360 minutes. Optionally, the release within about 360 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm in the Phase 2 paddle method.

[0064] In criterion c) of the method, the composition can further meet the following requirement: at least about 90% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 480 minutes, such as at least about 95%, for example, about 95% to about 100% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 480 minutes. Optionally, the release within about 480 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm in the Phase 2 paddle method.

[0065] In criterion c) of the method, the composition can further meet the following requirement: at least about 90% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 600 minutes, such as at least about 95%, for example, about 95% to about 100% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 600 minutes. Optionally, the release within about 600 minutes is in the absence of a surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm in the Phase 2 paddle method.

[0066] In one embodiment, in criterion a) of step (i) of the method, the dissolution in the acid-resistant medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-3 of 2.9.3 of the European Pharmacopoeia.

[0067] In one embodiment, in criterion b) of step (i) of the method, dissolution in a pharmaceutically relevant dissolution medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-3 of 2.9.3 of the European Pharmacopoeia.

[0068] In one embodiment, in criterion c) of step (i) of the method, dissolution in a pharmaceutically relevant dissolution medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 4 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-4 of 2.9.3 of the European Pharmacopoeia.

[0069] For the avoidance of doubt, the amount of budesonide released at 30 minutes in criterion b) and at 120 minutes in criterion c) is achieved in the presence and absence of an added surfactant, such as added polysorbate 80 (e.g., Tween 80), at a concentration of about 0.5 mg / mL in a pharmaceutically relevant dissolution medium. Additionally, the amount of budesonide released at 37.5 minutes, 60 minutes, 75 minutes, 90 minutes, and 150 minutes is achieved in the presence and absence of an added surfactant, such as added polysorbate 80 (e.g., Tween 80), at a concentration of about 0.5 mg / mL in a pharmaceutically relevant dissolution medium.

[0070] In one embodiment, the method comprises:

[0071] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide for a modified release of the budesonide after administration to the gastrointestinal tract, which composition meets the following requirements: in a standard in vitro USP <711> / European Pharmacopoeia 2.9.3 dissolution test, using a dissolution apparatus according to the 2nd method (paddle method) of the test operating at 50 rpm (as described hereinafter);

[0072] (a) the composition meets the requirement that not more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 1.2;

[0073] (b) the composition meets the requirement that not more than about 10% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 30 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant;

[0074] (c) the composition meets the requirement that not more than about 10% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 37.5 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant;

[0075] (d) the composition meets the requirement that about 23% to about 74% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 75 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant;

[0076] (e) the composition meets the requirement that at least about 77% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 150 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant; optionally

[0077] (f) wherein the composition meets the requirement that at least about 70% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 120 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant.

[0078] In another embodiment, the method comprises:

[0079] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide for a modified release of the budesonide after administration to the gastrointestinal tract, which composition meets the following requirements: in a standard in vitro USP <711> / European Pharmacopoeia 2.9.3 dissolution test, using a dissolution apparatus according to the 2nd method (paddle method) of the test operating at 100 rpm (as described hereinafter);

[0080] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0081] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 30 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant;

[0082] (c) the composition satisfies the requirement that no more than about 10% of the budesonide is released into a pharmaceutically relevant dissolution medium within about 60 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant;

[0083] (d) the composition meets the requirement of releasing about 50% to about 90% of the budesonide into a pharmaceutically relevant dissolution medium within about 90 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant; and

[0084] (e) The composition meets the requirement that at least about 70%, such as at least 75%, of the budesonide is released into a pharmaceutically relevant dissolution medium within about 120 minutes when the pharmaceutically relevant dissolution medium does not have a surfactant.

[0085] Pharmacokinetic studies of drug absorption in the fasting state indicate that, with the ingestion of 200 to 250 mL of water with a dosage form, a maximum total volume of approximately 300 to 500 mL is available in the proximal small intestine (see Klein, AAPS J., 12, 397, (2010)). Therefore, the dissolution test employed in the present method should employ a volume of at least approximately 500 mL (e.g., approximately 900 mL) of dissolution medium. The initial volume of dissolution medium used for criteria a), b), and c) can be approximately 900 mL.

[0086] The procedures for testing the compositions can be essentially based on USP <711> / Delayed-release solid dosage forms according to European Pharmacopoeia 2.9.3 Method B.

[0087] The temperature of the dissolution medium in criteria a), b) and c) may be maintained at about 37°C ± 0.5°C.

[0088] The number of compositions tested may be 6, or greater than 6, such as 12 or 24.

[0089] In criteria (a), (b), and (c), the amount of volume extracted from the dissolution medium at each time point can be 10 mL or 15 mL, optionally without replacing the extracted volume. The extraction of the dissolution medium does not affect the overall dissolution profile of the composition. In other words, preferably, the dissolution test is performed under sink conditions, and the amount of solvent exceeds the amount of solute, which means that small amounts extracted for analytical purposes do not affect dissolution.

[0090] Release in media at pH 6.8

[0091] According to an alternative aspect of the present invention, there is provided a method for treating IgA nephropathy, the method comprising:

[0092] (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements: <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to Method 2 (Paddle Method) of said test (as described below) was used;

[0093] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0094] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and

[0095] (c) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 6.8, at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes, and thereafter

[0096] (ii) administering the composition to a patient with IgA nephropathy in need of such treatment, and the method is hereinafter referred to as "the method of the present invention".

[0097] The method of the present invention may comprise (I) combining budesonide with one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract to produce a pharmaceutically acceptable composition intended for use in treating IgA nephropathy, and then (II) conducting a standard in vitro USP test as described above. <711> / European Pharmacopoeia 2.9.3 dissolution test, and if the composition meets requirements (a) to (c) as described above (ie with respect to release in a medium at pH 6.8), administering the composition to a patient with IgA nephropathy in need of such treatment.

[0098] As an alternative embodiment, a composition is provided comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide following administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above (i.e., with respect to release in the pharmaceutically relevant medium described above at pH 6.8) for use in treating IgA nephropathy.

[0099] As another alternative embodiment, there is provided the use of a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide following administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above (i.e., with respect to release in a pharmaceutically relevant medium at pH 6.8) for the manufacture of a medicament for the treatment of IgA nephropathy.

[0100] For the avoidance of doubt, step (ii) of administering the composition to a patient will only proceed if the average (average / mean) of the tested composition meets all criteria (a), (b) and (c) of step (i).

[0101] The paddle method of method 2 can be operated at about 50 revolutions per minute (rpm), about 75 rpm, or about 100 rpm. Preferably, the paddle method of method 2 is operated at about 100 rpm or about 50 rpm.

[0102] The aqueous dissolution medium of criteria b) and c) may contain a surfactant in an amount of about 0.5 mg / mL (0.05% w / v). The surfactant may be a polysorbate, preferably wherein the surfactant is polysorbate 80 (e.g., Tween 80).

[0103] The aqueous dissolution medium of criteria b) and c) may be a phosphate buffer medium, such as a sodium phosphate buffer solution having a concentration of about 50 mM.

[0104] A phosphate buffered medium having a pH of about 6.8 can be prepared by first preparing a 0.2 M trisodium phosphate solution, then adding one part 0.2 M trisodium phosphate solution to three parts 0.1 N hydrochloric acid solution. After the two solutions are mixed together, the pH can be checked and adjusted to a pH of about 6.8 by adding hydrochloric acid or sodium hydroxide, if necessary.

[0105] In criterion a) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 120 minutes.

[0106] In criterion a) of step (i) of the method, the amount of budesonide released may be from about 0% to about 10%, such as from about 0% to about 5%, for example from about 0% to about 2.5% within about 120 minutes.

[0107] In criterion b) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 30 minutes.

[0108] In criterion b) of step (i) of the method, the amount of budesonide released may be from about 0% to about 10%, such as from about 0% to about 5%, for example from about 0% to about 2.5% within about 30 minutes.

[0109] In criterion c) of step (i) of the method, the amount of budesonide released may be at least about 75%, for example about 80%, such as about 84% or about 85% within about 120 minutes.

[0110] In criterion c) of step (i) of the method, the amount of budesonide released may be about 70% to about 100%, such as about 75% to about 100%, for example about 84% to about 100%, such as about 85% to 100% within about 120 minutes.

[0111] In criterion b) of step (i) of the method, the composition may further meet the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of the budesonide is released into the dissolution medium within about 37.5 minutes, such as when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 5%, for example, no more than about 2.5% of the budesonide is released within about 37.5 minutes. For example, when the dissolution medium is aqueous and has a pH of about 6.8, the amount of budesonide released within about 37.5 minutes may be from about 0% to about 10%, such as from about 0% to about 5%, for example, from about 0% to about 2.5%. Optionally, the release within about 37.5 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 50 rpm for a Phase 2 paddle method.

[0112] In criterion b) of step (i) of the method, the composition may further meet the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, at least about 20% of the budesonide is released into the dissolution medium within about 75 minutes, such as when the dissolution medium is aqueous and has a pH of about 6.8, at least about 21%, for example, at least about 22% or 23% of the budesonide is released within about 75 minutes. For example, when the dissolution medium is aqueous and has a pH of about 6.8, the amount of budesonide released within about 75 minutes may be about 23% to about 74%. Optionally, the release within about 75 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 50 rpm for a Phase 2 paddle method.

[0113] In criterion c) of the method, the composition can further meet the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, at least about 75% of the budesonide is released into the dissolution medium within about 150 minutes, such as when the dissolution medium is aqueous and has a pH of about 6.8, at least about 76%, for example, at least about 77%, of the budesonide is released within about 150 minutes. For example, when the dissolution medium is aqueous and has a pH of about 6.8, the amount of budesonide released within about 150 minutes can be about 77% to about 100%. Optionally, the release within about 150 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 50 rpm in a Phase 2 paddle method.

[0114] In criterion b) of the method, the composition may further meet the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of the budesonide is released into the dissolution medium within about 60 minutes, such as when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 5%, such as no more than about 2.5%, of the budesonide is released within about 60 minutes. For example, when the dissolution medium is aqueous and has a pH of about 6.8, the amount of budesonide released within about 60 minutes may be from about 0% to about 10%, such as from about 0% to about 5%, for example, from about 0% to about 2.5%. Optionally, the release within about 60 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 100 rpm in a Phase 2 paddle method.

[0115] In criterion b) of the method, the composition may further meet the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, 50 to 90% of the budesonide is released within about 90 minutes. Optionally, the release within about 90 minutes is in the absence of a surfactant in the dissolution medium and at a paddle rotation speed of 100 rpm in a Phase 2 paddle method.

[0116] In one embodiment, in criterion a) of step (i) of the method, the dissolution in the acid-resistant medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-3 of 2.9.3 of the European Pharmacopoeia.

[0117] In one embodiment, in criterion b) of step (i) of the method, the dissolution in the buffer stage medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-3 of 2.9.3 of the European Pharmacopoeia.

[0118] In one embodiment, in criterion c) of step (i) of the method, the dissolution in the buffer stage medium may be carried out according to USP <711> Acceptance criteria in Acceptance Table 2 and / or Acceptance Table 4 of 2.9.3 / Evaluation in accordance with Table 2.9.3-2 and / or Table 2.9.3.-4 of 2.9.3 of the European Pharmacopoeia.

[0119] For the avoidance of doubt, the amount of budesonide released at 30 minutes in criterion b) and at 120 minutes in criterion c) is achieved in the presence and absence of an added surfactant, such as polysorbate 80 (e.g., Tween 80), at a concentration of approximately 0.5 mg / mL in the dissolution medium. Additionally, the amount of budesonide released at 37.5 minutes, 60 minutes, 75 minutes, 90 minutes, and 150 minutes is achieved in the presence and absence of an added surfactant, such as polysorbate 80 (e.g., Tween 80), at a concentration of approximately 0.5 mg / mL in the dissolution medium.

[0120] In one embodiment, the method comprises:

[0121] (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements: <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to Method 2 (paddle method) of said test (described below) operated at 50 rpm was used;

[0122] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0123] (b) the composition meets the requirement that no more than about 10% of the budesonide is released into the dissolution medium in about 30 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8;

[0124] (c) the composition meets the requirement that no more than about 10% of the budesonide is released into the dissolution medium in about 37.5 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8;

[0125] (d) the composition meets the requirement that about 23% to about 74% of the budesonide is released into the dissolution medium in about 75 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8;

[0126] (e) the composition meets the requirement that at least about 77% of the budesonide is released into the dissolution medium in about 150 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8; optionally

[0127] (f) wherein the composition meets the requirement that at least about 70% of the budesonide is released into the dissolution medium in about 120 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8.

[0128] In another embodiment, the method comprises:

[0129] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide for the modified release of the budesonide after administration to the gastrointestinal tract, the composition meeting the following requirements in a standard in vitro USP <711> / European Pharmacopoeia 2.9.3 dissolution test using a dissolution apparatus according to the 2nd method (paddle method) of the test operating at 100 rpm (as described below);

[0130] (a) the composition meets the requirement that no more than about 10% of the budesonide is released into the dissolution medium in about 120 minutes when the dissolution medium is aqueous and has a pH of about 1.2;

[0131] (b) the composition meets the requirement that no more than about 10% of the budesonide is released into the dissolution medium in about 30 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8;

[0132] (c) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 60 minutes when the dissolution medium is aqueous, without a surfactant, and has a pH of about 6.8;

[0133] (d) the composition satisfies the requirement that when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8, about 50% to about 90% of the budesonide is released into the dissolution medium within about 90 minutes; and

[0134] (e) The composition satisfies the requirement that at least about 70%, such as at least 75%, of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous, has no surfactant, and has a pH of about 6.8.

[0135] Pharmacokinetic studies of drug absorption in the fasting state indicate that, with the ingestion of 200 to 250 mL of water with a dosage form, a maximum total volume of approximately 300 to 500 mL is available in the proximal small intestine (see Klein, AAPS J., 12, 397, (2010)). Therefore, the dissolution test employed in the present method should employ a volume of at least approximately 500 mL (e.g., approximately 900 mL) of dissolution medium. The initial volume of dissolution medium used for criteria a), b), and c) can be approximately 900 mL.

[0136] The procedures for testing the compositions can be essentially based on USP <711> / Long-release and / or delayed-release solid dosage forms according to Ph. Eur. 2.9.3 Method B.

[0137] The temperature of the dissolution medium in criteria a), b) and c) may be maintained at about 37°C ± 0.5°C.

[0138] The number of compositions tested may be 6, or greater than 6, such as 12 or 24.

[0139] In criteria (a), (b), and (c), the volume withdrawn from the dissolution medium at each time point can be 10 mL or 15 mL, optionally without replacing the withdrawn volume. Withdrawal of the dissolution medium does not affect the overall dissolution profile of the composition. In other words, preferably, the dissolution test is performed under sink conditions with the amount of solvent exceeding the amount of solute, meaning that small withdrawals for analytical purposes do not affect dissolution.

[0140] Release in fasted simulated intestinal fluid at pH 6.5

[0141] According to another alternative aspect of the present invention, there is provided a method for treating IgA nephropathy, the method comprising:

[0142] (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements: <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to method 2 (paddle method) of the test was used;

[0143] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes; and

[0144] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released in about 30 minutes into a dissolution medium comprising a level 1 fasted state simulated intestinal fluid at a pH of about 6.5; and

[0145] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into a dissolution medium comprising a level 1 fasted state simulated intestinal fluid at a pH of about 6.5 within about 120 minutes; and thereafter

[0146] (ii) administering said composition to a patient with IgA nephropathy in need of said treatment,

[0147] And said method is hereinafter referred to as "the method of the present invention".

[0148] The method of the present invention may comprise (I) combining budesonide with one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract to produce a pharmaceutically acceptable composition intended for use in treating IgA nephropathy, and then (II) conducting a standard in vitro USP test as described above. <711> / European Pharmacopoeia 2.9.3 dissolution test, and if the composition meets requirements (a) to (c) as described above (i.e. with respect to release in a stage 1 fasting state simulated intestinal fluid at a pH of about 6.5), administering the composition to a patient with IgA nephropathy in need of such treatment.

[0149] As an alternative embodiment, a composition is provided comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, wherein the composition meets the dissolution profile of step (i) outlined above (i.e., with respect to release in a first fasting state simulated intestinal fluid at a pH of about 6.5) for use in treating IgA nephropathy.

[0150] As another alternative embodiment, there is provided the use of a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide following administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above (i.e., with respect to release in a first fasting state simulated intestinal fluid at a pH of about 6.5) for the manufacture of a medicament for the treatment of IgA nephropathy.

[0151] As referred to herein, the term "treatment" of IgA nephropathy further includes, in addition to therapeutic, symptomatic and / or palliative treatment of the relevant condition, the prevention or diagnosis thereof.

[0152] It will be understood by those of ordinary skill in the art that the term "Level 1 fasted state simulated intestinal fluid" (Level 1 FaSSIF-V1) includes biorelevant dissolution media that have a lower pH and buffering capacity than standard simulated intestinal fluid (the medium typically used in standard USP / European Pharmacopoeia testing; pH 6.8) and are specifically developed to simulate fasting conditions in the proximal small intestine (see, e.g., Markopoulous et al., In-vitrosimulation of luminal conditions for evaluation of performance of oral drug products: Choosing the appropriate test media, European Journal of Pharmaceutics and Biopharmaceutics, 93, 2015, 173-182).

[0153] Level 1 FaSSIF-V1 comprises a phosphate buffer system, such as a system comprising NaH2PO4 (at a concentration of approximately 28.5 mM), NaOH (at a concentration of approximately 13.8 mM), HCl (q.s.), and deionized water (q.s.), which produces a medium with an osmolality of approximately 270 mOsmol / kg and a buffering capacity of approximately 12 mEq / pH / L.

[0154] In the methods of the present invention, a surfactant may be added to the FaSSIF medium. The surfactant may be a polysorbate, such as polysorbate 80 (e.g., Tween 80). The surfactant may be present at a concentration of about 0.05% w / v (0.5 mg / mL) to facilitate analysis.

[0155] Therefore, it is further provided that:

[0156] ● the method of the invention as defined above;

[0157] a composition for treating IgA nephropathy that satisfies the dissolution profile of step (i) outlined above; and

[0158] Use of a composition that satisfies the dissolution profile of step (i) outlined above for the manufacture of a medicament for the treatment of IgA nephropathy,

[0159] Provided that in each case the FaSSIF medium employed in step (i) comprises a surfactant, such as a polysorbate, including polysorbate 80 (e.g. Tween 80), optionally present at a concentration of about 0.05% w / v (0.5 mg / mL) to facilitate analysis.

[0160] For the avoidance of doubt, step (ii) of administering the composition to a patient will only proceed if the average (average / mean) of the tested composition meets all criteria (a), (b) and (c) of step (i).

[0161] In criterion a) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 120 minutes.

[0162] In criterion a) of step (i) of the method, the amount of budesonide released may be from about 0% to about 10%, such as from about 0% to about 5%, for example from about 0% to about 2.5% within about 120 minutes.

[0163] In criterion b) of step (i) of the method, the amount of budesonide released may not exceed about 5%, such as not exceed about 2.5%, within about 30 minutes.

[0164] In criterion b) of step (i) of the method, the amount of budesonide released may be from about 0% to about 10%, such as from about 0% to about 5%, for example from about 0% to about 2.5% within about 30 minutes.

[0165] In criterion c) of step (i) of the method, the amount of budesonide released may be at least about 75%, for example about 80%, such as about 84% or about 85% within about 120 minutes.

[0166] In criterion c) of step (i) of the method, the amount of budesonide released may be from about 70% to about 99%, such as from about 70% to about 90%, within about 120 minutes.

[0167] For the avoidance of doubt, the amount of budesonide released at 30 minutes in criterion b) and at 120 minutes in criterion c) is achieved in the presence and absence of added polysorbate 80 (e.g. Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in FaSSIF medium.

[0168] In criterion b) of step (i) of the method, the composition can further meet the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 60 minutes, such as no more than about 5%, for example no more than about 2.5% of the budesonide is released within about 60 minutes. For example, the amount of budesonide released within about 60 minutes can be about 0% to about 10%, such as about 0% to about 5%, for example about 0% to about 2.5%. The amount of budesonide released within about 60 minutes is achieved in the presence and absence of added polysorbate 80 (e.g. Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in FaSSIF medium.

[0169] In criterion c) of step (i) of the method, in the presence of added polysorbate 80 (e.g. Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in FaSSIF medium, the composition can further meet the requirement that at least about 20%, for example 25%, or 30%, such as 35% of the budesonide is released into the dissolution medium within about 90 minutes, for example at least about 40% of the budesonide is released, such as about 30 to about 65% of the budesonide is released, such as about 35% to about 65% of the budesonide is released, including about 40% to about 60% of the budesonide is released, for example about 45% to about 55% of the budesonide is released into the dissolution medium.

[0170] In criterion c) of step (i) of the method, in the absence of added polysorbate 80 (e.g. Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in FaSSIF medium, the composition can further meet the requirement that at least about 10%, for example at least about 15% of the budesonide is released within about 90 minutes, such as about 10 to about 50% of the budesonide is released, such as about 10% to about 40% of the budesonide is released, including about 10% to about 30% of the budesonide is released, for example about 15% to about 30% of the budesonide is released into the dissolution medium.

[0171] In criterion c) of step (i) of the method, the composition can further meet the following requirement: at least about 90% of the budesonide is released into the dissolution medium within about 180 minutes, for example, at least about 95% of the budesonide is released within about 180 minutes. The amount of budesonide released within about 180 minutes is achieved in the presence and absence of added polysorbate 80 (e.g., Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in FaSSIF medium.

[0172] The paddle method of method 2 can be operated at about 50 rpm, about 75 rpm, or about 100 rpm. Preferably, the paddle method of method 2 is operated at about 100 rpm.

[0173] From pharmacokinetic studies of drug absorption in the fasting state, it is known that with the ingestion of 200 to 250 mL of water with the dosage form, a maximum total volume of approximately 300 to 500 mL is available in the proximal small intestine (see Klein, AAPS J., 12, 397, (2010)). Therefore, the dissolution test used in the method of the present invention should use a volume of dissolution medium (including FaSSIF) of at least about 500 mL (e.g., about 900 mL). Preferably, the initial volume of the dissolution medium used in criteria a), b), and c) is about 900 mL.

[0174] The procedures for testing the compositions can be essentially based on USP <711> / Delayed-release solid dosage forms according to European Pharmacopoeia 2.9.3 Method B.

[0175] The temperature of the dissolution medium in criteria a), b) and c) may be maintained at about 37°C ± 0.5°C.

[0176] The number of compositions tested may be at least 3, such as 6, or greater than 6, such as 12 or 24.

[0177] In criteria (a), (b), and (c), the volume withdrawn from the dissolution medium at each time point can be 10 mL or 15 mL, optionally without replacing the withdrawn volume. Withdrawal of the dissolution medium does not affect the overall dissolution profile of the composition. In other words, preferably, the dissolution test is performed under sink conditions with the amount of solvent exceeding the amount of solute, meaning that small withdrawals for analytical purposes do not affect dissolution.

[0178] Specifically, but not exclusively, when the composition of the invention is a core-shell composition as defined below, the method of the invention as defined herein may comprise the following additional steps:

[0179] (1) providing budesonide with the same extended-release excipient, but in the absence of a delayed-release excipient as described herein; and

[0180] (2) discriminating compositions that meet the following requirements in the standard in vitro USP <711> / European Pharmacopoeia 2.9.3 dissolution test using a dissolution apparatus according to the 2nd method (paddle method) of the test: about 20% to about 60%, such as about 25% to about 50% of budesonide is released into a dissolution medium comprising simulated intestinal fluid in fasted state simulated intestinal fluid, grade 1, pH about 6.5 in about 15 minutes.

[0181] In one embodiment, in the absence of a delayed release coating (e.g. no enteric coating), about 70 to about 90% of budesonide is released into grade 1 FaSSIF-V1 dissolution medium as defined herein in about 30 minutes, and more preferably, about 75 to about 85% of budesonide is released into the dissolution medium in about 45 minutes, in the absence of a delayed release (e.g. enteric) coating.

[0182] In the absence of a delayed release (e.g. enteric) coating, about 80% to about 90% of budesonide can be released into grade 1 FaSSIF-V1 dissolution medium as defined herein in about 60 minutes, more particularly, about 90%, such as about 95% (including about 97% and about 100%) of budesonide can be released into the dissolution medium in about 90 minutes, such as in about 120 minutes, including in about 180 minutes.

[0183] Budesonide compositions having dissolution profiles as described above in the absence of a delayed release excipient are a further confirmation that the majority of budesonide will be released in vivo to the ileum.

[0184] Effects on biomarkers

[0185] In all aspects outlined above, the method of the application can result in a statistically significant reduction in serum B cell activating factor (BAFF) (also known as tumour necrosis factor ligand superfamily member 13B (TNFSF13B)) levels in the subject relative to baseline levels of serum BAFF in the subject prior to treatment.

[0186] “Statistically significant reduction” includes the meaning of a statistically significant reduction using a p-value of <0.05 upon Analysis of Variance (ANOVA) when comparing the change seen in the treated patient group to the change seen in patients receiving placebo.

[0187] "Relative to the baseline level" includes the meaning that the measured level of a molecule (e.g., BAFF) is lower than the level measured at the beginning of the study (i.e., before drug administration). The baseline level is the level immediately before treatment begins and serves as a comparator for subsequently measured levels (e.g., at a time point immediately after or after the end of a course of treatment). Thus, such a decrease is specific to the subject or group of subjects in question and is not an absolute value.

[0188] Relative to the baseline serum level of BAFF in the subject before treatment, the reduction in serum BAFF levels in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. For example, the reduction in serum BAFF levels in the subject can be at least about 5%. Specifically, the reduction in serum BAFF levels in the subject can be at least about 10%, such as the reduction in serum BAFF levels in the subject can be at least about 14%.

[0189] The reduction in BAFF serum levels in a subject may be from about 1% to about 70%. For example, the reduction in BAFF serum levels in a subject may be from about 5% to about 50%. Specifically, the reduction in BAFF serum levels in a subject may be from about 5% to about 25%, such as from about 10% to about 25%. For example, the reduction in BAFF serum levels in a subject may be from about 14% to about 23%.

[0190] A statistically significant decrease in serum BAFF levels observed following the methods of the present invention can be associated with a statistically significant decrease in serum levels of one or more biomarkers associated with B cell activation and / or proliferation relative to the serum baseline levels of one or more biomarkers in the subject prior to treatment. A decrease in serum levels of one or more biomarkers associated with B cell activation and / or proliferation will indicate a beneficial effect in the treatment of diseases in which excessive activity, excess, and / or excessive proliferation of B cells is associated with pathogenesis. For example, a decrease in serum levels of a biomarker produced by active and / or proliferating B cells can indicate a decrease in the activity / proliferation of B cells and can indicate a beneficial effect in the treatment of diseases in which excessive activity, excess, and / or excessive proliferation of B cells is associated with pathogenesis, such as IgAN.

[0191] The one or more biomarkers may include: transmembrane activator and CAML interactor (TACI) (also known as tumor necrosis factor receptor superfamily member 13B (TNFRSF13B)); B-cell maturation antigen (BCMA) (also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17)); BAFF-R (also known as tumor necrosis factor receptor superfamily member 13C (TNFRSF13C)); CD27; CD30; CXC motif chemokine 12 (CXCL12); CXC motif chemokine 13 (CXCL13); chemokine (CC motif) ligand 19 (CCL19); interleukin 2 (IL-23); 2, IL-2); interleukin 6 (IL-6); chemokine (C-C motif) ligand 3 (CCL3); chemokine (C-C motif) ligand 4 (CCL4); soluble CD23 (sCD23); secretory IgA; IgA-IgG immune complex; poorly O-galactosylated IgA1; or a combination thereof.

[0192] The term "biomarker" (also called "biological marker") encompasses any measurable indicator of a biological state or condition. Biomarkers are typically naturally occurring biological molecules, such as proteins, amino acids, antibodies, nucleic acids (e.g., RNA or DNA), nucleotides, lipids, carbohydrates / sugars, primary metabolites, or secondary metabolites. Such biomarkers can be associated with specific pathological or physiological processes, diseases, or pharmacological responses to drugs, and can be used to predict disease incidence and prevalence, or the outcomes of disease and therapeutic interventions.

[0193] Relative to the baseline serum level of the biomarker in the subject before treatment, the reduction in the serum level of the biomarker in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. For example, the reduction in the serum level of a biomarker can be from about 1% to about 90%, or from about 5% to about 70%, or from about 10% to about 50%.

[0194] Relative to the baseline serum level of TACI in the subject before treatment, the reduction in serum level of TACI in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, or 75%. For example, the reduction in serum level of TACI in the subject can be at least about 5%. Specifically, the reduction in serum level of TACI in the subject can be at least about 11%.

[0195] The reduction in serum levels of TACI in a subject may be from about 1% to about 70%. For example, the reduction in serum levels of TACI in a subject may be from about 5% to about 50%. Specifically, the reduction in serum levels of TACI in a subject may be from about 5% to about 20%, such as from about 10% to about 20%. For example, the reduction in serum levels of TACI in a subject may be from about 11% to about 17%.

[0196] Relative to the baseline serum level of BCMA in the subject before treatment, the reduction in serum levels of BCMA in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60%. For example, the reduction in serum levels of BCMA in the subject can be at least about 1%. Specifically, the reduction in serum levels of BCMA in the subject can be at least about 6%.

[0197] Furthermore, the reduction in serum levels of BCMA in a subject may be from about 1% to about 60%. For example, the reduction in serum levels of BCMA in a subject may be from about 1% to about 20%. Specifically, the reduction in serum levels of BCMA in a subject may be from about 1% to about 10%, such as from about 5% to about 10%. For example, the reduction in serum levels of BCMA in a subject may be from about 6% to about 7%.

[0198] The reduction in serum levels of BAFF-R in a subject may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% relative to the subject's baseline serum level of BAFF-R before treatment.

[0199] Relative to the baseline serum level of CD27 in the subject before treatment, the reduction in serum levels of CD27 in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65%. For example, the reduction in serum levels of CD27 in the subject can be at least about 5%. Specifically, the reduction in serum levels of CD27 in the subject can be at least about 10%, such as the reduction in serum levels of CD27 in the subject can be at least about 15%.

[0200] The reduction in serum levels of CD27 in a subject may be from about 1% to about 60%. For example, the reduction in serum levels of CD27 in a subject may be from about 1% to about 25%. Specifically, the reduction in serum levels of CD27 in a subject may be from about 5% to about 25%, such as from about 10% to about 20%. For example, the reduction in serum levels of CD27 in a subject may be from about 15% to about 19%.

[0201] Relative to the baseline serum level of CD30 in the subject prior to treatment, the reduction in serum levels of CD30 in the subject may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%. For example, the reduction in serum levels of CD30 in the subject may be at least about 1%. Specifically, the reduction in serum levels of CD30 in the subject may be at least about 3%, such as the reduction in serum levels of CD30 in the subject may be at least about 5%.

[0202] The reduction in serum CD30 levels in a subject may be from about 1% to about 75%. For example, the reduction in serum CD30 levels in a subject may be from about 1% to about 25%. Specifically, the reduction in serum CD30 levels in a subject may be from about 1% to about 10%, such as from about 5% to about 10%. For example, the reduction in serum CD30 levels in a subject may be from about 5% to about 8%.

[0203] Relative to the baseline serum level of secretory IgA in the subject before treatment, the reduction in serum levels of secretory IgA in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%. For example, the reduction in serum levels of secretory IgA in the subject can be at least about 1%. Specifically, the reduction in serum levels of secretory IgA in the subject can be at least about 2%, such as the reduction in serum levels of secretory IgA in the subject can be at least about 3%.

[0204] The reduction in serum secretory IgA levels in a subject may be from about 1% to about 75%. For example, the reduction in serum secretory IgA levels in a subject may be from about 1% to about 25%. Specifically, the reduction in serum secretory IgA levels in a subject may be from about 1% to about 10%, such as the reduction in serum secretory IgA levels in a subject may be from about 1% to about 5%. For example, the reduction in serum secretory IgA levels in a subject may be from about 1% to about 3%.

[0205] Relative to the baseline serum level of IgA-IgG immune complexes in the subject before treatment, the reduction in serum levels of IgA-IgG immune complexes in the subject can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%. For example, the reduction in serum levels of IgA-IgG immune complexes in the subject can be at least about 1%. Specifically, the reduction in serum levels of IgA-IgG immune complexes in the subject can be at least about 5%, such as the reduction in serum levels of IgA-IgG immune complexes in the subject can be at least about 8%.

[0206] The decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 75%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 25%. In particular, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 20%, such as the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 20%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 15%.

[0207] The decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 75%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 25%. In particular, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 20%, such as the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 20%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 15%.

[0208] The decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 75%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 25%. In particular, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 1% to about 20%, such as the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 20%. For example, the decrease in serum levels of IgA-IgG immune complexes in the subject can be from about 2% to about 15%.

[0209] The above-mentioned biomarker reduction is relevant to the method of the present application, which requires that the budesonide-containing composition exhibits the specified in vitro release profile and is intended for the treatment of IgAN and / or is capable of treating IgAN.

[0210] As we have found, the composition exhibiting the in vitro release profile also exhibits an appropriate reduction of the relevant biomarker, which indicates that:

[0211] • budesonide is released into the region of the gastrointestinal tract (e.g. ileum) where the Peys patches are mainly located; and

[0212] • Such compositions are therefore able to safely and effectively treat IgAN at appropriate doses of budesonide.

[0213] Composition containing budesonide

[0214] The compositions useful in the methods of the present invention may comprise any combination of budesonide and one or more excipients that produce the desired in vitro release profile in all aspects as described herein. This may be a combination of an extended release coating and / or a delayed release coating, which may be applied by a variety of formulation principles as described below.

[0215] In any case, we prefer that the composition comprises at least one delayed release coating, preferably located on the exterior of the composition to ensure that the active ingredient is not released in the stomach and / or until it reaches the small intestine.

[0216] Such delayed-release coatings may thus comprise so-called "enteric coatings," which refer to materials having gastroresistant properties, ie, materials that prevent dissolution or disintegration in the gastric environment, thereby allowing the composition to pass through the stomach into the ileum region of the small intestine.

[0217] Enteric coatings can include azo polymers, disulfide polymers, cellulose acetate, cellulose acetate succinate, cellulose acetate phthalate, tetrahydrocellulose acetate phthalate, polyvinyl acetate phthalate, hydroxyethyl ethylcellulose phthalate, methacrylic acid copolymers, polymethacrylic acid / acrylic acid copolymers, styrene maleic acid copolymers, hydroxypropyl methylcellulose phthalate, acrylic resins, cellulose acetate trimellitate, hydroxypropyl methylcellulose trimellitate, shellac, hydroxyethyl ethylcellulose phthalate, carboxymethyl cellulose, and hydroxypropyl methylcellulose acetate succinate.

[0218] Specific enteric coating materials include cellulose acetate, cellulose acetate succinate, cellulose acetate phthalate, tetrahydrocellulose acetate phthalate, polyvinyl acetate phthalate, hydroxyethyl ethylcellulose phthalate, methacrylic acid copolymers, polymethacrylic acid / acrylic acid copolymers, styrene maleic acid copolymers, hydroxypropyl methylcellulose phthalate, acrylic resins, cellulose acetate trimellitate, hydroxypropyl methylcellulose trimellitate, shellac, hydroxyethyl ethylcellulose phthalate, carboxymethyl cellulose, and hydroxypropyl methylcellulose acetate succinate.

[0219] Preferred enteric coating materials include polyvinyl acetate phthalate and, especially, methacrylic acid copolymers.

[0220] It will be understood by those skilled in the art that the enteric coating may contain other commonly used materials such as talc (as a plasticizer), dibutyl sebacate (as a plasticizer), and a blend of HMPC and PEG as a secondary coating agent.

[0221] The composition may comprise one or more cores comprising budesonide encapsulated by a combination of delayed and extended release excipients to substantially prevent release of the contents of the composition prior to reaching the distal region of the small intestine (e.g., the ileum, such as the distal ileum). Such compositions are hereinafter referred to as "core-shell compositions of the invention," encompassing "beads" and "encapsulated cores."

[0222] The core containing budesonide can be loaded into a capsule. When capsules are used, a delayed release coating (eg, an enteric coating) can be on the capsule and not directly on the core.

[0223] When the enteric coating is on capsules, such as size 1 capsules, the enteric coating may be present in an amount of about 34 to about 46 mg per capsule, such as about 34 to about 42 mg per capsule, for example about 36 to about 40 mg per capsule.

[0224] To ensure that in such core-shell compositions of the invention the majority of the budesonide is released substantially to the distal region of the small intestine (eg the ileum, such as the distal ileum), they may (or may further) be separately coated with a prolonged release polymer coating.

[0225] For the avoidance of doubt, an extended release polymer coating is distinct from a delayed release coating.

[0226] Such extended-release coatings can ensure that the majority of the budesonide is adequately released throughout the ileum, and in combination with delayed-release coatings (eg, enteric coatings) can further ensure that such release is achieved substantially and / or predominantly to the ileal region of the small intestine.

[0227] "Substantially released to the ileum region" includes at least about 51%, such as at least about 60%, including at least about 70% or at least about 75%, such as at least about 80%, including at least about 90% of the initial content of the active ingredient in the composition being released to said region.

[0228] It will be understood by those skilled in the art that any pharmaceutical composition should be taken according to the prescription instructions, and if taken in a manner different from the prescription information, the desired effect may not be achieved. In the context of the present invention, the composition is preferably taken orally at least one hour before a meal, and more preferably, in the morning at least one hour before the first meal of the day.

[0229] The extended-release coating may comprise a pharmaceutically acceptable polymer blend comprising a water-insoluble polymer and a pore-forming polymer applied directly to the budesonide-containing core. The resulting core or composition comprising a plurality of cores may then be encapsulated in a delayed-release coating, the combination of excipients of which substantially prevents release of the contents of the composition prior to reaching the ileal region of the small intestine.

[0230] As used herein, the term "water insoluble polymer" refers to a polymer that has a water insoluble content of less than about 0.1 mg / mL in an aqueous solvent, such as water, at about 25°C. -1 The presence of a water-insoluble polymer enables the release rate of budesonide in the composition to be controlled.

[0231] The water-insoluble polymer may be an alkyl cellulose or a derivative thereof, for example, the water-insoluble polymer may be ethyl cellulose (or a derivative thereof).

[0232] The term "alkylcellulose or derivatives thereof" refers to compounds derived from cellulose in which the protons on at least some of the cellulose hydroxyl groups have been replaced by alkyl groups.

[0233] As used herein, the term "pore forming polymer" refers to a polymer that is more water soluble than a water insoluble polymer and therefore dissolves first, leaving pores in the coating, thereby allowing a certain amount of water to penetrate toward the core.

[0234] Thus, the porogenic polymer can be defined as "water-soluble." In other words, the porogenic polymer has a water-soluble capacity of at least about 10 mg mL-1 in an aqueous solvent, such as water, at 25°C. -1 dissolution.

[0235] The nominal viscosity of the porogenic polymer may be from about 1 to about 300 mPa*s, for example from about 1 to about 50 mPa*s, such as from about 1 to about 30 mPa*s, for example from about 1 to about 20 mPa*s, such as from about 2 to about 9 mPa*s, for example from about 2 to about 7 mPa*s, preferably from about 2 to about 6 mPa*s. The nominal viscosity of the porogenic polymer can be measured at 20° C. as a 2 wt % solution of the polymer in water by the standard European Pharmacopoeia 2.2.9 capillary viscometer method.

[0236] Furthermore, the porogenic polymer may have a gelling temperature of about 35 to about 65°C, for example about 55 to about 65°C, such as about 58 to about 64°C.

[0237] The porogenic polymer may comprise a polymer selected from the list consisting of polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC) and hydroxypropyl cellulose (HPC). Preferably, the porogenic polymer is hydroxypropyl methylcellulose.

[0238] The degree of substitution of HPMC with methoxy groups may be from about 15 to about 35 weight %, for example, from about 25 to about 35 weight %, or from about 27 to about 31 weight %, such as from about 27 to about 30 weight %. In addition, the degree of substitution of HPMC with hydroxypropyl groups may be from about 4 to about 32 weight %, for example, from about 4 to about 20 weight %, or from about 5 to about 15 weight %, such as from about 7 to about 12 weight %.

[0239] The term "degree of substitution of the HPMC" refers to the average level of substitution of hydroxyl groups on the cellulose chains and is expressed herein in percentage terms, ie, the percentage of hydroxyl groups that have been substituted with the moiety in question.

[0240] The water-insoluble polymer can be present in an amount of about 45% to about 90% by weight of the total extended-release coating and the pore-forming polymer can be present in an amount of about 35% to about 5% by weight of the total extended-release coating. For example, the water-insoluble polymer can be present in an amount of about 45% to about 65% by weight of the total extended-release coating and the pore-forming polymer can be present in an amount of about 35% to about 15% by weight of the total extended-release coating. For example, the water-insoluble polymer can be present in an amount of about 47% to about 56% by weight of the total extended-release coating and the pore-forming polymer can be present in an amount of about 32% to about 22% by weight of the total extended-release coating.

[0241] The pharmaceutically acceptable polymeric blend of the extended release coating may comprise fatty acids in an amount of about 2 wt% to about 8 wt%, such as about 3 wt% to about 7 wt% of the total extended release coating.

[0242] The fatty acids may be unsaturated fatty acids, such as C4 to C28 Unsaturated fatty acids, such as C 13 to C 22 Unsaturated fatty acids. For example, the unsaturated fatty acid can be selected from the group consisting of myristoleic acid, palmitoleic acid, hexadecenoic acid, oleic acid, elaidic acid, vaccenic acid, and erucic acid. Preferably, the unsaturated fatty acid is oleic acid.

[0243] The pharmaceutically acceptable polymeric blend of the extended release coating may comprise medium chain triglycerides in an amount of about 3 wt % to about 12 wt %, such as about 5 wt % to about 12 wt %, for example about 5 to about 8 wt % of the total extended release coating.

[0244] The term "medium-chain triglyceride" refers to a triglyceride having an aliphatic tail of 6 to 12 carbons. For example, the medium-chain triglyceride can be selected from the list consisting of caproic acid, caprylic acid, capric acid, and lauric acid.

[0245] The pharmaceutically acceptable polymer blend of the extended-release coating may comprise another water-soluble polymer in an amount of about 1% to about 5% by weight, such as about 2% to about 3% by weight, of the total extended-release coating. For the avoidance of doubt, the other water-soluble polymer is different from the porogenic polymer. Preferably, the other water-soluble polymer is a poly(ethylene glycol) having a molecular weight in the range of about 200 to about 1000 g / mol.

[0246] The polymeric blend coating the one or more cores may be coalescable.The term "coalescable" when referring to an extended release polymeric blend means that the polymers of the extended release blend are capable of being blended to form a single polymeric phase coating.

[0247] Thus, the extended release polymeric blend coating the one or more cores may comprise one or more coalescing polymers.The one or more coalescing polymers may comprise a water-insoluble polymer.

[0248] The extended release polymer blend may be present in an amount of about 5 to about 18 wt % of the total bead / core, for example about 6 to about 16 wt % of the total bead / core, for example about 6 to about 13 wt %, such as about 6 to about 12 wt % of the total bead / core.

[0249] It has been found that in the polymer blend of the extended release coating, the pore-forming polymer dissolves first in the aqueous solution before the water-insoluble polymer, leaving pores in the coating, thereby allowing a certain amount of water to permeate towards the core in a controlled manner.

[0250] The average size of the core may range from about 0.5 to about 3 mm, for example from about 0.5 to about 2 mm, such as from about 0.8 to about 1.5 mm.

[0251] The core-shell composition of the present invention can be prepared by the following operations:

[0252] (a) providing one or more cores comprising budesonide;

[0253] (b) the one or more cores are individually coated with a pharmaceutically acceptable extended release polymeric blend comprising a water-insoluble polymer in an amount of from about 45 wt % to about 90 wt % and a porogenic polymer in an amount of from about 35 wt % to about 5 wt %; and

[0254] wherein the composition is encapsulated in a delayed-release coating so as to substantially prevent release of the contents of the composition prior to reaching the ileum region of the small intestine.

[0255] The core-shell composition of the above operation may comprise any of the features as outlined above in relation to the method of the invention.

[0256] The core may be prepared by providing inert (eg sugar) beads and coating them with the aqueous budesonide suspension. The size of the inert beads may be from about 1 to about 2 mm, such as from about 1 to about 1.5 mm, for example from about 1 to about 1.2 mm.

[0257] As referred to herein, the term "inert bead" includes a single pharmaceutically inert bead that provides the starting material for preparing the core-shell composition of the present invention.

[0258] The inert beads are preferably commercially available sugar spheres (commonly known as Non-pareil). Sugar spheres primarily contain sucrose, with smaller amounts of other materials such as starch added. Suppliers of sugar spheres include Paulaur Corporation (USA), Chr. Hansen (Denmark), NP Pharm (France), Emilio Castelli (Italy), and JRS Pharma (Germany).

[0259] Prior to adding the extended release polymeric blend, the core may be coated with a seal coat comprising a stabilizing and water-soluble polymer. The stabilizer may be an acid, preferably citric acid, and the soluble polymer is the same polymer used as the pore-forming polymer in the polymeric blend. Alternative stabilizers for inclusion in the seal coat include poly(vinyl pyrrolidone) (PVP), and the soluble polymer is the same polymer used as the pore-forming polymer in the polymeric blend.

[0260] The extended release polymeric blend can be applied to the core as an aqueous polymeric suspension and the suspension sprayed onto the core. The aqueous polymeric blend can be sprayed onto the core at a temperature of from about 30°C to about 65°C, such as from about 30°C to about 50°C.

[0261] At the upper end of this range, such as at a temperature of from about 50 to about 65°C, spraying can avoid the need for a separate solidification / coalescing step as outlined below.

[0262] The core-shell composition of the present application can be obtained by the operation of coating the core in a fluid bed apparatus as defined above. That is, the coating of the extended release polymeric blend can be carried out in a fluid bed apparatus. Suitable fluid bed apparatuses are readily available from suppliers such as Glatt GmbH.

[0263] Following coating with the polymeric blend, the polymer can be coalesced, wherein coalescing can be carried out by solidification.

[0264] Solidification can be carried out at a temperature of from about 55°C to about 75°C, such as from about 60°C to about 70°C, for example from about 63°C to about 66°C. Further, solidification can be carried out for from about 1 hour to about 10 hours, for example from about 1 to about 5 hours, such as from about 2 to about 4 hours.

[0265] It was found that the extended release polymeric coating blend allows the use of a fluid bed apparatus to produce budesonide cores in a cost effective manner, while obtaining a composition with the desired release profile.

[0266] Accordingly, additionally, solidification of the core-shell composition of the present application can be carried out in a fluid bed apparatus.

[0267] According to another aspect of the present application, there is provided a composition producing the desired in vitro release profile in all aspects as described herein, comprising a plurality of beads, the beads comprising:

[0268] (a) a budesonide-containing core, the budesonide in the core being present as a coating on one of the more inert core substrates (e.g. sugar beads) as described herein;

[0269] (b) an extended release coating present on the budesonide-containing core in an amount of between about 6 and about 12 wt% of the total bead weight, and the coating comprising a coalesced blend of at least two polymers (i) and (ii):

[0270] (i) any one of the water-insoluble polymers described herein (e.g. ethyl cellulose), and

[0271] (ii) any one of the pore-forming polymers described herein (e.g. hydroxypropyl methyl cellulose which is substituted to the extent of from about 27 to about 30 wt% by methoxy groups and / or from about 7 to about 12 wt% by hydroxypropyl groups),

[0272] The blend of polymers (i) and (ii) is in any of the above mentioned ratios (e.g. water-insoluble polymer (i) in an amount of about 47% to about 56% by weight of the total extended release coating and pore-forming polymer (ii) in an amount of about 32% to about 22% by weight of the total extended release coating, respectively),

[0273] The beads are subsequently loaded into capsules which are coated with a delayed release coating (e.g. any of the enteric coatings described herein, such as polyvinyl acetate phthalate or especially methacrylic acid copolymers).

[0274] The above composition can comprise one or more of the other preferred features disclosed in relation to the core-shell composition and / or beads as described herein, such as:

[0275] • any of the fatty acids described herein, e.g. in an amount of about 3% to about 7% by weight of the total extended release coating;

[0276] • any of the medium chain triglycerides described herein, e.g. in an amount of about 5 to about 8% by weight of the total extended release coating;

[0277] • one or more other water-soluble polymers, such as poly(ethylene glycol) having a molecular weight in the range of about 200 to about 1000 g / mol, in an amount of about 2% to about 3% by weight of the total extended release coating.

[0278] The above composition can further comprise a seal coating solution of a suitable acid (e.g. citric acid) between the budesonide-containing core and the extended release coating.

[0279] Each capsule can comprise about 4 mg of budesonide and can deliver a total of about 16 mg when four capsules are taken orally by a patient when administered to a patient.

[0280] The coalesced blend of at least two polymers (i) and (ii) obtained prior to loading the coated beads into capsules is preferably obtained by coating the budesonide-containing core in a fluid bed apparatus by applying an aqueous dispersion comprising polymers (i) and (ii) as defined above at a temperature between about 30°C to about 65°C, such as about 30°C to about 50°C, especially about 50 to about 65°C, and subsequently, if necessary, solidifying it by applying a temperature of about 60°C to about 70°C (e.g. about 63°C to about 66°C) for a suitable time (e.g. about 2 to about 4 hours) to further coalesce the polymeric extended release coating thus coated within the fluid bed apparatus.

[0281] Alternatively, the polymeric blend can be sprayed as an organic solution. In this embodiment, no further solidification step is necessary.

[0282] As mentioned above, the core coated by the aforementioned extended release polymer blend may be loaded into a capsule, and the capsule may be coated with the delayed release coating.

[0283] Regardless of how they are made, the compositions useful in the methods of the present invention are useful for treating IgAN because they meet the in vitro release characteristics defined above.

[0284] Therefore, it is further provided that:

[0285] Any one of the compositions as defined herein, for use in treating IgAN;

[0286] ● Use of any of the compositions as defined herein for the manufacture of a medicament for the treatment of IgAN; and

[0287] - A method of treating IgAN, the method comprising administering any one of the compositions as defined herein to a patient in need of such treatment.

[0288] According to another aspect of the present invention, there is provided a core-shell composition of the present invention as described above.

[0289] As an alternative to core-shell compositions, the composition may comprise a tablet comprising budesonide encapsulated within one or more excipients that substantially prevent release of the contents of the composition prior to reaching the ileum region of the small intestine. Such compositions are hereinafter referred to as "encapsulated tablet compositions of the invention".

[0290] The one or more excipients that encapsulate the tablet to substantially prevent release of the contents of the composition prior to reaching the ileal region of the small intestine may be an enteric coating as defined above.

[0291] The enteric coating in the tablet composition may be present in an amount from about 5% to about 15% by weight of the total tablet, such as from about 7% to about 13% by weight, for example from about 8% to about 12% by weight.

[0292] The tablet composition of the present invention may comprise wet granules of budesonide and a filler. Optionally, the filler comprises dibasic calcium phosphate, microcrystalline cellulose, mannitol, or a mixture thereof.

[0293] The filler may be present in an amount from about 50% to about 80% by weight of the total tablet, such as from about 60% to about 75% by weight, for example from about 65% to about 75% by weight.

[0294] The tablet composition of the present invention may be a compressed tablet further comprising a lubricant, optionally wherein the lubricant is magnesium stearate, aluminum stearate, calcium stearate, sodium stearate, zinc stearate, stearic acid, capric acid, lauric acid, sodium stearyl fumarate, or a mixture thereof.

[0295] The lubricant may be present in an amount from about 0.1% to about 2% by weight of the total tablet, such as from about 0.1% to about 1% by weight.

[0296] The tablet composition of the present invention may further comprise a disintegrant, optionally wherein the disintegrant is selected from crospovidone, croscarmellose sodium, or sodium starch glycolate. Alternatively, the tablet may not comprise a disintegrant.

[0297] The disintegrant may be present in an amount from about 0.5% to about 5% by weight of the total tablet, such as from about 0.5% to about 4% by weight, for example from about 0.8% to about 3.5% by weight.

[0298] The tablet composition of the present invention may further comprise a binder, optionally wherein the binder is selected from hydroxyethyl cellulose, hydroxypropyl cellulose, copovidone, or a mixture thereof.

[0299] The binder may be present in an amount from about 5% to about 10% by weight of the total tablet, such as from about 6% to about 9% by weight, for example from about 7% to about 9% by weight.

[0300] Such tablet compositions of the present invention may be gel matrix tablets further comprising a gel matrix material, such as low molecular weight HPMC (eg, hypromellose).

[0301] The gelling matrix material may be present in an amount of about 10 to about 25 weight %, such as about 10 to about 20 weight %, for example about 15 to about 20 weight % of the total tablet.

[0302] The gelling matrix material may be diluted with a water-soluble filler, which may include lactose, dextrose, mannitol, and combinations thereof.

[0303] The tablet may comprise an enteric coating comprising any of the materials as outlined above. The enteric coating may be present in an amount of about 5 to about 15 weight % of the total tablet, such as about 8 to about 12 weight % of the total tablet.

[0304] Each tablet may contain from about 2 to about 20 mg budesonide, such as from about 4 to about 16 mg budesonide. Preferably, each tablet contains about 4 mg budesonide.

[0305] Compositions, when characterized by the methods of the invention (in all aspects), are useful because they can more effectively treat IgAN. Such compositions, pharmaceutical formulations, uses, and methods described herein may also have the following advantages: in the treatment of IgAN, they may be more convenient for the physician and / or patient, more effective, less toxic, have a broader spectrum of activity, be more potent, produce fewer side effects, have lower inter-patient variability, or may have other useful pharmacological properties that are superior to similar formulations or methods (treatments) known in the prior art, whether for the treatment of IgAN or otherwise.

[0306] When the word "about" is used herein in the context of quantities, e.g., absolute quantities such as weight, volume, size, diameter, etc., or relative quantities (e.g., percentages) of individual ingredients in a composition or component of a composition (including concentrations and ratios), time frames, and parameters such as temperature, it is understood that such variations are approximate and, therefore, may vary from the actual numerical value specified herein by ±10%, e.g., ±5%, and preferably ±2% (e.g., ±1%). This is true even if such numerical values ​​are first expressed as a percentage (e.g., "about 10%" may mean approximately 10 ± 10%, i.e., anywhere between 9% and 11%). BRIEF DESCRIPTION OF THE DRAWINGS

[0307] Figure 1 A flow chart detailing the preparation of cured polymer-coated beads.

[0308] Figure 2 Shown are in vitro dissolution profiles of budesonide modified-release capsules in pH 6.8 buffered medium in the presence of added surfactant at a paddle rotation speed of 100 rpm; data are shown for different capsule batches prepared according to the present invention.

[0309] Figure 3 repeat Figure 2 The in vitro dissolution profiles presented in , but only focused on dissolution at pH 6.8.

[0310] Figure 4 : The in vitro dissolution profiles of seven separate batches of budesonide modified-release capsules according to the present invention are relatively shown, wherein the dissolution in a pH 6.8 buffer medium is in the absence of a surfactant and at a paddle rotation speed of 50 rpm.

[0311] Figure 5 : Shows the in vitro dissolution profiles of the budesonide modified-release capsules according to the present invention and three other commercially available budesonide-containing formulations in a pH 6.8 buffer medium in the presence of added surfactant and at a paddle rotation speed of 100 rpm.

[0312] Figure 6: Shows the in vitro dissolution profiles of the budesonide modified-release capsules according to the present invention and three other commercially available budesonide-containing formulations in a pH 6.8 buffer medium in the absence of added surfactant and at a paddle rotation speed of 100 rpm.

[0313] Figure 7 : Percent change in BAFF levels relative to baseline levels after treatment. Percent change in BAFF relative to baseline levels measured in patients after 9 months of treatment with: (a) placebo; (b) nafoam-budesonide (8 mg / day); and (c) nafoam-budesonide (16 mg / day). Dashed lines indicate no percent change after intervention with placebo or nafoam-budesonide.

[0314] Figure 8 : Percent change in BAFF levels after the follow-up period relative to the end-of-treatment level. Percent change in BAFF was measured in patients treated with 16 mg / day of budesonide for 9 months: (a) compared to baseline at the end of the 9-month treatment period; and (b) compared to the end of treatment after the 12-month follow-up period. The dotted line indicates the percentage of no change after intervention with placebo or budesonide.

[0315] Figure 9 : Percent change in APRIL levels relative to baseline levels after treatment. Percent change in APRIL levels relative to baseline levels measured in patients after 9 months of treatment with: (a) placebo; (b) nefocon-budesonide (8 mg / day); and (c) nefocon-budesonide (16 mg / day). The dotted line indicates the percentage of no change after intervention with placebo or nefocon-budesonide.

[0316] Figure 10 : Percent change in TACI levels from baseline after treatment. Percent change in TACI levels from baseline measured in patients after 9 months of treatment with: (a) placebo; (b) nafoam-budesonide (8 mg / day); and (c) nafoam-budesonide (16 mg / day). The dotted line indicates no percentage change after intervention with placebo or nafoam-budesonide.

[0317] Figure 11 : Percent change in BCMA levels relative to baseline levels after treatment. Percent change in BCMA levels relative to baseline levels measured in patients after 9 months of treatment with: (a) placebo; (b) nafocon-budesonide (8 mg / day); and (c) nafocon-budesonide (16 mg / day). The dotted line indicates no percentage change after intervention with placebo or nafocon-budesonide.

[0318] Figure 12 : Percent change in CD27 levels from baseline following treatment. Percent change in CD27 levels measured in patients following 9 months of treatment with: (a) placebo; (b) Resochin-Budesonide (8 mg / day); and (c) Resochin-Budesonide (16 mg / day). Dotted line indicates no percent change following intervention with placebo or Resochin-Budesonide.

[0319] Figure 13 : Percent change in CD27 levels from treatment end following follow-up period. Percent change in CD27 measured in patients administered Resochin-Budesonide (16 mg / day) for 9 months in: (a) comparison to baseline, at the end of the 9 month treatment period; and (b) comparison to treatment end, following the 12 month follow-up period. Dotted line indicates no % change following intervention with placebo or Resochin-Budesonide.

[0320] Figure 14 : Percent change in CD30 levels from baseline following treatment. Percent change in CD30 levels measured in patients following 9 months of treatment with: (a) placebo; (b) Resochin-Budesonide (8 mg / day); and (c) Resochin-Budesonide (16 mg / day). Dotted line indicates no percent change following intervention with placebo or Resochin-Budesonide.

[0321] Figure 15 : Serum levels of secretory IgA were shown to have a significant (p<0.05) Budesonide capsule dependent decrease. However, serum levels of IgA were unchanged.

[0322] Figure 16 : Serum levels of IgA-IgG immune complexes were shown to have a significant (p<0.05) Budesonide capsule dose dependent decrease.

[0323] Figure 17 : Levels of poorly O-galactosylated IgAl were shown to have a significant (p<0.05) Budesonide capsule dose dependent decrease.

[0324] Figure 18 : No differences in total IgA, IgAl and IgG levels were observed with Budesonide capsule treatment.

[0325] Figure 19 : In vitro dissolution profile of Budesonide modified release capsules in simulated intestinal fluid at pH ~ 6.5 in a fasted state, grade 1, with added surfactant.

[0326] Figure 20: Presents the in vitro dissolution profiles of budesonide modified-release capsules in FaSSIF compared to three other commercially available budesonide-containing formulations in the presence of added surfactants in a fasted-state simulated intestinal fluid level 1 at a pH of approximately 6.5.

[0327] Figure 21 : Shows the in vitro dissolution profile of a core-shell bead formulation containing budesonide in the absence of an enteric-coated capsule.

[0328] Figure 22 : Presents the in vitro dissolution profiles of budesonide modified-release capsules in FaSSIF compared to three other commercially available budesonide-containing formulations in a fasted state simulated intestinal fluid level 1 at pH approximately 6.5 in the absence of added surfactant.

[0329] Figure 23: (a) shows the results of the PBPK model based on the dissolution of modified-release capsules; (b) shows the results of the PBPK model based on the comparative product Results of the PBPK model of dissolution.

[0330] Figure 24 : Coronal T2* / T1-weighted TRUFI MRI images of the iron oxide-loaded capsules at 15 and 90 minutes, and the iron oxide dispersion in the ileum at 270 minutes. DETAILED DESCRIPTION

[0331] Examples

[0332] Example 1: Preparation of core-shell compositions

[0333] Opadry OY-7240, referred to below, is a dry powder polymer blend having the following components:

[0334]

[0335] Surelease is a polymer dispersion with the following components:

[0336] test Minimum maximum Oleic acid, % 1.6 2.2 Ethyl cellulose, % 17.0 20.0 Oleic acid to ethyl cellulose ratio 0.00 0.14 Medium chain triglycerides, % 0.80 4.00 MCT to ethylcellulose ratio 0.00 0.24 glycerin,% 0.0 0.6 solid,% 23.0 26.0 Ph 9.5 11.5 Brookfield viscosity, cps 400.00 1500.00

[0337] For the avoidance of doubt, the minimum and maximum values ​​in the above table refer to the minimum and maximum amounts of these components in the different batches of Surelease.

[0338] A flow chart detailing the preparation of budesonide beads of a core-shell composition according to the present invention is provided in Figure 1 and are explained in further detail below.

[0339] The budesonide coating suspension was prepared by dissolving Opadry OY-7240 Clear (2.29 kg) in purified water (26.5 kg) and then adding micronized budesonide (0.640 kg) to the solution while continuing to mix.

[0340] Sugar spheres (40.3 kg) with a mesh size of 16 to 18 were loaded into the preheated product tank of the fluidized bed. When the product temperature reached the target of 45°C, the active coating suspension was sprayed onto the sugar spheres / inert cores. The operation was monitored and controlled by a process computer. After the desired amount of active coating suspension had been sprayed, the active coated beads were dried and cooled.

[0341] A seal coating solution was prepared by dissolving citric acid monohydrate (0.093 kg) and Opadry OY-7240 Clear (2.26 kg) in purified water (21.8 kg) while continuously mixing.

[0342] The seal coating solution is applied to the pre-warmed active coated beads. The operation is monitored and controlled by a processing computer. When the required amount of solution has been sprayed, the coating is stopped and the seal coated beads are dried and cooled. The fluidized bed is emptied and the beads are screened using 1.4mm (14 mesh) and 0.5mm (35 mesh) screens to remove any oversized and undersized particles. The beads are weighed and the yield of the accepted fraction is calculated.

[0343] The extended release coating solution was prepared by adding Opadry OY-7240 (1.37 kg) to purified water (16.3 kg) while continuing to mix. Ethylcellulose type B suspension dispersion (Surelease, 12.8 kg) was added to the Opadry solution during continued mixing.

[0344] The qualified portion of the sealed coated beads is loaded into the preheated product tank of the fluidized bed with Wurster column. When the product temperature reaches the target temperature, the extended release polymer coating suspension is applied to the beads. By processing computer monitoring and control operation. The spraying amount is calculated by the amount of the qualified portion of the sealed coated beads from the previous step.

[0345] The resulting polymer blend for the extended release coating on the beads contained approximately 27.3 wt% HPMC in the total blend and approximately 51.8 wt% ethylcellulose in the total blend. Ethylcellulose is a water-insoluble polymer as defined above and HPMC serves as a pore-forming polymer.

[0346] After spraying is complete, the beads are dried and cooled. The fluidized bed is emptied and the beads are screened using 1.4 mm (14 mesh) and 0.5 mm (35 mesh) sieves to remove any oversized and undersized particles. The beads are weighed and the yield of the qualified portion is calculated.

[0347] An acceptable portion of the polymer-coated beads was loaded into the preheated drying tank of the fluidized bed apparatus. The beads were cured at a target temperature of 65°C for 3 hours. The operation was monitored and controlled by a processing computer. The fluidized bed was emptied and a sample of the cured beads was obtained for analysis and dissolution testing. The cured beads were weighed and the yield was calculated. The beads were filled into a hopper in a stainless steel bin.

[0348] The solidified beads were then filled into size 1 capsules using an automated encapsulator and then coated with an enteric coating. The enteric coating used for the capsules was a 1:1 and 1:2 blend of methacrylic acid and methyl methacrylate copolymer. The amount of enteric coating applied to each capsule ranged from about 34 to about 42 mg per capsule. The total amount of budesonide in each capsule was about 4 mg.

[0349] Example 2: In the presence of a surfactant and at a paddle rotation speed of 100 rpm in the buffer stage according to USP <711> / General method for in vitro dissolution test according to European Pharmacopoeia 2.9.3

[0350] As described in Ph. Eur. 2.9.3 Dissolution test for solid dosage forms (using Apparatus 2) and USP <711> The in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed as described in Dissolution (using Method 2). The measurements were performed as follows.

[0351] Dissolution Equipment Setup

[0352]

[0353]

[0354] Budesonide release was measured using ultra performance liquid chromatography (UPLC).

[0355] Reagents and standards

[0356] Standards and reference materials:

[0357] Budesonide, European Pharmacopoeia CSR, or a suitable secondary standard.

[0358] Other reagents:

[0359] Tween 80, (polyethylene oxide (20), polysorbate (80), Fisher Scientific, or equivalent.

[0360] Dissolution media and diluents

[0361] Acid resistant media

[0362] 0.1 N HCl solution. To prepare 6 L of acid-resistant medium, 50 mL of concentrated HCl was mixed with 6000 mL of water and the resulting solution was mixed thoroughly.

[0363] 0.2 M trisodium phosphate buffer solution

[0364] To prepare 1 L of 0.2 M trisodium phosphate buffer solution, approximately 76.02 g of trisodium phosphate was added and dissolved in 1000 mL of water, followed by mixing.

[0365] Buffered dissolution medium: 50 mM sodium phosphate buffer containing Tween 80, pH 6.8.

[0366] To make 6 L of buffered dissolution medium, combine 4500 mL of acid-resistant medium with 1500 mL of 0.2 M trisodium phosphate buffer solution and 3 g of Tween 80, followed by mixing. Check the pH and, if necessary, adjust the pH to 6.8 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0367] Analytical Operations

[0368] Acid-resistant operation

[0369] Notes: Care should be taken not to scratch or damage the capsule when placing it in the sinker.

[0370] Place 900 mL of preheated, degassed, acid-resistant medium in each of the six dissolution vessels. Maintain the medium at a temperature of 37°C ± 0.5°C.

[0371] According to USP <711> / Pharmacopoeia Test No. 2.9.3 Rotating Paddle Apparatus Method The apparatus was operated at 100 rpm.

[0372] The 6 capsules were then each placed in a separate coil sinker and then placed in a separate container.

[0373] At 2 hours, a 15 mL aliquot of the acid-resistant solution was withdrawn using a syringe.

[0374] The test solution was filtered through a Whatman GF / F GMF filter, wherein the first 5 mL was discarded and the remaining solution was collected in a test tube.

[0375] The following two steps are completed after the buffer phase dissolution has been initiated.

[0376] 5.0 mL of the filtered acid-resistant sample solution was pipetted into a 10 mL volumetric flask and diluted with acetonitrile to make up the volume.

[0377] The solution was mixed thoroughly and aliquots were transferred to HPLC vials and analyzed.

[0378] The release of budesonide during the acid-resistant phase was based on USP <711> / Evaluation of the acceptance criteria in European Pharmacopoeia 2.9.3.

[0379] Buffer stage dissolution operation

[0380] After drawing the acid-resistant sample, use forceps to transfer each coil sinker containing budesonide capsules to a different set of dissolution vessels containing 900 mL of buffered dissolution medium at a temperature of 37°C ± 0.5°C.

[0381] According to USP <711> / Pharmacopoeia Test No. 2.9.3 Rotating Paddle Apparatus Method The apparatus was operated at 100 rpm.

[0382] Sampling at designated time points: At 0.5 and 2 hours, 15 mL aliquots of the dissolution solution were withdrawn using a syringe. The withdrawn fluid was not replaced.

[0383] The test solution was filtered through a Whatman GF / F GMF filter, wherein the first 5 mL was discarded and the remaining solution was collected in a test tube.

[0384] 5.0 mL of the filtered dissolution sample solution was transferred to a 10 mL volumetric flask and diluted with acetonitrile to make up the volume.

[0385] The solution was mixed thoroughly and aliquots were transferred to HPLC vials.

[0386] The release of budesonide during the buffer phase was based on USP <711> / Evaluation of the acceptance criteria in European Pharmacopoeia 2.9.3.

[0387] Example 3: In the presence of surfactant and at a paddle rotation speed of 100 rpm in the buffer stage External USP <711> / Dissolution curve analysis of Pharmacopoeia Test No. 2.9.3

[0388] The capsules prepared in Example 1 ("budesonide capsules" or "nefecon budesonide") were tested under the dissolution conditions outlined in Example 2.

[0389] The overall dissolution profiles for three separate batches can be found in Figure 2The 0 to 2 hour period was at acidic pH (pH 1.2) and the 2 to 4 hour period was at buffered pH 6.8. Figure 3 repeat Figure 2 , but only showed dissolution at buffer pH 6.8, Figure 3 The time from 0 to 120 minutes corresponds to Figure 2 The time is 2 to 4 hours. Twelve capsules are tested in each batch.

[0390] The quantitative results of budesonide dissolution in various media at the time points of 2 hours at pH 1.2, 0.5 hours at pH 6.8, and 2 hours at pH 6.8 are provided in Table 1 below.

[0391] Table 1

[0392]

[0393] *SD = Standard Deviation

[0394] The release of budesonide in the acid-resistant and buffered phases is based on <711> / Evaluation of the acceptance criteria in European Pharmacopoeia 2.9.3.

[0395] Example 4: In the absence of surfactant and at a paddle rotation speed of 50 rpm in the buffer stage according to USP <711> / General method for in vitro dissolution test according to European Pharmacopoeia 2.9.3

[0396] As described in Ph. Eur. 2.9.3 Dissolution Tests for Solid Dosage Forms (Using Apparatus 2) and USP <711> The in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed as described in Dissolution (using Method 2). The measurement was performed as follows.

[0397] Dissolution Equipment Setup

[0398]

[0399] Budesonide release was measured using ultra-performance liquid chromatography (UPLC).

[0400] Reagents and standards

[0401] Standards and reference materials:

[0402] Budesonide, European Pharmacopoeia CSR, or a suitable secondary standard.

[0403] Dissolution media and diluents

[0404] Acid resistant media

[0405] 0.1 N HCl solution. To prepare 6 L of acid-resistant medium, 50 mL of concentrated HCl was mixed with 6000 mL of water and the resulting solution was mixed thoroughly.

[0406] 0.2 M trisodium phosphate buffer solution

[0407] To prepare 1 L of 0.2 M trisodium phosphate buffer solution, add approximately 76.02 g of trisodium phosphate and dissolve in 1000 mL of water, then mix.

[0408] Buffered dissolution medium: 50 mM sodium phosphate buffer with Tween 80, pH 6.8.

[0409] To make 6 L of buffered dissolution medium, combine 4500 mL of acid resistant medium with 1500 mL of 0.2 M trisodium phosphate buffer solution. Check the pH and adjust the pH to 6.8 ± 0.05 using hydrochloric acid or sodium hydroxide, as necessary.

[0410] Analytical Operations

[0411] Acid resistant handling

[0412] Notes: Care should be taken not to scratch or damage the capsules when placed in the sinker basket.

[0413] Place 900 mL of preheated, degassed acid resistant medium into each of the 6 dissolution cups. Maintain the medium at a temperature of 37 °C ± 0.5 °C.

[0414] Operate the equipment according to USP <711> / Pharmacopeial Test Number 2.9.3, Rotating Paddle Method Method at 50 rpm.

[0415] Subsequently place 6 capsules each into a separate coil sinker basket and then into a separate container.

[0416] At 2 hours, withdraw a 15 mL aliquot of the acid resistant solution using a syringe.

[0417] Filter the test solution with a Whatman GF / F GMF filter, discarding the first 5 mL and collecting the remaining solution in a test tube.

[0418] Complete the following two steps after the buffer phase dissolution has been initiated.

[0419] Pipette 5.0 mL of the filtered acid resistant sample solution into a 10 mL volumetric flask and dilute with acetonitrile to volume.

[0420] Mix the solution thoroughly and transfer an aliquot to an HPLC vial and analyze.

[0421] Budesonide release in the acid resistant phase is evaluated based on accepted standards in USP <711> / European Pharmacopoeia 2.9.3.

[0422] The capsules prepared in Example 1 ("Budesonide Capsules" or "Nevconazole Budesonide") were tested under the dissolution conditions outlined in this example.

[0423] The overall dissolution profiles for seven individual batches can be found in Figure 4 The 0 to 2 hour period was at acidic pH (pH 1.2) and the 2 to 4.5 hour period was at buffered pH 6.8.

[0424] For all samples, no more than 10% of the budesonide was released at the time point of 0.625 hours.

[0425] The quantitative results of budesonide dissolution at pH 6.8 at time point 1.25 hours (75 minutes) are provided in the table below.

[0426] Capsule batch number average(%) Min(%) Max(%) SD 3160733R 71 66 74 3.2 3166190R 48 43 53 4.1 3181645R 34 23 39 5.6 3197634R 61 57 71 5.1 3197635R 56 46 63 5.7 3197636R 55 49 60 3.9 3197637R 51 48 53 1.7

[0427] *SD = Standard Deviation

[0428] At 1.25 hours (75 minutes), 23% to 74% of the budesonide was released.

[0429] The quantitative results of budesonide dissolution at pH 6.8 at time point 2.5 hours (150 minutes) are provided in the table below.

[0430] Capsule batch number average(%) Min(%) Max(%) SD 3160733R 101 99 103 1.5 3166190R 90 87 93 2.2 3181645R 80 77 83 2.1 3197634R 93 90 95 2.1 3197635R 93 88 98 3.9 3197636R 92 88 95 2.4 3197637R 89 87 92 1.7

[0431] The lowest release observed was 77% at 2.5 hours.

[0432] Example 5: In the presence of a surfactant and at a paddle rotation speed of 100 rpm in the buffer stage according to USP <711> / Comparative test according to European Pharmacopoeia 2.9.3

[0433] The following is an overview of a variation of the in vitro test of Example 2. In this test, budesonide modified-release capsules according to the present invention and three other commercially available budesonide-containing formulations were analyzed. (TillottsPharma), (Dr Falk Pharma GmbH) and (Ferring Pharmaceuticals, CH).

[0434]

[0435]

[0436] Standards and reference materials

[0437] Budesonide, European Pharmacopoeia CSR.

[0438] Other reagents:

[0439] Tween 80, (Polysorbate (80)), Fisher Scientific, or equivalent.

[0440] Dissolution media, mobile phase, and diluents

[0441] acid resistant media

[0442] 0.1N HCI solution. For example, to prepare 10L, combine 82mL of concentrated HCI with 10000mL of water, mix well.

[0443] buffered dissolution media

[0444] Sodium Phosphate pH 6.80 dissolution media was prepared by diluting one bottle (961.5mL) of Reagecon DBC09-960 concentrate to a total volume of 25L.

[0445] For more details see Sodium Phosphate pH 6.80 Dissolution Media 6x961.5ml (reagecon.com).

[0446] Check the pH of the buffer solution after preparation.

[0447] After the acid resistant sample has been withdrawn, the capsule is removed from the solution using forceps and set aside, while the vessel is emptied, cleaned, and filled with pre-heated buffer media. Add 0.05% Tween 80 (or equivalent) to each dissolution cup; for example, add 450mg of Tween 80 to the dissolution cup after filling the dissolution cup with 900mL of pre-heated buffer to obtain a surfactant concentration of 0.05% w / v.

[0448] After all vessels have reached the target temperature, the experiment is started by adding the capsule to each vessel.

[0449] Operational modifications for Budenofalk

[0450] A different procedure is used when the capsule breaks during the acid phase. Carefully decant most of the acid phase and then carefully pipette out the remaining acid in order to remove as little of the pellets as possible from the vessel. Start the buffer phase by adding 900mL of pre-heated buffer media followed by the addition of Tween.

[0451] Two-stage sampling

[0452] Pipette 10mL, discard 8mL (via Whatman filter (0.7pm)), take 1mL in an HPLC vial.

[0453] The changes in this test compared to Example 2 had no effect on the overall dissolution profile of the tested product, and comparative dissolution profiles of three other commercially available budesonide-containing formulations can be found in Figure 5 .

[0454] Example 6: In the absence of surfactant and at a paddle rotation speed of 100 rpm in the buffer stage according to USP <711> / Comparative test according to European Pharmacopoeia 2.9.3

[0455] The following summarizes a variation of the in vitro test of Examples 2 and 5. In this test, enteric-coated capsules filled with the core-shell beads of Example 1 and three other commercially available budesonide-containing formulations were analyzed in a buffered phase solution at pH 6.8 and a paddle rotation speed of 100 rpm. The three formulations were (Tillotts Pharma), (Dr Falk Pharma GmbH) and (Ferring Pharmaceuticals, CH).

[0456]

[0457] Standards and reference materials

[0458] Budesonide, European Pharmacopoeia CSR.

[0459] Dissolution media, mobile phase, and diluents

[0460] Acid resistant media

[0461] 0.1N HCl solution. For example, to prepare 10 L, combine 82 mL of concentrated HCl with 10,000 mL of water and mix thoroughly.

[0462] Buffered dissolution medium

[0463] Sodium phosphate pH 6.80 dissolution medium was prepared by diluting one bottle (961.5 mL) of Reagecon DBC09-960 concentrate to 25 L total volume.

[0464] For further details see Sodium Phosphate pH 6.80 Dissolution Medium 6 x 961.5 ml (reagecon.com).

[0465] Check the pH of the buffer solution after preparation.

[0466] After withdrawing the acid-resistant sample, the capsule was removed from the solution with forceps and set aside while the container was emptied, cleaned, and filled with 900 mL of pre-warmed buffered medium.

[0467] After all containers reached the target temperature, the experiment was started by adding capsules to each container.

[0468] Operational modifications for Budenofalk

[0469] A different procedure is used when the capsules break during the acid phase. Carefully decant the majority of the acid phase and then carefully remove the remaining acid with a pipette in order to remove as few pellets as possible from the container. The buffer phase is started by adding 900 mL of preheated buffer medium.

[0470] Two-stage sampling

[0471] 10 mL was aspirated, 8 mL was discarded (through a Whatman filter (0.7 μm)), and 1 mL was sampled into an HPLC vial.

[0472] The dissolution profiles of the tested products as well as comparative dissolution profiles of the budesonide core-shell beads of Example 1 and three other commercially available budesonide-containing formulations can be found in Figure 6 .

[0473] The following table shows the f2 values ​​for comparing the budesonide capsules according to the present invention and each of other commercially available products tested under this method. An f2 value of 50 or greater is required to show similarity of the curves (FDA SUPAC Guidances 1995, 1997).

[0474] Naifukan Entocort EC Budenofalk Cortiment F2 value (USP) 18.1 16.0 15.8

[0475] It is clear that the release profiles of budesonide vary significantly among the four marketed products. An f2 comparison between Nafucon and the other products shows no similarity, which would require an f2 value of 50 or greater. Indeed, based on the f2 evaluation and visual inspection of the graphical curves, their release profiles must be considered very different.

[0476] Example 7: Administration of Bead-Filled Capsules and Measurement of Biomarkers

[0477] For the studies detailed in Examples 7 to 16, enteric coated capsules were used that were filled with solidified beads as described in Example 1. The capsules are hereinafter referred to interchangeably as "budesonide capsules" or "Nevconazole budesonide".

[0478] Study Design

[0479] A randomized, double-blind, placebo-controlled trial was conducted in which budesonide capsules were administered to subjects with biopsy-confirmed primary IgA nephropathy and significant proteinuria, and the levels of various biomarkers were measured in blood taken from those patients: before the start of treatment; at the completion of treatment; and at time points after the completion of treatment.

[0480] patient

[0481] Men or women at least 18 years of age with biopsy-confirmed primary IgA nephropathy and overt proteinuria were enrolled into the run-in phase. All patients provided written informed consent prior to enrollment. Inclusion criteria for treatment randomization included an estimated GFR (eGFR) of at least 45 mL / min / 1.73 m 2 and a urine protein creatinine ratio (UPCR) of greater than 0.5 g / g, or total urine protein of at least 0.75 g / day, levels considered to increase the risk of progression to end-stage renal disease. The use of 24-hour protein excretion or UPCR from 24-hour urine collection was used to determine the applicability of overcoming possible collection errors and bias from normal creatinine excretion (e.g., physically active and muscular men) to minimize the risk of inadvertently excluding patients.

[0482] operate

[0483] The drug product was an oral capsule formulation (budesonide capsules) as described in Example 1 or placebo, designed to have an in vitro dissolution profile as outlined above to provide sustained release of the active compound, delayed until the capsule reaches the ileum, especially the distal ileum, targeting the site with high density of Peyer's patches.

[0484] After screening, eligible patients were enrolled into a 6-month run-in phase, a 9-month treatment phase, and a 3-month follow-up phase; patients were assessed for eligibility prior to the run-in and treatment phases. During the run-in period, RAS blockade was optimized by up-titration of ACE inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) to the maximum recommended dose or maximum tolerated dose (consistent with established clinical practice), to a target blood pressure of less than 130 / 80 mm Hg, UPCR of less than 0.5 g / g, and urine protein of less than 0.75 g / day. At the end of the run-in, patients with persistent proteinuria (UPCR > 0.5 g / g or proteinuria > 0.75 g / day), eGFR (estimated by the Chronic Kidney Disease Epidemiology Collaboration [CKD-EPI] serum creatinine equation) > 45 mL / min, or measured GFR > 45 mL / min / 1.73 m 2 and blood pressure 160 / 100 mm Hg or less despite optimization of RAS blockade were eligible for treatment randomization.

[0485] An independent Data and Safety Monitoring Board (DSMB) monitored all safety issues and reviewed data in the interim analysis.

[0486] Randomization and Blinding

[0487] Patients were stratified according to their baseline UPCR (≤0.9 g / g and >0.9 g / g) at month 0 (baseline). Patients were randomly assigned to treatment groups using a permuted pool computer algorithm. Within each pool, patients were assigned in a 1:1:1 ratio to 16 mg / day budesonide capsules, 8 mg / day budesonide capsules, or placebo. All patients continued optimized RAS blockade therapy throughout the treatment period.

[0488] A total of 50 patients received placebo, 51 patients received 8 mg / day budesonide capsules, and 48 patients received 16 mg / day budesonide capsules. Randomization was performed by Pharma Consulting Group AB (Uppsala, Sweden). The demographic data and baseline characteristics of the enrolled patients are shown in Table 2.

[0489] The experiment was double-blind. Therefore, throughout the experiment and analysis, each patient, all experimental staff (including investigators and other staff who performed randomization and analysis), the sponsor, and the DSMB were unaware of the allocation of treatment groups (blinded safety data reviewed by the DSMB and unblinded data were available in the event of a problem).

[0490] To ensure blinding, placebo capsules provided by the sponsor had the same appearance and route of administration as the active capsules. Patients self-administered the blinding capsules once daily, one hour before breakfast, during the treatment period. During follow-up (Months 9 to 12), patients who received 16 mg / day budesonide capsules during Months 0 to 9 were transitioned to 8 mg / day for 2 weeks, while all other patients (i.e., those who received 8 mg / day budesonide capsules or placebo during Months 0 to 9) received placebo to maintain blinding. No other study drug was administered after the transition.

[0491]

[0492]

[0493] Table 2. Patient demographics and baseline characteristics. Data are presented as n (%), mean (SD), or median (IQR). Acronyms: BMI = body-mass index; CKD-EPI = chronic kidney disease epidemiology collaboration equation; eGFR = estimated glomerular filtration rate; UACR = urine albumin creatinine ratio; UPCR = urine protein creatinine ratio.

[0494] Blood samples were obtained from patients at the beginning of the treatment period (month 0, before any treatment was given), at the end of the treatment period (month 9), and at the end of the follow-up period (month 12). The obtained samples were tested for levels of multiple biomarkers, including: BAFF; APRIL; TACI; BCMA; CD27, CD30; secretory IgA; IgA-IgG immune complexes; and poorly O-galactosylated IgA1.

[0495] Each randomized patient will be provided with a treatment-coded envelope. In the event of an emergency, the coded envelope may be opened. Any patient whose blind is unmasked must withdraw from the trial.

[0496] Example 8: Treatment of patients with budesonide capsules results in a decrease in serum levels of BAFF

[0497] Materials and methods

[0498] Biomarkers were detected using custom-designed bead-based multiplexing according to the manufacturer's instructions. Analytical (R&D Systems) measurements.

[0499] The assay concept is based on fluorescently labeled microspheres that selectively bind to molecules of interest, thus allowing the simultaneous detection and quantification of multiple biomarkers in extremely small volumes of serum.

[0500] In this study, biomarkers were selected and divided into panels based on the analytical dynamic range (Panel 1: BAFF; APRIL; Panel 2: TACI; BCMA; CD27; CD30).

[0501] The panel 1 specifically labeled microparticle mixture or the panel 2 specifically labeled microparticle mixture was diluted 1:10 with reagent diluent (microparticles provided by R&D Systems), serum samples were diluted 1:2 in reagent diluent, and serial dilutions of standard solutions (microparticles provided by R&D Systems) were used to generate a standard curve.

[0502] Towards The assay plate was applied with 50 μl of the microparticle mix, followed by 50 μl of the standard or sample. The plate was incubated for 2 hours at room temperature on a microplate shaker at 800 rpm. Protein not bound to the microbeads was removed by washing with wash buffer (provided with the microparticles by R&D Systems).

[0503] To each well was added a biotin-labeled antibody mix (provided with the microparticles by R&D Systems) and incubated for 1 hour at room temperature. The plate was washed with wash buffer (provided with the microparticles by R&D Systems) and then incubated with 50 μl streptavidin-phycoerythrin (provided with the microparticles by R&D Systems) per well for 30 minutes, followed by a final wash step.

[0504] The microparticles were resuspended in 100 μl wash buffer and incubated for 10 minutes at room temperature on a plate shaker at 800 rpm. Fluorescence in each well was read on a flow cytometer within 90 minutes.

[0505] Statistical tests using one-way analysis of variance (ANOVA) were used to compare differences in biomarker levels at a p-value < 0.05.

[0506] result

[0507] As Figure 7 As can be seen in Table 3, in samples taken at the end of the 9-month treatment period, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules resulted in a statistically significant decrease in serum levels of BAFF compared to placebo-treated patients.

[0508]

[0509] Table 3. Percent change in serum levels of BAFF from start to end of treatment. Mean values (placebo, n=50; 8 mg / day budesonide capsules, n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and value ranges are shown.

[0510] In addition, Figure 8 Table 4 shows that serum levels of BAFF in patients previously treated with 16 mg / day budesonide capsules again increased in serum samples taken after completion of the follow-up period (i.e. 3 months after the end of the treatment period), indicating that the observed decrease was dependent on budesonide capsule exposure.

[0511]

[0512] Table 4. Percent change in serum levels of BAFF from the end of treatment to the end of the follow-up period. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0513] The reduction in serum levels of BAFF after budesonide-naïve treatment is consistent with a disease-modifying effect in IgAN.

[0514] Example 9: Treatment of patients with budesonide capsules does not result in a decrease in serum levels of APRIL

[0515] Materials and methods

[0516] The same materials and methods as described in Example 8 above were used.

[0517] Comparison of differences in APRIL levels was performed using one-way analysis of variance (ANOVA) statistical test with a p value < 0.05.

[0518] result

[0519] like Figure 9 As can be seen in Table 5, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules did not result in observable changes in serum levels of APRIL in samples obtained at the end of the 9-month treatment period, indicating that the effects produced by treatment with budesonide capsules are specific to BAFF. Similarly, no changes were observed in the placebo group.

[0520]

[0521] Table 5. Percent change in serum levels of APRIL from the start to the end of treatment. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0522] Example 10: Decreased serum levels of BAFF correlate with decreased serum levels of TACI following budesonide capsule treatment close

[0523] Materials and methods

[0524] The same materials and methods as described in Example 8 above were used.

[0525] The differences in TACI levels were compared using one-way analysis of variance (ANOVA) statistical test with a p value < 0.05.

[0526] result

[0527] like Figure 10 As can be seen in Table 6, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules resulted in statistically significant reductions in serum levels of TACI compared to placebo-treated patients in samples obtained at the end of the 9-month treatment period. In the placebo group, a slight increase in TACI levels was actually seen relative to baseline levels.

[0528]

[0529] Table 6. Percent change in serum levels of TACI from the start to the end of treatment. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0530] Example 11: Decreased serum levels of BAFF correlate with decreased serum levels of BCMA following budesonide capsule treatment close

[0531] Materials and methods

[0532] The same materials and methods as described in Example 6 above were used.

[0533] The comparison of BCMA level differences was performed using one-way analysis of variance (ANOVA) statistical test with a p value < 0.05.

[0534] result

[0535] like Figure 11 As can be seen in Table 7, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules resulted in statistically significant reductions in serum levels of BCMA compared to placebo-treated patients in samples obtained at the end of the 9-month treatment period. In the placebo group, a very slight increase in BCMA levels was actually seen relative to baseline levels.

[0536]

[0537] Table 7. Percent change in serum levels of BCMA from the start to the end of treatment. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0538] Example 12: Decreased serum levels of BAFF correlate with decreased serum levels of CD27 following budesonide capsule treatment close

[0539] Because the study was limited to evaluating changes in circulating rather than tissue levels of each biomarker, and therefore the actual site of modulation was uncertain, pathway analysis was performed to determine whether biomarkers significantly modulated by budesonide, including those surrogate biomarkers of immune cell activation (including sCD27 and sCD30 (see Example 13)), were associated with any specific biological processes and pathways.

[0540] Materials and methods

[0541] The same materials and methods as described in Example 8 above were used.

[0542] Comparison of differences in CD27 levels was performed using one-way analysis of variance (ANOVA) statistical test with a p value < 0.05.

[0543] result

[0544] like Figure 12 As can be seen in Table 8, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules resulted in statistically significant reductions in serum levels of CD27 compared to placebo-treated patients in samples obtained at the end of the 9-month treatment period. In the placebo group, a slight increase in CD27 levels was actually seen relative to baseline levels.

[0545]

[0546] Table 8. Percent change in serum levels of CD27 from the start to the end of treatment. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0547] also, Figure 13 and Table 9 show that serum levels of CD27 increased again in patients previously treated with 16 mg / day budesonide capsules in samples obtained after completion of the follow-up period (ie, 3 months after the end of the treatment period), indicating that the observed reduction was dependent on budesonide capsule exposure.

[0548]

[0549] Table 9. Percent change in serum levels of CD27 from the end of treatment to the end of the follow-up period. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0550] Example 13: Decreased serum levels of BAFF correlate with decreased serum levels of CD30 following budesonide capsule treatment close

[0551] Materials and methods

[0552] The same materials and methods as described in Example 8 above were used.

[0553] Comparison of differences in CD30 levels was performed using one-way analysis of variance (ANOVA) statistical test with a p value < 0.05.

[0554] result

[0555] like Figure 14 As can be seen in Table 10, treatment of patients with 8 mg / day and 16 mg / day budesonide capsules resulted in a small but statistically significant decrease in serum levels of CD30 compared to placebo-treated patients in samples obtained at the end of the 9-month treatment period. In the placebo group, a slight increase in CD30 levels was actually seen relative to baseline levels.

[0556]

[0557] Table 10. Percent change in serum levels of CD30 from the start to the end of treatment. Mean values ​​(placebo, n=50; 8 mg / day budesonide capsules n=51; 16 mg / day budesonide capsules, n=48; 8 mg / day and 16 mg / day budesonide capsules, n=99) and ranges are shown.

[0558] Consistent with large-scale integrated analyses of genome-wide association (GWA) studies (Gesualdo L, Di Leo V, Coppo R. The mucosal immune system and IgA nephropathy. Semin Immunopathol. 2021; 43: 657-668; Coppo R. The gut-renal connection in IgAnephropathy. Semin Nephrol. 2018; 38: 504-512.), this study identified the intestinal immune network for IgA production as one of the most enriched Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathways, indicating that the mechanism of action of budesonide is at least partially driven by actions within the gut-associated lymphoid tissue (GALT).

[0559] Example 14: Analysis of Secretory IgA Levels After Budesonide Capsule Treatment

[0560] Materials and methods

[0561] Monoclonal mouse anti-human secretory component (Sigma) diluted 1 : 10,000 in coating buffer was applied to the wells of the immunoplate and incubated overnight at 4°C. The plate was then washed and blocked with 2% BSA for 1 hour at room temperature. Serum samples and standards (high, medium, and low) were diluted 1 : 10 in PBS and applied to the plate after washing and incubated overnight at 4°C. After this, the plate was washed and polyclonal rabbit anti-human IgA HRP (Sigma, 1 : 2000) was added to each well and the plate was incubated for 90 minutes at room temperature. The plate was then washed again and the level of secretory IgA was observed with the o-phenylenediamine dihydrochloride substrate. The values of the plate were normalized to the standard plate using the OD492 on each plate with the high, medium, and low standards.

[0562] result

[0563] There was a significant (p < 0.05) budesonide capsule-dependent decrease in serum levels of secretory IgA. Serum levels of secretory IgA did not change Figure 15 ) indicating local release and local intestinal effects of targeted release of budesonide capsules, rather than systemic exposure of budesonide capsules

[0564] Example 15: IgA-IgG immune complexes and poorly O-galactosylated IgA1 levels after budesonide capsule treatment Flat analysis

[0565] Materials and methods

[0566] IgA-IgG immune complexes: Wells from a 96-well immunoplate were coated with AffiniPure F(ab')2 fragment goat anti-human serum IgA (alpha chain specific) (Jackson Immunology) diluted to 5 pg / mL in coating buffer. After overnight incubation at 4°C, the plate was washed and blocked for 1 hour at room temperature with 2% BSA in PBS to block non-specific protein binding. Test serum samples and standards (high, medium, and low) were diluted 1 : 500 in PBS, added to the replicates, and incubated overnight at 4°C. The plate was then washed and incubated for 90 minutes with polyclonal rabbit anti-human IgG-HRP (Dako) diluted 1 : 2000 in PBS. The plate was washed again for 4 cycles and the level of IgA / IgG ICs in the serum samples was observed using the o-phenylenediamine dihydrochloride substrate. The values of the plate were normalized to the standard plate using the OD492 on each plate with the high, medium, and low standards.

[0567] Poorly O-galactosylated IgA: The levels of poorly O-galactosylated IgA1 were measured using the commercially available KM55 ELISA (Cat. No. 27600, Immuno-Biological Laboratories, Inc. Minneapolis, MN 55432, USA).

[0568] result

[0569] At the end of treatment, there was a significant (p < 0.05) dose-dependent decrease in serum levels of IgA-IgG immune complexes with budesonide capsules. After cessation of budesonide capsule treatment, IgA-IgG immune complex levels returned to baseline levels within 3 months ( Figure 16 There were similar but less pronounced changes in the levels of adversely O-galactosylated IgA1 ( Figure 17 ).

[0570] Of particular interest is that treatment of IgAN with systemic glucocorticoids has been shown to reduce total serum IgA and O-galactosylated IgA1 (Kosztyu P et al., Glucocorticoids Reduce Aberrant O-Glycosylation of IgA1 in IgA Nephropathy Patients. Kidney Blood Press Res 2018; 43: 350-359. However, no differences were observed in the levels of total IgA, IgA1, and IgG under budesonide capsule treatment by the present invention. Figure 18 ), concluded that local ileal treatment with budesonide capsules has a selective effect on pathogenic antibodies but is ineffective against the total pool of IgA, IgA1, and IgG.

[0571] These results demonstrate that treatment with nafoetida supports a direct effect of nafoetida on underlying pathogenic pathways in IgAN and that the budesonide payload has primarily local rather than systemic effects, resulting in fewer side effects in patients treated with nafoetida.

[0572] Example 16: Comparison of Lag Time to Onset of Plasma Curve Between Budesonide Capsules and a Reference Marketed Product

[0573] In a randomized crossover clinical study, the test product (obtained according to Example 1 above) and the reference commercial product ( EC; AstraZeneca) was administered to 24 subjects in the fasting state.

[0574] The lag time in hours to the onset of blood levels for each subject following administration of the test formulation on two different occasions, and administration of the reference (REF) product (with an appropriate washout period of 7 to 14 days) is shown in Table 11 below.

[0575] Table 11. Lag time of plasma curve onset

[0576] Statistical analysis

[0577] Data were analyzed using Sigmaplot version 11.0 for Windows.

[0578] Descriptive statistics

[0579] The median lag time to onset of plasma levels was calculated for each arm of the study and is presented below in Table 12. (Because these values ​​are discrete data, i.e., lag times can only correspond to sampling times within the study, medians are reported rather than means).

[0580] Group N median 25th percentile 75th percentile F1 24 4.000 3.000 4.500 F2 24 4.000 3.500 4.500 REF 24 1.000 0.670 1.000

[0581] Table 12. Median lag time values

[0582] It was concluded that the median (50th percentile) lag time to onset of plasma levels for both administrations was 4 hours for the test formulation and 1 hour for the reference commercial formulation.

[0583] To determine whether the lag time observed for the test formulation was statistically different from the lag time of the reference product, two repeated measures analysis of variance (ANOVA) tests were applied to the data, one assuming normal distribution of the data and one assuming no distribution of the data.

[0584] Repeated measures ANOVA assuming a normally distributed data set yielded an F value of 51.815 and a P value of P < 0.001, which are highly statistically significant. Post hoc comparisons using the Tukey test to determine which arms of the study differed from each other yielded the results presented in Table 13 below:

[0585] Compare Mean difference q statistic P-value P<0.050 F2 relative to REF 3.151 18.005 <0.001 yes F2 relative to F1 0.261 1.517 0.820 none F1 relative to REF 2.890 16.770 <0.001 yes

[0586] Table 13: Post hoc comparisons between arms of the study

[0587] This indicates that although there was no difference in the lag time to onset of plasma levels between administrations of the test formulations, the lag time to onset of plasma levels after administration of the test formulations was statistically highly different from that after administration of the commercial reference product (P<0.001).

[0588] Repeated measures ANOVA was also applied to the ranking of the individual lag time values ​​using the Friedman test (see, eg, Stanton. A Glanz, Primer of Biostatistics, 5th edition, McGraw Hill 2002, ISBN 0-07-137946-0, pp. 370-380).

[0589] Using this test, the chi-square value was 52.950, resulting in a P value of <0.001, which is highly statistically significant. Post hoc comparisons using the Tukey test to determine which arms of the study differed from each other resulted in the results presented in Table 14 below:

[0590] Compare Rank Difference q statistic P<0.05 F2 relative to REF 68.500 9.237 yes F2 relative to F1 9.500 1.281 none F1 relative to REF 59.000 7.956 yes

[0591] Table 14: Post hoc comparisons between arms of the study

[0592] This test also demonstrated that, although the lag time to onset of plasma levels did not differ between the test formulations, the lag time to onset of plasma levels was statistically different (P<0.05) when the test formulations were administered compared to the commercial reference formulation.

[0593] In conclusion, regardless of the type of statistical analysis applied, the The test formulations had significantly different lag times to plasma level onset compared to EC. The median lag time to plasma level onset was 4 hours in each of the study arms where Naifencon F was administered, while The median lag time for EC was 1 hour.

[0594] This analysis clearly demonstrates that, unlike the reference product, the test formulation as described and claimed herein does not release the majority of its active ingredient until it reaches the distal portion of the small intestine (eg, the ileum, such as the distal ileum).

[0595] Example 17: In the presence of added surfactant Tween 80 according to USP <711> / European Pharmacopoeia 2.9.3 General Method for Quasi-In Vitro Dissolution Testing

[0596] As described in Ph. Eur. 2.9.3 Dissolution Tests for Solid Dosage Forms (Using Apparatus 2) and USP <711> The in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed as described in Dissolution (using Method 2). The measurement was performed as follows.

[0597] Three commercially available budesonide-containing formulations were also analyzed in this test. (TillottsPharma), (Dr Falk Pharma GmbH) and Budesonide (Rhinolast®) (Ferring Pharmaceuticals, CH).

[0598] Dissolution Equipment Setup

[0599]

[0600] Budesonide release was measured using ultra-performance liquid chromatography (UPLC).

[0601] Reagents and standards

[0602] Standard and reference substances:

[0603] Budesonide, European Pharmacopoeia CSR, or suitable secondary standard.

[0604] Other reagents:

[0605] Tween 80, (Polysorbate (80)), Fisher Scientific, or equivalent.

[0606] Dissolution media and diluents

[0607] Acid resistant medium

[0608] 0.1 N HCI solution. To prepare 6 L of acid resistant medium, 50 mL of concentrated HCI is combined with 6000 mL of water and the resulting solution is mixed thoroughly.

[0609] Buffered dissolution medium

[0610] The buffered solution was prepared using FaSSIF buffer concentrate (from Biorelevant.com, product code FASBUF01).

[0611] 0.05 w / v % (0.5 mg / mL) Tween 80 to the buffered solution: for example to make 6 L of buffered dissolution medium, 3 g of Tween 80 is added to 6 L of the buffered solution to obtain a Tween 80 concentration of 0.05 w / v %.

[0612] The resulting solution is mixed thoroughly and the pH is checked. If necessary, the pH is adjusted to 6.5 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0613] Budesonide release was evaluated based on the acceptance criteria in USP <711> / European Pharmacopoeia 2.9.3.

[0614] Example 18: In vitro USP <711> / Pharmacopoeia Test No. Dissolution curve analysis of budesonide capsules No. 2.9.3

[0615] Enteric coated capsules ("Budesonide Capsules") filled with solidified beads prepared as described in Example 1 above were tested under the dissolution conditions outlined in Example 17.

[0616] Figure 19 The overall average dissolution profile of the three samples in the buffer phase can be seen in . No budesonide release was observed in the acid-resistant phase at the 2-hour sampling time point.

[0617] The quantitative results of budesonide dissolution in different media at time points 2 hours at pH 1.2 and 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 120 minutes and 180 minutes at pH 6.5 in the buffer phase are provided in the table below.

[0618]

[0619]

[0620] *SD = Standard Deviation

[0621] Based on USP <711> Budesonide release was evaluated using the acceptance criteria in Ph. Eur. 2.9.3.

[0622] For comparison, dissolution profiles of budesonide capsules and three other budesonide-containing formulations were obtained according to the protocol outlined below. The three other budesonide-containing formulations are (Tillots Pharma), (DrFalk Pharma GmbH) and (Ferring Pharmaceuticals, CH).

[0623] Method for capsules

[0624]

[0625] Standards and reference materials

[0626] Budesonide, European Pharmacopoeia CSR.

[0627] Other reagents:

[0628] Tween 80, (polysorbate (80)), Fisher Scientific, or equivalent.

[0629] Dissolution media, mobile phase, and diluents

[0630] Acid resistant media

[0631] 0.1N HCl solution. For example, to prepare 10 L, combine 82 mL of concentrated HCl with 10,000 mL of water and mix thoroughly.

[0632] Buffered dissolution medium

[0633] Prepare the buffer solution using FaSSIF buffer concentrate from Biorelevant.com according to the provided instructions. Mix the resulting solution thoroughly. Check the pH of the buffer solution after preparation. If necessary, adjust the pH to 6.5 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0634] After withdrawing the acid-resistant sample, remove the capsule from the solution with forceps and set aside while the container is emptied, cleaned, and filled with preheated buffer. Add 0.05 w / v% Tween 80 (or equivalent) to each dissolution vessel; for example, add 450 mg of Tween 80 to each vessel after filling it with 900 mL of preheated buffer to achieve a surfactant concentration of 0.05%.

[0635] After all containers reached the target temperature, the experiment was started by adding capsules to each container.

[0636] Operational modifications for Budenofalk

[0637] A different procedure is used when the capsules rupture during the acid phase. Carefully decant the majority of the acid phase and then carefully remove the remaining acid with a pipette to remove as few pellets as possible from the container. The buffer phase is initiated by adding 900 mL of preheated buffer medium followed by Tween.

[0638] Two-stage sampling

[0639] 10 mL was aspirated, 8 mL was discarded (via Whatman filter), and 1 mL was sampled into an HPLC vial.

[0640] For the avoidance of doubt, this test change compared to Example 2 had no impact on the overall dissolution profile of the product tested.

[0641] Figure 20 The dissolution profile of budesonide capsules compared to three other budesonide-containing formulations is shown. From this figure, it can be clearly seen that in FaSSIF medium, which simulates the environment in the small intestine, budesonide capsules have a release profile that is different from all other commercially available budesonide-containing formulations.

[0642] Example 19: In vitro USP in the absence of enteric-coated capsules <711> / Pharmacopoeia Test No. 2.9.3 Dissolution curve analysis of core-shell beads

[0643] The solidified core-shell beads as prepared in Example 1 were also (only) tested under the buffer phase dissolution conditions as outlined in Example 17 in the absence of enteric coated capsules.

[0644] Figure 21 The overall average dissolution profile of the three samples in the buffer phase can be seen in Figure 1 1.

[0645] The quantitative results of budesonide dissolution from the core-shell beads in the buffer phase at pH 6.5 at time points 15, 30, 45, 60, 90, 120 and 180 minutes are provided in the table below.

[0646]

[0647]

[0648] SD = standard deviation

[0649] The release profile of the core-shell beads in the absence of enteric coated capsules in FaSSIF buffer concentrate further confirms that the majority of budesonide will be released in vivo to the ileum. In other words, the majority of release from the beads occurs in the 90 minute period and in combination with the release delay of the enteric coating achieves the desired dissolution profile of the overall formulation, with the majority of budesonide being released in vivo to the ileum, as evidenced by the biomarker data obtained and the modelling results provided in Example 25 below.

[0650] Example 20: In the absence of added surfactant Tween 80 according to USP <711> / European Pharmacopoeia 2.9.3 General Method for Quasi-In Vitro Dissolution Testing

[0651] Example 1 were analysed for in vitro dissolution as described in European Pharmacopoeia 2.9.3 Dissolution Test for Solid Dosage Forms (using Method 2) and as described in USP <711> Dissolution (using Method 2). Measurements were made as described below.

[0652] Three commercially available budesonide containing formulations were also analysed in this test. The three formulations were (Tillotts Pharma), (Dr Falk Pharma GmbH) and (Swiss-Care Pharma AG).

[0653] Dissolution apparatus settings

[0654]

[0655] Budesonide release was measured using Ultra Performance Liquid Chromatography (UPLC).

[0656] Reagents and standards

[0657] Standards and reference materials:

[0658] Budesonide, European Pharmacopoeia CSR, or a suitable secondary standard.

[0659] Dissolution media and diluents

[0660] Acid resistant media

[0661] 0.1N HCl solution. To prepare 10 L of acid-resistant medium, 82 mL of concentrated HCl was mixed with 10,000 mL of water and the resulting solution was mixed thoroughly.

[0662] Buffered dissolution medium

[0663] Prepare the buffer solution using FaSSIF buffer concentrate from Biorelevant.com according to the provided instructions. Mix the resulting solution thoroughly. Check the pH of the buffer solution after preparation. If necessary, adjust the pH to 6.5 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0664] After withdrawing the acid-resistant sample, the capsule was removed from the solution with forceps and set aside while the container was emptied, cleaned, and filled with 900 mL of pre-warmed buffered medium.

[0665] After all containers reached the target temperature, the experiment was started by adding capsules to each container.

[0666] Operational modifications for Budenofalk

[0667] A different procedure is used when the capsules break during the acid phase. Carefully decant the majority of the acid phase and then carefully remove the remaining acid with a pipette in order to remove as few pellets as possible from the container. The buffer phase is started by adding 900 mL of preheated buffer medium.

[0668] Two-stage sampling

[0669] 10 mL was aspirated, 8 mL was discarded (via Whatman filter), and 1 mL was sampled into an HPLC vial.

[0670] Based on USP <711> Budesonide release was evaluated using the acceptance criteria in Ph. Eur. 2.9.3.

[0671] Example 21: In vitro USP in the absence of added surfactant Tween 80 <711> / Pharmacopoeia Test No. Dissolution curve analysis of budesonide capsules No. 2.9.3

[0672] Enteric coated capsules ("Budesonide Capsules") filled with solidified beads prepared as described in Example 1 above were tested under the dissolution conditions outlined in Example 20.

[0673] The overall mean dissolution profile of budesonide capsules can be seen in Figure 22 middle. Figure 22 The dissolution profiles of three other budesonide-containing formulations tested under the same conditions are also included for comparison. From this figure, it can be clearly seen that in FaSSIF medium, which simulates the environment in the small intestine, the budesonide capsules have a release profile that is different from all other commercially available budesonide-containing formulations.

[0674] For budesonide capsules and Cortiment, no budesonide release was observed during the acid-resistant phase at the 2-hour sampling time point. For the 2-hour sampling time point, a total of 0.8% budesonide release was observed for Entocort and 0.6% budesonide release was observed for Budenofalk during the acid-resistant phase.

[0675] The quantitative results of budesonide dissolution in different media at time points of 2 hours at pH 1.2 and 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 120 minutes and 180 minutes at pH 6.5 in the buffer phase for budesonide capsules and three comparative formulations are provided in the table below.

[0676]

[0677] *SD = Standard Deviation

[0678] Based on USP <711> Budesonide release was evaluated using the acceptance criteria in Ph. Eur. 2.9.3.

[0679] The following table shows the f2 values ​​for comparing the budesonide capsules according to the present invention and each of other commercially available products tested under this method. An f2 value of 50 or greater is required to show similarity of the curves (FDA SUPAC Guidances 1995, 1997).

[0680] Naifukan Entocort EC Budenofalk Cortiment F2 value (biologically relevant) 11.7 16.1 15.8

[0681] It is clear that the release profiles of budesonide vary significantly among the four marketed products. An f2 comparison between Nafucon and the other products shows no similarity, which would require an f2 value of 50 or greater. Indeed, based on the f2 evaluation and visual inspection of the graphical curves, their release profiles must be considered very different.

[0682] Example 22: First Tablet Formulation

[0683] Tablets are manufactured by the following steps:

[0684] 1. Wet granulation. Budesonide, mannitol, hydroxyethylcellulose, hydroxypropylcellulose, and sodium starch glycolate are blended. An ethylene-water mixture is then sprayed onto the powder during blending. The resulting granules are then dried.

[0685] 2. Final blending: Blend the dried granules with sodium stearoyl fumarate.

[0686] 3. Tablet compression: Use a tablet compression machine to compress tablets.

[0687] 4. Enteric coating: Methacrylic acid and methyl methacrylate copolymer, talc, and dibutyl sebacate are dispersed in an isopropyl alcohol-water mixture while mixing. The coating dispersion is then sprayed onto the tablets using a fluidized bed apparatus.

[0688] Examples of tablet compositions according to the invention are shown in the table below.

[0689] Tablet composition.

[0690] <![CDATA[ Components ]]> <![CDATA[ Amount (mg) ]]> <![CDATA[ Function ]]> <![CDATA[ budesonide ]]> <![CDATA[ 4.00 ]]> <![CDATA[ API (intragranular) ]]> <![CDATA[ Mannitol ]]> <![CDATA[ 81.50 ]]> <![CDATA[ Filler (intragranular) ]]> Hydroxyethyl cellulose ]]> ​ <![CDATA[ 5.00 ]]> <![CDATA[ Binder (intragranular) ]]> <![CDATA[ Hydroxypropyl cellulose ]]> <![CDATA[ 5.00 ]]> <![CDATA[ Binder (intragranular) ]]> <![CDATA[ Sodium starch glycolate ]]> <![CDATA[ 4.00 ]]> <![CDATA[ Disintegrant (intragranular) ]]> * ]]> ​ <![CDATA[ * ]]> <![CDATA[ Wet granulation solvent ]]> <![CDATA[ ethanol ]]> <![CDATA[ * ]]> <![CDATA[ Wet granulation solvent ]]> <![CDATA[ Sodium stearoyl fumarate ]]> <![CDATA[ 0.50 ]]> <![CDATA[ Lubricant (extragranular) ]]> <![CDATA[ Total (uncoated tablets) ]]> <![CDATA[ 100.00mg ]]> Methacrylic acid and methyl methacrylate copolymer (1:1) <![CDATA[ 9.85 ]]> <![CDATA[ Enteric coating polymers ]]> Methacrylic acid and methyl methacrylate copolymer (1:2) <![CDATA[ 3.31 ]]> <![CDATA[ Enteric coating polymers ]]> talc <![CDATA[ 3.31 ]]> <![CDATA[ Slip agent, coating uniformity ]]> Dibutyl sebacate <![CDATA[ 2.55 ]]> <![CDATA[ plasticizers ]]> Isopropyl alcohol <![CDATA[ * ]]> purified water ]]> * ​ purified water * ]]> ​ <![CDATA[ Coating solvent ]]> Total (coated tablets) ]]> ​ <![CDATA[ 119.02mg ]]>

[0691] *Removed during operation.

[0692] Example 23: Second Tablet Formulation

[0693] Tablets are manufactured by the following steps:

[0694] 1. Wet granulation. Budesonide is initially blended with colloidal silicon dioxide. Microcrystalline cellulose and dicalcium phosphate are then added and blended again. A solution of hydroxypropylcellulose in an ethylene-water mixture is then sprayed onto the powder while blending. The resulting granules are then dried.

[0695] 2. Blending. Blend the dry granules with microcrystalline cellulose, sodium starch glycolate, and copovidone. As a final blending step, add magnesium stearate to the blend, followed by final blending.

[0696] 3. Tablet compression: Use a tablet compression machine to compress tablets.

[0697] 4. Enteric coating: Methacrylic acid and methyl methacrylate copolymer, talc, and dibutyl sebacate are dispersed in an isopropyl alcohol-water mixture while mixing. The coating dispersion is then sprayed onto the tablets using a pan coater.

[0698] Examples of tablet compositions according to the invention are shown in the table below.

[0699] Tablet composition.

[0700] <![CDATA[ Components ]]> <![CDATA[ Amount (mg) ]]> <![CDATA[ Function ]]> <![CDATA[ Budesonide, micronized ]]> <![CDATA[ 4.00 ]]> <![CDATA[ API (intragranular) ]]> <![CDATA[ colloidal silica ]]> <![CDATA[ 0.50 ]]> <![CDATA[ Sliding agent (intragranular) ]]> Dicalcium phosphate ]]> ​ <![CDATA[ 16.00 ]]> <![CDATA[ Filler (intragranular) ]]> <![CDATA[ microcrystalline cellulose ]]> <![CDATA[ 15.50 ]]> <![CDATA[ Filler (intragranular) ]]> <![CDATA[ Hydroxypropyl cellulose ]]> <![CDATA[ 4.00 ]]> <![CDATA[ Binder (intragranular) <!-- 51 -->]]> <![CDATA[ ethanol ]]> * ]]> ​ <![CDATA[ Wet granulation solvent ]]> purified water ]]> ​ <![CDATA[ * ]]> <![CDATA[ Wet granulation solvent ]]> <![CDATA[ microcrystalline cellulose ]]> <![CDATA[ 53.50 ]]> <![CDATA[ Filler (extragranular) ]]> <![CDATA[ Copolyvidone ]]> <![CDATA[ 5.00 ]]> <![CDATA[ Binder (extragranular) ]]> <![CDATA[ Sodium starch glycolate ]]> <![CDATA[ 1.00 ]]> <![CDATA[ Disintegrant (extragranular) ]]> <![CDATA[ magnesium stearate ]]> Lubricant (extragranular) ]]> ​ <![CDATA[ Lubricant (extragranular) ]]> <![CDATA[ Total (uncoated tablets) ]]> ]]> Methacrylic acid and methyl methacrylate copolymer (1:1) ​ Methacrylic acid and methyl methacrylate copolymer (1:1) <![CDATA[ 9.85 ]]> <![CDATA[ Enteric coating polymers ]]> Methacrylic acid and methyl methacrylate copolymer (1:2) <![CDATA[ 3.31 ]]> <![CDATA[ Enteric coating polymers ]]> talc <![CDATA[ 3.31 ]]> Slip agent, coating uniformity ]]> ​ Dibutyl sebacate <![CDATA[ 2.55 ]]> plasticizers ]]> ​ Isopropyl alcohol <![CDATA[ * ]]> <![CDATA[ Coating solvent ]]> purified water <![CDATA[ * ]]> <![CDATA[ Coating solvent ]]> 119.02mg ]]> ​ <![CDATA[ 119.02mg ]]>

[0701] *Removed during operation.

[0702] Example 24: Third Tablet Formulation

[0703] Tablet ingredients:

[0704]

[0705] *Removed during manufacturing.

[0706] Uncoated tablet manufacturing (200g batch)

[0707] 1. Blend budesonide and all excipients except sodium stearyl fumarate using a Turbula mixer (blending at 46 rpm for 75 minutes).

[0708] 2. The powder blend was granulated by spraying water onto the powder during blending. The amount of water sprayed was 15% of the weight of the dry powder.

[0709] 3. Dry the granules at 50°C overnight.

[0710] 4. Mix the dry granules with sodium stearyl fumarate (lubricant) using a Turbula mixer at 46 rpm for 10 minutes.

[0711] 5. Compress the tablets to obtain an uncoated tablet weight of 100 mg.

[0712] Coating dispersion preparation

[0713] The coating dispersion was prepared using the composition shown in the table below. The dispersion was prepared according to the following steps:

[0714] 1. Prepare a diluent mixture by blending isopropyl alcohol, water, and dibutyl sebacate in a container (Container A).

[0715] 2. Prepare Eudragit suspension by transferring approximately half of the diluent mixture (in container A) to another container (container B). Thereafter, slowly add Eudragit L100 and Eudragit S100 to container B while mixing. Thereafter, mix the Eudragit suspension in container B for an additional 30 to 60 minutes.

[0716] 3. While mixing with the high shear mixer, slowly add talc to the remaining diluent mixture in container A. The talc suspension was then mixed with the high shear mixer for an additional 10 minutes.

[0717] 4. Thereafter the talc suspension in container A was slowly poured into the Eudragit suspension (container B) during mixing.

[0718] 5. Thereafter, the mixture in vessel B was stirred at room temperature for 24 hours.

[0719] 6. Pass the mixture through a 0.5 mm sieve.

[0720] 7. The final coating dispersion was then stored at room temperature until coating and stirring was continued during coating.

[0721] Product Name Amount (g) supplier Eudragit L100 (Methacrylic A Copolymer) 39.4 Evonik Eudragit S100 (Methacrylic Acid Copolymer Type B) 13.22 Evonik Talc, pharmaceutical grade M 13.22 Imerys Dibutyl Sebacate USP / NF 10.18 Merck Isopropyl alcohol 594.36 - Purified water 20.44 - total 690.82

[0722] Tablet coating (50g batch)

[0723] Tablets were coated using a stainless steel pan coater with a diameter of 12 cm and a standard spray nozzle. Coating weight gain was determined by weighing tablet samples after different coating durations. The following spray parameters were used:

[0724] ●Coating pan rotation speed: 20 to 30 rpm

[0725] ●Product temperature: 24 to 27°C.

[0726] Pump flow rate: 250 to 260 μl / min

[0727] ●Nozzle air pressure: 0.25 bar

[0728] In vitro dissolution of tablets

[0729] The in vitro dissolution profile of the tablets was prepared according to the protocol outlined in Example 6 above (in the phosphate buffer stage in the absence of surfactant and at a paddle rotation speed of 100 rpm according to USP <711> / European Pharmacopoeia 2.9.3) and the protocol outlined in Example 20 above (at 100 rpm in FaSSIF buffer in the absence of added surfactant Tween 80 according to USP <711> The dissolution values ​​at certain time points can be found below.

[0730]

[0731] The budesonide release from these tablets conformed to the dissolution profile required to achieve the majority of release in the ileum. Particularly unexpectedly, the combination of a gelling agent (hydroxypropyl methylcellulose) and a disintegrant (crospovidone) allowed for the achievement of an appropriate in vitro dissolution profile. However, it is anticipated that other tablet formulations may also achieve the desired release profile.

[0732] Example 25: Computer simulation modeling of the release site of budesonide capsules

[0733] use The software (Simulations Plus, CA; version 9.8.3002) ran a physiologically based pharmacokinetic (PBPK) model on the capsules prepared in Example 1 ("Budesonide Capsules" or "Nevcon Budesonide").

[0734] According to the protocol outlined in Example 2, the in vitro release of budesonide capsules during the acidic phase (first 2 hours) followed by the buffered phase was uploaded to the PBPK software. In vitro in vivo correlation (IVIVC) was obtained to correlate the PBPK model predictions with the measured pharmacokinetic results. The predicted Cmax was faster and higher than the observed one. However, budesonide is known to undergo intestinal wall metabolism in the small intestine. J et al. Presystemic elimination of budesonide in man when administered locally at different levels in the gut, with and without local inhibition by ketoconazole. Eur J Pharm Sci. 2008 Nov 15; 35(4): 264-70; Raje et al. Evaluation of separate roles of intestine and liver in first-pass metabolism of budesonide in rat Xenobiotica. 2018 Dec; 48(12): 1206-1214). After introducing gut wall metabolism into the PBPK model, well-fitting data were achieved.

[0735] As can be seen in Figure 23(a), the IVIVC PBPK model demonstrates that the budesonide capsule begins to release budesonide only upon reaching the ileum, and releases at least about 90% of the budesonide throughout the ileum, with very little remaining being released in the intestinal portion following the ileum (i.e., the cecum). Therefore, this model demonstrates that compositions having an in vitro release profile as defined herein achieve release of the budesonide payload to the ileum, the site of highest concentration of Peyer's patches in the intestine, and therefore any composition meeting this release profile will be effective in treating IgA nephropathy.

[0736] Also used Software modeling comes from (Tillotts Pharma), and the results showed that all the budesonide was released before the formulation entered the ileum portion of the small intestine (see Figure 23(b)).

[0737] Example 26: Study on the in vivo release of capsule contents

[0738] A study was conducted to evaluate where and when capsules coated with the same enteric coating as the capsules described in Example 1 release their contents in the gastrointestinal tract.

[0739] In this study, VCaps plus size 1 HPMC capsules were filled with 75 mg of caffeine, 10 mg of black iron oxide, and 87.5 mg of manganese gluconate dehydrate. 140.9 mg of sugar beads (also referred to as pellets) were added to replicate the total weight of the core in the capsules described in Example 1. The capsules were then coated with the same enteric coating using the same coating procedure and the same equipment and facilities as the capsules described in Example 1.

[0740] Caffeine is placed in the capsule to be used as a marker, to determine when the contents of the capsule are released by measuring the appearance of caffeine in saliva at different time points. Caffeine is rapidly absorbed after being released in the intestinal tract, and therefore, the appearance of caffeine in saliva provides a sensitive marker (Sager et al. Low dose caffeine asasalivary tracer for the determination of gastric water emptying in fed and fasted state: A MRI validation study. Eur J Pharm Biopharm 127: 443-452 (2018)) that the capsule is opened. Iron oxide is placed inside the capsule so that magnetic resonance imaging (MRI) can be used to locate the position of the capsule when the capsule moves through the gastrointestinal tract by observing iron oxide.

[0741] The study was conducted as an open-label, single-center study in 12 healthy young human subjects at the University Clinic in Greifswald, Germany. Subjects abstained from caffeine-containing food and beverages for three days prior to their participation in the study and fasted overnight for at least 10 hours prior to the study.

[0742] MRI scans were performed and saliva samples were obtained, followed by each participant taking enteric-coated capsules with a glass of water. Subsequently, MRI scans were performed every 15 minutes for the first four hours, and every 30 minutes thereafter, until the study was completed. Saliva samples were obtained one minute after each MRI scan. MRI imaging was performed using a Siemens MAGNETOM Avanto MR scanner (Siemens Healthcare, Erlangen, Germany), with a field strength of 1.5 Tesla, and imaging data were analyzed using Horos 2.2.0 (The Horos Project). All measurements were performed with the subject in a supine position (lying on his back, raising his head). Saliva samples were analyzed using an LCMS 8060 system (Shimadzu Corporation, Kyoto, Japan) and were suitably prepared for this purpose.

[0743] Figure 24 MRI images show different locations within the gastrointestinal tract. At 15 and 90 minutes, the capsule is intact and located in the stomach and jejunum, respectively, while at 270 minutes, the capsule has released its contents and the iron oxide has dispersed in the ileum.

[0744] The mean gastric emptying time (the time it takes for the capsule to exit the stomach) of the capsules was 58 ± 30 minutes (the maximum observed gastric emptying time was 112.5 minutes). These values ​​are consistent with typical gastric emptying times for large, non-disintegrating dosage forms (Wilson et al., Chapter 3. Gastrointestinal Transit and Drug Absorption (pp. 41-65), in "Oral Drug Absorption: Prediction and Assessment," J. Dressman and C. Reppas, eds., 2nd ed. Drugs and the Pharmaceutical Sciences, Vol. 193, Marcel Dekker, NY, NY ISBN-13: 978-1-4200-7733-9 (2010)). None of the capsules showed signs of disintegration in the stomach.

[0745] The first time point at which a concentration of ≥10 ng / ml caffeine was measured in undiluted saliva was considered "salivary caffeine appearance." As shown in the table below, the mean time of first appearance of caffeine in saliva was 238 minutes after capsule ingestion, with a standard deviation of 47 minutes and a range of 158 to 345 minutes. After subtracting the subject's gastric emptying time from the time of first appearance of caffeine in the subject's saliva, the time of caffeine release after entering the small intestine was calculated to be an average of 181 ± 31 minutes (range 120 to 233 minutes). These values ​​indicate that enteric-coated capsule opening and caffeine release fall well within the typical small intestinal transit time range of 3.5 to 4.5 hours (210 to 270 minutes) (Wilson et al. Chapter 3. Gastrointestinal Transit and Drug Absorption (pp. 41-65), in "Oral Drug Absorption: Prediction and Assessment", J. Dressman and C. Reppas, eds. - 2nd ed. Drugs and the Pharmaceutical Sciences Vol. 193 Marcel Dekker, NY, NY ISBN-13:978-1-4200-7733-9 (2010)).

[0746] The table below shows the individual and mean results for gastric emptying time (as determined by MRI), salivary caffeine appearance time, and salivary caffeine appearance time after gastric emptying.

[0747]

[0748] Based on the time of first appearance of caffeine in saliva and the location of iron oxide in the corresponding MRI images, the enteric-coated capsules opened in the ileum and released their contents in 11 of the 12 subjects. This study thus demonstrates that the enteric coating and capsule described in Example 1 consistently release the capsule contents into the ileum. The beads contained in Example 1 begin to release budesonide after the capsule is opened, with the majority of the release occurring within one hour. Comparing the average small intestinal transit time of the beads of 3.5 to 4.5 hours (Wilson et al. Chapter 3. Gastrointestinal Transit and Drug Absorption (pp. 41-65), in "Oral Drug Absorption: Prediction and Assessment", J. Dressman and C. Reppas, eds. - 2nd ed. Drug and the Pharmaceutical Sciences, Vol. 193, Marcel Dekker, NY, NY ISBN-13: 978-1-4200-7733-9 (2010)) with the average time it takes for the capsule to open after reaching the small intestine (181 minutes in this study), plus the time required for most of the budesonide to be released from the beads (approximately one hour), it can be concluded that most of the budesonide will be released from the beads into the distal ileum and thus targeted to the Peyer's patches located there.

[0749] The present disclosure relates to the following embodiments:

[0750] 1. A method for treating IgA nephropathy, comprising:

[0751] (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition being responsive to standard in vitro USP <711> / In the European Pharmacopoeia (Ph.Eur.) 2.9.3 dissolution test, the dissolution apparatus using method 2 (paddle method) according to the test meets the following requirements:

[0752] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0753] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and

[0754] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8;

[0755] (ii) wherein the method comprises the step of administering the composition to a patient with IgA nephropathy in need of such treatment.

[0756] 2. Use of a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, wherein the composition meets the following requirements:

[0757] (i) In standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to method 2 (paddle method) of the test was used;

[0758] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0759] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and

[0760] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8;

[0761] It is used to manufacture a medicament for treating IgA nephropathy.

[0762] 3. A composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements:

[0763] (i) In standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to method 2 (paddle method) of the test was used;

[0764] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0765] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and

[0766] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8;

[0767] It is used to treat IgA nephropathy.

[0768] 4. A composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition being soluble in water under standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, the dissolution apparatus using the second method (paddle method) according to the test meets the following requirements:

[0769] (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes;

[0770] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and

[0771] (c) The composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8.

[0772] 5. The method of claim 1 , the use of claim 2, the composition for use of claim 3 or the composition of claim 4, wherein the composition comprises one or more cores comprising budesonide coated with a pharmaceutically acceptable extended release polymer blend comprising a water-insoluble polymer and a pore-forming polymer.

[0773] 6. The method, the use, the composition for use or the composition according to item 5, wherein the core is encapsulated by a delayed release coating.

[0774] 7. The method, use, composition for use or composition according to item 5 or item 6, wherein the encapsulated core is loaded into a capsule, and wherein the delayed-release coating is on the capsule.

[0775] 8. The method, use, composition for use or composition according to claim 6 or 7, wherein the extended release polymer blend coating and the delayed release coating allow for substantially preventing the release of the contents of the composition before reaching the ileum region of the small intestine.

[0776] 9. The method, use, composition for use or composition of any one of clauses 5 to 8, wherein the water-insoluble polymer is present in an amount of about 45% to about 90% by weight of the total extended release coating, and the pore-forming polymer is present in an amount of about 35% to about 5% by weight of the total extended release coating, such as the water-insoluble polymer is present in an amount of about 45% to about 65% by weight of the total extended release coating, and the pore-forming polymer is present in an amount of about 35% to about 15% by weight of the total extended release coating, for example wherein the water-insoluble polymer is present in an amount of about 47% to about 55% by weight of the total extended release coating, and the pore-forming polymer is present in an amount of about 32% to about 22% by weight.

[0777] 10. The method, use, composition for use or composition of any one of clauses 5 to 9, wherein the pore-forming polymer is water-soluble.

[0778] 11. The method, use, composition for use or composition of any one of clauses 5 to 10, wherein the extended release polymeric blend coating the one or more cores is coalescible.

[0779] 12. The method, use, composition for use or composition of any one of clauses 5 to 11, wherein the extended release polymeric blend coating the one or more cores comprises one or more coalescible polymers.

[0780] 13. The method, use, composition for use or composition of clause 11 or clause 12, wherein the one or more coalescible polymers comprise the water-insoluble polymer.

[0781] 14. The method, use, composition for use or composition of one of clauses 5 to 13, wherein the extended release polymeric blend comprises the water-insoluble polymer in an amount of between about 45% to about 65% by weight, for example about 47% to about 56% by weight.

[0782] 15. The method, use, composition for use or composition of any one of clauses 5 to 14, wherein the extended release polymeric blend comprises the pore-forming polymer in an amount of between about 35% and about 15% by weight, for example about 32% to about 22% by weight.

[0783] 16. The method, use, composition for use or composition of any one of clauses 5 to 15, wherein the water-insoluble polymer is ethyl cellulose.

[0784] 17. The method, use, composition for use or composition according to any one of items 5 to 16, wherein the porogenic polymer has a nominal viscosity of about 1 to about 300 mPa*s, for example about 1 to about 50 mPa*s, such as about 1 to about 30 mPa*s, for example about 1 to about 20 mPa*s, such as about 1 to about 10 mPa*s, such as about 2 to about 9 mPa*s, for example about 2 to about 7 mPa*s, preferably about 2 to about 6 mPa*s.

[0785] 18. The method, use, composition for use or composition according to any one of items 5 to 17, wherein the porogenic polymer has a gelling temperature of about 35 to about 65°C, for example about 55 to about 65°C, such as about 58 to about 64°C.

[0786] 19. The method, use, composition for use or composition according to any one of items 5 to 18, wherein the porogenic polymer comprises hydroxypropylmethylcellulose (HPMC).

[0787] 20. The method, use, composition for use or composition of claim 19, wherein the HPMC is methoxy-substituted to an extent of from about 15 to about 35 wt%, for example from about 25 to about 35 wt%, or from about 27 to about 31 wt%, such as from about 27 to about 30 wt%.

[0788] 21. The method, use, composition for use or composition according to claim 19 or claim 20, wherein the degree of substitution of the HPMC with hydroxypropoxy groups is from about 4 to about 32 wt%, for example from about 4 to about 20 wt%, or from about 5 to about 15 wt%, such as from about 7 to about 12 wt%.

[0789] 22. The method, use, composition for use or composition according to any one of items 5 to 21, wherein the extended release polymer blend is present in an amount of about 5 to about 18 wt % of the total bead composition, such as about 6 to about 16 wt % of the total bead composition, such as about 6 to about 12 wt % of the total bead composition.

[0790] 23. A method of preparing a composition according to any one of items 5 to 22, wherein the core is coated in a fluidized bed apparatus.

[0791] 24. A method according to item 23 (when dependent on any one of items 11 to 22), wherein agglomeration of the polymeric material is also carried out in the fluidised bed apparatus.

[0792] 25. A composition obtainable by the method according to item 23 or item 24.

[0793] 26. The method, use, composition for use or composition of any one of items 6 to 22, wherein the delayed-release coating is an enteric coating.

[0794] 27. The method, use, composition for use or composition according to claim 26, wherein the enteric coating comprises a polymer selected from the list consisting of: azo polymers, disulfide polymers, cellulose acetate, cellulose acetate succinate, cellulose acetate phthalate, tetrahydrocellulose acetate phthalate, polyvinyl acetate phthalate, hydroxyethyl ethylcellulose phthalate, methacrylic acid copolymers, polymethacrylic acid / acrylic acid copolymers, styrene maleic acid copolymers, hydroxypropyl methylcellulose phthalate, acrylic resins, cellulose acetate trimellitate, hydroxypropyl methylcellulose trimellitate, shellac, hydroxyethyl ethylcellulose phthalate, carboxymethyl cellulose and hydroxypropyl methylcellulose acetate succinate.

[0795] 28. The method, use, composition for use or composition of any of the preceding items, wherein treatment of IgA nephropathy is manifested by a statistically significant reduction in the level of one or more biomarkers associated with B cell activation and / or proliferation relative to the serum baseline level of the one or more biomarkers in the subject before treatment.

[0796] 29. The method, use, composition for use or composition according to claim 28, wherein the one or more biomarkers are serum B cell activation factor, or an immunoglobulin or immunoglobulin complex associated with the pathogenesis of IgA nephropathy.

[0797] 30. The method, use, composition for use or composition according to item 28, wherein the reduction in serum baseline level is at least 5%, such as at least 10%.

Claims

1. A method for treating IgA nephropathy, comprising: (i) Identifying a pharmaceutically acceptable composition intended for use in the treatment of IgA nephropathy comprising budesonide and one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition being responsive to standard in vitro USP <711> / In the European Pharmacopoeia (Ph.Eur.) 2.9.3 dissolution test, the dissolution apparatus using method 2 (paddle method) according to the test meets the following requirements: (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes; (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8; (ii) wherein the method comprises the step of administering the composition to a patient with IgA nephropathy in need of such treatment.

2. Use of a composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, wherein the composition meets the following requirements: (i) In standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to method 2 (paddle method) of the test was used; (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes; (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8; It is used to manufacture a medicament for treating IgA nephropathy.

3. A composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of the budesonide after administration to the gastrointestinal tract, said composition meeting the following requirements: (i) In standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, a dissolution apparatus according to method 2 (paddle method) of the test was used; (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes; (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8; It is used to treat IgA nephropathy.

4. A composition comprising budesonide in combination with one or more pharmaceutically acceptable excipients that provide modified release of said budesonide after administration to the gastrointestinal tract, said composition being soluble in water under standard in vitro USP <711> / In the European Pharmacopoeia 2.9.3 dissolution test, the dissolution apparatus using the second method (paddle method) according to the test meets the following requirements: (a) the composition satisfies the requirement that when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of the budesonide is released into the dissolution medium within about 120 minutes; (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) The composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.

8.

5. The method of claim 1 , the use of claim 2 , the composition for use of claim 3 , or the composition of claim 4 , wherein the composition comprises one or more cores comprising budesonide coated with a pharmaceutically acceptable extended release polymeric blend comprising a water-insoluble polymer and a pore-forming polymer.

6. The method, the use, the composition for use or the composition according to claim 5, wherein the core is encapsulated by a delayed release coating.

7. The method, use, composition for use or composition according to claim 5 or claim 6, wherein the encapsulated cores are loaded into a capsule, and wherein the delayed release coating is on the capsule.

8. The method, use, composition for use or composition according to claim 6 or claim 7, wherein the extended release polymeric blend coating and the delayed release coating allow for substantially preventing release of the contents of the composition before reaching the ileum region of the small intestine.

9. The method, use, composition for use or composition according to any one of claims 5 to 8, wherein the amount of the water-insoluble polymer is from about 45% to about 90% by weight of the total extended-release coat and the pore-forming polymer is present in an amount from about 35% to about 5% by weight of the total extended-release coat, such as from about 45% to about 65% by weight of the total extended-release coat and the pore-forming polymer is present in an amount from about 35% to about 15% by weight of the total extended-release coat, for example, wherein the water-insoluble polymer is present in an amount from about 47% to about 55% by weight of the total extended-release coat and the pore-forming polymer is present in an amount from about 32% to about 22% by weight.

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