Method for treating membranous nephropathy with taci fc fusion protein

Abstract

This invention relates to drugs, dosage regimens, dosing intervals, and administration methods for treating membranous nephropathy using TACI-Fc fusion protein. Results show that the TACI-Fc fusion protein provided by this invention exhibits good clinical efficacy and safety in treating patients with membranous nephropathy.

Description

Technical Field

[0001] The present invention relates to a TACI-Fc fusion protein drug for treating membranous nephropathy, a dosage regimen, a dosing interval and an administration method. Prior Art

[0002] Membranous nephropathy (MN) is an antibody-mediated glomerular disease characterized by the formation of immunoprecipitates containing antigens, IgG, and complement components beneath the epithelial cells in the glomerular vascular loops. Sublethal damage to the epithelial cells leads to cellular simplification and destruction of the glomerular filtration barrier, resulting in proteinuria and other manifestations of nephrotic syndrome.

[0003] Clinically, MN can be divided into primary and secondary MN based on etiology. Most cases of MN are primary, also known as idiopathic MN, an organ-specific autoimmune disease that develops without any identifiable cause or initiating event. Approximately 20-30% of MN cases are secondary, with common causes including autoimmune diseases (such as lupus), viruses (hepatitis B, hepatitis C, syphilis), tumors, certain medications (such as painkillers), and toxins (such as exposure to heavy metals like lead and mercury). MN is one of the most common causes of nephrotic syndrome in non-diabetic adults, accounting for up to one-third of nephrotic syndrome biopsy findings. Its main clinical symptoms are edema, proteinuria, hypoalbuminemia, and hyperlipidemia. It primarily affects the function of the glomerular basement membrane and increases the risk of thrombosis, infection, and cardiovascular disease.

[0004] Currently, the global incidence of MN is approximately 1 case per 100,000 people per year. Whether the incidence varies by region or ethnicity is unclear, but it is known that MN affects patients of all ages and races. In the United States, the incidence of MN is approximately 12 cases per million people per year, with the average age of onset being between 50 and 60 years old. In China, the incidence of MN has been steadily increasing over the past 20 years, and other studies suggest that the high incidence may be related to increasing levels of air pollution. According to statistics, among patients with MN who remain untreated for a long time, nearly 60% experience renal impairment, and 30-40% will eventually progress to end-stage renal disease within 10 years. Even with immunosuppressant therapy, 20-40% of patients will experience persistent heavy proteinuria and progress to end-stage renal disease within 10 years, requiring renal replacement therapy or transplantation.

[0005] The treatment of membranous nephropathy depends on its type and cause. Currently, the treatment of membranous nephropathy mainly includes the following treatment methods: 1. Conventional treatment: If the 24-hour urine protein quantity of membranous nephropathy is less than 3.5g; or between 3.5g-8g, and the renal function is normal and there is no high-risk phenomenon, pril drugs and sartans are usually used to reduce urine protein, and supportive treatment such as diuresis, antihypertensive, and anticoagulant treatment is given; 2. Hormone combined with immunosuppressive drugs: If the 24-hour urine protein quantity of membranous nephropathy exceeds 3.5g and is accompanied by renal impairment, or the urine protein exceeds 8g, a combined hormone + immunosuppressant method is used for treatment; 3. Monoclonal antibody drugs: such as rituximab.

[0006] Table 1. Recommendations for treatment of membranous nephropathy in the 2021 KDIGO guidelines Cyclophosphamide (Periodic use) l In the first few days of the first, third, and fifth months, give 1 gram of methylprednisolone intravenously for three consecutive days. l Prednisone 0.5 mg / kg orally once a day. l In the second, fourth, and sixth months, the dose is changed to oral cyclophosphamide, 2.5 mg per kilogram of body weight per day. Cyclophosphamide (Continuous use) l In the first few days of the first, third, and fifth months, give 1 gram of methylprednisolone intravenously for three consecutive days. l Prednisone 0.5 mg / kg orally every other day for 1-6 months, then gradually reduce the dose. l Cyclophosphamide is taken orally, 1.5 mg / kg body weight per day, for 16 months. Rituximab Rituximab 1000 mg intravenously twice within 2 weeks l Rituximab 375 mg per square meter of body surface area, once a week for 1 to 4 times. Tacrolimus l Tacrolimus is administered at a dose of 0.05 mg to 0.1 mg per kilogram of body weight per day, with a target blood concentration of 3 to 8 ng per milliliter for 12 months. Cyclosporine l Cyclosporine is 3.5 mg / kg body weight per day, and its target blood concentration is 125-225 mg / ml.

[0007] The 2021 KDIGO guidelines recommend five treatment options for membranous nephropathy (see Table 1): cyclical or continuous use of cyclophosphamide, rituximab, tacrolimus, and cyclosporine. The following are: 1. Cyclic or continuous use of cyclophosphamide requires the combined use of hormonal drugs (such as prednisone), which have significant side effects. Cyclophosphamide also has adverse reactions such as reproductive toxicity, bone marrow suppression, infection, and hemorrhagic cystitis. Furthermore, nearly half of patients still do not achieve remission after treatment with a hormonal + cyclophosphamide regimen, and patients who achieve remission are likely to experience relapse upon discontinuation of the drug. 2. Tacrolimus and cyclosporine are calcineurin inhibitors (CNIs), a large class of immunosuppressive drugs. CNIs can be used for patients who are unwilling to receive glucocorticoids + cyclophosphamide or have contraindications to treatment. Among them, the advantages of cyclosporine treatment are rapid onset of action, superior efficacy compared to cyclophosphamide, and fewer and milder adverse reactions than cyclophosphamide. However, the disadvantages are that the overall remission rate is not higher than that of the cyclophosphamide regimen and the short-term relapse rate is significantly higher than that of cyclophosphamide. Tacrolimus has a 10-100-fold higher immunosuppressive potency than cyclosporine, but its disadvantages are similar to those of cyclosporine, with a high relapse rate. In addition, tacrolimus's more prominent adverse effects are impaired glucose tolerance and new-onset diabetes. 3. Currently, rituximab alone or in combination with a calcineurin inhibitor is recommended for the initial treatment of primary MN at intermediate, high, or very high risk. It has a remission rate of 67% and is suitable for patients with MN who are either positive or negative for anti-PLA2R antibodies. However, rituximab is expensive, not widely available to patients, and has severe infusion reactions. Various studies have reported that the incidence of rituximab-related infusion-related reactions is 26% to 85%. In patients with hematological malignancies, rituximab has also been reported to cause rare adverse events such as hepatitis B virus reactivation and severe skin reactions (such as toxic epidermal necrolysis and Stevens-Jansen syndrome) (Reference 1: Alsharhan L, Beck LH Jr. Membranous Nephropathy: Core Curriculum 2021. Am J Kidney Dis. 2021 Mar;77(3):440-453.). In general, the clinical course of membranous nephropathy is often indolent and existing treatments are toxic. Existing treatment options have certain limitations and risks, and the treatment effects are not satisfactory. However, from a clinical application perspective, if hormone combined with immunosuppressant treatment is ineffective, monoclonal antibodies (such as rituximab) are usually used for treatment. Therefore, the development of new biological drugs (such as monoclonal antibodies) has become an urgent need for the current treatment of membranous nephropathy.

[0008] Table 2. Approved / Phase III clinical trials of biological drugs for the treatment of membranous nephropathy Drug name Target R&D institutions Highest research stage Rituximab CD20 Roche Approved for marketing Obtenuzumab CD20 Glycart Biotechnology (Roche) Approved for marketing MIL62 CD20 Tian Guangshi Stage III Fizetuzumab CD38 I-Mab Stage III

[0009] As of September 22, 2022, only two biologic drugs for MN have been approved for marketing worldwide: rituximab and obinutuzumab. Two other drugs are in Phase III clinical trials (see Table 2). Although rituximab was included in the recommended treatment options for MN in the 2021 KDIGO guidelines, it still faces challenges such as low remission rates, high rates of infusion reactions, severe adverse reactions, and a high price. While obinutuzumab, which also targets CD20, has achieved some clinical efficacy in MN patients who have failed treatment, this effect is only partial, not complete, because it cannot completely eliminate antibodies and immune complexes, leaving the kidney's basement membrane damaged and allowing some urinary protein to leak. In addition, because obinutuzumab has stronger cytotoxicity and a stronger ability to consume B cells than rituximab, its adverse reactions may face greater challenges (Reference 2: Hudson, R., Rawlings, C., Mon, SY et al. Treatment resistant M-type phospholipase A2 receptor associated membranous nephropathy responds to obinutuzumab: a report of two cases. BMC Nephrol 23, 134 (2022).). Therefore, there is a huge unmet clinical need in the treatment of membranous nephropathy both in China and globally. Summary of the Invention

[0010] The present invention surprisingly discovered that the TACI-Fc fusion protein produced a significant therapeutic effect when treating patients with membranous nephropathy.

[0011] Specifically, the present invention provides a method for treating membranous nephropathy, comprising administering a therapeutically effective amount of TACI-Fc fusion protein to a patient with the membranous nephropathy.

[0012] Specifically, the present invention also provides a method for treating a patient with membranous nephropathy who has received a membranous nephropathy treatment regimen, the method comprising (1) determining whether the patient has received a membranous nephropathy treatment regimen, and (2) if the patient has previously received a membranous nephropathy treatment regimen, administering an effective amount of TACI-Fc fusion protein to the patient with the membranous nephropathy.

[0013] Specifically, the present invention also provides a use of a TACI-Fc fusion protein in the preparation of a drug for treating patients with membranous nephropathy.

[0014] Furthermore, the TACI-Fc fusion protein described in any of the above items comprises: (i) the TACI extracellular region or a fragment thereof that binds to Blys and / or APRIL; and (ii) a human immunoglobulin constant region fragment.

[0015] Preferably, the TACI extracellular region or a fragment thereof comprises the amino acid sequence shown in SEQ ID NO: 1.

[0016] Preferably, the human immunoglobulin is IgG1.

[0017] Furthermore, the human immunoglobulin constant region fragment comprises the amino acid sequence of SEQ ID NO: 2 or comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2.

[0018] Furthermore, the human immunoglobulin constant region fragment comprises amino acid modifications at one or more positions corresponding to positions 3, 8, 14, 15, 17, 110, 111 or 173 of SEQ ID NO: 2.

[0019] Furthermore, the modification is the substitution, deletion or insertion of amino acids.

[0020] Furthermore, the substitution is selected from the following group: P3T, L8P, L14A, L15E, G17A, A110S, P111S and A173T.

[0021] Preferably, the human immunoglobulin constant region fragment comprises the amino acid sequence of SEQ ID NO: 3.

[0022] Preferably, the TACI-Fc fusion protein has the amino acid sequence shown in SEQ ID NO:4.

[0023] Preferably, the TACI-Fc fusion protein is Telitacicept.

[0024] Further, the single dose of the TACI-Fc fusion protein is about 0.1 to 10 mg / kg, further including 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8,5.9,6.0,6.1,6.2,6.3,6.4,6.6,6.6,6.7,6.8,6.9,7.0,7.1,7.2 ,7.3,7.4,7.7,7.6,7.7,7.8,7.9,8.0,8.1,8.2,8.3,8.4,8.8,8.6,8.7,8.8,8.9,9.0,9.1,9.2,9.3,9.4,9.9,9.6,9.7,9.8,9.9,10 mg / kg.

[0025] Furthermore, the single administration dose of the TACI-Fc fusion protein is 160-240 mg, more preferably 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, or 240 mg.

[0026] Furthermore, the fusion protein content was determined by UV-visible spectrophotometry, where the absorbance of the tetasip sample at 280 nm was measured, based on the protein's maximum UV absorption at that wavelength. After correcting the absorbance at 320 nm, the absorbance at 280 nm was found to be proportional to the protein concentration. Protein concentration was calculated according to the Lambert-Beer law to determine the protein content. The protein content calculation formula is as follows: Protein content (mg / ml) = Where, ε is the extinction coefficient of tetasip, the unit is (mg / ml) -1·cm -1; A280 is the average absorbance of the sample solution at 280 nm; A280 (corrected) is the average corrected absorbance of the sample solution at 280 nm.

[0027] Furthermore, the TACI-Fc fusion protein is used 2-4 times at intervals of one month, that is, the administration frequency of the TACI-Fc fusion protein is 2 times per month, 3 times per month, or 4 times per month.

[0028] Furthermore, the TACI-Fc fusion protein is administered once a week.

[0029] More preferably, the treatment lasts for about 2-50 weeks. More preferably, the treatment lasts for 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, or 50 weeks.

[0030] Further preferably, the TACI-Fc fusion protein is administered subcutaneously, intramuscularly, or intravenously, preferably into the thigh, abdomen, or upper arm. In some specific embodiments, the TACI-Fc fusion protein is administered by subcutaneous injection, intramuscular injection, or intravenous injection.

[0031] Further preferably, the TACI-Fc fusion protein is injected into the same or different sites each time. In some specific embodiments, the TACI-Fc fusion protein is injected into the same site each time; in other specific embodiments, the TACI-Fc fusion protein is injected into different sites each time.

[0032] Furthermore, the membranous nephropathy is idiopathic membranous nephropathy or secondary membranous nephropathy.

[0033] Furthermore, the membranous nephropathy is manifested as PLA2R positive or PLA2R negative.

[0034] Furthermore, the patient is an adult patient or a pediatric patient.

[0035] Furthermore, the patient had previously received a treatment regimen for membranous nephropathy.

[0036] Furthermore, the membranous nephropathy treatment regimen includes: conventional treatment for membranous nephropathy, hormone combined with immunosuppressant treatment regimen, and monoclonal antibody drug treatment regimen.

[0037] Furthermore, the membranous nephropathy treatment regimen comprises: periodically administering cyclophosphamide combined with hormone drugs to the patient, or continuously administering cyclophosphamide combined with hormone drugs to the patient, or administering tacrolimus to the patient, or administering cyclosporine to the patient, or administering rituximab to the patient, or administering obinutuzumab to the patient, or administering MIL62 to the patient, or administering fezinomab to the patient.

[0038] The TACI-Fc fusion protein provided by the present invention shows unexpected clinical efficacy and good safety in the treatment of membranous nephropathy patients. Simple diagram description

[0039] Figure 1 shows the change in albumin values ​​during treatment of the subjects in Example 1;

[0040] Figure 2 is a diachronic analysis of the rate of change of immunoglobulin IgG levels compared to baseline during treatment in the subjects of Example 1;

[0041] Figure 3 is a diachronic analysis of the rate of change of immunoglobulin IgM levels compared to baseline during treatment in the subjects of Example 1;

[0042] FIG4 is a diachronic analysis of the rate of change of immunoglobulin IgA levels compared to baseline during treatment of the subjects in Example 1. Implementation Method

[0043] Unless otherwise defined, all technical terms used herein have the same meanings as understood by those of ordinary skill in the art. For definitions and terms in this field, professionals can refer to Current Protocols in Molecular Biology (Ausubel).

[0044] The three-letter and one-letter amino acid codes used in the present invention are as described in J. biol. chem, 243, p3558 (1968).

[0045] The term "TACI" as used herein stands for transmembrane activator and CAML interactor, a member of the tumor necrosis factor receptor superfamily. The term "BLys" as used herein refers to B lymphocyte stimulator, a member of the TNF ligand superfamily that exists in both membrane-bound and soluble forms. It is specifically expressed on the surface of bone marrow cells and selectively stimulates B lymphocyte proliferation and immunoglobulin production. The term "APRIL" (a proliferation-inducing ligand) as used herein refers to a tumor necrosis factor (TNF) analog that stimulates the proliferation of naive B and T cells in the body, promoting B cell accumulation and increasing splenic levels. APRIL specifically binds to TACI and BCMA, preventing APRIL from binding to B cells and inhibiting APRIL-stimulated naive B cell proliferation. Furthermore, APRIL competes with BLys for receptor binding (BCMA, TACI).

[0046] The term "TACI-Fc fusion protein" referred to in the present invention refers to a transmembrane activator, calcium regulator and cyclophilin ligand interactor (TACI)-immunoglobulin fusion protein (i.e., TACI-Fc fusion protein). The TACI-immunoglobulin fusion protein provided by the present invention includes: (i) a TACI extracellular region or a fragment thereof that binds to Blys and / or APRIL; and (ii) a human immunoglobulin constant region fragment.

[0047] The term "TACI extracellular region or a fragment thereof that binds to Blys and / or APRIL" can be specifically referred to the extracellular domain of TACI disclosed in U.S. Patent Nos. 5,969,102, 6,316,222 and 6,500,428 and U.S. patent applications 09 / 569,245 and 09 / 627,206 (the contents of which are incorporated herein by reference), as well as specific fragments of the TACI extracellular domain that can interact with the TACI ligand, or the amino acid fragment at positions 13-118 of the TACI extracellular domain disclosed in Chinese Patent Publication No. CN101323643A.

[0048] In the term "human immunoglobulin constant region fragment," the immunoglobulin portion is preferably IgG1, which may include a heavy chain constant region, such as a human heavy chain constant region. Preferred "human immunoglobulin constant region fragments" of the present invention are amino acid fragments comprising a portion of the hinge domain, CH2 domain, and CH3 domain. In some more preferred embodiments, the amino acid sequence of the "human immunoglobulin constant region fragment" of the present invention is as shown in SEQ ID NO:2, or comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:2. In some more preferred embodiments, the amino acid sequence of the "human immunoglobulin constant region fragment" is as shown in SEQ ID NO:3.

[0049] The term "treatment" as used herein is related to a given disease or condition, including but not limited to: inhibiting the disease or condition, such as preventing the development of the disease or condition; alleviating the disease or condition, such as causing the disease or condition to regress; or alleviating the symptoms caused by the disease or condition, such as alleviating, preventing or treating the symptoms of the disease or condition.

[0050] The term "amino acid" as used herein is understood in the broadest sense and is a general term for a class of organic compounds containing amino and carboxyl groups. Preferably, the amino acids used herein are the main units that make up proteins in living organisms, including but not limited to glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, and histidine.

[0051] The three-letter and single-letter amino acid codes used in the present invention are as described in J. Biol. Chem, 243, p3558 (1968). There are various numbering schemes for amino acid positions, such as the Kabat numbering system, the EU numbering system, and sequential numbering. In the present invention, amino acid positions are numbered using a sequential numbering scheme. For example, "position 3, 8, 14, 15, 17, 110, 111, or 173 of SEQ ID NO: 2" as used herein refers to amino acid position 3 or position 8 of SEQ ID NO: 2, and so on. For example, "P3T" as used herein refers to a mutation of amino acid position 3 of SEQ ID NO: 2 from a previous "P" to a "T." For example, "L8P" refers to a mutation of amino acid position 8 of SEQ ID NO: 2 from a previous "L" to a "P," and so on.

[0052] As an optional embodiment, the constant region of the immunoglobulin provided by the present invention can introduce one or more amino acid changes, such as substitution (ie, mutation), addition (ie, insertion) or deletion (ie, deletion).

[0053] The term "Telitacicept" (or "Telitacicept", which can be used interchangeably in the present invention) in the present invention is a TACI-Fc fusion protein, its INN name is Telitacicept, and its amino acid sequence is shown in SEQ ID NO: 4, or see https: / / extranet.who.int / soinn / mod / page / view.php?id=137&inn_n=10932.

[0054] The TACI-Fc fusion protein of the present invention can be administered by any of a variety of routes, including but not limited to oral, intravenous, intramuscular, intraarterial, intramedullary, intraperitoneal, intrathecal, intracardiac, transdermal, transdermal, topical, subcutaneous, intranasal, enteral, sublingual, vaginal, or rectal routes.

[0055] The term "membranous nephropathy" in the present invention represents a group of diseases with a common histopathological pattern, namely the presence of immunoglobulins and complement-containing immune precipitates in a subepithelial location.

[0056] The term "idiopathic membranous nephropathy" in the present invention refers to an organ-specific autoimmune disease that occurs without any identifiable cause or initiating event, including but not limited to PLA2R-related idiopathic membranous nephropathy, THS7A-related idiopathic membranous nephropathy, NELL-1-related idiopathic membranous nephropathy, Sema3B-related idiopathic membranous nephropathy and other related idiopathic membranous nephropathy.

[0057] The term "secondary membranous nephropathy" as used herein refers to glomerular diseases characterized by renal involvement in a clear systemic disease, with pathological changes manifesting as membranous nephropathy. Causes of secondary membranous nephropathy include immune diseases (such as systemic lupus erythematosus, type 1 diabetes, rheumatoid arthritis, Hashimoto's thyroiditis, Graves' disease, mixed connective tissue disease, Henoch-Schönlein purpura, primary biliary cirrhosis, enteropathic syndrome, anti-glomerular basement membrane nephritis, ANCA-associated crescentic nephritis, graft-versus-host disease, bone marrow and stem cell transplantation, etc.), infections and parasitic diseases (such as HBV, HCV, syphilis, filariasis, cysticercosis, schistosomiasis, malarial parasites, leprosy, etc.), drugs and toxins (such as gold preparations, penicillamine, nonsteroidal anti-inflammatory drugs, mercury, captopril, formaldehyde, hydrocarbons), and others (such as tumors and kidney transplantation).

[0058] The term "PLA 2R positive" in the present invention means that PLA 2R is detected in the patient's blood, that is, PLA 2R is serologically positive.

[0059] The term "PLA 2R negative" in the present invention means that PLA 2R cannot be detected in the patient's blood.

[0060] The term "conventional treatment of membranous nephropathy" in the present invention exemplarily refers to if the 24-hour urine protein quantity of membranous nephropathy is below 3.5g; or between 3.5g-8g, and the renal function is normal and there are no high-risk phenomena, usually pril drugs and sartans are used to reduce urine protein, and supportive treatments such as diuretics, antihypertensives, and anticoagulation are given.

[0061] The term "hormone combined with immunosuppressant treatment regimen" in the present invention exemplarily refers to the use of a combined hormone + immunosuppressant method for treatment if the 24-hour urine protein quantity of membranous nephropathy exceeds 3.5g and is accompanied by renal impairment, or the urine protein exceeds 8g, wherein the immunosuppressant includes but is not limited to tacrolimus and cyclosporine.

[0062] The term "monoclonal antibody drug treatment regimen" in the present invention refers to, for example, the treatment recommended in the 2021 KDIGO guidelines, using but not limited to rituximab, 1000 mg of rituximab intravenously twice within two weeks; rituximab 375 mg per square meter of body surface area, once a week for a total of 1 to 4 times.

[0063] The term "cyclical administration of cyclophosphamide combined with hormone drugs" in the present invention refers to the recommendation in the 2021 KDIGO guidelines: intravenous injection of 1 gram of methylprednisolone for three consecutive days in the first few days of the 1st, 3rd and 5th months; oral administration of 0.5 mg / kg of prednisone per day; and oral administration of cyclophosphamide 2.5 mg per kilogram of body weight per day in the 2nd, 4th and 6th months.

[0064] The term "continuous administration of cyclophosphamide combined with hormone drugs" in the present invention refers to the recommendation in the 2021 KDIGO guidelines: intravenous injection of 1 gram of methylprednisolone for three consecutive days in the first few days of the 1st, 3rd, and 5th months; oral administration of 0.5 mg / kg of prednisone every other day for 1-6 months and then gradually reducing the dose; oral administration of cyclophosphamide, 1.5 mg per kilogram of body weight per day, for 16 months. [Example]

[0065] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.

[0066] Example 1: Clinical trial of Tetasip in the treatment of membranous nephropathy

[0067] 1. Research Methods

[0068] This prospective, single-center, open-label, single-arm clinical trial aims to evaluate the efficacy and safety of tetasipir in the treatment of MN. It also explores the side effects of tetasipir in refractory MN. Thirty patients with MN are enrolled (12 patients have been enrolled, interim analysis completed). They will receive tetasipir 160 mg subcutaneously once weekly for 48 weeks.

[0069] 2. Patient selection

[0070] Subject inclusion criteria: l Patients diagnosed with primary membranous nephropathy by renal pathological puncture and with positive PLA 2R antibody; l Age 18-70 years old; l Before enrollment, patients must have maintained a 24-hour urine protein (UTP) ≥ 3.5 g on the basis of adequate ACEI / ARB treatment for at least 3 months; l Stable blood pressure, ≤140 / 90 mmHg; l Estimated glomerular filtration rate (eGFR) ≥ 45 mL / min / 1.73 m2; l Agree to maintain a stable diet and sodium intake during the study; l Agree and sign the informed consent form.

[0071] Subjects were excluded by the following criteria: l Urine protein quantification has been unstable in the past two months, with fluctuations of 24-hour urine protein quantification >2g / d; l Combined with chronic liver disease, or liver enzyme levels exceeding 3 times the upper limit of normal; l New cardiovascular and cerebrovascular disease (acute coronary syndrome, heart failure, cerebral infarction, stroke) within 3 months; l Uncontrolled severe hypertension; l Malignant tumors; l Patients with severe infection; l Patients who have used Tetasip in the past, or who are allergic to Tetasip or have other contraindications to Tetasip; l Pregnancy or breastfeeding; l Life expectancy is less than 6 months; l Currently participating in other clinical studies; l Other circumstances that the researcher deems unsuitable for participation in this study.

[0072] 3. Endpoint indicators

[0073] (1) Primary endpoint Serial number index Evaluation time end 1 At week 48, 24-hour urine protein excretion level Week 48 Complete remission: urine protein content is less than 0.3g / 24h Partial remission: urine protein content is 0.3-3.5g / 24h or urine protein is reduced by 50% compared with baseline

[0074] (2) Secondary endpoints Serial number index Evaluation time 1 Changes in 24-hour urine protein content (or urine protein / creatinine ratio and urine albumin / creatinine ratio) Weeks 0, 4, 8, 12, 16, 20, 24, 36, and 48 2 Changes in estimated glomerular filtration rate (eGFR) Weeks 0, 4, 8, 12, 16, 20, 24, 36, and 48 3 Changes in serum albumin Weeks 0, 4, 8, 12, 16, 20, 24, 36, and 48 4 Changes in immunoglobulins IgA, IgG, and IgM Weeks 0, 4, 8, 12, 16, 20, 24, 36, and 48 5 Side effects of medications /

[0075] 5. Main results of the interim analysis

[0076] 5.1 Primary Efficacy Endpoint

[0077] The primary endpoint of this study was 24-hour urinary protein excretion at week 48. A complete response was considered when the 24-hour urinary protein level was less than 0.3 g / 24 hours. A partial response was considered when the 24-hour urinary protein level was between 0.3 and 3.5 g / 24 hours, or when the urinary protein level decreased by 50% from baseline. At week 48 or the end of treatment, 10 subjects (83.3% of the enrolled population) achieved a decrease in urinary protein from baseline. Five subjects had 24-hour urinary protein levels between 0.3 and 3.5 g / 24 hours, and three subjects achieved a decrease of greater than 50% from baseline (the percentages of decrease from baseline were 70.86%, 77.42%, and 94.17%, respectively). Therefore, five subjects achieved a partial response in this trial, representing 42% of the total enrolled population. Details of the 24-hour urinary protein levels at baseline and at week 48 or the end of treatment, as well as the percentage of decrease from baseline, are shown in Table 3.

[0078] Table 3. 24-hour urine protein content and the percentage of decrease compared with baseline Subjects 24-hour urine protein (baseline) Unit: ml / min 24-hour urine protein (at week 48 or at the end of treatment) Unit: ml / min Decrease ratio compared with baseline Subject 1 4.53 1.32 70.86% Subject 2 October 23 2.31 77.42% Subject 3 7.19 0.419 94.17% Subject 4 4.44 2.97 / Subject 5 3.72 3.02 /

[0079] 5.2 Secondary Efficacy Endpoints

[0080] 5.2.1. Analysis of Changes in Urine Protein / Creatinine Ratio and Urine Albumin / Creatinine Ratio

[0081] In this study, the subjects' average urine protein / creatinine ratio (UPCR) and urine albumin-to-creatinine ratio (UCAR) decreased compared to baseline. The decrease was statistically significant compared to baseline.

[0082] 5.2.2. Analysis of changes in estimated glomerular filtration rate (eGFR)

[0083] In this study, after the end of medication, the estimated glomerular filtration rate (eGFR) of 4 subjects increased. Among them, the subject with the highest increase had eGFR increase from 47.9 mL / min / 1.73m2 to 62.46 mL / min / 1.73m2, an increase of 30.4%.

[0084] 5.2.3. Chronic analysis of serum albumin

[0085] In this study, the average albumin level decreased during the treatment. After the end of medication, the average serum albumin level of the enrolled subjects decreased by 0.59 g / L compared with the baseline. The temporal changes in albumin are shown in Figure 1.

[0086] 5.2.4. Immunological indicators (IgG, IgM, IgA)

[0087] 5.2.4.1. Chronological analysis of immunoglobulins (IgG)

[0088] Compared with the baseline, after 48 weeks of administration, the average IgG level of the subjects showed an overall downward trend, with the average IgG level decreasing by approximately 2.60 g / L (a decrease of 38.8%). The temporal change rate of the average IgG level of the subjects is shown in Figure 2.

[0089] 5.2.4.2. Chronological Analysis of Immunoglobulin (IgM)

[0090] Compared with the baseline, the average IgM level of the enrolled subjects also showed an overall downward trend. After 48 weeks of administration, the average IgM level decreased by approximately 0.708 g / L (a decrease of 70.1%). The temporal change rate of the average IgM level of the enrolled subjects is shown in Figure 3.

[0091] 5.2.4.3. Chronological Analysis of Immunoglobulin (IgA)

[0092] Compared with the baseline, the change rate of the average IgA level of the enrolled subjects also showed an overall downward trend. After 48 weeks of administration, the average IgA level decreased by approximately 0.717 g / L (a decrease of 39.7%). The temporal change rate of the average IgA level of the enrolled subjects is shown in Figure 4.

[0093] 5.3 Effectiveness Results

[0094] In this study, available data demonstrate that tetasip demonstrates sustained clinical improvement in patients with primary membranous nephropathy, with mean urine protein-to-creatinine ratio (UPCR) and urine albumin-to-creatinine ratio (UCAR) levels decreasing compared to baseline. These decreases were statistically significant. Tetasip also significantly reduced mean IgG, IgA, and IgM levels compared to baseline.

[0095] 5.4 Safety Results

[0096] In this study, the current results demonstrate a favorable safety profile for once-weekly administration of 160 mg of tetasipir in patients with primary MN. Adverse events and reactions were mild in severity (CTCAE grade 1), with no CTCAE grade 2, 3, 4, or 5 adverse events or reactions occurring. No adverse events or reactions led to study withdrawal or death, and no serious adverse reactions occurred. Two subjects experienced adverse events during the study, one of which experienced abdominal pain and vomiting, but this was not considered related to tetasipir. The other experienced pruritus, which resolved after 2-3 days without medication.

[0097] 5.5 Conclusion

[0098] Based on the above results and analysis, in this study, the current experimental data showed that the use of tetasip in the treatment of patients with membranous nephropathy showed good clinical efficacy and safety.

[0099] The above description is merely a preferred embodiment, which is intended to be illustrative and non-limiting of the combinations of features necessary to practice the present invention. The headings provided are not intended to limit the various embodiments of the present invention. Terms such as "comprising," "including," and "include" are not intended to be limiting. Furthermore, unless otherwise indicated, unmodified numerals include plural forms, and "or" and "or" mean "and / or." Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art.

[0100] All publications and patents mentioned herein are incorporated herein by reference. Numerous modifications and variations of the described methods and compositions of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. While the present invention has been described in terms of specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various variations of the described modes for carrying out the invention that are apparent to those skilled in the relevant art are intended to be within the scope of the appended claims.

[0101] none

[0102] none

[0103] TW202421653A_112137435_SEQL.xml

Claims

1. The application of TACI-Fc fusion protein in the preparation of drugs for the treatment of primary membranous nephropathy, wherein, The TACI-Fc fusion protein comprises: (i) the extracellular region of TACI or a fragment thereof binding to Blys and / or APRIL; and (ii) a fragment of the human immunoglobulin constant region, wherein the TACI-Fc fusion protein has the amino acid sequence shown in SEQ ID NO:

4.

2. The application as described in claim 1, wherein, The extracellular region of TACI or the fragment thereof that binds to Blys and / or APRIL contains the amino acid sequence shown in SEQ ID NO:

1.

3. The application as described in claim 2, wherein, The human immunoglobulin is IgG1 or the human immunoglobulin constant region fragment contains the amino acid sequence of SEQ ID NO:2 or contains an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:

2.

4. The application as described in claim 3, wherein, The human immunoglobulin constant region fragment contains amino acid modifications at one or more sites corresponding to sites 3, 8, 14, 15, 17, 110, 111 or 173 of SEQ ID NO:

2.

5. The application as described in claim 4, wherein, The modification is the substitution of amino acids.

6. The application as described in claim 5, wherein, The substitutes are selected from the group consisting of: P3T, L8P, L14A, L15E, G17A, A110S, P111S and A173T.

7. The application as described in claim 1, wherein, The human immunoglobulin constant region fragment contains the amino acid sequence of SEQ ID NO:

3.

8. The application as described in claim 1, wherein, The TACI-Fc fusion protein is Telitacicept.

9. The application as described in claim 1, wherein, The primary membranous nephropathy described herein is either PLA2R positive or PLA2R negative.

10. The application as described in claim 9, wherein, The patients mentioned are either adult patients or pediatric patients.

11. The application as described in claim 10, wherein, The patient had previously received treatment for membranous nephropathy.

12. The application as described in claim 10 or 11, wherein, The single-dose dose of the TACI-Fc fusion protein is 0.1 to 10 mg / kg.

13. The application as described in claim 10 or 11, wherein, The single-dose dose of the TACI-Fc fusion protein is 160-240 mg.

14. The application as described in claim 10 or 11, wherein, The single-dose dose of the TACI-Fc fusion protein is 160 mg or 240 mg.

15. The application as described in claim 10 or 11, wherein, The TACI-Fc fusion protein is administered subcutaneously, intramuscularly, or intravenously, or at the thigh, abdomen, or upper arm.

16. The application as described in claim 11 or 12, wherein, The TACI-Fc fusion protein is used 2-4 times at one-month intervals and / or treatment is continued for 2-50 weeks.

17. The application as described in claim 16, wherein, The TACI-Fc fusion protein is administered once a week.

Images