Combination of endothelin receptor antagonists and glucocorticoids for treatment of IgA nephropathy

Abstract

A method of treating IgA nephropathy and a pharmaceutical composition comprising an endothelin receptor antagonist and a glucocorticoid.

Description

Combination of an endothelin receptor antagonist and a glucocorticoid for the treatment of IgA nephropathy Technical Field

[0001] The present invention relates to a pharmaceutical composition of an endothelin receptor antagonist and a glucocorticoid for treating IgA nephropathy, and uses of the endothelin receptor antagonist and the glucocorticoid in preparing a pharmaceutical composition or a medicine kit for treating IgA nephropathy. Background Art

[0002] IgA nephropathy (IgAN) is the most common primary glomerulonephritis worldwide and a major cause of chronic kidney disease and end-stage renal disease (ESRD). Within 10 years of onset, IgA nephropathy leads to end-stage renal disease (ESRD) in 15% to 20% of patients, and within 20 years, in 30% to 40% of patients. my country is a high-incidence region for IgAN. Its pathological hallmark is the deposition of circulating immune complexes composed of galactose-deficient IgA1 and glycan-specific IgG antibodies, which trigger glomerular mesangial cell activation and an inflammatory response, ultimately leading to renal tissue cell damage. IgG⁃Gd⁃IgA1 is deposited in the glomerular mesangial area, stimulating an inflammatory cascade, including local secretion of chemokines, cytokines, reactive oxygen species, and complement activation, leading to glomerular mesangial cell proliferation, inflammatory cell recruitment, crescent formation, and damage to proximal tubular epithelial cells, leading to tubular interstitial fibrosis. Patients present with a range of symptoms, usually including microscopic hematuria or gross hematuria and proteinuria. Due to persistent renal damage, patients may also develop hypertension. A considerable number of patients will develop end-stage renal disease (ESRD) within 10-20 years of being diagnosed with IgAN. During this period, in addition to decreased renal function, patients will also experience a variety of symptoms that will greatly reduce their quality of life.

[0003] As recorded in patent document WO2021126977A1, Atrasentan is a selective endothelin A receptor (ETA) small molecule antagonist, and research on its use in the treatment of immunoglobulin A nephropathy (IgAN) is being conducted by Chinook.

[0004] Although the introduction of alternative clinical endpoints has greatly accelerated the pace of research and development of innovative drugs, the selection of research endpoints for kidney disease is more difficult. It takes an average of 20 years for IgAN to develop into uremia. Observing the effectiveness of a drug requires a large sample size and a 20-year follow-up, which is obviously unrealistic. The emergence of alternative endpoints can solve this problem to a certain extent. The KDIGO guidelines recommend that reducing proteinuria to below 1g / d is a surrogate marker for improved renal outcomes in IgAN. Reducing proteinuria to below 1g / d is a reasonable treatment goal. Proteinuria levels are also often used as a surrogate endpoint in clinical studies and have been widely used in the design of new drug studies for the treatment of IgA nephropathy. In December 2021, the FDA approved Calliditas Therapeutics' new drug TARPEYO® (budesonide) for the treatment of IgA nephropathy for the first time based on the alternative clinical endpoint of decreased proteinuria. Summary of the Invention

[0005] The purpose of the present invention is to overcome the limitations of drug therapy and provide a combined therapy for treating IgA nephropathy with better effects, as well as corresponding combined drugs and drug kits.

[0006] In order to solve the technical problems of the present invention, the present invention is implemented by the following technical solutions:

[0007] A method of treating IgA nephropathy in a subject in need thereof comprises administering to the subject effective amounts of an endothelin receptor antagonist and a glucocorticoid, wherein the endothelin receptor antagonist and glucocorticoid can be administered as a single dosage form, separately, or sequentially.

[0008] In a preferred technical solution of the present invention, the endothelin receptor antagonist is selected from any one or any combination of tezosentan, sparsentan, bosentan, macitentan, ambrisentan, sitaxsentan, aprotentan and atrasentan or pharmaceutically acceptable salts thereof.

[0009] In a preferred technical solution of the present invention, the endothelin receptor antagonist is selected from atrasentan or a pharmaceutically acceptable salt thereof.

[0010] In a preferred technical solution of the present invention, the glucocorticoid is selected from any one of dexamethasone, betamethasone, fluorometholone, prednisone, prednisolone, methylprednisolone, hydrocortisone, fluocinolone acetonide, fluticasone, mometasone, loteprednol etabonate, ramesolone, fluticasone, beclomethasone, ciclesonide, budesonide, triamcinolone acetonide, prednicarbate, buticort, tipredan, tixocortol or pharmaceutically acceptable salts thereof, or any combination thereof.

[0011] In a preferred technical solution of the present invention, the glucocorticoid is selected from budesonide, methylprednisolone or pharmaceutically acceptable salts thereof.

[0012] In a preferred technical embodiment of the present invention, the endothelin receptor antagonist is administered in a dosage form for oral, buccal or extra-digestive tract administration, for example, the oral dosage form may be tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the dosage form for the extra-digestive tract administration may be, for example, a dosage form for intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal penetration, or transmucosal administration.

[0013] In a preferred technical solution of the present invention, the endothelin receptor antagonist is administered once or twice a day; or once every 2, 3, 4, 5, 6, 7, 8, 9, 10 days or every 1, 2 or 3 weeks; or once a day for 5 consecutive days a week, followed by a 2-day interval.

[0014] In a preferred embodiment of the present invention, the endothelin receptor antagonist is administered to the subject at an effective dose of about 0.5 to about 250 mg / kg, 1 to about 250 mg / kg, about 2 to about 200 mg / kg, about 3 to about 120 mg / kg, about 5 to about 250 mg / kg, about 10 to about 200 mg / kg, or about 20 to about 120 mg / kg.

[0015] In a preferred technical solution of the present invention, the glucocorticoid is administered in a dosage form for oral, buccal or extra-digestive tract administration, for example, the oral dosage form may be tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the dosage form for the extra-digestive tract administration may be, for example, a dosage form for intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal penetration, or transmucosal administration.

[0016] In a preferred technical solution of the present invention, the glucocorticoid is administered once or twice a day; or once every 2, 3, 4, 5, 6, 7, 8, 9, 10 days or every 1, 2 or 3 weeks; or once a day for 5 consecutive days per week, followed by a 2-day interval.

[0017] In preferred embodiments of the present invention, the glucocorticoid is administered to the subject at an effective dose of about 0.5 to about 250 mg / kg, 1 to about 250 mg / kg, about 2 to about 200 mg / kg, about 3 to about 120 mg / kg, about 5 to about 250 mg / kg, about 10 to about 200 mg / kg, or about 20 to about 120 mg / kg.

[0018] In a preferred embodiment of the present invention, the method may further optionally comprise administering to the subject an effective amount of a third therapeutically active substance.

[0019] In a preferred technical solution of the present invention, the third therapeutically active substance is selected from SGLT-2 inhibitors.

[0020] In a preferred technical solution of the present invention, the SGLT-2 inhibitor is selected from any one of empagliflozin, canagliflozin, repagliflozin, ispagliflozin, HM41322, dapagliflozin, bepagliflozin, erpagliflozin, sogliaflozin, rupagliflozin and togliflozin, or any combination thereof.

[0021] In a preferred technical embodiment of the present invention, the SGLT-2 inhibitor is administered in a dosage form for oral, buccal or extra-digestive tract administration, for example, the oral dosage form may be tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the dosage form for the extra-digestive tract administration may be, for example, a dosage form for intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal penetration, or transmucosal administration.

[0022] In a preferred technical solution of the present invention, the SGLT-2 inhibitor is administered once or twice a day; or once every 2, 3, 4, 5, 6, 7, 8, 9, 10 days or every 1, 2 or 3 weeks; or once a day for 5 consecutive days a week, followed by a 2-day interval.

[0023] In a preferred embodiment of the present invention, the SGLT-2 inhibitor is administered to the subject at an effective dose of about 0.5 to about 250 mg / kg, 1 to about 250 mg / kg, about 2 to about 200 mg / kg, about 3 to about 120 mg / kg, about 5 to about 250 mg / kg, about 10 to about 200 mg / kg, or about 20 to about 120 mg / kg.

[0024] In a preferred technical solution of the present invention, the third therapeutically active substance is selected from APRIL neutralizing antibodies.

[0025] In a preferred technical solution of the present invention, the APRIL neutralizing antibody is BION-1301 monoclonal antibody.

[0026] In a preferred technical solution of the present invention, the APRIL neutralizing antibody is formulated into a solution or a lyophilized agent.

[0027] In a preferred technical solution of the present invention, the APRIL neutralizing antibody is administered as an intravenous injection at a pH of 5-8, preferably 5.5; the concentration of the buffer solution is 1-50 mM, preferably 5-40 mM, more preferably 10-20 mM, wherein the buffer solution is preferably a histidine buffer solution; wherein the concentration of the APRIL neutralizing antibody is 10-60 mg / mL, preferably 20-50 mg / mL, more preferably 25 mg / mL; wherein the concentration of sucrose is 1-10% (w / v), preferably 2-8% (w / v); and the concentration of polysorbate 80 is 0.01%-1% (w / v), preferably 0.01%-1% (w / v).

[0028] In a preferred embodiment of the present invention, the APRIL neutralizing antibody is administered once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days or every 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks, or the endothelin receptor antagonist is administered once a day for 5 consecutive days per week, followed by a 2-day interval.

[0029] In a preferred embodiment of the present invention, the APRIL neutralizing antibody is administered at an effective dose of 0.05 mg / kg, 0.1 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg or 8 mg / kg.

[0030] In addition, the present invention also provides a pharmaceutical composition comprising an effective amount of an endothelin receptor antagonist and a glucocorticoid, wherein the endothelin receptor antagonist and the glucocorticoid can be administered as a single dosage form, separately or sequentially.

[0031] In a preferred embodiment of the present invention, the endothelin receptor antagonist can be formulated into a dosage form for oral, buccal or extra-digestive administration, for example, the oral dosage form can be tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the dosage form for extra-digestive administration can be, for example, a dosage form for intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal penetration, or transmucosal administration, for example, a solution or a lyophilized agent; and / or

[0032] The glucocorticoid can be formulated into a dosage form for administration by an extra-digestive route, for example, the extra-digestive route can be intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal permeation, or transmucosal administration, for example, it can be a solution or a lyophilized agent.

[0033] In a preferred technical solution of the present invention, the endothelin receptor antagonist is selected from any one or any combination of tezosentan, sparsentan, bosentan, macitentan, ambrisentan, sitaxsentan, aprotentan and atrasentan or pharmaceutically acceptable salts thereof.

[0034] In a preferred technical solution of the present invention, the endothelin receptor antagonist is selected from atrasentan or a pharmaceutically acceptable salt thereof.

[0035] In a preferred technical solution of the present invention, the glucocorticoid is selected from any one of dexamethasone, betamethasone, fluorometholone, prednisone, prednisolone, methylprednisolone, hydrocortisone, fluocinolone acetonide, fluticasone, mometasone, loteprednol etabonate, ramesolone, fluticasone, beclomethasone, ciclesonide, budesonide, triamcinolone acetonide, prednicarbate, buticort, tipredan, tixocortol or pharmaceutically acceptable salts thereof, or any combination thereof.

[0036] In a preferred technical solution of the present invention, the glucocorticoid is selected from budesonide, methylprednisolone or pharmaceutically acceptable salts thereof.

[0037] In a preferred embodiment of the present invention, each unit dosage form contains an endothelin receptor antagonist at a dose of 1-1000 mg, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90, 100, 110, 120, 125, 130, 140, 150, 160, 170, 175, 180, 190, 200, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 900, 1000 mg or a value between any two of the above values.

[0038] In a preferred embodiment of the present invention, each unit dosage form contains 1-5000 mg of glucocorticoid, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90, 100, 110, 120, 125, 130, 140, 150, 160, 170, 175, 180, 190, 200 , 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2200, 2400, 2500, 2600, 2700, 2750, 2800, 3000, 3500, 4000, 4500, 5000 mg or a value between any two of the above values.

[0039] In a preferred technical solution of the present invention, the pharmaceutical composition may optionally further comprise an effective amount of a third therapeutically active substance.

[0040] In a preferred technical solution of the present invention, the third therapeutically active substance is selected from SGLT-2 inhibitors.

[0041] In a preferred technical solution of the present invention, the SGLT-2 inhibitor is selected from any one of empagliflozin, canagliflozin, repagliflozin, ispagliflozin, HM41322, dapagliflozin, bepagliflozin, erpagliflozin, sogliaflozin, rupagliflozin and togliflozin, or any combination thereof.

[0042] In a preferred technical solution of the present invention, the SGLT-2 inhibitor is selected from dapagliflozin.

[0043] In a preferred embodiment of the present invention, each unit dosage form contains a dose of 1-1000 mg of SGLT-2 inhibitor, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90, 100, 110, 120, 125, 130, 140, 150, 160, 170, 175, 180, 190, 200, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 900, 1000 mg or a value between any two of the above values.

[0044] In a preferred technical solution of the present invention, the third therapeutically active substance is selected from APRIL neutralizing antibodies.

[0045] In a preferred technical solution of the present invention, the APRIL neutralizing antibody is BION-1301 monoclonal antibody.

[0046] In a preferred embodiment of the present invention, each unit dosage form comprises 1-5000 mg of APRIL neutralizing antibody, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90, 100, 110, 120, 125, 130, 140, 150, 160, 170, 175, 180, 190, 200, 250, 300, 350, 400, 500, 600, 700, 750, 800, 900, 1000, 1100, 1200, 1250, 1300, 1400, 1500, 1600, 1700, 1750, 1800, 1900, 2000, 2500, 3000, 3500 00, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2200, 2400, 2500, 2600, 2700, 2750, 2800, 3000, 3500, 4000, 4500, 5000 mg or a value between any two of the above values.

[0047] In a preferred technical solution of the present invention, the APRIL neutralizing antibody is administered as an intravenous injection at a pH of 5-8, preferably 5.5; the concentration of the buffer solution is 1-50 mM, preferably 5-40 mM, more preferably 10-20 mM, wherein the buffer solution is preferably a histidine buffer solution; wherein the concentration of the APRIL neutralizing antibody is 10-60 mg / mL, preferably 20-50 mg / mL, more preferably 25 mg / mL; wherein the concentration of sucrose is 1-10% (w / v), preferably 2-8% (w / v); and the concentration of polysorbate 80 is 0.01%-1% (w / v), preferably 0.01%-1% (w / v).

[0048] In addition, the present invention also provides a medicine box, which comprises any of the above-mentioned pharmaceutical compositions of the present invention and instructions for use. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1. Changes in glomeruli before and after establishment of IgAN model in mice;

[0050] Figure 2 Changes in hIgA1, IgG, and IgM in mice before and after IgAN modeling;

[0051] Figure 3-1, Figure 3-2 Comparison of changes in hlgA1 indices in IgAN mice after administration of blank control group (Ctrl), atrasentan (A), methylprednisolone (MP), combined administration of atrasentan (A) + methylprednisolone (MP), and combined administration of atrasentan (A) + methylprednisolone (MP) + dapagliflozin (Dapa);

[0052] Figure 4-1, Figure 4-2 Comparison of changes in IgG indicators in IgAN mice after blank control group (Ctrl), administration of atrasentan (A), administration of methylprednisolone (MP), combined administration of atrasentan (A) + methylprednisolone (MP), and combined administration of atrasentan (A) + methylprednisolone (MP) + dapagliflozin (Dapa);

[0053] Figure 5-1 and Figure 5-2 Comparison of changes in IgM indices in IgAN mice after blank control group (Ctrl), administration of atrasentan (A), administration of methylprednisolone (MP), combined administration of atrasentan (A) + methylprednisolone (MP), and combined administration of atrasentan (A) + methylprednisolone (MP) + dapagliflozin (Dapa). DETAILED DESCRIPTION

[0054] Definition of terms:

[0055] Unless otherwise indicated herein, terms used herein have their ordinary meaning in the art to which they belong.

[0056] "Treatment," "treatment," and "treatment" refer to alleviating, inhibiting, and / or reversing the progression of a disease in a subject in need thereof. The term "treatment" includes any sign of successful treatment or improvement of a disease, including any objective or subjective parameter, such as alleviation; alleviation; reduction of symptoms or making the injury, pathology, or condition more tolerable to the subject; delaying or slowing the rate of progression, etc. Measurement of treatment or improvement can be based on, for example, the results of physical examination, pathological examination, and / or diagnostic examinations known in the art. For example, in one embodiment, "treating" IgA nephropathy as contemplated by the present invention refers to the combined treatment of a subject with IgA nephropathy with an endothelin receptor antagonist and a glucocorticoid to achieve at least one positive therapeutic outcome.

[0057] Treatment can also mean reducing the risk of developing or experiencing a disease, or reducing the risk of recurrence of a disease (e.g., delaying the onset of a disease) compared to what would occur without the intervention. In medicine, this type of treatment is also known as "prevention."

[0058] The term "effective amount" or "therapeutically effective amount" refers to an amount that is effective in treating a disease, as documented by clinical testing and evaluation, patient observation, and the like. An "effective amount" may further refer to an amount that causes a detectable change in biological or chemical activity. The detectable change can be detected and / or further quantified by a person skilled in the art who is familiar with the relevant mechanism or method. In addition, an "effective amount" may refer to an amount that maintains a desired physiological state (i.e., reduces or prevents significant decline and / or promotes improvement of a condition). An "effective amount" may further refer to a therapeutically effective amount.

[0059] In some embodiments, the endothelin receptor antagonist is administered to a subject in an effective amount. An effective amount is typically 0.01 mg / kg to 500 mg / kg body weight per day. In some embodiments, the pharmaceutically acceptable compositions may be formulated so that a dose of 0.01 mg / kg to 200 mg / kg body weight or 0.01 mg / kg to 100 mg / kg body weight of the compound per day (e.g., based on a 75 kg person, a dose of 0.75 mg to 7.5 g or 15 g) can be administered to a subject receiving these compositions. In certain embodiments, the active pharmaceutical ingredient of the present invention is formulated to provide a dose of 0.01 mg / kg to 70 mg / kg (e.g., based on a 75 kg person, a dose of 0.75 mg to 5.25 g).

[0060] In some embodiments, an effective dose of an endothelin receptor antagonist is about 0.5 to about 250 mg / kg, 1 to about 250 mg / kg, about 2 to about 200 mg / kg, about 3 to about 120 mg / kg, about 5 to about 250 mg / kg, about 10 to about 200 mg / kg, or about 20 to about 120 mg / kg. In some embodiments, an effective dose includes about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 8 mg / kg, 10 mg / kg, 20 mg / kg, 25 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 75 mg / kg, 100 mg / kg, 120 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg, 225 mg / kg, 250 mg / kg, and 300 mg / kg. The dosage form can take various suitable forms, such as tablets or capsules, and the effective dose can be provided in one or more unit dosage forms (such as tablets, capsules) and provided 1, 2 or 3 times a day, or provided throughout the day at intervals of, for example, 4, 8 or 12 hours. Tablets or capsules can, for example, contain 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100 or 1,250 mg of compound. For example, in some embodiments, the administration of an endothelin receptor antagonist to a human subject can include a daily dose of an endothelin receptor antagonist in the range of 100-1,250, 150-1,000, 200-800 or 250-750 mg, which can be administered all at once a day, or divided into multiple portions administered throughout the day at certain intervals. It can also be prepared as a liquid formulation so that any dose can be dispensed easily and conveniently.

[0061] The antibody will typically be mixed with a pharmaceutically acceptable, non-toxic carrier substance (e.g., physiological saline or phosphate-buffered saline) prior to administration, and may be administered using any medically appropriate procedure including, but not limited to, intravenous or intraarterial administration and injection into the cerebrospinal fluid.

[0062] In some embodiments, the effective dose of the antibody is from about 5 to about 250 mg / kg, from about 10 to about 200 mg / kg, or from about 20 to about 120 mg / kg. In some embodiments, the effective dose includes 5 mg / kg, 10 mg / kg, 20 mg / kg, 25 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 75 mg / kg, 100 mg / kg, 120 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg, 225 mg / kg, 250 mg / kg, and 300 mg / kg. The dosage form can be in the form of, for example, a tablet or capsule, and the effective dose can be provided in one or more tablets, capsules, etc. and provided once a day or throughout the day at intervals of, for example, 4, 8, or 12 hours. Tablets or capsules can contain, for example, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1,000 mg of antibody. Liquid formulations can also be prepared so that any dose can be easily and conveniently dispensed.

[0063] In some embodiments, the antibody is administered to the study subject in an effective amount. An effective amount is typically 0.01mg / kg body weight to 500mg / kg body weight every day. In some embodiments, pharmaceutically acceptable compositions can be formulated so that the patient receiving these compositions can be administered 0.01mg / kg body weight to 200mg / kg body weight or 0.01mg / kg body weight to 100mg / kg body weight of the compound. In certain embodiments, the compositions of the present invention are formulated to provide a dosage of 0.01mg / kg to 70mg / kg (e.g., based on a 75kg person, a dosage of 0.75mg to 5.25g).

[0064] An effective amount of the antibody can be, for example, 0.05 mg / kg, 0.1 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg or 8 mg / kg per dose (e.g., based on a 75 kg human, a dose of 3.75 mg to 600 mg).

[0065] Doses of the antibody can be administered once, twice, three times, four times, five times or more per week, weekly, every two weeks, or even every three weeks during the course of treatment. Dosing schedules can be daily, every two days, every three days, every four days, every five days, weekly, every two weeks, or every three weeks. Formulations containing the antibody can be prepared so that any dosage can be easily and conveniently dispensed.

[0066] The term "subject" refers to a mammalian subject, and in particular a human subject, including male or female subjects, and including newborns, infants, young children, adolescents, adults or elderly subjects, and further includes various races and ethnicities, such as Caucasians, Africans and Asians. Herein, the subject suffers from IgA nephropathy.

[0067] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic acid salt of a compound. These salts can be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in free form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative acid salts include, but are not limited to, acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hyphenate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinafoate. In one embodiment, the pharmaceutically acceptable salt is a hydrochloride / chloride salt.

[0068] Herein, the terms "pharmaceutically acceptable" and "pharmaceutically acceptable" are used interchangeably and refer to the types generally accepted by those skilled in the pharmaceutical field, such as pharmaceutically acceptable salts, pharmaceutically acceptable carriers, etc.

[0069] "Oral dosage form" refers to a pharmaceutical preparation prepared for oral administration to a subject. Examples of known oral dosage forms include, but are not limited to, tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the like. In some methods, powders, pills, granules, capsules, and tablets may be coated with a suitable polymer or conventional coating material to achieve, for example, greater stability in the gastrointestinal tract or to achieve a desired release rate. In addition, the capsule shell of the powder, pill, or granule may be further coated. Tablets may be scored to facilitate split administration.

[0070] When the endothelin receptor antagonist or glucocorticoid of the present invention is administered as an oral formulation, it is preferably coated with a film. Suitable films are known in the art and can be purchased from the market or can be manufactured according to known methods. Typically, the film coating material is a hydrophilic polymer, such as polyethylene glycol, polyvinyl pyrrolidone, polyvinyl alcohol, hydroxypropyl cellulose, hydroxymethyl cellulose and hydroxypropyl methyl cellulose. The film coating composition ingredients may include a commonly used amount of plasticizer, such as polyethylene glycol, triethyl citrate, diethyl phthalate, propylene glycol, glycerol; and opacifiers, such as titanium dioxide and colorants, such as iron oxide, aluminum lake, etc. Typically, the film coating material is applied in an amount that can provide a film coating in the range of 1% to 6% of the solid oral dosage form.

[0071] When the endothelin receptor antagonist of the present invention is administered as an oral formulation, it may optionally further comprise at least one pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" refers to any pharmaceutically inert material that is substantially biologically inactive and constitutes a substantial portion of the formulation. Examples of such carriers include, but are not limited to, diluents and fillers, disintegrants, glidants, binders, stabilizers, colorants, flavor enhancers, and preservatives.

[0072] As diluents, for example, but not limited to, microcrystalline cellulose, mannitol, powdered sugar, compressible sugar, dextran, dextrin, dextrose, lactose, cellulose powder, sorbitol, sucrose, talc, or a combination thereof may be selected. The diluent may comprise 5% to 90% of the total weight of the oral formulation, preferably 10% to 80%, 20% to 70%, 30% to 60%, or 40% to 50%.

[0073] As disintegrants, for example, but not limited to, cellulose, alginate, gum, cross-linked polymers such as cross-linked polyvinyl pyrrolidone or crospovidone, cross-linked sodium carboxymethyl cellulose, cross-linked calcium carboxymethyl cellulose, soy polysaccharide, sodium starch glycolate, guar gum or any combination thereof may be selected.

[0074] The diluent may be present in an amount of about 1% to 15%, preferably 2% to 10%, based on the total weight of the oral preparation.

[0075] As binders, for example, but not limited to, starch, cellulose or derivatives thereof, such as microcrystalline cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose and hydroxypropyl methyl cellulose, sucrose, dextrose, corn syrup, polysaccharides, gelatin or any combination thereof may be selected. The binder may be present in an amount of 0.01 to 10%, preferably 1% to 10%, based on the total weight of the formulation.

[0076] As the glidant, for example, but not limited to, colloidal silicon dioxide, magnesium trisilicate, cellulose powder, talc or a combination thereof can be selected. The glidant can be present in an amount of 0.1% to 10%, preferably 0.1% to 0.5%, based on the total weight of the composition.

[0077] Unless otherwise indicated, the terms "comprising" and "including" as used herein are open-ended and non-restrictive. For example, in addition to the pharmaceutical combination formed by the endothelin receptor antagonist and the APRIL neutralizing antibody of the present invention, other therapeutic agents, such as "biological therapeutic agents" and "chemotherapeutic agents" may also be included.

[0078] The choice of a dosage regimen for the combination therapy of the present invention depends on a number of factors, such as the entity serum or tissue turnover rate, symptom level, overall immunogenicity, and accessibility of target cells, tissues, or organs in the individual being treated. Preferably, the dosage regimen maximizes the amount of each therapeutic agent delivered to the patient while maintaining an acceptable level of side effects. Thus, the dosage and frequency of each biotherapeutic and chemotherapeutic agent in the combination therapy will depend, in part, on the specific therapeutic agent, the severity of the IgA nephropathy being treated, and the characteristics of the patient.

[0079] Continuous administration is particularly useful when the therapeutic agents in the combination therapy are administered in different dosage forms (e.g., one active agent is administered orally and the other active agent is a sterile solution) and / or on different dosage schedules (e.g., the endothelin receptor antagonist is the active agent and is administered at least daily, while the APRIL neutralizing antibody is administered less frequently, e.g., once a week, once every two weeks, or once every three weeks).

[0080] WO2016110587A1 describes the sequence and preparation of the BION-1301 monoclonal antibody, which is incorporated herein by reference in its entirety. The heavy chain full-length amino acid sequence, heavy chain variable region sequence, and CDR sequence of BION-1301 correspond to SEQ ID NOs: 52, 40, 5, 6, and 7, respectively, and the light chain full-length amino acid sequence, light chain variable region sequence, and CDR sequence correspond to SEQ ID NOs: 50, 30, 8, 9, and 10, respectively, in that patent application.

[0081] The present invention is further illustrated by the following examples, which are not intended to limit the scope of protection of the present invention. For the determination methods of the various parameters in the examples, please refer to Wan F, Wang H, Wang M, Lv J, Zhao M, Zhang H. Sustained release of Lactobacillus casei cell wall extract can induce a continuous and stable IgA deposition model. J Pathol. 2022;257(3):262-273. doi:10.1002 / path.5884; Xie X, Li J, Liu P, et al. Chimeric Fusion between Clostridium Ramosum IgA Protease and IgG Fc Provides Long-Lasting Clearance of IgA Deposits in Mouse Models of IgA Nephropathy. J Am Soc Nephrol. 2022;33(5):918-935. doi:10.1681 / ASN.2021030372. The above documents are incorporated into this application in their entirety.

[0082] Example 1 Construction of IgAN Animal Model

[0083] (a) Construction of a humanized IGHA1 mouse model;

[0084] (b) Construction of a renal IgA deposition model with high serum IgA expression. The mouse model described in this invention effectively expresses human IgA1 protein, ensuring the functionality of human IgA1 molecules in mice. This model is more specific than existing conventional construction methods and more closely resembles the natural properties of human IgA1. Furthermore, the CFA stimulation model is simple to operate, has a high success rate, is reproducible, and minimizes damage to the body. This provides an effective animal experimental platform for exploring the pathogenesis of IgA nephropathy and new treatment strategies.

[0085] Specific steps:

[0086] 1. Construction of a Humanized IGHA1 Mouse Model (For detailed methods, see patent document: CN115197942A, which is incorporated into this application in its entirety)

[0087] 1. The entire human IGHA1 gene sequence was introduced into the Igha gene region of C57BL / 6Jju mice, replacing the heavy chain region of mouse IgA with the heavy chain region of human IgA1, thereby constructing humanized IgA1 mice with an O-glycan-modified hinge region.

[0088] 2. After genetic sequencing and verification of the founder mice, homozygous humanized IGHA1 mice were obtained through multiple generations of backcrossing and self-crossing.

[0089] 2. Construction of a renal IgA deposition model

[0090] 1. Thoroughly mix complete Freund's adjuvant (CFA) and phosphate buffered saline (PBS) at a 1:1 ratio to produce an emulsified CFA injection. Intraperitoneally inject into 8-week-old humanized IGHA1 mice constructed in Step 1. The intraperitoneal injection volume is 4 times the mouse body weight (4 μl). Administer 6 injections every two weeks for a total of approximately 2.5 months.

[0091] 2. In this study, 8-week-old human IgA1 mice were given the last CFA injection and then underwent an induction period of 2-5 months.

[0092] Table 1

[0093]

[0094] The experimental results showed that the hIgA1, IgG, and IgM levels of the modeled IgAN mice were significantly higher than those of the unstimulated humanized IGHA1 transgenic mice (non-IgAN transgenic mice).

[0095] Example 2 Study on the efficacy and safety of atrasentan (A) combined with methylprednisolone (MP) and dapagliflozin (Dapa) in the treatment of IgA nephropathy

[0096] Methods: Forty-one IgA nephropathy model mice were randomly divided into an atrasentan + steroid + dapagliflozin group (n=9), an atrasentan + steroid group (n=8), a steroid group (n=8), an atrasentan group (n=8), and a treatment control group (n=8). Models were established in each group using intraperitoneal injection of CFA. After model establishment, atrasentan, steroids, and / or dapagliflozin were administered according to the following regimen. The control group received an equal volume of solvent (0.5% (w / v) methylcellulose, 0.5% (v / v) Tween 80 in water) by oral gavage for 30 days.

[0097] Dosage: Atrasentan (15 mg / kg / day), methylprednisolone (5 mg / kg / day), and dapagliflozin (1 mg / kg / day) were administered by oral gavage every morning for 30 days. For combination therapy, the required drugs (the dosage of each drug was calculated based on the mouse's body weight) were mixed together in proportion and administered by oral gavage.

[0098] Body weight and blood pressure of mice were measured before administration, once a week after administration, and 30 days after administration. Blood levels of hlgA1, Gd-IgA1, IgA-containing complexes, IgG, IgM, C3, BUN, Scr, Hb, BNP, and ET-1 were measured. Urine creatinine, urine protein, and albumin-to-creatinine ratio (ACR) were measured before administration, once every two weeks after administration, and 30 days after administration. The presence of hematuria was also detected. Renal tissue pathology was scored according to the Oxford classification before administration and 30 days after administration. IgA and C3 deposition in renal tissue were measured by immunofluorescence. Cardiac pathology in each group was also observed.

[0099] Characteristics of IgAN mouse model:

[0100] •IGHA1+ / + mice challenged with intraperitoneal CFA:

[0101] • Elevated levels of serum and intestinal mucus IgA1, Gd-IgA1, and IgA1 complexes

[0102] • IgAN renal pathological phenotype occurs:

[0103] -IgA1 and C3 co-deposited in the mesangial area, and the degree of complement activation was stronger than that of wild-type mice.

[0104] - Increased mesangial matrix, mesangial and endothelial cells, and some mice had more severe lesions, with focal sclerosis and tubulointerstitial fibrosis.

[0105] Analysis of the test results shows that, as shown in Table 2 and Figures 3-1 and 3-2, although there was no significant statistical difference in the changes of hIgA1 among the groups, compared with the treatment control group (hIgA1 showed an upward trend after 30 days), the hIgA1 in the MP alone group showed a downward trend after medication, and the downward trend of hIgA1 in the A+MP group after medication was more significant than that in the MP alone group, and the downward trend in the A+Dapa+MP group was even more obvious.

[0106] Table 2

[0107]

[0108] From the analysis of the test results, it can be seen from the data in Table 3 and Figures 4-1 and 4-2 that although there was no significant statistical difference in the changes of IgG among the groups, compared with the treatment control group (IgG showed an increasing trend after 30 days), the IgG of the MP alone group showed a decreasing trend after medication, and the decreasing trend of IgG in the A+MP group after medication was more significant than that of the MP alone group.

[0109] Table 3

[0110]

[0111] Analysis of the test results (Table 4 and Figures 5-1 and 5-2) showed that although there was no significant statistical difference in the changes in IgM among the groups, the downward trend of IgM after medication in the A+MP group was more significant compared with the MP alone group (IgM showed a slight downward trend after 30 days).

[0112] Table 4

[0113] . .

Claims

1. A method for treating IgA nephropathy in a subject in need thereof, comprising administering to the subject an effective amount of an endothelin receptor antagonist and a glucocorticoid, wherein the endothelin receptor antagonist and the glucocorticoid can be administered as a single dosage form, separately, or sequentially.

2. The method according to claim 1, wherein the endothelin receptor antagonist is selected from any one of tezosentan, sparsentan, bosentan, macitentan, ambrisentan, sitaxsentan, aprotentan and atrasentan or pharmaceutically acceptable salts thereof, or any combination thereof.

3. The method according to claim 1 or 2, wherein the endothelin receptor antagonist is selected from atrasentan or a pharmaceutically acceptable salt thereof.

4. The method according to claim 1, wherein the glucocorticoid is selected from any one of dexamethasone, betamethasone, fluorometholone, prednisone, prednisolone, methylprednisolone, hydrocortisone, fluocinolone acetonide, fluticasone, mometasone, loteprednol etabonate, ramesolone, fluticasone, beclomethasone, ciclesonide, budesonide, triamcinolone acetonide, prednicarbate, buticort, tipredan, tixocortol or pharmaceutically acceptable salts thereof, or any combination thereof.

5. The method of claim 4, wherein the glucocorticoid is selected from budesonide, methylprednisolone, or a pharmaceutically acceptable salt thereof.

6. The method according to claim 1, wherein the endothelin receptor antagonist is administered in a dosage form for oral, buccal or extra-digestive route, for example, the oral dosage form may be tablets, capsules, powders, pills, granules, suspensions, solutions and solution preconcentrates, emulsions and emulsion preconcentrates, and the dosage form for the extra-digestive route may be, for example, a dosage form for intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, transdermal penetration, or transmucosal administration.

7. The method according to claim 1, wherein the method further optionally comprises administering to the subject an effective amount of a third therapeutically active substance.

8. The method of claim 7, wherein the third therapeutically active substance is selected from SGLT-2 inhibitors.

9. The method according to claim 8, wherein the SGLT-2 inhibitor is selected from any one of empagliflozin, canagliflozin, repagliflozin, ispagliflozin, HM41322, dapagliflozin, bepagliflozin, erpagliflozin, sogliaflozin, rupagliflozin and togliflozin, or any combination thereof.

10. A pharmaceutical composition comprising an effective amount of an endothelin receptor antagonist and a glucocorticoid, wherein the endothelin receptor antagonist and the glucocorticoid can be administered as a single dosage form, separately or sequentially.

11. The pharmaceutical composition according to claim 10, wherein the endothelin receptor antagonist is selected from any one of tezosentan, sparsentan, bosentan, macitentan, ambrisentan, sitaxsentan, aprotentan and atrasentan or pharmaceutically acceptable salts thereof, or any combination thereof.

12. The pharmaceutical composition according to claim 10, wherein the endothelin receptor antagonist is selected from atrasentan or a pharmaceutically acceptable salt thereof.

13. The pharmaceutical composition according to claim 10, wherein the glucocorticoid is selected from any one of dexamethasone, betamethasone, fluorometholone, prednisone, prednisolone, methylprednisolone, hydrocortisone, fluocinolone acetonide, fluticasone, mometasone, loteprednol etabonate, ramesolone, fluticasone, beclomethasone, ciclesonide, budesonide, triamcinolone acetonide, prednicarbate, buticort, tipredan, tixocortol or pharmaceutically acceptable salts thereof, or any combination thereof.

14. The pharmaceutical composition according to claim 13, wherein the glucocorticoid is selected from budesonide, methylprednisolone or a pharmaceutically acceptable salt thereof.

15. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition may further optionally contain an effective amount of a third therapeutically active substance.

16. A pharmaceutical composition according to claim 15, wherein the third therapeutically active substance is selected from SGLT-2 inhibitors.

17. The pharmaceutical composition according to claim 16, wherein the SGLT-2 inhibitor is selected from any one of empagliflozin, canagliflozin, repagliflozin, ispagliflozin, HM41322, dapagliflozin, bepagliflozin, erpagliflozin, sogliflozin, rupagliflozin and topagliflozin, or any combination thereof.

18. The pharmaceutical composition according to claim 17, wherein the SGLT-2 inhibitor is selected from dapagliflozin.

Images