Pharmaceutical composition comprising budesonide for treating iga nephropathy

Abstract

The present invention provides a method of treating IgA nephropathy, the 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 the budesonide following administration to the gastrointestinal tract, the composition comprising budesonide and one or more pharmaceutically acceptable excipients in a standard in vitro dissolution test, the one or more excipients comprising budesonide and one or more pharmaceutically acceptable excipients comprising budesonide and one or more pharmaceutically acceptable excipients comprising budesonide and one or more pharmaceutically acceptable excipients comprising budesonide and one or more pharmaceutically acceptable excipients comprising budesonide; the following requirements are met; (a) 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) a requirement for no more than about 10% of the budesonide to release into the dissolution medium within about 30 minutes when the dissolution medium is aqueous and has a pH of about 6.8; and (c) a requirement for release of at least about 70% of the budesonide into the dissolution medium within about 120 minutes when the dissolution medium is aqueous and has a pH of about 6.8; (ii) a step of administering said composition to a patient suffering from IgA nephropathy in need of said treatment.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202510935869.3 (22) Application Date 2023.01.24 (30) Priority Data 2217146.6 2022.11.16 GB 2217150.8 2022.11.16 GB 63 / 302,216 2022.01.24 US 63 / 302,226 2022.01.24 US (62) Divisional Application Data 202380018644.0 2023.01.24 (71) Applicant: Calititas Therapeutics Company; Address: Stockholm, Sweden (72) Inventors: E.K. Risel, L.M. Pervetov-Moras, K. Sandward, C.O.A. Pedersen (74) Patent Agency: Beijing Kunrui Law Firm, 11494; Patent Attorney: Feng Xinqin (51) Int.Cl. A61K 31 / 58 (2006.01) A61K 9 / 52 (2006.01) A61P 13 / 12 (2006.01) (54) Title of Invention: Pharmaceutical Composition Containing Budesonide for Treating IgA Nephropathy (57) Abstract: This invention provides a method for treating IgA nephropathy, the 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 upon administration to the gastrointestinal tract, said composition meeting the following requirements in a standard in vitro dissolution test: (a) when the dissolution medium is aqueous and has a pH of about 1.2, no more than about 10% of said budesonide is released into said dissolution medium within about 120 minutes; (b) when said dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of said budesonide is released into said dissolution medium within about 30 minutes; and (c) when said dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of said budesonide is released into said dissolution medium within about 120 minutes. The requirement that at least about 70% of the budesonide be released into the dissolution medium within minutes; (ii) the step of administering the composition to a patient with IgA nephropathy who requires the treatment. Claims 2 pages Description 59 pages Drawings 25 pages CN 120754112 A 2025.10.10 CN 1 20 75 41 12 A 1. A method for treating IgA nephropathy, the method comprising: (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable methods for regulating the release of the budesonide after administration to the gastrointestinal tract.The excipients, the composition in standard in vitro USP <711> In the dissolution test according to the European Pharmacopoeia (Ph.Eur.) 2.9.3, the dissolution apparatus using the second method (paddle method) of the test shall meet the following requirements: (a) the composition shall meet 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 shall be released into the dissolution medium within about 120 minutes; (b) the composition shall meet the requirement that, when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of the budesonide shall be released into the dissolution medium within about 30 minutes; and (c) the composition shall meet the requirement that, when the dissolution medium is aqueous and has a pH of about 6.8, at least about 70% of the budesonide shall be released into the dissolution medium within about 120 minutes; (ii) the method comprises the step of administering the composition to a patient with IgA nephropathy who requires the treatment. 2. Use of a composition comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of the budesonide upon administration to the gastrointestinal tract, the composition satisfying the following requirement: (i) USP in standard in vitro <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus according to method 2 (paddle method) of the test is 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, 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 30 minutes; and (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; It is used to manufacture a medicament for the treatment of IgA nephropathy. 3. A composition comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of said budesonide upon administration to the gastrointestinal tract, said composition satisfying the following requirement: (i) USP in standard in vitro <711> In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus according to Method 2 (paddle method) of the test is 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, 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 30 minutes; and (c) the composition satisfies the requirement that, when the dissolution medium is aqueous and has a pH of about 6.8, at least [amount missing] is released into the dissolution medium within about 120 minutes.The requirement is that approximately 70% of the budesonide is released into the dissolution medium; it is used to treat IgA nephropathy. 4. A composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide upon administration to the gastrointestinal tract, the composition being tested in standard in vitro USP <711> In the dissolution test according to European Pharmacopoeia 2.9.3, the dissolution apparatus using method 2 (paddle method) of the test shall meet the following requirements: (a) the composition shall meet 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 shall be released into the dissolution medium within about 120 minutes; (b) the composition shall meet the requirement that, when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of the budesonide shall be released into the dissolution medium within about 30 minutes; and (c) the composition shall meet the requirement that, when the dissolution medium is aqueous and has a pH of about 6.8, at least about 70% of the budesonide shall be released into the dissolution medium within about 120 minutes. 5. The method of claim 1, the use of claim 2, the composition of claim 3, or the composition of claim 4, wherein the composition comprises one or more cores containing budesonide, the cores being coated with a pharmaceutically acceptable extended-release polymeric blend comprising a water-insoluble polymer and a porous polymer. 6. The method of claim 5, the use of claim 5, the composition of claim 5, wherein the core is encapsulated by a delayed-release coating. 7. The method of claim 5 or claim 6, the use of claim 6, the encapsulated core being loaded into a capsule, and wherein the delayed-release coating is applied to the capsule. 8. The method of claim 6 or claim 7, the use of claim 6, the extended-release polymeric blend coating, and the delayed-release coating allow for substantial prevention of release of the contents of the composition before reaching the ileal region of the small intestine. 9. The method, use, composition, 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 coating, and the amount of the porogenic polymer is from about 35% to about 5% by weight of the total extended-release coating, such as the amount of the water-insoluble polymer being from about 45% to about 65% by weight of the total extended-release coating, and the amount of the porogenic polymer being from about 35% to about 15% by weight of the total extended-release coating, for example, wherein the amount of the water-insoluble polymer is from about 47% to about 55% by weight of the total extended-release coating, and the amount of the porogenic polymer is about 32% by weight.From 10% to 22% by weight. 10. The method, use, composition or composition used according to any one of claims 5 to 9, wherein the extended release polymer blend encapsulating the one or more cores comprises one or more polymerizable polymers. Claims 2 / 2 Page 3 CN 120754112 A Pharmaceutical composition containing budesonide for treating IgA nephropathy

[0001] This application is a divisional application of the invention application filed on January 24, 2023, with Chinese application number 202380018644.0 and entitled "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] In this specification, the enumeration or discussion of obviously previously published literature should not be construed as an admission that such literature is part of the prior art or common knowledge.

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

[0005] IgAN is a rare disease and it is estimated that approximately 130,000 to 150,000 people in the United States and approximately 200,000 people in Europe are affected by it. A 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. Patches of lymphoid tissue, called Peyer's patches, are primarily found in the ileum, where they produce secreted IgA antibodies. IgA antibodies play a crucial role in the immune system by protecting the body from foreign substances such as foodborne agents, bacteria, and viruses.

[0007] Patients with IgAN have elevated levels of a subclass of IgA antibodies produced in the intestine that lacks a galactose unit at its hinge region. The hinge region is a flexible amino acid segment in the central part of the heavy chain of an IgA antibody. In IgAN patients, it is presumed that a combination of genetic predisposition and environmental, bacterial, or dietary factors leads to the production of these galactose-deficient IgA antibodies.Increased levels, potentially combined with increased intestinal permeability, can lead to the presence of these antibodies in the blood. Galactose-deficient IgA antibodies (also referred to herein as undesirable O-galactosylated IgA1) are immunogenic when found in 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 deposit in the membranes of the glomeruli, the kidney's filtration apparatus. These trapped immune complexes trigger an inflammatory cascade that damages the membrane, resulting in the leakage of proteins and blood into the urine. Ultimately, the glomeruli are destroyed, reducing the kidney's ability to remove waste products from the blood. As the disease progresses, the waste products that are normally removed from the blood accumulate, leading to potentially life-threatening complications in many patients that will require dialysis or kidney transplantation.

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

[0009] Despite the need for new therapies, few new drugs have been developed for chronic kidney disease in the past decade, and until recently, no therapy was approved to directly treat IgAN itself. Generally, antihypertensive medications are initially given to patients with IgAN. This treatment regimen initially attempted to manage the symptoms of IgAN by lowering blood pressure and reducing proteinuria, but has not been shown to address the underlying cause of IgAN (IgAN product information, page 1 / 59, 4 CN 120754112 A). Over time, physicians attempted to control disease progression with various off-label treatments, such as statins, ω-3-acids, and diuretics, but a significant proportion of patients experienced continued deterioration of renal function, and until recently, no approved treatment options were available.

[0010] For patients with progressive IgAN, clinicians can treat them with systemic immunosuppressants, primarily consisting of high-dose systemic corticosteroids such as prednisone, prednisolone, and methylprednisolone. Although some published reports suggest that these agents can reduce proteinuria, this high dose of systemic corticosteroids is also associated with a wide range of adverse events, including hypertension, weight gain, diabetes, serious infections, and osteoporosis. Furthermore, no potential effect on underlying disease has been confirmed in terms of renal function as measured by estimated glomerular filtration rate (eGFR).

[0011] Therefore, in attempting to meet the current clinical need for an effective treatment for IgAN, there is a clear need for new and / or modified treatments for IgAN in which effective local therapy utilizing immunosuppressants is obtained without such unwanted side effects.

[0012] Peyre's patches (aggregated lymphoid nodules) are small patches of lymphoid tissue found in the ileal region of the small intestine. Because of their monitoring...The gut microbiota, which prevents the growth of pathogenic bacteria in the gut, is an important part of the immune system.

[0013] Since Peyre's plaque is responsible for most of the synthesis of IgA in the body, a dose of locally acting immunosuppressant targeting the ileum (especially the terminal / distal ileum), where Peyre's plaque is mainly located, can be used to reduce the formation of IgA molecules that ultimately drive the formation of immune complexes in IgA by reducing the formation of secretory galactose-deficient IgA antibodies and their presence in the blood. Such targeted release can also limit systemic exposure to locally acting immunosuppressants, such as certain corticosteroids, to avoid unwanted side effects.

[0014] Peyre's plaque is a site of strong activation of B cells in the human body. Therefore, monitoring survival factors associated with B cell activation can be considered to provide an indication of the efficacy of locally acting immunosuppressant therapy.

[0015] Tumor necrosis factor (TNF) family members, B cell activating factor (BAFF) and its homolog, 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 to BAFF, while TACI and BCMA also interact with...APRIL binding (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 regulation of B cell selection. Excessive BAFF has been shown to be associated with the development of autoimmune diseases such as IgAN in animal models, and high levels of BAFF have been detected in the serum of patients with various autoimmune conditions. Increased levels of BAFF have been associated with the upregulation of humoral immunity by increased levels of B cells and immunoglobulins (Steri et al., 2017. N. Engl. J. Med., 376:1615-1626).

[0018] Increased serum levels of BAFF and APRIL have been found in patients with IgAN, and this has led to the development of drugs attempting to inhibit those molecules. These drugs focus on blocking 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 its interaction with the BAFF receptor. Similarly, the combination BAFF / APRIL antagonist atcicept (Merck Serono, licensed by Vera Therapeutics) is also a recombinant fusion protein that combines the BAFF and APRIL binding domains with the antibody's Fc region and blocks its interaction with TACI. The APRIL antagonist VIS649 (Visterra, a subsidiary of Otsuka) and the BAFF inhibitor belimumab (GlaxoSmithKline) are monoclonal antibodies that bind directly 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] Current understanding of the pathogenesis and current treatment of IgAN is summarized in J. Barratt et al., Treatment of IgA Nephropathy: Evolution Over Half a Century, Seminars in Nephrology, 2018, 38(5), 531-540, and also see Boyd et al., Kidney International, 2012, 81, 833-843. Current understanding of IgANThe understanding of the pathogenesis is also summarized in Seikrit et al., The Immune Landscape of IgA Induction in the Gut, Seminars in Immunopathology, 2021, 43, 627-637.

[0020] Unexpectedly, we found that oral administration of budesonide formulations with unique in vitro release profiles resulted in a significant reduction in serum BAFF levels in those subjects relative to levels observed before budesonide administration. Furthermore, the observed reduction in serum BAFF levels occurred concurrently with a decrease in the levels of biomarkers associated with B cell activation and proliferation. Therefore, the in vitro release profile can indicate successful targeted release in the intestine of the subjects (i.e., successful targeted release to the distal ileum).

[0021] In addition, it is of particular interest that systemic glucocorticoid treatment of IgAN has been shown to reduce total serum IgA and adverse 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 budesonide formulations as defined herein, no difference in the total levels of functional IgA antibodies, including IgA1, and IgG was observed with budesonide capsules, but serum levels of galactose-deficient IgA (adverse O-galactosylated IgA) were indeed reduced. This finding leads to the conclusion that topical ileal treatment with budesonide capsules has a selective effect on pathogenic antibodies but is ineffective on the total pool of IgA, IgA1, and IgG.

[0022] These results demonstrate that treatment with budesonide formulations as defined herein supports a direct action on the underlying pathogenic pathway in IgAN, and that the budesonide loading drug has a predominantly local rather than systemic effect, resulting in fewer side effects in patients treated with Nefecon budesonide.

[0023] To support this view, computer simulation modeling of budesonide formulations with unique in vitro release profiles shows that the loading drug is primarily released into the ileum, particularly the distal ileum. This is significant given that budesonide is primarily metabolized through the intestinal wall in the small intestine, resulting in 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] In summary, the results described herein demonstrate that budesonide formulations exhibiting unique in vitro release profiles as defined herein are an effective treatment for IgAN. Due to the targeted local release and action of the local corticosteroid, lower levels of unwanted side effects were obtained.

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

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

[0027] (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 upon administration to the gastrointestinal tract, said composition satisfying the following requirement: USP in standard in vitro <711> In the dissolution test according to the European Pharmacopoeia (Ph.Eur.) 2.9.3, a dissolution apparatus according to the second method (paddle method) of the test (described below) is used;

[0028] (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;

[0029] (b) the composition satisfies the requirement that no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 30 minutes; and

[0030] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 120 minutes, subsequently

[0031] (ii) the composition is administered to a patient with IgA nephropathy who requires the treatment,

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

[0033] The term "pharmaceutically relevant dissolution medium""medium" includes media suitable for in vitro dissolution analysis, the results of which indicate in vivo release at the intestinal relevant portion. For example, pharmaceutically relevant dissolution media may be used instead of "enterically pharmaceutically relevant dissolution medium" or "pharmaceutically relevant enteric dissolution medium," and may be any such medium simulating dissolution and release in the small intestine or its relevant portion.

[0034] Pharmaceutically relevant dissolution media are preferably aqueous.

[0035] The pH of pharmaceutically relevant dissolution media 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 may be a phosphate buffer medium with a pH of about 6.2, or a Fasted State Simulated Intestinal Fluid (FaSSIF) with a pH of about 6.5 (e.g., hereinafter titled "Release in Fasted State Simulated Intestinal Fluid with a pH of about 6.5"). The methods of the present invention may comprise (I) combining budesonide with one or more pharmaceutically acceptable excipients that provide modulated release of said budesonide upon administration to the gastrointestinal tract to manufacture a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, and subsequently (II) as described above in standard in vitro USP <711> The composition is tested in the dissolution test of the European Pharmacopoeia 2.9.3, and if the composition meets the requirements (a) to (c) as described above, the composition is administered to a patient with IgA nephropathy who requires the treatment.

[0038] As an alternative embodiment of the invention, a composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide upon administration to the gastrointestinal tract is provided, wherein the composition meets the dissolution profile of step (i) outlined above for the treatment of IgA nephropathy.

[0039] As another alternative embodiment of the invention, a use of a composition is provided, the composition comprising budesonide and one or more pharmaceutically acceptable excipients providing a modulated release of the budesonide upon administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above for use in the manufacture of an agent for treating IgA nephropathy.

[0040] As mentioned herein, the term “treatment” for IgA nephropathy further includes, in addition to therapeutic, symptomatic, and / or remissionary treatment of the associated symptoms, prevention or diagnosis.

[0041] For the avoidance of doubt, when referring to USP <711> "At that time" refers to the test published on May 1, 2016, and when referring to European Pharmacopoeia 2.9.3, it refers to Chapter 2.9.3 of 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 European Pharmacopoeia, such as the Chinese Pharmacopoeia.

[0042] To avoid ambiguity, step (ii) of administering the composition to a patient will only be performed if the average / mean of the tested composition meets all 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)-butylidenebis(oxy)]-11β,21-dihydroxypregn-1,4-diene-3,20-dione).

[0046] Although the compositions of the present invention contain budesonide, it should be understood that the compositions may alternatively contain different corticosteroids capable of having a local effect in a manner similar to budesonide. 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 containing budesonide.

[0047] The paddle motion of the second method can be about 50 revolutions per minute (rpm), about 75 rpm, or about100 rpm operation. Preferably, the paddle method of the second method is operated at about 100 rpm or about 50 rpm.

[0048] The pharmaceutically relevant 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 polysorbate, preferably polysorbate 80 (e.g., Tween 80). Specification 5 / 59 pages 8 CN 120754112 A

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

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

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

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

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

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

[0055] In criterion b) of step (i) of the method, the composition may further satisfy the requirement that no more than about 10% of budesonide is released into the pharmaceutically relevant dissolution medium within about 37.5 minutes, such as no more than about 5%, for example, no more than about 2.5% of budesonide is released into the pharmaceutically relevant dissolution medium within about 37.5 minutes. For example, the amount of budesonide released into the pharmaceutically relevant dissolution medium 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 under the condition that there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0056] In step (i) of the method, in criterion b), the composition may further satisfy the requirement that at least about 20% of budesonide is released into the pharmaceutically relevant dissolution medium within about 75 minutes, such as at least about 21% within about 75 minutes.For example, at least about 22% or 23% of budesonide is released into the pharmaceutically relevant dissolution medium. For instance, the amount of budesonide released into the pharmaceutically relevant dissolution medium within about 75 minutes may be about 23% to about 74%. Optionally, the release within about 75 minutes occurs when there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0057] In criterion c) of the method, the composition may further satisfy the requirement that at least about 75% of 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 budesonide within about 150 minutes. For instance, the amount of budesonide released into the pharmaceutically relevant dissolution medium within about 150 minutes may be about 77% to about 100%. Optionally, the release within approximately 150 minutes occurs when there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0058] In criterion b) of the method, the composition may further satisfy the requirement that no more than about 10% of budesonide is released into the pharmaceutically relevant dissolution medium within approximately 45 minutes, such as no more than about 5%, such as no more than about 2.5% of budesonide is released into the pharmaceutically relevant dissolution medium within approximately 45 minutes. For example, the amount of budesonide released into the pharmaceutically relevant dissolution medium within approximately 45 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 approximately 45 minutes occurs when there is no surfactant in the pharmaceutically relevant dissolution medium and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0059] In criterion b) of the method, the composition may further satisfy the requirement that no more than about 10% of 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 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 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% as per the specification page 6 / 59 9 CN 120754112 A. Optionally, the release within about 60 minutes is achieved when there is no surfactant in the pharmaceutically relevant dissolution medium and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0060] In guideline b) of the method, the composition may further satisfy the requirement that 50 to 90% of budesonide is released into a pharmaceutically relevant dissolution medium within about 90 minutes. Optionally, the release within about 90 minutes is pharmaceutically relevant.The dissolution medium is free of surfactant and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0061] In criterion c) of the method, the composition may further satisfy the requirement that at least about 80% of 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 budesonide is released into the pharmaceutically relevant dissolution medium within about 180 minutes. Optionally, the release within about 180 minutes is achieved when the pharmaceutically relevant dissolution medium is free of surfactant and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0062] In criterion c) of the method, the composition may further satisfy the requirement that at least about 85% of 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 budesonide is released into the pharmaceutically relevant dissolution medium within about 240 minutes. Optionally, the release within about 240 minutes is achieved in the absence of surfactant in the pharmaceutically relevant dissolution medium and at a paddle rotation speed of 50, 75, or 100 rpm in the second paddle method.

[0063] In criterion c) of the method, the composition may further satisfy the requirement that at least about 90% of 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 budesonide is released into the pharmaceutically relevant dissolution medium within about 360 minutes. Optionally, the release within approximately 360 minutes occurs when there is no surfactant in the pharmaceutically relevant dissolution medium and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0064] In criterion c) of the method, the composition may further satisfy the requirement that at least about 90% of budesonide is released into the pharmaceutically relevant dissolution medium within approximately 480 minutes, such as at least about 95%, for example, about 95% to about 100% of budesonide is released into the pharmaceutically relevant dissolution medium within approximately 480 minutes. Optionally, the release within approximately 480 minutes occurs when there is no surfactant in the pharmaceutically relevant dissolution medium and the blade rotation speed of the second paddle method is 50, 75, or 100 rpm.

[0065] In criterion c) of the method, the composition may further satisfy the requirement that at least about 90% of 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 budesonide is released into the pharmaceutically relevant dissolution medium within about 600 minutes. Optionally, the release within about 600 minutes is achieved by a second paddle method using a paddle at 50, 75, or 100 rpm, in the absence of a surfactant in the pharmaceutically relevant dissolution medium.Under the condition of blade rotation speed.

[0066] In one embodiment, in criterion a) of step (i) of the method, leaching in the acid-resistant medium can be determined according to USP. <711> The acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 / European Pharmacopoeia 2.9.3 Table 2.9.3-2 and / or Table 2.9.3.-3 are used for evaluation.

[0067] In one embodiment, in criterion b) of step (i) of the method, dissolution in the pharmaceutically relevant dissolution medium can be evaluated according to USP <711> The acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 / Tables 2.9.3-2 and / or 2.9.3-3 of European Pharmacopoeia 2.9.3 are used for evaluation.

[0068] In one embodiment, in criterion c) of step (i) of the method, dissolution in the pharmaceutically relevant dissolution medium can be evaluated according to USP <711> The acceptance criteria in Acceptance Form 2 and / or Acceptance Form 4 / Table 2.9.3-2 of European Pharmacopoeia 2.9.3 and / Instructions for Use 7 / 59 pages 10 CN 120754112 A or Table 2.9.3-4 are evaluated.

[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 with and without the addition of a surfactant, such as polysorbate 80 (e.g., Tween 80), at a concentration of about 0.5 mg / mL in a pharmaceutically relevant dissolution medium. In addition, the amount of budesonide released at 37.5 minutes, 60 minutes, 75 minutes, 90 minutes and 150 minutes is achieved with and without the addition of a surfactant, such as 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 providing modulated release of the budesonide upon administration to the gastrointestinal tract, the composition meeting the following requirement: USP in standard in vitro <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus (described below) according to Method 2 (paddle method) of the test is used, operating at 50 rpm;

[0072] (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;

[0073] (b) the composition satisfies the requirement that, when the pharmaceutically relevant dissolution medium does not contain a surfactant, no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 30 minutes;

[0074] (c) The composition satisfies the requirement that, when the pharmaceutically relevant dissolution medium is not containing a surfactant, no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 37.5 minutes;

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

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

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

[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 providing modulated release of the budesonide upon administration to the gastrointestinal tract, the composition meeting the following requirement: USP in standard in vitro <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus (described below) according to Method 2 (paddle method) of the test is used, operating at 100 rpm;

[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, when the pharmaceutically relevant dissolution medium does not contain a surfactant, no more than about 10% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 30 minutes;

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

[0083] (d) The composition satisfies the requirement that, when the pharmaceutically relevant dissolution medium is not surfactant-containing, about 50% to about 90% of the budesonide is released into the pharmaceutically relevant dissolution medium within about 90 minutes; and

[0084] (e) The composition satisfies the requirement that, when the pharmaceutically relevant dissolution medium is not surfactant-containing, at least about 70%, for example at least 75%, of the budesonide is released into the pharmaceutically relevant dissolution medium within about 120 minutes.

[0085] Pharmacokinetic studies of drug absorption from a fasting state are known, with dosage form intake of 200 to 250 mL of water, most...The total volume is approximately 300 to 500 mL, and it is usable 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 dissolution medium of at least about 500 mL (e.g., about 900 mL). The initial volume of the dissolution medium used for criteria a), b), and c) can be about 900 mL.

[0086] The operation of the test composition can be substantially in accordance with USP <711> / European Pharmacopoeia 2.9.3 Delayed-release solid dosage form method B.

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

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

[0089] In criteria (a), (b) and (c), at each time point, the volume of the dissolution medium drawn off can be 10 mL or 15 mL, optionally without replacement of the drawn volume. The drawing off 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, meaning that a small amount of drawing off for analytical purposes does not affect dissolution.

[0090] Release in a medium at pH 6.8

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

[0092] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of the budesonide upon administration to the gastrointestinal tract, the composition satisfying the following requirement: USP in standard in vitro <711> / In the dissolution test of the European Pharmacopoeia 2.9.3, a dissolution apparatus according to the second method (paddle method) of the test (described below) is used;

[0093] (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;

[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 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, and then

[0096] (ii) the composition is administered to a patient with IgA nephropathy who requires the treatment, and the method is referred to below as "the method of the invention".

[0097] The method of the present invention may include (I) combining budesonide with one or more ingredients that provide post-administration regulation of release into the gastrointestinal tract.The pharmaceutically acceptable excipients of budesonide are placed to manufacture a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, and subsequently (II) as described above, in standard in vitro USP <711> The composition is tested in the dissolution test of the European Pharmacopoeia 2.9.3, and if the composition meets the requirements (a) to (c) as described above (i.e., release in a medium at pH 6.8), the composition is administered to a patient with IgA nephropathy who requires the treatment.

[0098] As an alternative embodiment, a composition comprising budesonide and one or more pharmaceutically acceptable excipients providing a modulated release of the budesonide upon administration to the gastrointestinal tract is provided, wherein the composition meets the dissolution profile of step (i) outlined above (i.e., release in a pharmaceutically relevant medium at pH 6.8) for the treatment of IgA nephropathy.

[0099] As another alternative embodiment, use of a composition comprising budesonide and one or more pharmaceutically acceptable excipients providing a modulated release of the budesonide upon administration to the gastrointestinal tract is provided, wherein the composition meets the dissolution profile of step (i) outlined above (i.e., release in a pharmaceutically relevant medium at pH 6.8) for the manufacture of a medicament for the treatment of IgA nephropathy is provided. Instructions for Use, Page 9 / 59, CN 120754112 A

[0100] To avoid ambiguity, step (ii) of administering the composition to the patient will only be performed if the average / mean of the tested composition satisfies all criteria (a), (b), and (c) of step (i).

[0101] The paddle method of the second method can be operated at about 50 rpm, about 75 rpm, or about 100 rpm. Preferably, the paddle method of the second method 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 polysorbate, preferably 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 with a concentration of about 50 mM.

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

[0105] In criterion a) of step (i) of the method, the amount of budesonide released may not exceed approximately 5%, such as not exceeding approximately 2.5%, over approximately 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% over about 120 minutes, such as from about 0% to about 5%, for example from about 0% to about 2.5%.

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

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

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

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

[0111] In criterion b) of step (i) of the method, the composition may further satisfy the requirement that when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of budesonide is released into the dissolution medium over 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 budesonide is released over 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 in about 37.5 minutes may 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 under the condition that there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0112] In criterion b) of step (i) of the method, the composition may further satisfy the requirement that when the dissolution medium is aqueous and has a pH of about 6.8, at least about 20% of budesonide is released into the dissolution medium in 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 budesonide is released in 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 can be about 23% to about 74%. Optionally, the release within about 75 minutes is achieved when there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0113] In criterion c) of the method, the composition may further satisfy the requirement that the dissolution medium is aqueous.When the pH is approximately 6.8, at least approximately 75% of budesonide is released into the dissolution medium within approximately 150 minutes. For example, when the dissolution medium is aqueous and has a pH of approximately 6.8, at least approximately 76%, such as at least approximately 77% (see specification page 10 / 59, CN 120754112 A), is released within approximately 150 minutes. For instance, when the dissolution medium is aqueous and has a pH of approximately 6.8, the amount of budesonide released within approximately 150 minutes can be approximately 77% to approximately 100%. Optionally, the release within approximately 150 minutes occurs when there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 50 rpm.

[0114] In criterion b) of the method, the composition may further satisfy the following requirement: when the dissolution medium is aqueous and has a pH of about 6.8, no more than about 10% of budesonide is released into the dissolution medium within about 60 minutes, such as no more than about 5%, such as no more than about 2.5%, of budesonide being released within about 60 minutes when the dissolution medium is aqueous and has a pH of about 6.8. 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 about 0% to about 10%, such as about 0% to about 5%, for example about 0% to about 2.5%. Optionally, the release within about 60 minutes is under the condition that there is no surfactant in the dissolution medium and the blade rotation speed of the second paddle method is 100 rpm.

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

[0116] In one embodiment, in criterion a) of step (i) of the method, dissolution in an acid-resistant medium may be performed according to USP <711> The acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 / European Pharmacopoeia 2.9.3 Table 2.9.3-2 and / or Table 2.9.3.-3 are used for evaluation.

[0117] In one embodiment, in criterion b) of step (i) of the method, dissolution in the buffer stage medium can be evaluated according to USP <711> The acceptance criteria in Acceptance Table 2 and / or Acceptance Table 3 / European Pharmacopoeia 2.9.3 Table 2.9.3-2 and / or Table 2.9.3.-3 are used for evaluation.

[0118] In one embodiment, in criterion c) of step (i) of the method, dissolution in the buffer stage medium can be evaluated according to USP <711> The acceptance criteria in Acceptance Form 2 and / or Acceptance Form 4 / European Pharmacopoeia 2.9.3 Table 2.9.3-2 and / or Table 2.9.3.-4 shall be evaluated.

[0119] To avoid ambiguity, the amounts of budesonide released at 30 minutes in criterion b) and at 120 minutes in criterion c) were achieved with and without the addition of a surfactant, such as polysorbate 80 (e.g., Tween 80), at a concentration of about 0.5 mg / mL in the dissolution medium. Additionally, the amounts of budesonide released at 37.5 minutes, 60 minutes, 75 minutes, 90 minutes, and 150 minutes were achieved with and without the addition of a surfactant, such as polysorbate 80 (e.g., Tween 80), at a concentration of about 0.5 mg / mL in the dissolution medium.

[0120] In one embodiment, the method comprises:

[0121] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of the budesonide upon administration to the gastrointestinal tract, the composition meeting the following requirement: USP in standard in vitro <711> / In the dissolution test according to the European Pharmacopoeia 2.9.3, a dissolution apparatus (described below) according to Method 2 (paddle method) of the test is used at 50 rpm;

[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 satisfies the requirement that when the dissolution medium is aqueous, without a surfactant, and has a pH of about 6.8, no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes;

[0124] (c) the composition satisfies the requirement that when the dissolution medium is aqueous, without a surfactant, 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; Specification 11 / 59 pages 14 CN 120754112 A

[0125] (d) The composition satisfies the requirement that, when the dissolution medium is aqueous, surfactant-free, and has a pH of about 6.8, about 23% to about 74% of the budesonide is released into the dissolution medium within about 75 minutes;

[0126] (e) The composition satisfies the requirement that, when the dissolution medium is aqueous, surfactant-free, and has a pH of about 6.8, at least about 77% of the budesonide is released into the dissolution medium within about 150 minutes; optionally

[0127] (f) The composition satisfies the requirement that, when the dissolution medium is aqueous, surfactant-free, 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.

[0128] In another embodiment, the method comprises:

[0129] (i) Identify pharmaceutically acceptable compositions intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of said budesonide upon administration to the gastrointestinal tract, wherein said compositions meet the following requirements: [The last part, "USP," appears to be incomplete and requires further context.] <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus (described below) according to Method 2 (paddle method) of the test is used, operating at 100 rpm;

[0130] (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;

[0131] (b) the composition satisfies the requirement that, when the dissolution medium is aqueous, without a surfactant, and has a pH of about 6.8, no more than about 10% of the budesonide is released into the dissolution medium within about 30 minutes;

[0132] (c) the composition satisfies the requirement that, when the dissolution medium is aqueous, without a surfactant, 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;

[0133] (d) The composition satisfies the requirement that, when the dissolution medium is aqueous, surfactant-free, 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, when the dissolution medium is aqueous, surfactant-free, and has a pH of about 6.8, at least about 70%, for example, at least 75%, of the budesonide is released into the dissolution medium within about 120 minutes.

[0135] Pharmacokinetic studies of drug absorption from a fasting state are known to be accompanied by the intake of 200 to 250 mL of water, with a maximum total volume of about 300 to 500 mL, 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 dissolution medium with a volume of at least about 500 mL (e.g., about 900 mL). The initial volume of the dissolution medium used for criteria a), b), and c) may be approximately 900 mL.

[0136] The operation of the test composition may be substantially in accordance with USP. <711> / European Pharmacopoeia 2.9.3 Long-release and / or delayed-release solid dosage form 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, such as 12 or 24.

[0139] In criteria (a), (b) and (c), at each time point, the volume drawn from the dissolution medium may be 10 mL or15 mL, optionally, without replacing the volume drawn. The drawing of the dissolution medium does not affect the overall dissolution profile of the composition. In other words, preferably, the dissolution test is performed under drain conditions and the amount of solvent exceeds the amount of solute, meaning that a small amount drawn for analytical purposes does not affect dissolution.

[0140] Release in intestinal fluid simulated by a level 1 fasting state at pH about 6.5

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

[0142] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of said budesonide upon administration to the gastrointestinal tract, said composition meeting the following requirements: USP in standard in vitro <711> / In the dissolution test of the European Pharmacopoeia 2.9.3, a dissolution apparatus according to the method (paddle method) of the second specification 12 / 59 page 15 CN 120754112 A according to the test described therein is used;

[0143] (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; and

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

[0145] (c) the composition satisfies the requirement that at least about 70% of the budesonide is released into the dissolution medium containing simulated intestinal fluid of Grade 1 fasting state at pH of about 6.5 within about 120 minutes; subsequently

[0146] (ii) administering the composition to a patient with IgA nephropathy who requires the treatment,

[0147] and the method is referred to below as "the method of the invention".

[0148] The method of the invention may comprise (I) combining budesonide with one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide after administration to the gastrointestinal tract to manufacture a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, and subsequently (II) performing the treatment in a standard in vitro USP as described above. <711> The composition is tested in the dissolution test of the European Pharmacopoeia 2.9.3, and if the composition meets the requirements (a) to (c) as described above (i.e., regarding release in simulated intestinal fluid under a fasting state at pH about 6.5), the composition is administered to a patient with IgA nephropathy who requires the treatment.

[0149] As an alternative embodiment, a composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide upon administration to the gastrointestinal tract is provided, wherein the composition meets the dissolution profile of step (i) outlined above (i.e., regarding release in simulated intestinal fluid under a fasting state at pH about 6.5) for the treatment of IgA nephropathy.

[0150] As another alternative embodiment, there is provided a use of a composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide upon administration to the gastrointestinal tract, wherein the composition satisfies the dissolution profile of step (i) outlined above (i.e., release in simulated intestinal fluid under a fasting state at pH approximately 6.5) for use in the manufacture of an agent for the treatment of IgA nephropathy.

[0151] As mentioned herein, the term “treatment” for IgA nephropathy further includes its prevention or diagnosis in addition to therapeutic, symptomatic, and / or remissionary treatment of the associated symptoms.

[0152] Those skilled in the art will understand that the term “Level 1 fasted state simulated intestinal fluid” (Level 1 Fasted State Simulated Intestinal Fluid-V1) includes a biologically relevant dissolution medium with a lower pH and buffering capacity than standard simulated intestinal fluid (generally used in standard USP / European Pharmacopoeia tests; pH 6.8) and specifically developed to simulate fasting conditions in the proximal small intestine (see, for example, Markopoulous et al., In-vitro simulation 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] The Grade 1 FaSSIF-V1 comprises a phosphate buffer system, such as a system comprising NaH2PO4 (concentration of about 28.5 mM), NaOH (concentration of about 13.8 mM), HCl (appropriate amount), and deionized water (appropriate amount), which produces a medium having a weight molar osmotic concentration of about 270 mOsmol / kg and a buffering capacity of about 12 mEq / pH / L.

[0154] In the method 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) for analysis.

[0155] Therefore, the following are further provided:

[0156] ● The method of the invention as defined above; Specification 13 / 59 pages 16 CN 120754112 A

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

[0158] ● The use of a composition that satisfies the dissolution curve of step (i) outlined above for the manufacture of a pharmaceutical agent for the treatment of IgA nephropathy,

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

[0160] To avoid ambiguity, step (ii) of administering the composition to a patient will only be performed if the average / mean of the tested composition satisfies 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 exceeding about 2.5%, over 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% over about 120 minutes, such as from about 0% to about 5%, for example from about 0% to about 2.5%.

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

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

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

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

[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 polysorbate 80 (e.g., Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in the FaSSIF medium.

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

[0169] In criterion c) of step (i) of the method, the composition may further satisfy the following requirement: at least about 20%, such as 25%, or 30%, such as 35%, of budesonide is released into the dissolution medium within about 90 minutes, such as at least about 40%, such as about 30% to about 65%, such as about 35% to about 65%, including about 40% to about 60%, such as about 45% to about 55%, of budesonide is released into the dissolution medium.

[0170] In step (i) of the method, in the absence of polysorbate 80 (e.g., Tween 80) at a concentration of about 0.05% w / v (about 0.5 mg / mL) in the added FaSSIF medium, the composition may further satisfy the following requirement: at least about 10%, for example at least about 15%, of budesonide is released within about 90 minutes, such as about 10% to about 50% of budesonide is released, such as about 10% to about 40% of budesonide is released, including about 10% to about 30% of budesonide released by the specification page 14 / 59 17 CN 120754112 A, for example about 15% to about 30% of budesonide is released into the dissolution medium.

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

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

[0173] Pharmacokinetic studies of drug absorption from a fasting state are known, with the dosage form taking in 200 to 250 mL of water, for a maximum total volume of about 300 to 500 mL, which is usable 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 dissolution medium (including FaSSIF) with a volume 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 composition can be largely based on the USP. <711> / European Pharmacopoeia 2.9.3 Delayed-release solid dosage form 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), at each time point, the volume of the dissolution medium drawn may be 10 mL or 15 mL, optionally without replacement of the drawn volume. The drawing of the dissolution medium does not affect the overall dissolution profile of the composition. In other words, preferably, the dissolution test is performed under drain conditions and the amount of solvent exceeds the amount of solute, meaning that a small amount of drawing for analytical purposes does not affect dissolution.

[0178] Specifically, but not exclusively, when the composition of the present invention is a core-shell composition as defined below, the method of the present invention as defined herein may include 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) identifying USP in a standard in vitro. <711> In the dissolution test according to the European Pharmacopoeia 2.9.3, using a dissolution apparatus according to Method 2 (paddle method) of the test, the composition meets the following requirement: about 20% to about 60%, such as about 25% to about 50%, of budesonide is released into a dissolution medium containing Grade 1 fasting state simulated intestinal fluid at pH about 6.5 within 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 the Grade 1 FaSSIF-V1 dissolution medium as defined herein within about 30 minutes, and more preferably, about 75% to about 85% of budesonide is released into the dissolution medium within about 45 minutes.

[0182] In the absence of a delayed-release (e.g., enteric) coating, approximately 80% to approximately 90% of budesonide is released into a Class 1 FaSSIF-V1 dissolution medium as defined herein within approximately 60 minutes, and more specifically, approximately 90% (e.g., approximately 95% (including approximately 97% and approximately 100%) of budesonide is released into said dissolution medium within approximately 90 minutes, such as within approximately 120 minutes, including within approximately 180 minutes.

[0183] A budesonide composition having the dissolution profile described above in the absence of a delayed-release excipient is further confirmation that the majority of budesonide will be released into the ileum in vivo.

[0184] Effects on Biomarkers

[0185] In all aspects outlined above, the method of the present invention enables serum B-cell activating factor (BAFF) (also known as tumor necrosis factor ligand superfamily member 13B) in subjects.The level of ligand superfamily member 13B (TNFSF13B) was statistically significantly reduced relative to the baseline level of serum BAFF in subjects prior to treatment.

[0186] "Statistically significant reduction" means a statistically significant reduction with a p-value <0.05 after one-way ANOVA when comparing changes seen in the treated patient group with changes seen in the placebo group.

[0187] "Relative to the baseline level" means that the measured level of the molecule (e.g., BAFF) is lower than the level measured at the start of the study (i.e., before drug administration). The baseline level is the level before the start of treatment and is used as a comparison for levels measured subsequently (e.g., immediately after the treatment process or after the end of the treatment process). Therefore, such reduction is specific to the subject or subject group in question and is not an absolute value.

[0188] The reduction in BAFF serum level in the subject relative to the baseline serum BAFF level before treatment 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%. For example, the reduction in BAFF serum level in the subject may be at least about 5%. More specifically, the reduction in serum BAFF level in the subject may be at least about 10%, such as a reduction in BAFF serum level in the subject of at least about 14%.

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

[0190] A statistically significant reduction in serum BAFF levels observed after the method of the present invention can be related to the serum levels of one or more biomarkers associated with B cell activation and / or proliferation relative to the levels of one or more of the biomarkers in the subject before treatment.A statistically significant decrease in the serum baseline levels of several biomarkers is associated with a reduction in the serum levels of one or more biomarkers related to B cell activation and / or proliferation. A decrease in the serum levels of one or more biomarkers associated with B cell activation and / or proliferation will indicate a favorable role in the treatment of diseases in which excessive B cell activity, excess, and / or excessive proliferation are related to the pathogenesis. For example, a decrease in the serum levels of biomarkers produced by active and / or proliferating B cells can indicate reduced B cell activity / proliferation and may indicate a favorable role in the treatment of diseases in which excessive B cell activity, excess, and / or excessive proliferation are related to the pathogenesis, such as IgAN.

[0191] One or more biomarkers may include: transmembrane activating factor and CAML interacting factor (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; C-X-C motif chemokine 12 (C-X-C motif chemokine 12, CXCL12); C-X-C motif chemokine 13 (CXCL13); chemokine (C-C motif) ligand 19 (CCL19); interleukin 2 (IL-2); interleukin 6 (IL-6); chemokine (C-C motif) Ligand 3 (CCL3); chemokine (C-C motif) ligand 4 (CCL4); soluble CD23 (sCD23); secreted IgA; IgA-IgG immune complex; undesirable O-galactosylated IgA1; or combinations thereof.

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

[0193] The reduction in serum levels of the biomarker in the subject relative to the baseline serum levels of the biomarker before treatment may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or 1% compared to the baseline serum levels of the biomarker in the subject before treatment.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, a decrease in serum levels of a biomarker can be about 1% to about 90%, or about 5% to about 70%, or about 10% to about 50%.

[0194] The reduction in serum TACI levels in the subject relative to the baseline serum TACI levels in the subject before treatment 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%, 60%, 65%, 70%, or 75%. For example, the reduction in serum TACI levels in the subject may be at least about 5%. More specifically, the reduction in serum TACI levels in the subject may be at least about 11%.

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

[0196] The reduction in serum BCMA levels in the subject relative to the baseline serum BCMA levels in the subject before treatment 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%, or 60%. For example, the reduction in serum BCMA levels in the subject may be at least about 1%. Specifically, the reduction in serum BCMA levels in the subject may be at least about 6%.

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

[0198] The reduction in serum BAFF-R levels in the subject relative to the baseline serum BAFF-R levels before treatment 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%.

[0199] The reduction in serum CD27 levels in the subject relative to the baseline serum CD27 levels before treatment 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%, or 65%. For example, the reduction in serum CD27 levels in the subject may be at least about 5%. More specifically, the reduction in serum CD27 levels in the subject may be at least about 10%, such as a reduction in serum CD27 levels in the subject of at least about 15%.

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

[0201] The reduction in serum CD30 levels in the subject relative to the baseline serum CD30 level before treatment 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 CD30 levels in the subject may be at least about 1%. Specifically, the reduction in serum CD30 levels in the subject may be at least about 3%, such as the reduction in CD30 levels in the subject.The decrease in serum levels may be at least about 5%.

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

[0203] The reduction in serum IgA levels in the subject relative to the baseline serum IgA levels before treatment 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 IgA levels in the subject may be at least about 1%. More specifically, the reduction in serum IgA levels in the subject may be at least about 2%, such as a reduction in serum IgA levels in the subject of at least about 3%.

[0204] The decrease in serum IgA-secreting levels in the subject may be from about 1% to about 75%. For example, the decrease in serum IgA-secreting levels in the subject may be from about 1% to about 25%. More specifically, the decrease in serum IgA-secreting levels in the subject may be from about 1% to about 10%, such as a decrease in serum IgA-secreting levels in the subject from about 1% to about 5%. For example, the decrease in serum IgA-secreting levels in the subject may be from about 1% to about 3%.

[0205] The reduction in serum IgA-IgG immune complex levels in a subject relative to baseline serum IgA-IgG immune complex levels before treatment 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 IgA-IgG immune complex levels in a subject can be at least about 1%. More specifically, the reduction in serum IgA-IgG immune complex levels in a subject can be at least about 5%, such as a reduction in serum IgA-IgG immune complex levels in a subject of 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...Specifically, the decrease in serum IgA-IgG immune complex levels in subjects can be from about 1% to about 25%. More precisely, the decrease in serum IgA-IgG immune complex levels in subjects can be from about 1% to about 20%, such as a decrease in serum IgA-IgG immune complex levels in subjects of about 2% to about 20%. For example, the decrease in serum IgA-IgG immune complex levels in subjects of about 2% to about 15%.

[0207] The reduction in serum levels of adverse O-galactosylated IgA1 in the subject relative to the baseline serum level of adverse O-galactosylated IgA1 before treatment 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 adverse O-galactosylated IgA1 in the subject may be at least about 1%. Specifically, the reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be at least about 5%, such as a reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be at least about 8%.

[0208] The reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be from about 1% to about 75%. For example, the reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be from about 1% to about 25%. Specifically, page 18 / 59 of the specification 21 CN 120754112 A states that the reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be from about 1% to about 20%, such as a reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be from about 2% to about 20%. For example, the reduction in serum levels of adverse O-galactosylated IgA1 in the subject may be from about 2% to about 15%.

[0209] The reduction of biomarkers mentioned above is related to the method of the present invention, which requires the budesonide-containing composition to exhibit a specified in vitro release profile and to be intended for the treatment of IgAN and / or to be able to treat IgAN.

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

[0211] ● budesonide is released into the gastrointestinal region (e.g., the ileum) in which Peyre's plaque is predominantly located; and

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

[0213] Budesonide-containing compositions

[0214] Compositions that can be used in the method of the present invention may contain budesonide and one or more of the biomarkers described herein.Any combination of excipients that produces the desired in vitro release profile. This can be a combination of extended-release coatings and / or delayed-release coatings, which can be applied by various formulation principles described below.

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

[0216] Such delayed-release coatings may therefore contain so-called “enteric coatings,” which refer to materials with gastroresistant properties, i.e., materials that prevent dissolution or disintegration in the gastric environment, thereby allowing the composition to pass through the stomach into the ileum of the small intestine.

[0217] The enteric coating may contain azo polymers, disulfide polymers, cellulose acetate, cellulose succinate, cellulose acetate tetrahydrophthalate, polyvinyl acetate phthalate, hydroxyethyl cellulose phthalate, methacrylic acid copolymer, polymethacrylic acid / acrylic acid copolymer, styrene-maleic acid copolymer, hydroxypropyl methyl cellulose phthalate, acrylic resin, cellulose benzotriglyceride, hydroxypropyl methyl cellulose benzotriglyceride, shellac, hydroxyethyl cellulose phthalate, carboxymethyl cellulose, and hydroxypropyl methyl cellulose acetate succinate.

[0218] Specific enteric coating materials include cellulose acetate, cellulose acetate succinate, cellulose acetate phthalate, cellulose acetate tetrahydrophthalate, polyvinyl acetate phthalate, hydroxyethyl ethyl cellulose phthalate, methacrylic acid copolymer, polymethacrylic acid / acrylic acid copolymer, styrene-maleic acid copolymer, hydroxypropyl methyl cellulose phthalate, acrylic resin, cellulose acetate benzoate, hydroxypropyl methyl cellulose benzoate, shellac, hydroxyethyl ethyl cellulose phthalate, carboxymethyl cellulose, and hydroxypropyl methyl cellulose acetate succinate.

[0219] Preferred enteric coating materials include polyvinyl acetate phthalate and, in particular, methacrylic acid copolymer.

[0220] Those skilled in the art will understand that enteric coatings may contain other commonly used materials, such as talc (as a plasticizer), dibutyl sebacate (as a plasticizer), and blends of HMPC and PEG as secondary coating agents.

[0221] The composition may comprise one or more cores containing budesonide that encapsulates the contents of the composition by delaying and prolonging the release of the excipients, thereby substantially preventing the release of the contents of the composition before reaching the distal region of the small intestine (e.g., the ileum, such as the distal ileum). Such compositions are referred to below as “core-shell composition of the invention”, which encompasses “beads”"(bead)" and "encapsulated core".

[0222] The core containing budesonide can be loaded into a capsule. When using a capsule, a delayed-release coating (e.g., enteric coating) may be on the capsule and not directly on the core. Specification 19 / 59 pages 22 CN 120754112 A

[0223] When the enteric coating is on a capsule, such as a size 1 capsule, 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 present invention, most of the budesonide is released substantially into the distal region of the small intestine (e.g., the ileum, such as the distal ileum), it may (or may further) be individually coated with an extended-release polymer coating.

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

[0226] Such extended-release coatings ensure that most of budesonide is adequately released throughout the ileum, and in combination with delayed-release coatings (e.g., enteric coatings) further ensure that such release is substantially and / or primarily achieved for the ileum region of the small intestine.

[0227] “Substantially released to the ileum region” includes at least about 51% of the initial amount of the active ingredient in the composition, 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% released to said region.

[0228] Those skilled in the art will understand 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 invention, preferably, the composition is taken orally at least one hour before a meal, and more preferably, at least one hour before the first meal of the day in the morning.

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

[0230] As used herein, the term "water-insoluble polymer" refers to a polymer having a solubility of less than about 0.1 mg mL⁻¹ in an aqueous solvent, such as water, at about 25°C. The presence of a water-insoluble polymer allows for control of the release rate of budesonide from the composition.

[0231] The water-insoluble polymer may be 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 derivative thereof” refers to a compound derived from cellulose in which at least some of the protons on the hydroxyl groups of the cellulose 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 is therefore able to dissolve first, leaving pores in the coating to allow a certain amount of water to permeate toward the core.

[0234] Thus, a pore-forming polymer can be defined as “water-soluble”. In other words, a pore-forming polymer has a solubility of at least about 10 mg mL⁻¹ in an aqueous solvent, such as water, at 25°C.

[0235] The nominal viscosity of a pore-forming polymer can 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, such as from about 1 to about 20 mPa*s, such as from about 2 to about 9 mPa*s, such as from about 2 to about 7 mPa*s, preferably from about 2 to about 6 mPa*s. The nominal viscosity of the porous polymer can be measured at 20°C as a 2% by weight solution of the polymer in water using the capillary viscometer method of the standard European Pharmacopoeia 2.2.9.

[0236] Furthermore, the gelation temperature of the porous polymer can be from about 35 to about 65°C, for example from about 55 to about 65°C, such as from about 58 to about 64°C.

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

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

[0239] The term "degree of substitution of the HPMC" refers to the average level of substitution of hydroxyl groups on the cellulose chain and is expressed herein in percentage terms, i.e., the percentage of hydroxyl groups that have been partially substituted in discussion.

[0240] The water-insoluble polymer may be present in an amount from about 45% by weight to about 90% by weight of the total extended release coating, and the porogenic polymer may be present in an amount from about 35% by weight to about 5% by weight of the total extended release coating. For example, the water-insoluble polymer may be present in an amount from about 45% by weight to about 65% by weight of the total extended release coating, and the porogenic polymer...The polymer may 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 may be present in an amount of about 47% to about 56% by weight of the total extended-release coating and the porogenic polymer may 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 contain fatty acids in an amount of about 2% to about 8% by weight of the total extended-release coating, such as about 3% to about 7% by weight.

[0242] The fatty acids may be unsaturated fatty acids, such as C4 to C28 unsaturated fatty acids, such as C13 to C22 unsaturated fatty acids. For example, the unsaturated fatty acids may be selected from the group consisting of: myristoleic acid, palmitoleic acid, hexadecenoic acid, oleic acid, transoleic acid, isoleic acid and sinapic acid. Preferably, the unsaturated fatty acid is oleic acid.

[0243] The pharmaceutically acceptable polymeric blend with extended-release coating may contain a medium-chain triglyceride in an amount of about 3% to about 12% by weight of the total extended-release coating, such as about 5% to about 12% by weight, for example about 5% to about 8% by weight.

[0244] The term "medium-chain triglyceride" refers to a triglyceride having an aliphatic tail with 6 to 12 carbon atoms. For example, medium-chain triglycerides may be selected from the list of the following compositions: hexanoic acid, caprylic acid, capric acid, and lauric acid.

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

[0246] The polymeric blends coating one or more cores may be coalescing. When referring to extended-release polymeric blends, the term "coalescable" means that the polymers in the extended-release blend are capable of being blended to form a monopolymeric phase coating.

[0247] Thus, extended-release polymeric blends coating one or more cores may comprise one or more coalescing polymers. One or more coalescing polymers may comprise water-insoluble polymers.

[0248] The extended-release polymeric blend may be present in amounts such as about 5 to about 18% by weight of the total beads / cores, for example, about 6 to about 16% by weight of the total beads / cores, for example, about 6 to about 13% by weight of the total beads / cores, such as about 6 to about 12% by weight.

[0249] It has been found that in extended-release coated polymeric blends, the porous polymer is first dissolved in an aqueous solution before the water-insoluble polymer, leaving pores in the coating to allow a controlled amount of water to permeate toward the cores.

[0250] The average size of the core can be 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 comprising the following operations:

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

[0253] (b) said one or more cores individually coated with a pharmaceutically acceptable extended-release polymer blend comprising about 45% to about 90% by weight of a water-insoluble polymer and about 35% to about 5% by weight of a porous polymer; and

[0254] wherein said composition is encapsulated in a delayed-release coating to substantially prevent the release of the contents of said composition before reaching the ileal region of the small intestine.

[0255] The core-shell composition of the above operations may comprise any of the features outlined above with respect to the method of the present invention.

[0256] The core can be prepared by providing inert (e.g., sugar) beads and coating them with an aqueous budesonide suspension. The size of the inert beads can 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 mentioned 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 beads (commonly referred to as non-pareil). Sugar beads mainly contain sucrose and contain small amounts of other materials, such as starch. Suppliers of sugar beads include Paulaur Corporation (USA), Chr. Hansen (Denmark), NP Pharm (France), Emilio Castelli (Italy), and JRS Pharma (Germany).

[0259] The core can be coated with a sealant containing a stabilizing and water-soluble polymer before the addition of the extended-release polymer blend. The stabilizer may be an acid, preferably citric acid, and the soluble polymer is the same polymer used as the porogen in the polymeric blend. Alternative stabilizers used in the sealing coating include poly(vinylpyrrolidone) (PVP), and the soluble polymer is the same polymer used as the porogen in the polymeric blend.

[0260] The extended-release polymeric blend can be applied to the core as an aqueous polymer suspension by spraying the suspension onto the core. The aqueous polymeric blend can be sprayed onto the core at temperatures ranging from about 30°C to about 65°C, such as from about 30°C to about 50°C.

[0261] Spraying towards the upper end of this range, such as from about 50°C to about 65°C, avoids the need for a separate curing / coalescing step as outlined below.

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

[0263] After coating with the polymer blend, the polymer can be polymerized, wherein the polymerization can be carried out by curing.

[0264] Curing can be carried out at temperatures of about 55°C to about 75°C, such as about 60°C to about 70°C, for example about 63°C to about 66°C. Furthermore, curing can be carried out for about 1 hour to about 10 hours, for example about 1 to about 5 hours, such as about 2 to about 4 hours.

[0265] It has been found that the extended-release polymer-coated blend allows for the preparation of the budesonide core in a low-cost manner using a fluidized bed apparatus, while obtaining a composition having the desired release profile.

[0266] Therefore, in addition, the curing of the core-shell composition of the present invention can be carried out in a fluidized bed apparatus.

[0267] According to another aspect of the invention, a composition is provided that produces a desired in vitro release profile in all aspects described herein, comprising a plurality of beads, the beads comprising:

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

[0269] (b) an extended release coating, which is presented on the budesonide-containing core in an amount between about 6 and about 12% by weight of the total bead weight, and the coating comprises a polymeric blend of at least two polymers (i) and (ii):

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

[0271] (ii) any of the porogenic polymers described herein (e.g., hydroxypropyl methylcellulose, which is substituted with methoxy groups to a degree of about 27 to about 30% by weight and / or substituted with hydroxypropyl groups to a degree of about 7 to about 12% by weight),

[0272] the blends of polymers (i) and (ii) are in any of the proportions mentioned above (e.g., about 47% to about 56% by weight of the water-insoluble polymer (i) and about 32% to about 22% by weight of the porogenic polymer (ii) in total extended-release coating),

[0273] the beads are then encapsulated in capsules coated with a delayed-release coating (e.g., any of the enteric coatings described herein, such as polyvinyl acetate phthalate or, in particular, methacrylic acid copolymers).

[0274] The above compositions may contain one or more of the other preferred features disclosed herein with respect to the core-shell compositions and / or beads, such as:

[0275] ● any of the fatty acids described herein, for example, 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, for example, 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) with 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 compositions may also contain a sealed coating solution of a suitable acid (e.g., citric acid) located between the budesonide core and the extended-release coating.

[0279] Each capsule may contain about 4 mg of budesonide and when administered to a patient, a total of about 16 mg may be delivered orally by the patient taking four capsules.

[0280] The polymeric extended-release coating of at least two polymers (i) and (ii) obtained before loading the coated beads into the capsule is preferably obtained by: coating the budesonide-containing core in a fluidized bed apparatus by applying an aqueous dispersion containing polymers (i) and (ii) as defined above at a temperature of about 30°C to about 65°C, such as about 30°C to about 50°C, especially about 50°C to about 65°C, and then, 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 an appropriate time (e.g., about 2 to about 4 hours).

[0281] Alternatively, the polymeric blend can be sprayed as an organic solution. In this embodiment, another curing step is not necessary.

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

[0283] Regardless of the manufacturing process, compositions that can be used in the methods of the present invention can be used to treat IgAN because they meet the in vitro release characteristics defined above.

[0284] Therefore, it is further provided that:

[0285] ● any of the compositions as defined herein, for the treatment of IgAN;

[0286] ● any of the compositions as defined herein, for the manufacture of a pharmaceutical agent for the treatment of IgAN; and specification 23 / 59 pages 26 CN 120754112 A

[0287] ● a method of treating IgAN, the method comprising administering any of the compositions as defined herein to a patient requiring such treatment.

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

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

[0290] One or more excipients used in the encapsulated tablets to substantially prevent the release of the contents of the composition before reaching the ileal region of the small intestine may be enteric coatings as defined above.

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

[0292] The tablet compositions of the present invention may contain wet granules of budesonide and a filler. Optionally, the filler comprises dicalcium phosphate, microcrystalline cellulose, mannitol, or mixtures thereof.

[0293] The filler may be present in an amount of about 50% to about 80% by weight of the total tablets, such as about 60% to about 75% by weight, for example 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, decanoic acid, dodecanoic acid, sodium stearoyl fumarate, or a mixture thereof.

[0295] The lubricant may be present in an amount of about 0.1% by weight to about 2% by weight of the total tablets, such as about 0.1% by weight 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, crospovidone sodium carboxymethyl cellulose, or sodium glycolate starch. Alternatively, the tablet may not contain a disintegrant.

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

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

[0299] The binder may be present in an amount of about 5% to about 10% by weight of the total tablets, such as about 6% to about 9% by weight, for example 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 (e.g., hydroxypropyl methylcellulose).

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

[0302] The gel matrix material may be diluted with a water-soluble filler, said water-soluble filler may comprise lactose, dextrose, mannitol, or combinations thereof.

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

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

[0305] The composition is useful when characterized in the method of the invention (in all respects) because it can more effectively treat IgAN. The compositions, pharmaceutical preparations, uses and methods described herein may also have the advantages of being more convenient, more effective, less toxic, having a wider range of activity, more potent, producing fewer side effects, having lower inter-patient variability, or having other useful pharmacological properties that are superior to those of other similar preparations or methods (treatments) known in the prior art, whether for treating IgAN or otherwise.

[0306] When the term “about” is used herein in the context of quantities, such as absolute amounts, such as weight, volume, size, diameter, etc., or relative amounts (e.g., percentages) of individual components in a composition or a component of a composition (including concentration and ratio), time ranges, and parameters such as temperature, it should be understood that such variables are approximate values ​​and can therefore differ from the actual values ​​specified herein by ±10%, for example ±5%, and preferably ±2% (e.g., ±1%). This is also true even if such values ​​are first expressed as percentages (e.g., “about 10%” can mean an approximate value of 10 ± 10%, i.e., any value between 9% and 11%).

[0307] Figure 1 is a flowchart detailing the preparation of cured polymer-coated beads.

[0308] Figure 2 shows the in vitro dissolution profiles of budesonide-regulated release capsules in a pH 6.8 buffer medium in the presence of an added surfactant at a paddle rotation speed of 100 rpm; the data show different batches of capsules prepared according to the invention.

[0309] Figure 3 repeats the in vitro dissolution curves shown in Figure 2, but focuses only on dissolution at pH 6.8.

[0310] Figure 4: In vitro dissolution curves of seven individual batches of the budesonide-regulated release capsules according to the invention are shown in contrast, wherein dissolution in a pH 6.8 buffer medium is performed in the absence of surfactant and at a paddle rotation speed of 50 rpm.

[0311] Figure 5: In vitro dissolution curves of the budesonide-regulated release capsules according to the 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: In the absence of added surfactant and at a paddle rotation speed of 100 rpm, at pH 6.8...In vitro dissolution profiles of the budesonide-modulated release capsules according to the invention in buffered media and three other commercially available budesonide-containing formulations.

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

[0314] Figure 8: Percentage change in BAFF levels relative to end-of-treatment levels after follow-up. Percentage change in BAFF measured in patients after 9 months of nivolumab-budesonide (16 mg / day) administration: (a) at the end of the 9-month treatment period compared to baseline; and (b) after the 12-month follow-up period compared to the end of treatment. Dashed lines indicate the percentage of no change after intervention with placebo or nivolumab-budesonide.

[0315] Figure 9: Percentage change in APRIL levels relative to baseline after treatment. Percentage change in APRIL levels relative to baseline measured in patients after 9 months of treatment with the following: (a) placebo; (b) nivolumab-budesonide (8 mg / day); and (c) nivolumab-budesonide (16 mg / day). Dashed lines indicate the percentage of no change after intervention with placebo or nivolumab-budesonide.

[0316] Figure 10: Percentage change in TACI levels relative to baseline after treatment. Percentage change in TACI levels relative to baseline measured in patients after 9 months of treatment with the following: (a) placebo; (b) nivolumab-budesonide (8 mg / day); and (c) nivolumab-budesonide (16 mg / day). Dashed lines indicate the percentage of no change after intervention with placebo or nivolumab-budesonide.

[0317] Figure 11: Percentage change in BCMA levels relative to baseline after treatment. Percentage change in BCMA levels relative to baseline measured after 9 months of treatment with the following: (a) placebo; (b) nivolumab-budesonide (8 mg / day); and (c) nivolumab-budesonide (16 mg / day). Dashed lines indicate the percentage of no change after intervention with placebo or nivolumab-budesonide.

[0318] Figure 12: Percentage change in CD27 levels relative to baseline after treatment. Percentage change in CD27 levels relative to baseline measured after 9 months of treatment with the following: (a) placebo; (b) nivolumab-budesonide (8 mg / day); and (c) nivolumab-budesonide (16 mg / day). Dashed lines indicate the percentage of no change after intervention with placebo or nivolumab-budesonide.

[0319] Figure 13: Percentage change in CD27 levels relative to baseline levels after follow-up. The percentage change in CD27 was measured in patients treated with nivolumab-budesonide (16 mg / day) for 9 months at: (a) at the end of the 9-month treatment period compared to baseline; and (b) after the 12-month follow-up period compared to the end of treatment. Dashed lines indicate no change after intervention with placebo or nivolumab-budesonide.

[0320] Figure 14: Percentage change in CD30 levels relative to baseline levels after treatment. The percentage change in CD30 levels relative to baseline levels was measured in patients treated for 9 months at: (a) placebo; (b) nivolumab-budesonide (8 mg / day); and (c) nivolumab-budesonide (16 mg / day). Dashed lines indicate no change after intervention with placebo or nivolumab-budesonide.

[0321] Figure 15: Shows a significant (p<0.05) dose-dependent decrease in serum IgA secretion levels with budesonide capsules. However, serum IgA levels did not change.

[0322] Figure 16: Shows a significant (p<0.05) dose-dependent decrease in serum IgA-IgG immune complex levels with budesonide capsules.

[0323] Figure 17: Shows a significant (p<0.05) dose-dependent decrease in adverse O-galactosylated IgA1 levels with budesonide capsules.

[0324] Figure 18: Shows no difference in total IgA, IgA1, and IgG levels observed after budesonide capsule treatment.

[0325] Figure 19: Shows the in vitro dissolution curves of budesonide-regulated release capsules in simulated intestinal fluid at pH approximately 6.5 under a fasting state, in the presence of added surfactants.

[0326] Figure 20: Showing the in vitro dissolution profiles of budesonide-regulated release capsules in FaSSIF in the presence of added surfactants in a Grade 1 fasting state simulated intestinal fluid at pH 6.5, compared to three other commercially available budesonide-containing formulations.

[0327] Figure 21: Showing the in vitro dissolution profiles of core-shell bead formulations containing budesonide in the absence of enteric-coated capsules.

[0328] Figure 22: Showing the in vitro dissolution profiles of budesonide-regulated release capsules in FaSSIF in the absence of added surfactants in a Grade 1 fasting state simulated intestinal fluid at pH 6.5, compared to three other commercially available budesonide-containing formulations.

[0329] Figure 23: (a) Showing the results of a PBPK model based on the dissolution of regulated release capsules; (b) Showing the results of a PBPK model based on the dissolution of comparative products.

[0330] Figure 24: Detection of iron oxide-loaded capsules at 15 and 90 minutes, and coronal MRI images of T2* / T1-weighted TRUFI of iron oxide dispersion in the ileum at 270 minutes. Specification 26 / 59 pages 29 CN 120754112 A Detailed Description

[0331] Examples

[0332] Example 1: Preparation of Core-Shell Composition

[0333] The Opadry OY-7240 mentioned below is a dry powder polymer blend having the following components:

[0334]

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

[0336] Minimum and 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 ethyl cellulose ratio 0.00 0.24 Glycerol, % 0.0 0.6 Solids, % 23.0 26.0 Ph 9.5 11.5 Brookfield viscosity, cps 400.00 1500.00

[0337] To avoid ambiguity, the minimum and maximum values ​​in the table above refer to the minimum and maximum amounts of these components in different batches of Surelease.

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

[0339] A budesonide-coated suspension was prepared by dissolving Opadry OY-7240Clear (2.29 kg) in purified water (26.5 kg) and subsequently adding micronized budesonide (0.640 kg) to the solution while continuously mixing.

[0340] Glycospheres (40.3 kg) with a sieve aperture size of 16 to 18 were loaded into a preheated product tank in a fluidized bed. When the product temperature reached the target of 45°C, the active coating suspension was sprayed onto the glycospheres / inert core. The operation was monitored and controlled by a processing computer. After the required amount of active coating suspension had been sprayed, the active coated beads were dried and cooled.

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

[0342] The sealing coating solution was applied to preheated active coated beads. The operation was monitored and controlled by a processing computer. When the required amount of solution had been sprayed, the coating was stopped, and the sealed coated beads were dried and cooled. The fluidized bed was emptied, and the beads were screened using 1.4 mm (14 mesh) and 0.5 mm (35 mesh) sieves to remove any oversized or undersized particles. The beads were weighed, and the yield of the accepted fraction was calculated.

[0343] The extended release coating solution was prepared by adding Opadry OY-7240 (1.37 kg) to purified water (16.3 kg) while continuously mixing. Add type B ethyl cellulose suspension dispersion to the Opadry solution during continuous mixing.(Surelease, 12.8 kg). Instructions 27 / 59 pages 30 CN 120754112 A

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

[0345] The resulting polymer blend of the extended-release coating on the beads contains approximately 27.3% by weight of HPMC and approximately 51.8% by weight of ethyl cellulose in the total blend. Ethyl cellulose is a water-insoluble polymer as defined above and HPMC acts as a pore-forming polymer.

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

[0347] The qualified portion of the polymer-coated beads was loaded into the preheating 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 the hopper in the stainless steel bin.

[0348] The cured beads were then filled into size 1 capsules using an automated encapsulator, and the capsules were subsequently coated with 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 was in the range of about 34 to about 42 mg per capsule. The total amount of budesonide in each capsule is approximately 4 mg.

[0349] Example 2: During the buffering phase, in the presence of a surfactant and at a blade rotation speed of 100 rpm, according to USP <711> / General method for standard in vitro dissolution test as per European Pharmacopoeia 2.9.3

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

[0351] Dissolution Equipment Setup

[0352]

[0353]

[0354] Budesonide release was measured using ultra-high performance liquid chromatography (Ultra Performance Liquid Chromatography).Chromatography (UPLC) measurement. Instructions for use 28 / 59 pages 31 CN 120754112 A

[0355] Reagents and standards

[0356] Standards and reference substances:

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

[0358] Other reagents:

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

[0360] Dissolution medium and diluent

[0361] Acid-resistant medium

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

[0363] 0.2M trisodium phosphate buffer solution

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

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

[0366] To prepare 6L of buffer dissolution medium, 4500mL of acid-resistant medium was mixed with 1500mL of water. Combine 0.2M trisodium phosphate buffer solution and 3g Tween 80, then mix. Check the pH and, if necessary, adjust the pH to 6.8 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0367] Analytical Procedure

[0368] Acid-resistant Procedure

[0369] Note: Care should be taken not to scratch or damage the capsules when placing them in the settling basket.

[0370] Place 900mL 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> The Rotating Paddle Apparatus method (Pharmacopoeia Test No. 2.9.3) was used, with the apparatus operated at 100 rpm.

[0372] Subsequently, each of the six capsules was placed in a separate coil settling basket and then in a separate container.

[0373] After 2 hours, 15 mL aliquots of the acid-resistant solution were aspirated using a syringe.

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

[0375] The following two steps were performed after dissolution in the initial buffer phase.

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

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

[0378] Budesonide release in the acid-resistant phase was based on USP. <711> Assessment according to the acceptance criteria in European Pharmacopoeia 2.9.3.

[0379] Buffer dissolution procedure

[0380] After extracting the acid-resistant sample, the coil settling baskets containing budesonide capsules were transferred using tweezers to different groups of dissolution cups containing 900 mL of buffer dissolution medium at 37°C ± 0.5°C.

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

[0382] Sampling at specified time points: At 0.5 and 2 hours, 15 mL aliquots of the dissolution solution were drawn using a syringe. The drawn fluid was not replaced. Instructions for Use 29 / 59 pages 32 CN 120754112 A

[0383] The test solution was filtered using a Whatman GF / F GMF filter, with the first 5 mL discarded and the remaining solution 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 thoroughly mixed and the aliquots were transferred to HPLC vials.

[0386] Budesonide release in the buffer phase is based on USP <711> Evaluation according to the acceptance criteria in European Pharmacopoeia 2.9.3.

[0387] Example 3: Evaluation according to in vitro USP in the presence of surfactant and at a blade rotation speed of 100 rpm during the buffer phase. <711> Dissolution profile analysis for Pharmacopoeia test number 2.9.3

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

[0389] The overall dissolution profiles for three individual batches are shown in Figure 2, where the time from 0 to 2 hours is at an acidic pH (pH 1.2) and the time from 2 to 4 hours is at a buffered pH 6.8. Figure 3 repeats Figure 2 but only shows dissolution at a buffered pH 6.8, where the time from 0 to 120 minutes in Figure 3 corresponds to the time from 2 to 4 hours in Figure 2. Twelve capsules were tested per batch.

[0390] Quantitative results of budesonide dissolution in various media at 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] Budesonide release in the acid-resistant and buffered phases is based on <711> Evaluation according to the acceptance criteria in European Pharmacopoeia 2.9.3.

[0395] Example 4: Evaluation according to USP in the absence of surfactant and at a blade rotation speed of 50 rpm during the buffering phase. <711> / General method for standard in vitro dissolution test as per European Pharmacopoeia 2.9.3

[0396] As described in European Pharmacopoeia 2.9.3 Dissolution test for solid dosage forms (using method 2) and USP <711> As described in Dissolution (using Method 2), the in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed. Measurements were performed as follows.

[0397] Dissolution Equipment Setup Instructions 30 / 59 pages 33 CN 120754112 A

[0398]

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

[0400] Reagents and Standards

[0401] Standards and Reference Substances:

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

[0403] Dissolution Medium and Diluent

[0404] Acid-resistant Medium

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

[0406] 0.2M Trisodium Phosphate Buffer Solution

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

[0408] Buffer dissolution medium: 50mM sodium phosphate buffer solution containing Tween 80, pH 6.8.

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

[0410] Analytical Procedure

[0411] Acid-resistant Procedure

[0412] Note: When placing in the settling basket, be careful not to scratch or damage the capsules.

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

[0414] According to USP <711> The rotary paddle method, pharmacopoeia test number 2.9.3, was operated at 50 rpm.

[0415] Subsequently, each of the six capsules was placed in a separate coil settling basket and then in a separate container.

[0416] After 2 hours, 15 mL aliquots of the acid-resistant solution were drawn using a syringe.

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

[0418] After dissolution in the initial buffer stage, the following two steps were performed.

[0419] 5.0 mL of the filtered acid-resistant sample solution was transferred to a 10 mL volumetric flask and diluted with acetonitrile to make up the volume.

[0420] The solution was thoroughly mixed and the aliquots were transferred to HPLC vials for analysis.

[0421] Budesonide release during the acid-resistant phase is based on USP. <711> Evaluation according to the acceptance criteria in European Pharmacopoeia 2.9.3.

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

[0423] Overall dissolution profiles for seven individual batches are shown in Figure 4, with the time from 0 to 2 hours at acidic pH (pH 1.2) and the time from 2 to 4.5 hours at buffered pH 6.8.

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

[0425] Quantitative results of budesonide dissolution at time point 1.25 hours (75 minutes) at pH 6.8 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 budesonide was released.

[0429] 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 at 2.5 hours was 77%.

[0432] Example 5: During the buffering phase, in the presence of a surfactant and at a blade rotation speed of 100 rpm, according to USP <711> / Comparative Tests according to European Pharmacopoeia 2.9.3

[0433] The following summarizes variations of the in vitro test for Example 2. In this test, budesonide-modulated release capsules according to the invention and three other commercially available budesonide-containing formulations were analyzed. The three formulations are (Tillotts Pharma), (Dr Falk Pharma GmbH), and (Ferring Pharmaceuticals, CH). DescriptionPage 32 / 59, CN 120754112 A

[0434]

[0435]

[0436] Standards and reference substances

[0437] Budesonide, European Pharmacopoeia CSR.

[0438] Other reagents:

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

[0440] Dissolution medium, mobile phase and diluent

[0441] Acid-resistant medium

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

[0443] Buffer dissolution medium

[0444] Sodium phosphate pH 6.80 dissolution medium is 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 6 × 961.5 ml (reagecon.com).

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

[0447] After extracting the acid-resistant sample, remove the capsule from the solution with forceps and set aside, while emptying the container, cleaning it, and filling it with preheated buffer medium. Add 0.05% Tween 80 (or equivalent) to each dissolution vessel; for example, add 450 mg Tween 80 to the dissolution vessel after filling it with 900 mL of preheated buffer to obtain a surfactant concentration of 0.05% w / v.

[0448] After all containers have reached the target temperature, begin the experiment by adding capsules to each container.

[0449] Modification of operation for Budenofalk

[0450] A different operation was used when the capsule ruptured during the acid phase. Most of the acid phase was carefully decanted and the remaining acid was then carefully removed with a pipette so that as few pellets as possible were removed from the container. The buffer phase was started by adding 900 mL of preheated buffer medium, followed by adding Tween.

[0451] Sampling instructions for both phases 33 / 59 pages 36 CN 120754112 A

[0452] 10 mL was aspirated, 8 mL was discarded (via a Whatman filter (0.7 μm)), and 1 mL was sampled into an HPLC vial.

[0453] The changes in this test compared to Example 2 did not affect the overall dissolution profile of the tested product, and comparative dissolution profiles of three other commercially available budesonide-containing formulations can be seen in Figure 5.

[0454] Example 6: In the buffer phase in the absence of surfactant and at a paddle rotation speed of 100 rpm, according toUSP <711> / Comparative Tests of European Pharmacopoeia 2.9.3

[0455] The variations of the in vitro tests of Examples 2 and 5 are summarized below. In this test, enteric-coated capsules filled with core-shell beads from Example 1 and three other commercially available budesonide formulations were analyzed in a buffer 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 References

[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, 82 mL of concentrated HCl was combined with 10000 mL of water and mixed thoroughly.

[0462] Buffer dissolution medium

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

[0464] See Sodium phosphate pH 6.80 dissolution medium 6 × 961.5 mL (reagecon.com) for more details.

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

[0466] After extracting the acid-resistant sample, remove the capsule from the solution with forceps and set aside, while emptying the container, cleaning it, and filling it with 900 mL of preheated buffer medium.

[0467] After all containers have reached the target temperature, begin the experiment by adding capsules to each container. Instructions 34 / 59 pages 37 CN 120754112 A

[0468] Modification of operation for Budenofalk

[0469] Use different operation when the capsule breaks during the acidic phase. Carefully decanter most of the acid phase and then carefully remove the remaining acid with a pipette to remove as few pellets as possible from the container. Begin the buffering phase by adding 900 mL of preheated buffer medium.

[0470] Sampling for both phases

[0471] Aspirate 10 mL, discard 8 mL (via a Whatman filter (0.7 μm)), and take 1 mL 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, are shown in Figure 6.

[0473] The table below shows the f2 values ​​used to compare the budesonide capsules according to the invention tested under this method with those of other commercially available products. A similarity in the profiles requires an f2 value of 50 or greater (FDA SUPAC Guidances 1995, 1997).

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

[0475] It is evident that the release profiles of budesonide differ significantly among the four commercially available products. Comparisons of f2 values ​​between Entocort EC and the other products do not show similarity; similarity would require an f2 value of 50 or greater. In fact, based on f2 evaluation and visual inspection of the graphical profiles, 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 through 16, enteric-coated capsules filled with solidified beads as described in Example 1 were used. The capsules are referred to interchangeably below as “budesonide capsules” or “Entocort budesonide”.

[0478] Study Design

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

[0480] Patients

[0481] Males or females aged at least 18 years with biopsied primary IgA nephropathy and significant proteinuria were recruited for the induction period. 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 m2 and a urinary protein creatinine ratio (UPCR) greater than 0.5 g / g, or total urinary protein of at least 0.75 g / day, levels considered to increase the risk of progression to end-stage renal disease. UPCR using 24-hour protein excretion or 24-hour urine collection was used to determine the applicability of overcoming potential collection errors and deviations from normal creatinine excretion (e.g., in physically active and muscular men) to minimize the risk of unintentionally excluding patients.

[0482] Procedure

[0483] The drug was an oral capsule formulation (budesonide capsules) or placebo as described in Example 1, designed with an in vitro dissolution profile as outlined above to provide sustained release of the active compound, which was delayed until the capsule reached the ileum, particularly the distal ileum, targeting sites with high-density Peyre's plaques.

[0484] Following screening, eligible patients were enrolled for a 6-month induction period, a 9-month treatment period, and a 3-month follow-up period; patient eligibility was assessed prior to the induction and treatment periods. During induction, RAS blockade was performed by combining an ACE inhibitor (ACEI) and an angiotensin II receptor blocker (ACEI).The blocker (ARB) was titrated up 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, less than 0.5 g / g of UPCR, and less than 0.75 g / day of urinary protein to optimize. At the end of the induction, 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 a measured GFR ≥ 45 mL / min / 1.73 m2 and blood pressure 160 / 100 mm Hg or less, despite optimized RAS blockade, were eligible for treatment randomization.

[0485] An independent Data and Safety Monitoring Board (DSMB) monitored all safety issues and reviewed the 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 permutation set computational method. Within each set, patients were assigned in a 1:1:1 ratio to budesonide capsules 16 mg / day, budesonide capsules 8 mg / day, or placebo. All patients continued optimized RAS blockade therapy throughout the treatment period.

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

[0489] The experiment was double-blind. Therefore, throughout the experiment and analysis, the allocation of treatment groups was unknown to each patient, all experimental staff (including the researchers and other staff who performed the randomization and analysis), the sponsor, and the DSMB (the occlusion safety data and unblinding data reviewed by the DSMB were available where issues arose).

[0490] To ensure occlusion, the placebo capsules provided by the sponsor had the same appearance and route of administration as the active capsules. Patients self-administered the occlusion capsules once daily, one hour before breakfast, during the treatment period. During the follow-up period (9 to 12 months), patients who received budesonide capsules at 16 mg / day during months 0 to 9 were transitioned to 8 mg / day for 2 weeks, while all others...Patients (i.e., those who received budesonide capsules 8 mg / day or placebo during months 0 to 9) received placebo to maintain blindness. No other investigational drugs were administered after the transition. Instructions for Use 36 / 59 pages 39 CN 120754112 A

[0491]

[0492]

[0493] Table 2. Patient demographics and baseline characteristics. Data are n (%), mean (SD), or median (IQR). Acronyms: BMI = body mass index; CKD-EPI = Chronic Kidney Disease Epidemiological Collaborative 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 start of the treatment period (month 0, before any treatment administration), at the end of the treatment period (month 9), and at the end of the follow-up period (month 12) as per the instructions on page 37 / 59 of the manual. Samples obtained were tested for levels of multiple biomarkers, including: BAFF; APRIL; TACI; BCMA; CD27, CD30; secreted IgA; IgA-IgG immune complexes; and adverse O-galactosylated IgA1.

[0495] Treatment-coded envelopes were provided to each randomized group of patients. In case of emergency, the coded envelopes may be opened. Any unblinded patient must withdraw from the experiment.

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

[0497] Materials and Methods

[0498] Biomarkers were measured using a multiplex analysis (R&D Systems) based on custom-designed beads, according to the manufacturer's instructions.

[0499] The concept of the analysis is based on the selective binding of molecules of interest, thus allowing for the simultaneous detection and quantification of fluorescently labeled microspheres containing multiple biomarkers in extremely small volumes of serum.

[0500] In this study, biomarkers were selected and grouped according to their analytical dynamic range (Group 1: BAFF; APRIL; Group 2: TACI; BCMA; CD27; CD30).

[0501] The microsphere mixtures specifically labeled in Group 1 or Group 2 were diluted 1:10 with reagent diluent (microspheres supplied by R&D Systems), and serum samples were diluted 1:2 with reagent diluent. Continuous dilutions of standard solutions (microspheres supplied by R&D Systems) were used to generate standard curves.

[0502] 50 μl of microsphere mixture was applied to the analytical plate, followed by 50 μl of standard or sample. The plate was vibrated on a microplate.The plates were incubated at 800 rpm at room temperature for 2 hours on an agitator. Proteins not bound to the microbeads were removed by washing with wash buffer (microparticles provided by R&D Systems).

[0503] Biotinylated antibody mixture (microparticles provided by R&D Systems) was added to each well and incubated at room temperature for 1 hour. The plates were washed with wash buffer (microparticles provided by R&D Systems) and then incubated for 30 minutes with 50 μl of streptococcal biotin-phycoerythrin (microparticles provided by R&D Systems) per well, followed by a final wash.

[0504] The microparticles were resuspended in 100 μl of wash buffer and the fluorescence in each well was read over 90 minutes on a flow cytometer.

[0505] Differences in biomarker levels were compared using one-way ANOVA statistical tests with p < 0.05.

[0506] Results

[0507] As shown in Figure 7 and Table 3, in the samples obtained at the end of the 9-month treatment period, the serum levels of BAFF induced by treatment with 8 mg / day and 16 mg / day budesonide capsules were statistically significantly lower than those in patients treated with placebo.

[0508]

[0509] Table 3. Percentage change in serum BAFF levels from the start of treatment to the end of treatment. Shows mean (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 range of values.

[0510] Furthermore, Figure 8 and Table 4 show that, in serum samples obtained after the completion of the follow-up period (i.e., 3 months after the end of the treatment period), the serum levels of BAFF in patients previously treated with 16 mg / day budesonide capsules increased again, indicating the observed reduction in budesonide capsule exposure as described on pages 38 / 59 of the product manual, CN 120754112 A.

[0511]

[0512] Table 4. Percentage change in serum BAFF levels from the end of treatment to the end of the follow-up period. Means (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.

[0513] The reduction in serum BAFF levels after budesonide treatment is consistent with the disease remission effect in IgAN.

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

[0515] Materials and Methods

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

[0517] Differences in APRIL levels were compared using one-way ANOVA statistical tests with p-values ​​< 0.05.

[0518] Results

[0519] As shown in Figure 9 and Table 5, treatment with budesonide capsules at 8 mg / day and 16 mg / day did not result in an observable change in serum APRIL levels in samples taken at the end of the 9-month treatment period, indicating that the effect produced by treatment with budesonide capsules was specific to BAFF. Similarly, no change was observed in the placebo group.

[0520]

[0521] Table 5. Percentage change in serum APRIL levels from the start to the end of treatment. Shows mean (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 range of values.

[0522] Example 10: Decrease in serum BAFF levels after budesonide capsule treatment was associated with a decrease in serum TACI levels

[0523] Materials and Methods

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

[0525] Differences in TACI levels were compared using one-way ANOVA with a p-value < 0.05.

[0526] Results

[0527] As shown in Figure 10 and Table 6, in the samples obtained at the end of the 9-month treatment period, the serum TACI levels induced by treatment with 8 mg / day and 16 mg / day budesonide capsules were statistically significantly lower than those treated with placebo. In the placebo group, a slight increase in TACI levels was actually observed relative to baseline.

[0528]

[0529] Table 6. Percentage change in serum TACI levels from the start to the end of treatment. Shows mean (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 range of values.

[0530] Example 11: Decreased serum BAFF levels after budesonide capsule treatment were associated with decreased serum BCMA levels

[0531] Materials and Methods

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

[0533] Differences in BCMA levels were compared using one-way ANOVA statistical tests with p-values ​​< 0.05.

[0534] Results

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

[0536]

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

[0538] Example 12: Decrease in BAFF serum levels after budesonide capsule treatment was associated with a decrease in CD27 serum levels

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

[0540] Materials and Methods

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

[0542] One-way ANOVA was used to compare differences in CD27 levels with a p-value < 0.05.

[0543] Results

[0544] As shown in Figure 12 and Table 8, in the samples obtained at the end of the 9-month treatment period, the serum CD27 levels induced by treatment with 8 mg / day and 16 mg / day budesonide capsules were statistically significantly lower than those in the placebo group. In the placebo group, a slight increase in CD27 levels was actually observed relative to baseline.

[0545]

[0546] Table 8. Percentage change in serum CD27 levels from the start to the end of treatment. Means (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] Furthermore, Figure 13 and Table 9 show that in samples obtained after the completion of the follow-up period (i.e., 3 months after the end of the treatment period), the serum CD27 level in patients previously treated with 16 mg / day budesonide capsules increased again, indicating the observed reduction in budesonide capsule exposure as described on pages 40 / 59 of the product manual, CN 120754112 A.

[0548]

[0549] Table 9. Percentage change in serum CD27 levels from the end of treatment to the end of the follow-up period. Shows mean (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 range of values.

[0550] Example 13: Decrease in BAFF serum levels after budesonide capsule treatment is associated with a decrease in serum CD30 levels

[0551] Materials and Methods

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

[0553] Differences in CD30 levels were compared using one-way ANOVA statistical tests with p-values ​​< 0.05.

[0554] Results

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

[0556]

[0557] Table 10. Percentage change in serum CD30 levels from the start to the end of treatment. Means (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 of values ​​are shown.

[0558] Consistent with large-scale ensemble analyses of genome-wide association (GWA) studies (Gesualdo L, Di Leo V, Coppo R. The mucosal immune system and IgAnephropathy. Semin Immunopathol. 2021; 43:657-668; Coppo R. The gut-renal connection in IgAnephropathy. Semin Nephrol. 2018; 38:504-512.), this study identified the gut 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 action within the gut-associated lymphoid tissue (GALT).

[0559] Example 14: Analysis of secreted IgA levels after budesonide capsule treatment

[0560] Materials and Methods

[0561] Monoclonal mouse anti-human secretory fraction (Sigma) diluted 1:10,000 in coating buffer was applied to the wells of an immunoassay plate 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 at room temperature for 90 minutes.Minutes. The plates were then washed again, and the level of secreted IgA was observed using o-phenylenediamine dihydrochloride substrate. The plate values ​​were normalized relative to the standard plate using the OD492 of high, medium, and low standards on each plate. Instructions for use 41 / 59 pages 44 CN 120754112 A

[0562] Results

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

[0564] Example 15: Analysis of IgA-IgG immune complexes and adverse O-galactosylated IgA1 levels after budesonide capsule treatment

[0565] Materials and Methods

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

[0567] Adverse galactosylated IgA: The level of adverse O-galactosylated IgA1 was measured using a commercially available KM55 ELISA (catalog number 27600, Immuno-Biological Laboratories, Inc. Minneapolis, MN 55432, USA)

[0568] Results

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

[0570] Of particular interest is the evidence that systemic glucocorticoid treatment of IgAN has reduced 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 difference was observed in the levels of total IgA, IgA1, and IgG under budesonide capsule treatment with the present invention (Figure 18), leading to the conclusion that local ileal treatment with budesonide capsules has a selective effect on pathogenic antibodies but is ineffective on the total pool of IgA, IgA1, and IgG.

[0571] These results demonstrate that treatment with budesonide supports the direct action of budesonide on the basic pathogenic pathway in IgAN, and that the budesonide loading drug has a primarily local rather than systemic effect, resulting in fewer side effects in patients treated with budesonide.

[0572] Example 16: Comparison of lag time for plasma curve onset between budesonide capsules and reference commercially available product

[0573] In a randomized crossover clinical study, 24 subjects were administered a test product (obtained according to Example 1 above) containing the same active ingredient, budesonide capsules, and a reference commercially available product (EC; AstraZeneca) to fasting subjects.

[0574] The lag time in hours for the onset of blood levels in each subject after administration of the test product and the reference (REF) product (with a suitable clearance period of 7 to 14 days) under two different conditions is shown in Table 11 below.

[0575] Instructions for Use 42 / 59 pages 45 CN 120754112 A

[0576]

[0577] Table 11. Lag time for plasma curve onset

[0578] Statistical analysis

[0579] Data were analyzed using Sigmaplot for Windows version 11.0.

[0580] Descriptive Statistics

[0581] The median lag time for plasma level onset in each arm of the study was calculated and is presented in Table 12 below. (Because these values ​​are discrete data, i.e., the lag time can only correspond to the sampling time in the study, the median is reported instead of the mean).

[0582] 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

[0583] Table 12. Median Lag Time Values ​​

[0584] It was concluded that the median (50th percentile) lag time for plasma level onset in both administrations of the test formulation was 4 hours, and 1 hour for the reference commercial formulation.

[0585] To determine whether the lag time observed in the test formulation is 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 the data were normally distributed and the other not assuming the data distribution.

[0586] The repeated measures ANOVA of the normally distributed dataset yielded an F-value of 51.815 and a P-value of P < 0.001, which is statistically highly significant (pages 43 / 59, CN 120754112 A). The Tukey test was used to determine which arms of the study differed from each other. The results of the post-hoc comparisons are presented in Table 13 below:

[0587] Comparison of mean difference q statistic P value P < 0.050 F2 vs REF 3.151 18.005 < 0.001 Yes F2 vs F1 0.261 1.517 0.820 No F1 vs REF 2.890 16.770 < 0.001 Yes

[0588] Table 13: Post-hoc comparisons between arms of the study

[0589] This indicates that although there was no difference in the onset lag time of plasma levels when the test formulation was administered under different conditions, the onset lag time of plasma levels after administration of the test formulation was statistically highly different compared with the administration of the commercially available reference product (P < 0.001).

[0590] Repeated measures ANOVA was also applied to the classification of individual lag time values ​​using the Friedman test (see, for example, Stanton. A.Glanz, Primer of Biostatistics, 5th edition, McGraw Hill 2002, ISBN 0-07-137946-0, pp. 370-380).

[0591] Using this test, the chi-square value was 52.950, producing a p-value <0.001, which was highly statistically significant. The Tukey test was used to determine which arms of the study differed from each other in a post-hoc comparison, the results of which are presented in Table 14 below:

[0592] Comparison of rank differences q statistic P < 0.05 F2 vs REF 68.500 9.237 Yes F2 vs F1 9.500 1.281 No F1 vs REF 59.000 7.956 Yes

[0593] Table 14: Post-hoc comparisons between arms of the study

[0594] This test (again) shows that although there was no difference in plasma level onset lag time when the test formulation was administered under different conditions, the plasma level onset lag time when the test formulation was administered was statistically different from that when the commercially available reference formulation was administered (P < 0.05).

[0595] In summary, regardless of the type of statistical analysis applied, the test formulation had a significantly different plasma level onset lag time compared to the commercially available reference formulation EC. In the study arm where Nexcom F was administered, the median plasma level onset lag time was 4 hours, while the median lag time for EC was 1 hour.

[0596] This analysis clearly demonstrates that, unlike the reference product, the test formulation, as described and claimed herein, does not release most of its active ingredient until it reaches the distal portion of the small intestine (e.g., the ileum, such as the distal ileum).

[0597] Example 17: According to USP, in the presence of the added surfactant Tween 80 <711> / General method for standard in vitro dissolution test as per European Pharmacopoeia 2.9.3

[0598] As described in European Pharmacopoeia 2.9.3 Dissolution test for solid dosage forms (using method 2) and USP <711> As described in Dissolution (using Method 2), the in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed. Measurements were performed as described below.

[0599] Three commercially available budesonide-containing formulations were also analyzed in this test. The three formulations were (Tillotts Pharma), (Dr Falk Pharma GmbH), and (Ferring Pharmaceuticals, CH).

[0600] Dissolution apparatus setup instructions 44 / 59 pages 47 CN 120754112 A

[0601]

[0602] Budesonide release was measured using ultra-high performance liquid chromatography (UPLC).

[0603] Reagents and standards

[0604] Standards and reference substances:

[0605] Budesonide, European Pharmacopoeia CSR, or suitable secondary standards.

[0606] Other reagents:

[0607] Tween 80 (polysorbate (80)), Fisher Scientific, or equivalent.

[0608] Dissolution medium and diluent

[0609] Acid-resistant medium

[0610] 0.1N HCl solution. To prepare 6L of acid-resistant medium, 50mL of concentrated HCl was hydrated with 6000mL of water and thoroughly mixed to obtain the solution.

[0611] Buffer dissolution medium

[0612] Buffer solutions were prepared using FaSSIF buffer concentrate (from Biorelevant.com, product code FASBUF01).

[0613] 0.05w / v% (0.5mg / mL) Tween 80 to buffer solution: For example, to prepare 6L of buffer dissolution medium, 3g of Tween 80 was added to 6L of buffer solution to obtain a Tween 80 concentration of 0.05w / v%.

[0614] Thoroughly mix the resulting solution and check the pH. If necessary, adjust the pH to 6.5 ± 0.05 using hydrochloric acid or sodium hydroxide.

[0615] Based on USP <711> Evaluation of budesonide release according to acceptance criteria in European Pharmacopoeia 2.9.3.

[0616] Example 18: Evaluation of budesonide release according to in vitro USP in the presence of surfactant Tween 80. <711> Dissolution curve analysis of budesonide capsules (Pharmacopoeia Test No. 2.9.3)

[0617] 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.

[0618] The overall average dissolution curves of the three samples in the buffer phase are visible in Figure 19. No budesonide release was observed in the acid-resistant phase at the 2-hour sampling time point. Specification 45 / 59 pages 48 CN 120754112 A

[0619] Quantitative results of budesonide dissolution in different media at 2 hours at pH 1.2 and at 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 120 minutes and 180 minutes in the buffer phase at pH 6.5 are provided in the table below.

[0620]

[0621]

[0622] *SD = Standard Deviation

[0623] Based on USP <711> The budesonide release was evaluated according to the acceptance criteria in European Pharmacopoeia 2.9.3.

[0624] For comparison, dissolution profiles of budesonide capsules and three other budesonide-containing formulations were obtained according to the protocols outlined below. The three other budesonide-containing formulations are (Tillots Pharma), (Dr Falk Pharma GmbH), and (Ferring Pharmaceuticals, CH).

[0625] Method for use in capsules

[0626]

[0627] Standards and reference substances

[0628] Budesonide, European Pharmacopoeia CSR.

[0629] Other reagents:

[0630] Tween 80, (polysorbate (80)), Fisher Scientific, or equivalent.

[0631] Dissolution media, mobile phase and diluents Instructions for use 46 / 59 pages 49 CN 120754112 A

[0632] Acid-resistant media

[0633] 0.1N HCl solution. For example, to prepare 10L, combine 82mL of concentrated HCl with 10000mL of water and mix thoroughly.

[0634] Buffer Dissolution Medium

[0635] 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.

[0636] After extracting the acid-resistant sample, remove the capsule from the solution with forceps and set aside, while emptying the container, cleaning it, and filling it with preheated buffer medium. Add 0.05 w / v% Tween 80 (or equivalent) to each dissolution vessel; for example, after filling the dissolution vessel with 900mL of preheated buffer, add 450mg Tween 80 to the dissolution vessel to obtain a surfactant concentration of 0.05%.

[0637] After all containers reached the target temperature, the experiment was started by adding capsules to each container.

[0638] Modification of operation for Budesonide

[0639] A different operation was used when the capsules ruptured during the acid phase. Most of the acid phase was carefully decanted and the remaining acid was then carefully removed with a pipette to remove as few pellets as possible from the container. The buffer phase was started by adding 900 mL of preheated buffer medium, followed by adding Tween.

[0640] Sampling of the two phases

[0641] 10 mL was aspirated, 8 mL was discarded (via a Whatman filter), and 1 mL was sampled into an HPLC vial.

[0642] To avoid ambiguity, the change in this test compared to Example 2 did not affect the overall dissolution profile of the tested product.

[0643] Figure 20 shows the dissolution profiles of budesonide capsules compared to three other budesonide-containing formulations. This figure clearly shows that, in the FaSSIF medium simulating the environment of the small intestine, budesonide capsules have release profiles that differ from all other commercially available budesonide-containing formulations.

[0644] Example 19: In vitro USP in the absence of enteric-coated capsules <711> / Analysis of dissolution profiles of core-shell beads for Pharmacopoeia test number 2.9.3

[0645] The solidified core-shell beads prepared as in Example 1 were also tested (only) under the buffer-stage dissolution conditions outlined in Example 17 in the absence of enteric-coated capsules.

[0646] The overall average dissolution profiles of the three samples in the buffer stage are shown in Figure 21.

[0647] Quantitative results of budesonide dissolution from the core-shell beads in the buffer stage at pH 6.5 at time points of 15, 30, 45, 60, 90, 120 and 180 minutes are provided in the table below.

[0648] *SD = Standard deviation Specification 47 / 59 pages 50 CN 120754112 A

[0649] The release profiles of the core-shell beads in the absence of enteric-coated capsules in the FaSSIF buffer concentrate further confirm that most of the budesonide will be released into the ileum in vivo. In other words, most of the release from the beads occurs within a 90-minute period, and combined with the release delay of the enteric coating, achieves the desired dissolution profile for the entire formulation. Most of the budesonide will be released into the ileum in vivo, as confirmed by the obtained biomarker data and the modeling results provided in Example 25 below.

[0650] Example 20: According to USP in the absence of surfactant Tween 80. <711> / General method for standard in vitro dissolution test as per European Pharmacopoeia 2.9.3

[0651] As described in European Pharmacopoeia 2.9.3 Dissolution test for solid dosage forms (using method 2) and USP <711> As described in Dissolution (using Method 2), the in vitro dissolution of the encapsulated budesonide core-shell beads of Example 1 was analyzed. Measurements were performed as follows.

[0652] Three commercially available budesonide formulations were also analyzed in this test. The three formulations were (Tillotts Pharma), (Dr Falk Pharma GmbH), and (Ferring Pharmaceuticals, CH).

[0653] Dissolution apparatus setup

[0654]

[0655] Budesonide release was measured using ultra-high performance liquid chromatography (UPLC).

[0656] Reagents and standards

[0657] Standards and reference substances:

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

[0659] Dissolution medium and diluent

[0660] Acid-resistant medium

[0661] 0.1N HCl solution. To prepare 10L of acid-resistant medium, 82mL of concentrated HCl was mixed thoroughly with 10000mL of water.

[0662] Buffer 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 extracting the acid-resistant sample, remove the capsule from the solution with forceps and set aside, while emptying the container, cleaning it, and filling it with 900 mL of preheated buffer medium.

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

[0666] Modification of Operation for Budenofalk

[0667] Use a different operation when the capsule breaks during the acid phase. Carefully decan 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. Begin the buffer phase by adding 900 mL of preheated buffer medium.

[0668] Two-stage sampling

[0669] Aspirate 10 mL, discard 8 mL (via Whatman filter), and take 1 mL into an HPLC vial.

[0670] Based on USP <711> Evaluation of budesonide release according to acceptance criteria in European Pharmacopoeia 2.9.3.

[0671] Example 21: Evaluation of budesonide release according to in vitro USP in the absence of surfactant Tween 80. <711> / Pharmacopoeia Test No. 2.9.3 Dissolution Profile Analysis of Budesonide Capsules

[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 average dissolution profile of the budesonide capsules is shown in Figure 22. Figure 22 also contains the dissolution profiles under the same test conditions.Dissolution profiles for three other budesonide-containing formulations are provided for comparison. It is clear from this figure that budesonide capsules exhibit release profiles distinct from all other commercially available budesonide-containing formulations in the FaSSIF medium, which simulates the environment of the small intestine.

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

[0675] Quantitative results of budesonide dissolution in different media at time points pH 1.2 for 2 hours and at buffer phases pH 6.5 for 15, 30, 45, 60, 90, 120, and 180 minutes are provided in the table below for budesonide capsules and the three comparative formulations. Instruction manual, pages 49 / 59, 52 CN 120754112 A

[0676]

[0677] *SD = Standard Deviation

[0678] Based on USP <711> / Acceptance criteria for budesonide release as per European Pharmacopoeia 2.9.3.

[0679] The table below shows the f2 values ​​used to compare the budesonide capsules according to the invention tested under this method with those of other commercially available products. A similarity in the curves requires an f2 value of 50 or greater (FDASUPAC Guidances 1995, 1997).

[0680] Entocort EC Budenofalk Cortiment F2 value (biorelevant) 11.7 16.1 15.8

[0681] It is evident that the release curves of budesonide differ significantly among the four commercially available products. F2 comparisons between Entocort and the other products do not show similarity, and a similarity would require an f2 value of 50 or greater. In fact, based on f2 evaluation and visual inspection of the graphical curves, their release curves must be considered very different.

[0682] Example 22: First Tablet Formulation

[0683] The tablets were manufactured via the following steps:

[0684] 1. Wet granulation. Budesonide, mannitol, hydroxyethyl cellulose, hydroxypropyl cellulose, and sodium glycolate starch were blended. An ethylene-water mixture was then sprayed onto the powder during blending. The resulting granules were then dried.

[0685] 2. Final blending. The dried granules were blended with sodium stearoyl fumarate.

[0686] 3. Tableting. Tablets were compressed using a tableting machine.

[0687] 4. Enteric coating. A copolymer of methacrylic acid and methyl methacrylate, talc, and dibutyl sebacate were dispersed in an isopropanol-water mixture under mixed conditions. The coating dispersion was 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. Specification 50 / 59 pages 53 CN 120754112 A

[0689] Tablet composition.

[0690]

[0691] *Removed during operation.

[0692] Example 23: Second tablet formulation

[0693] The tablets were manufactured via the following steps:

[0694] 1. Wet granulation. Budesonide was initially blended with colloidal silica. Microcrystalline cellulose and dicalcium hydrogen phosphate were then added, followed by further blending. A solution of hydroxypropyl cellulose in an ethylene-water mixture was then sprayed onto the powder while blending. The resulting granules were then dried.

[0695] 2. Blending. The dried granules were blended with microcrystalline cellulose, sodium glycolate starch, and copovidone. As a final blending step, magnesium stearate was added to the blend, followed by final blending.

[0696] 3. Tableting. Tablets were compressed using a tableting machine.

[0697] 4. Enteric coating. A copolymer of methacrylic acid and methyl methacrylate, talc, and dibutyl sebacate are dispersed in an isopropanol-water mixture under mixed conditions. The coating dispersion is then sprayed onto tablets using a pan coater.

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

[0699] Tablet Composition.

[0700] Component Amount (mg) Functional Budesonide, Micronized 4.00 API (Intragranule) Colloidal Silica 0.50 Lubricant (Intragranule) Dicalcium Hydrogen Phosphate 16.00 Filler (Intragranule) Instructions for Use 51 / 59 Page 54 CN 120754112 A Microcrystalline Cellulose 15.50 Filler (Intragranule) Hydroxypropyl Cellulose 4.00 Binder (Intragranule) Ethanol* Wet Granulation Solvent Purified Water* Wet Granulation Solvent Microcrystalline Cellulose 53.50 Filler (Outside Granules) Copovidone 5.00 Binder (Outside Granules) Sodium Glycoacetic Acid Starch 1.00 Disintegrant (Outside Granules) Magnesium Stearate 0.50 Lubricant (Outside Granules) Total (Uncoated Tablets) 100.00 mg Methacrylic Acid and Methyl Methacrylate Copolymer (1:1) 9.85 Enteric coating polymer: methacrylic acid and methyl methacrylate copolymer (1:2) 3.31; Enteric coating polymer: talc 3.31; Slime, coating uniformity: dibutyl sebacate 2.55; Plasticizer: isopropanol*; Coating solvent: purified water*; Total coating solvent (coated tablets): 119.02 mg

[0701] *Removed during operation.

[0702] Example 24: Third tablet formulation

[0703] Tablet components:

[0704] Instructions for use 52 / 59 pages 55 CN 120754112 A

[0705] *Removed during manufacturing.

[0706] Manufacturing of Uncoated Tablets (200g Batch)

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

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

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

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

[0711] 5. Compress tablets to obtain 100 mg of uncoated tablets.

[0712] Preparation of Coated Dispersion

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

[0714] 1. A diluent mixture was prepared by co-mixing isopropanol, water, and dibutyl sebacate in a container (container A).

[0715] 2. An Eudragit suspension was prepared by transferring approximately half of the diluent mixture (in container A) to another container (container B). Subsequently, Eudragit L100 and Eudragit S100 were slowly added to container B during mixing. The Eudragit suspension in container B was then further mixed for 30 to 60 minutes.

[0716] 3. Talc was slowly added to the remaining diluent mixture in container A during mixing with a high-shear mixer. The talc suspension was then further mixed with a high-shear mixer for 10 minutes.

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

[0718] 5. The mixture in container B was then stirred at room temperature for 24 hours.

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

[0720] 7. The final coating dispersion is then stored at room temperature until coating, with continuous stirring during coating.

[0721] Trade Name Amount (g) Supplier Eudragit L100 (Type A methacrylic acid copolymer) 39.4 Evonik Eudragit S100 (Type B methacrylic acid copolymer) 13.22 Evonik Talc, Pharmaceutical M Grade 13.22 Imerys Dibutyl Sebate USP / NF 10.18 Merck Isopropanol 594.36 - Purified Water 20.44 - Total 690.82

[0722] Coating of Tablets (50 g batch)

[0723] Tablets were coated using a 12 cm diameter stainless steel pan coating machine and a standard nozzle. The increase in coating weight was determined by weighing tablet samples after different coating durations. Use the following spraying parameters:

[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 pressure: 0.25 bar

[0728] In vitro dissolution of tablets

[0729] The in vitro dissolution profile of the tablets was based on the scheme outlined in Example 6 above (in the phosphate buffer stage, in the absence of surfactant, at a blade rotation speed of 100 rpm, according to USP) <711> (Comparative test according to European Pharmacopoeia 2.9.3) and the protocol outlined in Example 20 above (at 100 rpm in FaSSIF buffer with surfactant Tween 80 in absence, according to USP) <711> The analysis was performed using the general method of standard in vitro dissolution testing as per European Pharmacopoeia 2.9.3. Dissolution values ​​at certain time points are shown below.

[0730]

[0731] The budesonide release from these tablets conformed to the dissolution profile required to achieve a majority release into the ileum. Particularly unexpectedly, the combination of gelling agent (hydroxypropyl methylcellulose) and disintegrant (crospovidone) allowed for the achievement of a proper in vitro dissolution profile. However, it is expected 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] ​​The capsules prepared in Example 1 (“budesonide capsules” or “Nefok budesonide”) were run using software (Simulations Plus, CA; version 9.8.3002) in a physiologically based pharmacokinetic (PBPK) model.

[0734] Following the protocol outlined in Example 2, budesonide capsules were released in vitro during an acidic phase (first 2 hours) followed by a buffer phase and uploaded to PBPK software. In vitro-in vivo correlation (IVIVC) was obtained to correlate PBPK model predictions with measured pharmacokinetic results. The predicted Cmax was faster and higher than observed. 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 separateThe role of intestine and liver in first-pass metabolism of budesonide in rat Xenobiotica. Dec 2018; 48(12): 1206-1214). After introducing intestinal wall metabolism into the PBPK model, data with good fit were achieved.

[0735] As can be seen in Figure 23(a), the IVIVC PBPK model confirms that budesonide capsules only begin to release budesonide upon reaching the ileum, and at least about 90% of budesonide is released throughout the ileum, with very little remaining in the intestinal portion after the ileum (i.e., the cecum). Therefore, this model confirms that compositions having an in vitro release curve as defined in this invention achieve budesonide-loaded drug release to the site of highest Paecil plaque concentration in the intestine, i.e., the ileum, and therefore any composition that conforms to this release curve will be effective in treating IgA nephropathy.

[0736] The release of budesonide from (Tillotts Pharma) was also modeled using software, and the results showed that all budesonide was released before the formulation entered the ileum portion of the small intestine (see Figure 23(b)).

[0737] Example 26: Study on Intra-enteric Release of Capsule Contents

[0738] A study was conducted to evaluate the location and timing of release of the contents of capsules coated with the same enteric coating as those described in Example 1 in the gastrointestinal tract.

[0739] In this study, VCaps plus size 1HPMC capsules were filled with 75 mg of caffeine, 10 mg of black iron oxide, and 87.5 mg of dehydrated manganese gluconate. 140.9 mg of sugar beads (also called 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 operation and the same equipment and facilities as those described in Example 1.

[0740] Caffeine was placed inside the capsule as a marker to determine when the capsule contents were released by measuring the presence of caffeine in saliva at different time points. Caffeine is rapidly absorbed after release into the intestine, and therefore, the presence of caffeine in saliva provides a sensitive marker of capsule opening (Sager et al. Low dose caffeine as asalivary 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)).Iron oxide was placed inside the capsule so that magnetic resonance imaging (MRI) could be used to locate the capsule's position by observing the iron oxide as it moved through the gastrointestinal tract.

[0741] The study was conducted at the University Clinic in Greifswald, Germany, as an open-label, single-center study in 12 healthy young human subjects. Subjects stopped consuming caffeine-containing foods and beverages for three days prior to 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. Each participant then ingested the enteric-coated capsule 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 intensity of 1.5 Tesla, and the imaging data were analyzed using Horos 2.2.0 (The Horos Project). All measurements were performed with the subject in a supine position (supine, head up). Saliva samples were analyzed using an LCMS 8060 system (Shimadzu Corporation, Kyoto, Japan) and were appropriately prepared for this purpose.

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

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

[0745] The first time point at which a concentration of caffeine ≥10 ng / ml in undiluted saliva was measured was defined as "salivary caffeine appearance." As shown in the table below, the average time to 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. The time to release caffeine into the small intestine after the first appearance of caffeine in the subject's saliva, minus the subject's gastric emptying time, was calculated to be an average of 181 ± 31 minutes (range 120 to 233 minutes). These values ​​indicate that the opening of the enteric-coated capsule and the release of caffeine fall well within the typical small intestinal transit time range (3.5 to 4.5 hours (210 to 270 minutes)) (Wilson et al., Chapter 3. Gastrointestinal Transit and Drug Absorption (pp. 41 to 65), “Oral Drug Absorption: Prediction and Assessment”, J. Dressman and C. Reppas, eds., Drug and the Pharmaceutical Sciences, 2nd ed., Vol. 193, Marcel Dekker, NY, NY ISBN-13:978-1-4200-7733-, Instructions for Use, pp. 55 / 59, 58 CN 120754112 A 9 (2010)).

[0746] The table below shows individual and average results for gastric emptying time (as determined by MRI), time to salivary caffeine appearance, and time to salivary caffeine appearance 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, in 11 of the 12 subjects, the enteric-coated capsules opened and released their contents in the ileum. This study thus confirms that the enteric coating and capsules described in Example 1 consistently result in the release of capsule contents into the ileum. The beads contained in Example 1 will begin releasing budesonide after the capsule opens, with most of the release occurring within one hour. The mean small intestinal transit time of the beads was compared between 3.5 and 4.5 hours (Wilson et al., Chapter 3, Gastrointestinal Transit and Drug Absorption (pp. 41-65), “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)) and the average time 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 (about one hour), it can be concluded that most of the budesonide will be released from the beads into the distal ileum and thus target Peyre's plaque located there.

[0749] This disclosure relates to embodiments of the following:

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

[0751] (i) identifying a pharmaceutically acceptable composition intended for the treatment of IgA nephropathy, comprising budesonide and one or more pharmaceutically acceptable excipients providing modulated release of said budesonide upon administration to the gastrointestinal tract, said composition being tested in standard in vitro USP <711> In the dissolution test according to the European Pharmacopoeia (Ph.Eur.) 2.9.3, the following requirements are met using a dissolution apparatus according to Method 2 (paddle method) of the test:

[0752] (a) the composition meets 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 meets the requirement that, 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 30 minutes; and specification page 56 / 59 59 CN 120754112 A

[0754] (c) the composition meets 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;

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

[0756] 2. Use of a composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide upon administration to the gastrointestinal tract, the composition satisfying the following requirements:

[0757] (i) in standard in vitro USP <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus according to method 2 (paddle method) of the test is 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, 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 30 minutes; 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 and one or more pharmaceutically acceptable excipients that provide modulated release of the budesonide after administration to the gastrointestinal tract, the composition satisfying the following requirements:

[0763] (i) in standard in vitro USP <711> / In the dissolution test according to European Pharmacopoeia 2.9.3, a dissolution apparatus according to method 2 (paddle method) of the test is 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 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 30 minutes; and

[0766] (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;

[0767] It is used to treat IgA nephropathy.

[0768] 4. A composition comprising budesonide and one or more pharmaceutically acceptable excipients that provide modulated release of said budesonide upon administration to the gastrointestinal tract, said composition being tested in standard in vitro USP <711> In the dissolution test according to the European Pharmacopoeia 2.9.3, the dissolution apparatus used according to the second method (paddle method) of the test shall meet the following requirements:

[0769] (a) the composition shall meet 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 shall be released into the dissolution medium within about 120 minutes;

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

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

[0772] 5. The method according to claim 1, the use according to claim 2, the composition used according to claim 3, or the composition according to claim 4, wherein the composition comprises one or more cores containing budesonide, the cores being coated with a pharmaceutically acceptable extended-release polymeric blend comprising a water-insoluble polymer and a porous polymer.

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

[0774] 7. The method, use, composition, or composition according to claim 5 or 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, or composition according to claim 6 or 7, wherein the extended-release polymer blend coating and the delayed-release coating allow for substantial prevention of release of the contents of the composition before reaching the ileal region of the small intestine.

[0776] 9. The method, use, composition, 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 coating, and the amount of the porogenic polymer is from about 35% to about 5% by weight of the total extended release coating, such as the amount of the water-insoluble polymer being from about 45% to about 65% by weight of the total extended release coating, and the amount of the porogenic polymer being from about 35% to about 15% by weight of the total extended release coating, for example, wherein the amount of the water-insoluble polymer is from about 47% to about 55% by weight of the total extended release coating, and the amount of the porogenic polymer is from about 32% to about 22% by weight.

[0777] 10. The method, use, composition, or composition according to any one of claims 5 to 9, wherein the porogenic polymer is water-soluble.

[0778] 11. The method, use, composition, or composition of any one of claims 5 to 10, wherein the extended-release polymer blend coating the one or more cores is polymerizable.

[0779] 12. The method, use, composition, or composition of any one of claims 5 to 11, wherein the extended-release polymer blend coating the one or more cores comprises one or more polymerizable polymers.

[0780] 13. The method, use, composition, or composition of claim 11 or 12, wherein the one or more polymerizable polymers comprises the water-insoluble polymer.

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

[0782] 15. The method, use, composition or composition used according to any one of claims 5 to 14, wherein the extended release polymer blend comprises a porous polymer in an amount between about 35% by weight and about 15% by weight, for exampleAbout 32% by weight to about 22% by weight.

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

[0784] 17. The method, use, composition or composition used according to any one of items 5 to 16, wherein the porous 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, such as 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, such as about 2 to about 7 mPa*s, preferably about 2 to about 6 mPa*s.

[0785] 18. The method, use, composition or composition used according to any one of items 5 to 17, wherein the porous polymer has a gelation 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, or composition according to any one of items 5 to 18, wherein the porogenic polymer comprises hydroxypropylmethylcellulose (HPMC).

[0787] 20. The method, use, composition, or composition according to item 19, wherein the HPMC is methoxylated to a degree of about 15 to about 35% by weight, for example about 25 to about 35% by weight, or about 27 to about 31% by weight, such as about 27 to about 30% by weight.

[0788] 21. The method, use, composition, or composition according to item 19 or item 20, wherein the HPMC is hydroxypropoxylated to a degree of about 4 to about 32% by weight, for example about 4 to about 20% by weight, or about 5 to about 15% by weight, such as about 7 to about 12% by weight.

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

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

[0791] 24. The method according to claim 23 (when subordinate to any one of 11 to 22), wherein the coalescence of the polymeric material is further carried out in the fluidized bed apparatus.

[0792] 25. A composition which can be obtained by the method according to claim 23 or claim 24.

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

[0794] 27. The method, use, composition, or composition according to item 26, wherein the enteric coating comprises a polymer selected from the list of the following: azo polymers, disulfide polymers, cellulose acetate, cellulose succinate, cellulose acetate phthalate, cellulose tetrahydrophthalate, polyvinyl acetate phthalate, hydroxyethyl ethyl cellulose phthalate, methacrylic acid copolymer, polymethacrylic acid / acrylic acid copolymer, styrene-maleic acid copolymer, hydroxypropyl methyl cellulose phthalate, acrylic resin, cellulose benzotriester acetate, hydroxypropyl methyl cellulose benzotriester, shellac, hydroxyethyl ethyl cellulose phthalate, carboxymethyl cellulose, and hydroxypropyl methyl cellulose acetate-succinate.

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

[0796] 29. The method, use, composition, or composition of claim 28, wherein the one or more biomarkers are serum B cell activating factors, or immunoglobulins or immunoglobulin complexes associated with the pathogenesis of IgA nephropathy.

[0797] 30. The method, use, composition, or composition of claim 28, wherein the reduction in the baseline serum level is at least 5%, such as at least 10%. Instruction manual page 59 / 59, page 62, CN 120754112 A, Figure 1; Instruction manual figure 1 / 25, page 63, CN 120754112 A, Figure 2; Instruction manual figure 2 / 25, page 64, CN 120754112 A, Figure 3; Instruction manual figure 3 / 25, page 65, CN 120754112 A, Figure 4; Instruction manual figure 4 / 25, page 66, CN 120754112 A, Figure 5; Instruction manual figure 5 / 25, page 67, CN 120754112 A, Figure 6; Instruction manual figure 6 / 25, page 68, CN 120754112 A, Figure 7; Instruction manual figure 7 / 25, page 69, CN 120754112 A, Figure 8; Instruction manual figure 8 / 25, page 70, CN 120754112 A, Figure 9; Instruction manual figure 9 / 25, page 71, CN 120754112 A, Figure 10. Instruction manual, page 10 / 2572 CN 120754112 A Figure 11, Appendix to the Specification, Page 11 / 25 73 CN 120754112 A Figure 12, Appendix to the Specification, Page 12 / 25 74 CN 120754112 A Figure 13, Appendix to the Specification, Page 13 / 25 75 CN 120754112 A Figure 14, Appendix to the Specification, Page 14 / 25 76 CN 120754112 A Figure 15, Appendix to the Specification, Page 15 / 25 77 CN 120754112 A Figure 16, Appendix to the Specification, Page 16 / 25 78 CN 120754112 A Figure 17, Appendix to the Specification, Page 17 / 25 79 CN 120754112 A Figure 18, Appendix to the Specification, Page 18 / 25 80 CN 120754112 A Figure 19, Appendix to the Specification, Page 19 / 25 81 CN 120754112 A Figure 20 Appendix to the Instruction Manual, Page 20 / 25, 82 CN 120754112 A Figure 21 Appendix to the Instruction Manual, Page 21 / 25, 83 CN 120754112 A Figure 22 Appendix to the Instruction Manual, Page 22 / 25, 84 CN 120754112 A Figure 23a Appendix to the Instruction Manual, Page 23 / 25, 85 CN 120754112 A Figure 23b Appendix to the Instruction Manual, Page 24 / 25, 86 CN 120754112 A Figure 24 Appendix to the Instruction Manual, Page 25 / 25, 87 CN 120754112 A Abstract The present invention provides for a method of treatment of IgA nephropathy, which method comprises: (i) Identifying a pharmaceutically acceptable composition intended to treat IgA nephropathy comprising budesonide and one or more pharmaceutically-acceptable excipients that provide for a modified release of said budesonide after administration to the gastrointestinaltract, which composition fulfils the following requirements in the standard in vitro dissolution test; (a) the composition fulfils 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 fulfils 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 fulfils 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 said composition to a patient with IgA nephropathy in need of said treatment.

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.

10. The method, use, composition for use or composition according to any one of claims 5 to 9, wherein the extended release polymeric blend coating the one or more cores comprises one or more coalescible polymers.

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