Treatment of autoimmune disease using CD40l antibody

Abstract

Provided is a method relating to the prevention or treatment of an autoimmune disease in a patient, the method comprising administering an anti-CD40L antibody or an antigen-binding fragment thereof to a patient in need according to a dosing regimen, wherein the dosing regimen comprises administering at least four identical doses of the anti-CD40L antibody or the antigen-binding fragment thereof to the patient in need at regular intervals.

Description

Treatment of autoimmune diseases with CD40L antibodies

[0001] Cross-references to related applications

[0002] This application is based on and claims priority to Chinese Patent Application No. 202411509701.8, filed on October 28, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention generally relates to a method of treating autoimmune diseases such as Sjögren's syndrome with anti-CD40L antibodies, and particularly to a dosing regimen for treating said diseases. Background Technology

[0004] Primary Sjögren's syndrome Primary Sjögren's syndrome (pSS) is a chronic inflammatory autoimmune disease characterized by lymphocyte proliferation and progressive damage to exocrine glands. It is more common in middle-aged and elderly women, with a prevalence of 0.33%-0.77% in the Chinese population. It is one of the most common autoimmune diseases (Zhang Wen, Li Xiaomei, Xu Dong, et al. Diagnosis and treatment guidelines for primary Sjögren's syndrome [J]. Chinese Journal of Internal Medicine, 2020, 59(04):269-276).

[0005] Patients with primary Sjogren's syndrome (pSS) have multiple autoantibodies in their serum. The main clinical manifestations are dry mouth and eyes, body pain, and fatigue. In addition, 30%-50% of patients have systemic involvement (hematologic system, kidneys, lungs, etc.). Due to the lack of conclusive evidence of efficacy and the absence of satisfactory treatment measures, the current treatment of pSS is still mainly symptomatic and empirical, or based on the treatment of similar diseases (Seror R, Nocturne G and Mariette X. Current and future therapies for primary Sjogren syndrome. Nat Rev Rheumatol. 2021, 17(8):475-48). In patients with local symptoms of dry mouth and eyes, artificial saliva or artificial tears are often used. For patients with systemic involvement, guidelines recommend the use of glucocorticoids, immunosuppressants, and biologics. However, there is a lack of research data on the efficacy and safety of these treatments that can be directly compared. Currently, there are no biologics approved for primary Sjogren's syndrome.

[0006] Current research has found that innate immunity (especially the interferon pathway) is closely related to the pathogenesis of pSS. When the innate immunity of pSS patients is activated, it first activates salivary gland epithelial cells. These cells can produce B-cell activating factor (BAFF) to promote B-cell activation. At the same time, the interaction between T and B cells through the CD40 / CD40L pathway also promotes B-cell differentiation and maturation, secreting antibodies to form immune complexes, maintaining and prolonging the immune response, causing tissue damage in pSS patients. Therefore, although there are currently no approved biologics for pSS, guidelines still recommend the use of B-cell-targeting biologics in pSS patients who do not respond well to conventional treatment. A systematic review evaluating the efficacy of belimumab in treating pSS included three clinical studies. The results showed that after 28 weeks of belimumab treatment, 37% and 77% of patients reported improvements in their ESSDAI scores, which could be sustained until week 52. -Rivas N, Sang-Park H, Díaz Del Campo P, Fernández-Castro M, Corominas H, Andreu JL, Navarro-Compán V. Efficacy of belimumab in Primary syndrome: A systematic review. Reumatol Clin (Engl Ed). 2021Mar; 17(3):170-174. English, Spanish.).

[0007] As research progresses, CD40 / CD40L, a co-stimulatory molecule between B cells and T cells, has attracted researchers' interest.

[0008] CD40 ligand (CD40L), also known as CD154, gp39, TNF-associated activating protein (TRAP), 5c8 antigen, or T-BAM, is a 39 kDa type II transmembrane protein. CD40L belongs to the tumor necrosis factor (TNF) superfamily and exists on the cell membrane as a homotrimer. It is primarily expressed on activated T cells, but is also present on other types of immune and non-immune cells, such as epithelial cells, monocytes, dendritic cells, fibroblasts, smooth muscle cells, endothelial cells, and platelets. The CD40L / CD40 signaling pathway mediates various immune and inflammatory responses and plays a crucial role in normal immune responses. Existing research indicates that the CD40L / CD40 signaling pathway not only mediates signal transduction between various lymphocytes but also participates in interactions between non-immune cells. Therefore, the CD40L / CD40 signaling pathway plays an important pathogenic role in a variety of chronic inflammatory diseases, such as autoimmune diseases, neurodegenerative diseases, graft-versus-host disease, tumors, and atherosclerosis.

[0009] Currently, clinical trials are underway for CD40 / CD40L-related biologics in autoimmune diseases including psoriasis, rheumatoid arthritis, and SLE.

[0010] Early anti-CD40L antibody Ruplizumab (CN101072587B) showed good therapeutic effects in diseases such as systemic lupus erythematosus and lupus nephritis, but its development was discontinued due to serious safety events such as thrombosis (Nat. Med. 6(2)(2000) 114). Subsequent studies have shown that thrombosis mainly occurs because free CD40L forms an immune complex with the antibody, which then interacts with C1q and FcgR, especially with FcgRIIa expressed on platelets, inducing platelet activation and aggregation, leading to thrombosis. Therefore, completely removing the effector function of the Fc region is crucial for the safety of anti-CD40L antibodies. Subsequent CD40L-targeting antibodies that entered clinical trials all modified the Fc region, with two main technical approaches: one is to weaken the effector function of the Fc region through mutation; the other is to completely remove the Fc region while using other techniques to prolong the half-life of the molecule. However, both approaches have certain problems. In the first technical approach, taking frexalimab (AU2016294417B2) as an example, both frexalimab and frexalimab use C220S / C226S / C229S / P238S and E269R / K322A mutations, respectively, to weaken the effector function of the Fc region. However, subsequent studies have shown that these mutations cannot completely eliminate the effector function of the Fc region, thus posing certain safety risks. The second technical approach uses Dapirolizumab pegol (US8293237B2) and Dazodalibep (CN101679521B) as examples. Dapirolizumab pegol retains only the monovalent anti-CD40L Fab and extends its half-life by cross-linking with PEG. However, this monovalent molecular form weakens the molecule's biological activity, and PEG cross-linking also increases the difficulty of molecular CMC (cytotoxicity control). Dazodalibep uses a non-traditional antibody form of the Tn3 backbone protein as the molecule targeting and inhibiting CD40L, and extends the molecule's half-life by fusing with albumin. However, clinical studies have shown that this form of molecule has high immunogenicity, and therefore also poses certain safety risks.

[0011] CD40 monoclonal antibody drugs (Jobling K, Ng WF. CD40 as a therapeutic target in Sjögren's syndrome) have also been developed. Syndrome. Expert Rev Clin Immunol. 2018 Jul; 14(7):535-537). A phase II randomized, double-blind placebo study evaluated the efficacy and safety of the anti-CD40 monoclonal antibody CFZ533 in the treatment of pSS. A total of 44 subjects were enrolled in the study. Cohort 1 included 8 subjects who received 3 mg / kg CFZ533 and 4 subjects who received placebo; Cohort 2 included 21 subjects who received 10 mg / kg CFZ533 and 11 subjects who received placebo. The results showed that CFZ533 was well tolerated, and most adverse events (AEs) were mild or moderate. In cohort 1, after placebo adjustment, there was no significant difference in ESSDAI score improvement before and after treatment in the CFZ533 group (ΔESSDAI = 0.68, 95% CI = -4.71 to 6.46). However, in cohort 2, the ESSDAI improvement from baseline was 6.35 points in the CFZ533 group and 1.27 points in the placebo group, respectively, and remained statistically significant after placebo adjustment (ΔESSDAI = 5.64, 95% CI = 1.02 to 10.58), suggesting that the anti-CD40 monoclonal antibody CFZ533 may have a therapeutic effect on pSS (Jobling K, Ng WF. CD40 as a therapeutic target in...). Syndrome. Expert Rev Clin Immunol. 2018 Jul; 14(7): 535-537). Currently, CFZ533 is still undergoing longer-term clinical trials to observe the efficacy and safety of this target in pSS.

[0012] Despite the existence of CD40 monoclonal antibody drugs for Sjögren's syndrome, the utilization rate of biologics among patients with Sjögren's syndrome remains very low, and there is still a huge demand from patients for effective, safe, and low-cost domestically produced biologics.

[0013] Invention Overview

[0014] CD40L is primarily expressed on the surface of activated CD4 T cells. It interacts with CD40 expressed on the surface of B cells, playing a crucial role in various immune responses and inflammatory processes: mediating humoral immune responses and the release of inflammatory factors; acting as a co-stimulatory molecule in the interaction between T cells and antigen-presenting cells (APCs) and B cells; and playing an important role in germinal center (GC) formation and antibody class switching. Therefore, the CD40L / CD40 signaling pathway plays a significant pathogenic role in various B cell-related diseases, such as autoimmune diseases, neurodegenerative diseases, graft-versus-host disease, tumors, and atherosclerosis.

[0015] The inventors have independently developed an antibody or antigen-binding fragment that specifically binds to CD40L, which can block the interaction between CD40 and CD40L, reduce the formation of germinal centers, and / or decrease antibody production. The inventors have found that CD40L or CD40 blocking antibodies significantly alleviate and improve symptoms in various disease models. The antibody or antigen-binding fragment that specifically binds to CD40L in this invention adopts the traditional IgG molecule form, and utilizes a unique Fc modification technology to completely remove the effector function of the Fc region, avoiding platelet activation and resolving the thromboembolic risk associated with existing CD40 target molecules.

[0016] Based on this, the inventors further discovered that by administering anti-CD40L antibody or its antigen-binding fragment, especially according to the administration regimen of the present invention, it can exhibit good pharmacokinetic characteristics in healthy individuals, patients with primary Sjögren's syndrome, and patients with systemic lupus erythematosus, and has good tolerability and / or safety.

[0017] The present invention also addresses the above-mentioned needs by providing a method for treating autoimmune diseases using anti-CD40L antibodies.

[0018] Therefore, in a first aspect, a method for preventing or treating an autoimmune disease in a patient is provided, the method comprising administering the anti-CD40L antibody of the present invention or its antigen-binding fragment to a patient in need. In some embodiments, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0019] In a second aspect, a kit is provided comprising an effective amount of the anti-CD40L antibody or its antigen-binding fragment of the present invention, and a packaging insert printed with instructions for using the anti-CD40L antibody or its antigen-binding fragment for the prevention or treatment of autoimmune diseases. In some embodiments, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0020] In a third aspect, unit dosage forms are provided, comprising an effective amount of the anti-CD40L antibody of the present invention or its antigen-binding fragment.

[0021] In a fourth aspect, the present invention provides the use of the anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit for the prevention or treatment of autoimmune diseases. In some embodiments, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0022] In a fifth aspect, the present invention provides an anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit for the prevention or treatment of autoimmune diseases. In some embodiments, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0023] In a sixth aspect, the use of the anti-CD40L antibody of the present invention, or its antigen-binding fragment, unit dosage form, or kit, in the preparation of a medicament for the prevention or treatment of autoimmune diseases is provided. In some embodiments, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0024] Other embodiments of the present invention will become clear from the following detailed description. Attached Figure Description

[0025] Figure 1 shows the clinical study design of this invention.

[0026] Invention Details

[0027] Before describing the invention in detail, it should be understood that the invention is not limited to the specific methods and experimental conditions described herein, as these methods and conditions can be modified. Furthermore, the terminology used herein is for illustrative purposes only and is not intended to be restrictive.

[0028] definition

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For the purposes of this invention, the following terms are defined below.

[0030] Unless otherwise specified, terms used in the singular may also include the plural, and vice versa, where appropriate.

[0031] The term "about," when used in conjunction with a numerical value, means to encompass a range of numerical values ​​having a lower limit that is 5%, 4%, 3%, 2%, or 1% smaller than the specified numerical value and an upper limit that is 5%, 4%, 3%, 2%, or 1% larger than the specified numerical value. When the term "about" is used in conjunction with a numerical range, such as in "about 5.0-7.0," it is intended to encompass a numerical range whose lower limit is 5%, 4%, 3%, 2%, or 1% smaller than the lower limit of the numerical range associated with "about," and whose upper limit is 5%, 4%, 3%, 2%, or 1% larger than the upper limit of the numerical range associated with "about." When the term "about" is used in conjunction with multiple numerical values ​​or numerical ranges, it indicates that each of the stated numerical values ​​or numerical ranges is used in conjunction with the term "about." For example, the expression "approximately 1 mg / kg, 3 mg / kg, 5 mg / kg, 7.5 mg / kg, 15 mg / kg, 25 mg / kg, 30 mg / kg or 35 mg / kg" is intended to cover the case of "approximately 1 mg / kg, approximately 3 mg / kg, approximately 5 mg / kg, approximately 7.5 mg / kg, approximately 15 mg / kg, approximately 25 mg / kg, approximately 30 mg / kg or approximately 35 mg / kg", and the expression "approximately 0.3 mg / kg-35 mg / kg, 1 mg / kg-35 mg / kg" is intended to cover the case of "approximately 0.3 mg / kg-approximately 35 mg / kg, approximately 1 mg / kg-approximately 35 mg / kg".

[0032] When the term “and / or” is used to connect two or more options, it should be understood to mean any one of the options or any two or more of the options.

[0033] As used herein, the terms “comprising” or “including” mean to include the stated elements, integers, or steps, but do not exclude any other elements, integers, or steps. In this document, when the terms “comprising” or “including” are used, unless otherwise specified, they also cover situations consisting of the mentioned elements, integers, or steps. For example, when referring to an antibody variable region “comprising” a specific sequence, it is also intended to cover the antibody variable region consisting of that specific sequence.

[0034] The term "CD40L" refers to a ligand expressed on activated T cells. Other names for it are also known in the art, such as CD154, CD40 ligand (CD40L), CD40 anti-receptor (CD40CR), gp39, T-BAM, T-cell activation molecule, TRAF, TNF-associated activating protein (TRAP), and tumor necrosis factor ligand superfamily member 5 (TNFSF5). These terms are used interchangeably throughout the application. In some embodiments, the amino acid sequence of CD40L is shown in Uniport P29965. In some embodiments, CD40L comprises the sequence shown in SEQ ID NO: 17.

[0035] As used herein, the terms “anti-CD40L antibody,” “anti-CD40L,” “CD40L antibody,” or “antibody that binds to CD40L” refer to antibodies that are capable of binding with sufficient affinity to (human or cynomolgus monkey) CD40L or fragments thereof such that the antibody can be used as a diagnostic and / or therapeutic agent targeting (human or cynomolgus monkey) CD40L.

[0036] In this article, the term “antibody” is used in the broadest sense to refer to a protein that contains an antigen-binding site, encompassing natural and artificial antibodies of various structures, including but not limited to complete antibodies and antigen-binding fragments of antibodies, full-length antibodies, multispecific antibodies, or bispecific antibodies.

[0037] The terms “full-length antibody,” “complete antibody,” and “intact antibody” are used interchangeably herein to refer to glycoproteins comprising at least two heavy chains (H) and two light chains (L) linked by disulfide bonds. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (VL) and a light chain constant region. The light chain constant region consists of one domain: CL. The VH and VL regions can be further subdivided into hypervariable regions (complementarity-determining regions (CDRs) interspersed with more conserved regions (framework regions (FRs)). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The constant regions do not directly participate in antibody-antigen binding but exhibit various effector functions.

[0038] "Antibody fragment" refers to a molecule distinct from the intact antibody that contains a portion of the intact antibody and binds to the antigen bound by the intact antibody. The term "antigen-binding fragment" as used herein refers to a molecule distinct from the intact antibody that contains a portion of the intact antibody and binds to the antigen bound by the intact antibody.

[0039] The term "antigen-binding fragment" in antibody refers to a molecule distinct from a full-length antibody. It comprises a portion of the full-length antibody but can bind to the antigen of the full-length antibody or compete with the full-length antibody (i.e., the full-length antibody from which the antigen-binding fragment originates) for antigen binding. Antigen-binding fragments can be prepared using recombinant DNA technology or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, dAb (domain antibody), linear antibodies, single-chain antibodies (e.g., scFv); single-domain antibodies (sdAb) such as VHH, bivalent antibodies or fragments thereof, or camelid antibodies or diabody antibodies. For example, the Fab fragment can be obtained by digesting a full-length antibody with papain. Furthermore, digestion of a complete antibody with pepsin below the disulfide bonds in the hinge region produces F(ab')2, a dimer of Fab' and a divalent antibody fragment. F(ab')2 can be reduced under neutral conditions by breaking the disulfide bonds in the hinge region, thereby converting the F(ab')2 dimer into Fab' monomers. The Fv fragment consists of the VL and VH domains of the antibody single arm. The two domains, VL and VH, of the Fv fragment can be encoded by independent genes, but they can also be generated as a single protein chain by using a recombinant approach and connecting the two domains with a synthetic linker peptide. In this single protein chain, the VL and VH regions pair to form a single-chain Fv (scFv).

[0040] The complementarity-determining region (CDR) or CDR is a region within the antibody variable domain that is highly variable in sequence and forms a structurally defined loop ("hypervariant loop") and / or contains antigen contact residues ("antigen contact sites"). The CDR is primarily responsible for binding to antigen epitopes. The CDRs of the heavy and light chains are typically referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus. CDRs located within the antibody heavy chain variable domain are referred to as HCDR1, HCDR2, and HCDR3, while those located within the antibody light chain variable domain are referred to as LCDR1, LCDR2, and LCDR3. In a given light chain variable region or heavy chain variable region amino acid sequence, the precise amino acid sequence boundaries of each CDR can be determined using any one or a combination of many known antibody CDR assignment schemes, including, for example: Chothia (Chothia et al. (1989) Nature 342:877-883, Al-Lazikani et al., “Standard conformations for the canonical structures of immunoglobulins”, Journal of Molecular Biology, 273, 927-948 (1997)) based on antibody three-dimensional structure and CDR loop topology; Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, 4th ed., USDapartment of Health and Human Services, National Institutes of Health (1987)) based on antibody sequence variability; AbM (University of Bath); Contact (University College London); and the International ImMunoGeneTics. The database (IMGT) (on the World Wide Web at imgt.cines.fr / ) and the North CDR definition based on affinity propagation clustering using a large number of crystal structures.

[0041] Unless otherwise stated, in this invention, the term "CDR" or "CDR sequence" encompasses a CDR sequence determined in any of the foregoing methods or combinations thereof. A CDR may also be determined based on having the same Kabat numbering position as a reference CDR sequence (e.g., any of the exemplary CDRs of this invention). Unless otherwise stated, in this invention, when referring to the position of residues in the antibody variable region (including heavy chain variable region residues and light chain variable region residues), it means the numbering position according to the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

[0042] In some embodiments, the CDR of the heavy chain variable region of the antibody in this invention is determined according to Kabat or Chothia, or a combination of Kabat and Chothia. In some embodiments, the CDR of the light chain variable region of the antibody in this invention is determined according to Kabat.

[0043] In some embodiments, the CDR1 of the antibody heavy chain variable region in this invention is determined according to the Kabat and Chothia combination scheme (hereinafter referred to as "Kabat and Chothia combination", corresponding to H26-H32 in the Kabat numbering system), and the CDR2, CDR3 of the heavy chain variable region, and the LCDR1, LCDR2 and LCDR3 of the light chain variable region are determined according to the Kabat scheme.

[0044] A "humanized antibody" is an antibody that retains the antigen-specific reactivity of a non-human antibody (such as a mouse monoclonal antibody) while exhibiting low immunogenicity when administered to humans as a therapeutic agent. This can be achieved, for example, by retaining the non-human antigen-binding site and replacing the remaining portion of the antibody with its corresponding human portion (e.g., replacing the non-binding portions of the constant region and variable region with the corresponding portions of the human antibody).

[0045] The term "Fc domain" or "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of its constant region. This term includes native sequence Fc regions and variant Fc regions. A native immunoglobulin "Fc domain" contains two or three constant domains: a CH2 domain, a CH3 domain, and optionally a CH4 domain. For example, in native antibodies, an immunoglobulin Fc domain contains the second and third constant domains (CH2 and CH3 domains) of two heavy chains derived from IgG, IgA, and IgD antibodies; or it contains the second, third, and fourth constant domains (CH2, CH3, and CH4 domains) of two heavy chains derived from IgM and IgE antibodies. Unless otherwise stated herein, amino acid residues in the Fc region or heavy chain constant region are numbered according to the EU numbering system (also known as the EU index) described in Kabat et al., Sequences of Proteins of Immunological Interes, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, NIH Publication 91-3242. However, the C-terminal lysine (Lys447) in the Fc region may or may not be present. Two Fc regions can dimerize to form a dimer Fc, and two different Fc regions can heterodimerize to form a heterodimeric Fc. In this document, the terms "Fc region," "Fc portion," and "dimeric Fc" do not include the heavy chain variable region VH and light chain variable region VL of immunoglobulins, nor the heavy chain constant region CH1 and light chain constant region CL, but in some cases may include a hinge region or a partial hinge region at the N-terminus of the heavy chain constant region, such as the IgG1 hinge region or a partial IgG1 hinge region, for example, according to EU numbering, the sequence D221 to P230. In one embodiment, the human IgG heavy chain Fc region extends from Asp221 or from Cys226 or from Asp231 to the carboxyl terminus of the heavy chain.

[0046] In some implementations, amino acid substitutions are represented as (original amino acid, amino acid position, mutant amino acid). For example, when the substitution site is located in the C region, "L234A" means that leucine (L) at position 234 of EU is replaced by alanine (A). When referring to combinations of mutations, the mutations in the combination are connected by slashes. "L234A / L235A" indicates that both mutations L234A and L235A are included.

[0047] In some implementations, amino acid deletions are represented by △ + amino acid + mutation site, for example, △P329 represents the deletion of proline at position 329. Amino acid deletions can also be represented by "amino acid + mutation site + deletion", for example, the deletion of proline at position 329 can also be written as "P329 deletion".

[0048] The terms "pharmaceutical composition" or "formulation" refer to a composition suitable for administration to animals, preferably mammals (including humans), comprising at least one active ingredient and at least one inactive ingredient, such as a pharmaceutically acceptable excipient. The pharmaceutical compositions of the present invention are preferably in parenteral form, such as solutions, suspensions, lyophilized powders, concentrated solutions, etc.

[0049] The term "pharmacy kit" refers to a kit containing one or more individual pharmaceutical compositions, wherein at least one pharmaceutical composition contains an effective amount of an anti-CD40L antibody or its antigen-binding fragment, and a packaging insert containing instructions for the use of the anti-CD40L antibody or its antigen-binding fragment for the prevention or treatment of autoimmune diseases, particularly for administration according to the dosing regimen of the present invention. The pharmacy kit may also contain other therapeutic agents having the same or different activities. It is understood that when a pharmacy kit contains multiple individual pharmaceutical compositions, each of the pharmaceutical compositions may contain different doses and / or be administered via different routes.

[0050] As used herein, the term "unit dosage form" refers to a discrete pharmaceutical unit containing the antibody or antigen-binding fragment of the present invention, to be administered to a patient, such as an injection vial, ampoule, pre-filled syringe, or pre-filled syringe, containing a solution or lyophilized powder of the drug, preferably a lyophilized powder.

[0051] The terms "patient" and "subject" are used interchangeably to refer to animals. Preferably, the patient or subject is a mammal, such as a primate (e.g., human, monkey, orangutan), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, etc. More preferably, the patient or subject is a human.

[0052] The term "biopharmaceutical-naïve" refers to patients or subjects who have not previously received biologic therapy.

[0053] As used herein, the term “parenteral administration” means administration other than enteral and local administration, usually by injection or infusion, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subcutaneous, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions.

[0054] The term "pharmaceutical acceptable" means that the substance, composition, dosage form, etc. described thereafter does not have excessive toxicity, irritation, allergic reactions, or other unwanted properties in mammals, especially humans, and has a reasonable benefit / risk ratio when applied to animals or humans.

[0055] The term "pharmaceuticalally acceptable excipient" refers to a component in a pharmaceutical preparation other than the active ingredient that is non-toxic to an individual.

[0056] The term "effective dose" refers to the amount of medication required to achieve a desired therapeutic or preventative effect at the necessary dosage and for the required duration. It can be determined by the physician or veterinary practitioner involved and will vary depending on factors such as the compound, the state of the disease being treated, the severity of the disease, the individual's age and relevant health conditions, the route and form of administration, and the judgment of the attending physician or veterinary practitioner. Generally, the "preventive effective dose" will be less than the "therapeutic effective dose."

[0057] The term "treatment" refers to reducing or lessening the severity of at least one symptom or condition, slowing or halting its progression, or curing the condition.

[0058] The term "prevention" refers to delaying or preventing the occurrence of a disease or its symptoms.

[0059] The term "autoimmune disease" refers to a group of illnesses in which the body's immune system mistakenly attacks and damages its own tissues and organs. Under normal circumstances, the immune system recognizes and attacks foreign pathogens, such as bacteria, viruses, and parasites, as well as foreign substances, such as allergens. However, in autoimmune diseases, the immune system loses its ability to distinguish between self and non-self, and begins attacking the body's own cells and tissues. In some implementations, autoimmune diseases include systemic lupus erythematosus or Sjögren's syndrome.

[0060] The term "Sjögren's syndrome" Systemic lupus erythematosus (SS) is a chronic systemic autoimmune disease that primarily affects the body's exocrine glands, especially those that produce saliva and tears. It can be primary or secondary to other autoimmune diseases such as systemic lupus erythematosus.

[0061] The term "systemic lupus erythematosus (SLE)" is a typical autoimmune disease characterized by widespread organ involvement and the production of various autoantibodies. SLE can affect the skin, joints, kidneys, heart, lungs, blood vessels, brain, and other organs. The severity of the disease and the organs affected vary from person to person; some may experience only mild symptoms, while others may develop serious, life-threatening complications.

[0062] The term "week 0" refers to the time point at which CD40L antibody or its antigen-binding fragment is first administered. The term "week 4" refers to the fourth week from the time point at which CD40L antibody is first administered. The term "week 8" refers to the eighth week from the time point at which CD40L antibody is first administered. Other similar terms should be interpreted accordingly.

[0063] Detailed description

[0064] In a first aspect of the invention, a method for preventing or treating an autoimmune disease in a patient is provided, the method comprising administering the anti-CD40L antibody of the invention or an antigen-binding fragment thereof to a patient in need in accordance with a dosing regimen.

[0065] In some embodiments, the disease or condition for which the method or administration regimen of the present invention is suitable for prevention or treatment is an autoimmune disease, such as Sjögren's syndrome or systemic lupus erythematosus. In some specific embodiments, the Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to other autoimmune diseases such as systemic lupus erythematosus. In some embodiments, the disease or condition for which the method or administration regimen of the present invention is suitable for prevention or treatment is primary Sjögren's syndrome.

[0066] In some embodiments of the invention, the dosing regimen includes administering one or more doses of anti-CD40L antibody or its antigen-binding fragment to a patient in need, wherein the dose is approximately 0.1 mg / kg to 40 mg / kg, for example approximately 0.3 mg / kg to 35 mg / kg, 1 mg / kg to 35 mg / kg, 3 mg / kg to 35 mg / kg, 5 mg / kg to 35 mg / kg, 7.5 mg / kg to 35 mg / kg, 7.5 mg / kg to 30 mg / kg, 15 mg / kg to 35 mg / kg, 15 mg / kg to 30 mg / kg, or 25 mg / kg to 35 mg / kg. In some embodiments, the dose is, for example, about 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 5, 7.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5, 35, 37.5 or 40 mg / kg, and includes a range defined by any two of the said values.

[0067] In some specific embodiments, the anti-CD40L antibody or its antigen-binding fragment comprises a first heavy chain complementarity-determining region (HCDR1), a second heavy chain complementarity-determining region (HCDR2), and a third heavy chain complementarity-determining region (HCDR3), as well as a first light chain complementarity-determining region (LCDR1), a second light chain complementarity-determining region (LCDR2), and a third light chain complementarity-determining region (LCDR3). HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 respectively comprise the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, or are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9. The amino acid sequence composition shown in NO:9, for example, the CDR is determined by any CDR assignment scheme, such as Kabat, Chothia, IMGT, AbM or combinations thereof such as the Kabat and Chothia combination. For example, HCDR1 is determined according to the Kabat and Chothia combination scheme (hereinafter referred to as "Kabat and Chothia combination", corresponding to H26-H32 in the Kabat numbering system), and HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are determined according to the Kabat scheme.

[0068] In some specific embodiments, the anti-CD40L antibody or its antigen-binding fragment comprises a first heavy chain complementarity-determining region (HCDR1), a second heavy chain complementarity-determining region (HCDR2), and a third heavy chain complementarity-determining region (HCDR3), as well as a first light chain complementarity-determining region (LCDR1), a second light chain complementarity-determining region (LCDR2), and a third light chain complementarity-determining region (LCDR3), wherein HCDR1, HCDR2, and HCDR3 are those contained in VH as shown in SEQ ID NO:4, and LCDR1, LCDR2, and LCDR3 are those contained in SEQ ID NO:4. The VL shown in NO:10 contains LCDR1, LCDR2, and LCDR3, wherein the CDR is determined by any CDR assignment scheme, such as Kabat, Chothia, IMGT, AbM, or combinations thereof, such as the Kabat and Chothia combination. For example, HCDR1 is determined according to the Kabat and Chothia combination scheme (hereinafter referred to as "Kabat and Chothia combination", corresponding to H26-H32 in the Kabat numbering system), and HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are determined according to the Kabat scheme.

[0069] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use in the present invention comprises a heavy chain variable region (VH), wherein the heavy chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence, or comprises the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence; or comprises the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence; or comprises a light chain variable region (VL), wherein the light chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:10 or the nucleic acid sequence shown in SEQ ID NO:20, or is composed of said amino acid sequence, or comprises the amino acid sequence shown in SEQ ID NO:10, or comprises said amino acid sequence; or comprises the amino acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence; or comprises the amino acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence; or comprises the amino acid sequence shown in SEQ ID NO:10 ... The amino acid sequence shown in NO:10 or the amino acid sequence encoded by the nucleic acid sequence shown in SEQ ID NO:20, or the amino acid sequence thereof.

[0070] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment applicable to the present invention comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence, or is composed of the amino acid sequence, and the light chain variable region comprises an amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:10 or the nucleic acid sequence shown in SEQ ID NO:20, or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence, or is composed of the amino acid sequence.

[0071] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use with the present invention comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises or is composed of the amino acid sequence shown in SEQ ID NO:4, and the light chain variable region comprises or is composed of the amino acid sequence shown in SEQ ID NO:10. In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use with the present invention comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises or is composed of the amino acid sequence encoded by the nucleic acid sequence shown in SEQ ID NO:19, and the light chain variable region comprises or is composed of the amino acid sequence encoded by the nucleic acid sequence shown in SEQ ID NO:20.

[0072] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for the present invention further comprises an Fc region, for example, wherein the Fc region is or is derived from human IgG Fc, for example, is or is derived from human IgG1 Fc, is or is derived from human IgG2 Fc, is or is derived from human IgG3 Fc or is derived from human IgG4 Fc; preferably, the Fc region comprises a mutation that reduces or eliminates effector function, and / or eliminates the binding affinity to FcγR while retaining the binding affinity to FcRn, preferably, the mutation is an L234A / L235A and / or P329 deletion mutation, for example, an L234A / L235A and P329 deletion mutation. In some embodiments of the present invention, the Fc region

[0073] (i) Contains or consists of the amino acid sequence shown in SEQ ID NO:13 or 14;

[0074] (ii) Contains an amino acid sequence having at least 90% identity with the amino acid sequence shown in SEQ ID NO:13, for example, 95%, 96%, 97%, 99% or higher identity; or

[0075] (iii) Contains an amino acid sequence that has at least 90% identity with the amino acid sequence shown in SEQ ID NO:14, such as 95%, 96%, 97%, 99% or higher, and contains L234A / L235A and P329 deletion mutations.

[0076] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use in the present invention further comprises a heavy chain constant region, which is or is derived from a constant region of (human) IgG1, IgG2, IgG3, or IgG4. In some embodiments, the heavy chain constant region comprises or is composed of the Fc and CH1 regions described herein. In some embodiments, the CH1 region comprises or is composed of the amino acid sequence shown in SEQ ID NO:15 or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0077] In some embodiments, the heavy chain constant region suitable for the anti-CD40L antibody or its antigen-binding fragment of the present invention

[0078] (i) Contains or is composed of the amino acid sequence shown in SEQ ID NO:5 or 16;

[0079] (ii) comprising an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:16; or

[0080] (iii) Contains an amino acid sequence that has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequence shown in SEQ ID NO:5, and has L234A / L235A and P329 deletion mutations.

[0081] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use in the present invention further comprises a light chain constant region, said light chain constant region being or derived from the light chain constant region of (human) lambda or Kappa. Preferably, said light chain constant region comprises or consists of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 11; or comprises or consists of the amino acid sequence shown in SEQ ID NO: 11.

[0082] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for the present invention comprises a heavy chain, wherein the heavy chain...

[0083] (i) comprising or consisting of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:6; or

[0084] (ii) Contains or is composed of the amino acid sequence shown in SEQ ID NO:6; or

[0085] It contains a light chain, wherein the light chain

[0086] (i) comprising or consisting of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 12; or

[0087] (ii) Contains or consists of the amino acid sequence shown in SEQ ID NO:12.

[0088] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use in the present invention comprises a heavy chain and a light chain, wherein the heavy chain comprises or is composed of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:6; and the light chain comprises or is composed of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:12.

[0089] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment suitable for use in the present invention comprises a heavy chain and a light chain, wherein the heavy chain comprises or is composed of the amino acid sequence shown in SEQ ID NO:6, and the light chain comprises or is composed of the amino acid sequence shown in SEQ ID NO:12.

[0090] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment applicable to the present invention comprises two heavy chains and two light chains, wherein the heavy chains comprise or consist of the amino acid sequence shown in SEQ ID NO:6, and the light chains comprise or consist of the amino acid sequence shown in SEQ ID NO:12.

[0091] In some embodiments of the present invention, the anti-CD40L antibody suitable for use in the present invention is a monoclonal antibody. In some embodiments of the present invention, the anti-CD40L antibody suitable for use in the present invention is a humanized antibody or a chimeric antibody. In some embodiments of the present invention, the anti-CD40L antibody suitable for use in the present invention is a full-length antibody. In some embodiments of the present invention, the antigen-binding fragment of the anti-CD40L antibody suitable for use in the present invention is an antibody fragment selected from the following: Fv, Fab, Fab', Fab'-SH, F(ab')2, dAb (domain antibody), linear antibody, single-chain antibody (e.g., scFv); single-domain antibody (sdAb) such as VHH, bivalent antibody or fragment thereof, or camelid antibody or diabody antibody. In some embodiments of the present invention, the anti-CD40L antibody suitable for use in the present invention also encompasses multispecific antibodies, such as bispecific antibodies.

[0092] In some embodiments of the present invention, the anti-CD40L antibody or its antigen-binding fragment is recombinantly expressed in host cells such as mammalian cells like HEK 293 cells or CHO cells.

[0093] In some embodiments, the anti-CD40L antibody suitable for use in this invention is the anti-CD40L antibody disclosed in PCT application number PCT / CN2024 / 1090122, or its antigen-binding fragment, which is incorporated herein by reference in its entirety as if its entire contents were set forth herein. Preferably, the CD40L antibody is the anti-CD40L antibody Hz33B9C5 disclosed in PCT application number PCT / CN2024 / 1090122, which consists of two heavy chain sequences shown in SEQ ID NO:6 and two light chain sequences shown in SEQ ID NO:12.

[0094] In some specific embodiments, the antibody or its antigen-binding fragment is administered by injection, such as subcutaneous injection or intravenous injection (e.g., infusion), preferably by intravenous infusion or intravenous drip. In some specific embodiments, the antibody or its antigen-binding fragment is administered by intravenous injection. More preferably, the antibody or its antigen-binding fragment is administered by intravenous infusion.

[0095] In some embodiments, the antibody or its antigen-binding fragment may be administered once or multiple times, either continuously or discontinuously, for example, at least once, twice, three times, or four times, such as at least four consecutive administrations. In some embodiments, the number of administrations is determined based on patient assessment of efficacy. Therefore, in some embodiments, more than four doses of the antibody or its antigen-binding fragment may be given. It is understood that each dose may be equal or unequal, preferably equal. Preferably, the dose is given 1-8 times, for example, 1, 2, 3, 4, 5, 6, 7, or 8 times. More preferably, the dose is given at least 4 times, for example, 4 times. When multiple doses are given, the doses are administered at intervals of 2-8 weeks, for example, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, preferably at 4-week intervals. In some exemplary embodiments of the invention, when multiple doses are administered, the doses are administered at 4-week intervals. Preferably, the same dose of the antibody or its antigen-binding fragment is given each time. In some embodiments, the treatment phase lasts for at least 12 weeks from the first dose administration, such as 12, 16, or 20 weeks. In some specific embodiments, the method administers at least four (e.g., four) doses of the same anti-CD40L antibody or its antigen-binding fragment to the patient in need via intravenous infusion at 4-week intervals.

[0096] In some specific embodiments, the dosage of the antibody or its antigen-binding fragment in the method is 0.1 mg / kg to 40 mg / kg. In some embodiments, the dosage is, for example, about 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg, or 40 mg / kg. It is understood that the dosage can be 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg, or... The dose is 40 mg / kg or a range consisting of any two of these, such as approximately 0.3 mg / kg-35 mg / kg, 1 mg / kg-35 mg / kg, 3 mg / kg-35 mg / kg, 5 mg / kg-35 mg / kg, 7.5 mg / kg-35 mg / kg, 7.5 mg / kg-30 mg / kg, 15 mg / kg-35 mg / kg, 15 mg / kg-30 mg / kg, or 25 mg / kg-35 mg / kg, etc. In some embodiments, the dose is approximately 7.5 mg / kg-30 mg / kg. In some specific embodiments, the dose is approximately 0.3 mg / kg. In some specific embodiments, the dose is approximately 1 mg / kg. In some specific embodiments, the dose is approximately 3 mg / kg. In some specific embodiments, the dose is approximately 5 mg / kg. In some specific embodiments, the dose is approximately 7.5 mg / kg. In other specific embodiments, the dose is approximately 15 mg / kg. In some other embodiments, the dose is approximately 25 mg / kg. In some other embodiments, the dose is approximately 30 mg / kg. In some other embodiments, the dose is approximately 35 mg / kg.

[0097] In some specific implementation plans, safety follow-ups are conducted for 12 weeks starting from the last dose.

[0098] In some specific embodiments of the invention, the dosing regimen comprises administering four doses of anti-CD40L antibody or its antigen-binding fragment to a patient in need via intravenous infusion at 4-week intervals, each dose being approximately 0.1 mg / kg to 40 mg / kg, for example 0.3 mg / kg to 35 mg / kg, 1 mg / kg to 35 mg / kg, 3 mg / kg to 35 mg / kg, 5 mg / kg to 35 mg / kg, 7.5 mg / kg to 35 mg / kg, 7.5 mg / kg to 30 mg / kg, and 15 mg / kg to 35 mg / kg. , 15mg / kg-30mg / kg or 25mg / kg-35mg / kg, such as approximately 0.1mg / kg, 0.3mg / kg, 0.5mg / kg, 1mg / kg, 5mg / kg, 7.5mg / kg, 10mg / kg, 12.5mg / kg, 15mg / kg, 17.5mg / kg, 20mg / kg, 22.5mg / kg, 25mg / kg, 27.5mg / kg, 30mg / kg, 32.5mg / kg, 35mg / kg, 37.5mg / kg or 40mg / kg. In some embodiments, the four doses are fixed doses or the same doses, wherein the doses are approximately 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg, or 40 mg / kg.

[0099] In some specific embodiments of the invention, the dosing regimen includes administering the same dose of anti-CD40L antibody or its antigen-binding fragment to the patient in need four times at four-week intervals, each dose being approximately 7.5 mg / kg to 30 mg / kg.

[0100] In some specific embodiments of the invention, the dosing regimen includes administering the same dose of anti-CD40L antibody or its antigen-binding fragment to a patient in need at 4-week intervals, with each dose being 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg, 7.5 mg / kg, 15 mg / kg, 25 mg / kg, 30 mg / kg, or 35 mg / kg. In some specific embodiments of the invention, the dosing regimen includes administering the same dose of anti-CD40L antibody or its antigen-binding fragment to a patient in need at 4-week intervals, with each dose being approximately 7.5 mg / kg, approximately 15 mg / kg, or approximately 30 mg / kg.

[0101] In some embodiments of the invention, the patient is biologic-naïve. In an alternative embodiment of the method of the invention, the patient has previously received biologic treatment. In some embodiments of the method of the invention, the patient has received one or more prior treatments, such as treatment for primary Sjögren's syndrome, such as glucocorticoids, hydroxychloroquine, methotrexate, total glucosides of paeony, topical ophthalmic prescription medications (excluding artificial tears, gels, lubricants), or medications with secretory-stimulating properties.

[0102] In some further embodiments of the method of the present invention, the anti-CD40L antibody or its antigen-binding fragment shall not be administered in combination with hepatotoxic drugs.

[0103] The method of preventing or treating individual autoimmune diseases with the anti-CD40L antibody or its antigen-binding fragment of the present invention, administered once every 4 weeks, can increase patient compliance with medication, reduce patient fear of injection, alleviate the patient's physiological and psychological burden, and help maintain long-term remission.

[0104] The anti-CD40L antibody or its antigen-binding fragment of the present invention can be formulated into a dosage form for administration. In some embodiments, the anti-CD40L antibody of the present invention is formulated into a liquid dosage form or a lyophilized dosage form, such as a lyophilized powder for intravenous injection.

[0105] For example, the formulation of the anti-CD40L antibody of the present invention further comprises excipients such as histidine, histidine hydrochloride, sucrose, mannitol, and polysorbate 80, and the pH of the solution of the liquid formulation or lyophilized formulation of the present invention before lyophilization is between 5 and 6, for example, about 5.2. In some embodiments, the liquid formulation of the antibody of the present invention is formulated as follows: 50-100 mg / mL (e.g., 50, 60, 70, 80, 90, or 100 mg / mL) of the anti-CD40L antibody of the present invention or its antigen-binding fragment, 10 mM histidine (composed of 0.12 mg / mL histidine and 1.94 mg / mL histidine hydrochloride), 50.00 mg / mL sucrose, 20.00 mg / mL mannitol, and 0.3 mg / mL polysorbate 80.

[0106] In a second aspect of the invention, a unit dosage form is also provided, comprising: an effective amount of the anti-CD40L antibody of the present invention or its antigen-binding fragment, for example, about 100 mg to 1000 mg, such as 180 mg to 550 mg, 200 mg to 540 mg, 300 mg to 600 mg, 300 mg to 720 mg, 500 mg to 1000 mg, or 500 mg to 800 mg, such as about 180 mg, 300 mg, 540 mg, or 600 mg of the anti-CD40L antibody of the present invention or its antigen-binding fragment. The unit dosage form is intended for administration according to the dosing regimen of the present invention (e.g., the dosing regimen described in the first aspect) to prevent or treat autoimmune diseases. The anti-CD40L antibody of the present invention or its antigen-binding fragment is the anti-CD40L antibody or its antigen-binding fragment as defined in the first aspect.

[0107] The unit dosage form described herein is preferably for parenteral administration, such as for injection, such as intravenous injection (including intravenous infusion) or subcutaneous injection. In some embodiments, the unit dosage form of the present invention is a solution or lyophilized powder, preferably a lyophilized powder. The unit dosage form may be an injection vial, such as a vial, ampoule, or pre-filled syringe, containing a solution or lyophilized powder of CD40L antibody, preferably a lyophilized powder. More preferably, the unit dosage form described herein is a vial, such as a vial, containing a lyophilized powder of CD40L antibody or its antigen-binding fragment for parenteral administration.

[0108] In some embodiments, the formulation containing anti-CD40L antibody or its antigen-binding fragment, such as a lyophilized powder, is reconstituted prior to administration. In some embodiments, the required infusion volume is calculated based on the planned drug dosage and the solution, and the calculated volume of the reconstituted drug is diluted in an intravenous infusion bag, for example, containing 0.9% sodium chloride injection. In some embodiments, the final concentration is prepared in the range of approximately 1.0-15.0 mg / ml.

[0109] In a third aspect of the invention, there is also a kit containing an effective amount of the anti-CD40L antibody or its antigen-binding fragment described herein, such as one or more unit doses of the anti-CD40L antibody or its antigen-binding fragment; preferably, it also includes a packaging insert printed with instructions for the use of the anti-CD40L antibody or its antigen-binding fragment in an individual for the prevention or treatment of autoimmune diseases. In some specific embodiments, the prevention or treatment of autoimmune diseases comprises administering the anti-CD40L antibody or its antigen-binding fragment of the invention according to the dosing regimen described in the invention, such as the dosing regimen described in the first aspect. The anti-CD40L antibody or its antigen-binding fragment of the invention is the anti-CD40L antibody or its antigen-binding fragment as defined in the first aspect.

[0110] In some implementations, the kit is in the form of a drug dosage unit, thereby providing a single-dose dosing unit according to the dosing regimen or drug administration interval.

[0111] In some embodiments, the anti-CD40L antibody or its antigen-binding fragment is contained in a single-dose delivery unit, wherein the single-dose delivery unit may contain an effective amount of the anti-CD40L antibody or its antigen-binding fragment.

[0112] In some embodiments, a single-dose delivery unit comprising the anti-CD40L antibody or its antigen-binding fragment may contain approximately 0.1 mg / kg to 40 mg / kg, such as 0.3 mg / kg to 35 mg / kg, 1 mg / kg to 35 mg / kg, 3 mg / kg to 35 mg / kg, 5 mg / kg to 35 mg / kg, 7.5 mg / kg to 35 mg / kg, 7.5 mg / kg to 30 mg / kg, 15 mg / kg to 35 mg / kg, 15 mg / kg to 30 mg / kg, or 25 mg / kg. -35 mg / kg, for example, approximately 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg or 40 mg / kg of the said antiCD40L antibody or its antigen-binding fragment.

[0113] In some embodiments, the kit contains four single-dose delivery units, each of which delivers the same dose.

[0114] In some embodiments, the kit comprises four single-dose delivery units, each containing the same dose of the anti-CD40L antibody or its antigen-binding fragment, wherein the dose is approximately 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg, or 40 mg / kg.

[0115] In some embodiments, the kit contains four single-dose dosing units, each of which is the same dose and the dose is approximately 7.5 mg / kg to 30 mg / kg, such as 7.5 mg / kg, 15 mg / kg, or 30 mg / kg.

[0116] In a fourth aspect of the invention, the use of the anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit of the present invention for the prevention or treatment of autoimmune diseases is provided. In a fifth aspect of the invention, the use of the anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit of the present invention for the prevention or treatment of autoimmune diseases is provided. In a sixth aspect of the invention, the use of the anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit of the present invention in the preparation of a medicament for the prevention or treatment of autoimmune diseases is provided. In some specific embodiments, the prevention or treatment of autoimmune diseases includes administering the anti-CD40L antibody or its antigen-binding fragment of the present invention using the administration regimen described in the present invention, such as the administration regimen described in the first aspect. The anti-CD40L antibody or its antigen-binding fragment of the present invention is the anti-CD40L antibody or its antigen-binding fragment as defined in the first aspect.

[0117] In some embodiments of all aspects described herein, the autoimmune disease is Sjögren's syndrome or systemic lupus erythematosus. In some embodiments, Sjögren's syndrome is primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

[0118] In some embodiments of all aspects described herein, the anti-CD40L antibody or its antigen-binding fragment, unit dosage form or kit of the present invention, when used within the dosing regimen of the present invention for the prevention or treatment of autoimmune diseases, exhibits good safety and tolerability, for example, having one or more of the following properties:

[0119] (i) No SAE, AESI, or TEAE leading to death;

[0120] (ii) No unexpected serious safety signals;

[0121] (iii) It can reduce the formation of germinal centers and antibody production;

[0122] (iv) It does not increase the risk or severity of infection in the subjects; and / or

[0123] (v) The incidence and severity of various TRAEs, including upper respiratory tract infections, showed no clear dose-dependent trend with respect to the dose of the anti-CD40L antibody or its antigen-binding fragment of the present invention.

[0124] In some embodiments of all aspects described herein, the anti-CD40L antibody or its antigen-binding fragment, unit dosage form or kit of the present invention can effectively prevent or treat autoimmune diseases within the dosing regimen of the present invention, for example achieving one or more or all of the following effects:

[0125] (i) It has an ESSDAI-improving effect;

[0126] (ii) Improved ESSPRI scores, for example, better than Dazodalibep;

[0127] (iii) Significantly downregulates serum IgG, IgA, and / or RF levels, for example based on CRESS labeling, and the downregulation is clinically significant; and / or

[0128] (iv) Effectively inhibits the ability of B cells to produce antibodies.

[0129] In some embodiments of all aspects described herein, the anti-CD40L antibody or its antigen-binding fragment, unit dosage form, or kit of the present invention, within the dosing regimen of the present invention, exhibits low immunogenicity, high drug safety, and / or high drug stability in vivo, for example, having one or more of the following properties.

[0130] (i) The administration does not produce ADA, or produces very little ADA;

[0131] (ii) A long half-life, for example, ranging from about 16 to 25.2 days, thus supporting dosing at intervals of at least Q4W or longer.

[0132] The dosing regimens described herein (including the active agent CD40L antibody or its antigen-binding fragment, dosage, administration method, dosing interval, number of administrations, or duration of administration, etc.) and any of their features or indications can be applied to any aspect of the present invention, such as aspects one through six as described above. Furthermore, any embodiments, technical features, definitions, and any combinations thereof described herein with respect to one aspect can also be applied to other aspects, such as aspects one through eight as described above. For example, any embodiments, technical features, definitions, and any combinations thereof described herein with respect to the first aspect can be equivalently applied to aspects two through six as described above.

[0133] Any of the embodiments, technical features, and definitions described herein can be combined in any way to form embodiments that are not directly described in the specification but are consistent with the spirit of the present invention, and these embodiments are also included within the scope of the present invention. Example

[0134] The following embodiments are provided to illustrate the present invention. These embodiments are merely illustrative and do not limit the scope of the invention and the appended claims in any way. Similarly, the invention is not limited to any particular preferred embodiment described herein. Those skilled in the art can make modifications to the invention without departing from its spirit and scope.

[0135] Example 1. Preparation and purification of CD40L antibody and preclinical efficacy study

[0136] According to PCT application number PCT / CN2024 / 1090122, an antibody Hz33B9C5 specifically binding to CD40L was obtained, and its functions and activities were tested. This antibody has the heavy chain sequence of SEQ ID NO:6 and the light chain sequence of SEQ ID NO:12. PCT application number PCT / CN2024 / 1090122 is hereby incorporated by reference in its entirety.

[0137] 1.1 Expression and purification of antibodies in ExpiCHO cells:

[0138] The sequence of Hz33B9C5 was cloned into the pcDNA3.1 vector for later use. ExpiCHO cells were harvested, and the cell density was measured to be 8.0–10 x 10⁻⁶ cells / cm². 6 The viability of cells should be between 95-99%, and the cell density should be adjusted to 6.0 x 10⁶ cells / mL. 6Viable cells / mL. Take an appropriate-sized container and add Fectamine (source: Gibco) transfection buffer at 8% of the cell volume. Add the corresponding amount of plasmid at 0.8 μg / mL for cell transfection. Filter the transfection buffer containing DNA plasmid through a 0.22 μm filter to sterilize it into a new clean container. Add FectPro (source: Polyplus) transfection reagent at a ratio of 3.2 μL / mL to the filtered mixture and let it stand for 10 min. Quickly add the complex formed by the transfection reagent and plasmid DNA to the cells, gently shaking the flask while adding. Incubate at 36.5℃ on a shaker with 8% CO2. After 18-22 h of culture, add Enhancer (source: Gibco) at 6 μL / mL of cell volume and Feed (source: Gibco) at 300 μL / mL of cell volume. On day 7, collect the cell supernatant. Purify the protein using affinity chromatography to obtain the final antibody molecule. The purified antibody solution was centrifuged in a 15 ml ultrafiltration centrifuge tube at 4000 rpm for 10 minutes. The protein was then diluted with PBS and centrifuged again at 4000 rpm for 10 minutes. This process was repeated several times, changing the buffer solution each time. The antibodies were pooled after each buffer change, and the antibody concentration was measured. The components of the monoclonal antibody were further qualitatively analyzed using liquid chromatography-mass spectrometry (LC-MS) and size exclusion chromatography (SEC), and quantified using a UV-Vis spectrophotometer.

[0139] 1.2 Results of preclinical studies of the research drug

[0140] PCT application number PCT / CN2024 / 1090122 (in full text incorporated herein by reference) investigated and tested the activity, efficacy, pharmacokinetics and toxicology of Hz33B9C5.

[0141] 1.2.1 Preclinical efficacy studies

[0142] In vitro affinity studies showed that HZ33B9C5 exhibited a strong affinity (4.68 pM) for the CD40L protein in humans / cynomolgus monkeys (human CD40L and cynomolgus monkey CD40L share the same extracellular sequence). HZ33B9C5 did not bind to CD40L proteins in rats, mice, or rabbits. Furthermore, HZ33B9C5 did not bind to other human TNF family proteins such as TNF-alpha and OX40L, suggesting a low off-target risk for HZ33B9C5.

[0143] In vitro pharmacodynamic results showed that HZ33B9C5 could inhibit the binding of CD40L protein to CD40 protein and block the activity of CD40L and CD40 binding on the membrane. The inhibitory and blocking effects were comparable to those of INX-021, a drug that targets CD40L, indicating that HZ33B9C5 has clear target selectivity and significant inhibitory or blocking effects on the binding of CD40L and CD40.

[0144] The T-cell-dependent activation pathway of B cells involves antigen endocytosis and presentation of CD4 by the major histocompatibility complex (MHCII). + After T cells recognize the cells, B cells bind to the CD40L ligand via the CD40 receptor on their surface, thereby differentiating into memory B cells and plasma cells. HZ33B9C5 can inhibit the proliferation and differentiation of B cells into plasma cells in vitro, suggesting that HZ33B9C5 can inhibit B cell proliferation and differentiation by blocking the binding of CD40L to CD40.

[0145] HZ33B9C5 has an affinity of 10 for FcRn. -8 The affinity for FcγRI is on the order of M, and the affinity is in the range of 10. -7 The binding was on the order of M and showed no significant binding with FcγRIIa, FcγRIIIa(V158), FcγRIIb, FcγRIIIb, FcγRIIIa(F158), and C1q. No ADCC or CDC effects were detected in HZ33B9C5.

[0146] In a mouse model of experimental autoimmune encephalomyelitis (EAE), the HZ33B9C5 dosage groups showed slower weight loss and score increase compared to the h-IgG-10 mg / kg group, and a decrease in anti-MOG antibody titer. Compared to the h-IgG-10 mg / kg group, the HZ33B9C5 dosage groups exhibited dose-related inhibition of EAE pathogenesis. In a mouse model of KLH-induced in vivo T-cell-dependent antibody response, compared to the h-IgG-3 mg / kg group, all HZ33B9C5 groups showed a decrease in anti-KLH antibody titer, and the inhibitory effect of HZ33B9C5 on anti-KLH antibody production was dose-related. Pharmacokinetic studies further indicated that after a single dose of HZ33B9C5, exposure to HZ33B9C5 increased linearly within the dose range of 0.3–3 mg / kg.

[0147] Safety pharmacology studies showed that intravenous administration of HZ33B9C5 at doses of 50, 100, and 200 mg / kg every 2 weeks for 13 consecutive weeks had no significant effect on blood pressure, ECG parameters in lead II, or respiratory rate in any of the cynomolgus monkeys.

[0148] 1.2.2 Preclinical Pharmacokinetic Studies

[0149] Pharmacokinetic results from single-dose administration showed that after a single intravenous injection of HZ33B9C5 at doses of 3, 10, and 30 mg / kg in cynomolgus monkeys, the exposure level increased proportionally with increasing dose, and no significant differences were observed in the major pharmacokinetic parameters between males and females in any group. The individual confirmatory positivity rates for HZ33B9C5 in the low, medium, and high dose groups were 100% (6 / 6), 83.3% (5 / 6), and 50% (3 / 6), respectively, with corresponding titer ranges of 3–1834, 2–1732, and 7–702. The individual neutralizing antibody positivity rates for HZ33B9C5 were 100.0% (6 / 6), 83.3% (5 / 6), and 50.0% (3 / 6), respectively, with corresponding neutralizing activity titer ranges of 2–1471, 1–1279, and 1–2448, respectively.

[0150] Multiple-dose toxicokinetics studies of HZ33B9C5 showed that in cynomolgus monkeys, intravenous injections of HZ33B9C5 every two weeks at doses between 50 and 200 mg / kg resulted in... max and AUC 0-t The dosage increased proportionally to the dose increase, and no sex difference was observed in exposure across dosage groups. Compared to the first dose, the low-dose group showed a tendency to accumulate at the third administration, while no significant accumulation was observed in the medium and high-dose groups. Compared to the first dose, the low and medium-dose groups showed a tendency to accumulate at the sixth administration, while no significant accumulation was observed in the high-dose group. Pre-administration D levels in animals of each dosage group during the experiment were... 29 D 71 D 99 and D 113 All anti-drug antibody test results were negative.

[0151] 1.2.3 Preclinical Toxicology Studies

[0152] The no-observed-adverse-effect level (NOAEL) for cynomolgus monkeys administered intravenously for 13 consecutive weeks was 100 mg / kg. At this dose, after the 6th administration, the HZ33B9C5 exposure levels in female and male monkeys were 496,704 h·μg / mL and 670,164 h·μg / mL, respectively.

[0153] One female animal in the 200 mg / kg group was euthanized due to near-death experience; the cause of death was highly suspected to be hemorrhagic death due to chronic gastrointestinal bleeding. Since this animal was in the high-dose group, a link to HZ33B9C5 could not be ruled out. Furthermore, the main toxicities observed at the 200 mg / kg dose were a slight decrease in peripheral blood erythrocyte parameters (red blood cell count (RBC), hemoglobin content (HGB), and hematocrit (HCT)), a decrease in B lymphocytes, a decrease in spleen weight and coefficient, and a decrease in splenic lymphoid follicles / lymph node lymphocytes. In the 100 and 50 mg / kg groups, similar or slightly milder lesions were observed in the aforementioned immune-related organs (spleen, lymph nodes, etc.), along with a slight decrease in peripheral blood erythrocyte parameters (RBC, HGB, and HCT) and a decrease in B lymphocytes. Based on the mechanism of action of the HZ33B9C5 molecule, the decrease in spleen weight, the reduction in B lymphocytes, and the decrease in lymphocytes in the spleen and lymph nodes are considered to be due to the amplified pharmacological effects of HZ33B9C5. Combined with the negative results of pigment staining in the spleen and liver, these changes are considered to be not caused by erythrocyte fragmentation and no related organic changes were found; therefore, they are considered non-adverse reactions. The zona fasciculata of the adrenal gland primarily secretes glucocorticoids; no abnormal changes related to glucocorticoid secretion were observed in any dose group during the experiment, therefore, this is also considered a non-adverse reaction.

[0154] Based on the combined results of non-clinical studies, mechanism of action, and route of administration, HZ33B9C5 has a clearly defined target and a well-defined mechanism of action. HZ33B9C5 exhibits linear pharmacokinetic characteristics in cynomolgus monkeys, has a long half-life, and poses no risk of hemolysis or cytokine release in vitro.

[0155] Example 2. Preparation and administration of the formulation

[0156] The formulation for the injectable antibody preparation is as follows: 100 mg / mL of the anti-CD40L antibody HZ33B9C5 of the present invention, 10 mM histidine (composed of 0.12 mg / mL histidine and 1.94 mg / mL histidine hydrochloride), 50.00 mg / mL sucrose, 20.00 mg / mL mannitol, 0.3 mg / mL polysorbate 80, pH 5.2; the lyophilized powder preparation for injection of the antibody of the present invention was prepared according to this formulation (the stock solution was prepared first, and then filled and lyophilized), each 20R vial containing 600 mg of sterile lyophilized powder of the anti-CD40L antibody HZ33B9C5 of the present invention.

[0157] The placebo contains no active ingredient but only excipients, and the excipient composition and pH are consistent with those of the injectable antibody formulation.

[0158] The antibody preparation and excipients are both white to pale yellow lumpy loose bodies. The reconstituted sample is clear to slightly opalescent, colorless to slightly yellow liquid, and free of foreign matter.

[0159] Before injection, the antibody preparation / placebo needs to be reconstituted and further diluted, and appropriate aseptic techniques must be used during the procedure.

[0160] 1. Before preparation, bring the vial to room temperature and let it stand for at least 5 minutes. It can be left at room temperature for up to 24 hours before dilution. Before preparation, confirm that the antibody preparation / placebo is a white to pale yellow lumpy, loose substance that becomes clear after reconstitution, without any turbidity, precipitation, or other quality issues. From a microbiological point of view, this product must be used immediately after dilution and must not be frozen.

[0161] 2. Before dilution, calculate the planned drug dosage based on the subject's weight and the required infusion volume based on the solution. Dilute the calculated volume of the reconstituted HZ33B9C5 / placebo drug in an intravenous infusion bag containing 0.9% sodium chloride injection. Prepare a final concentration range of 1.0–15.0 mg / ml. Before administration, the drug should be shaken until completely dissolved, with no visible solids, and the liquid should be clear to slightly opalescent, colorless to slightly yellow, and free of foreign matter. Do not shake vigorously to avoid foaming. If a large amount of foam forms, allow the infusion bag to stand until the foam disappears. Ensure the homogeneity of the drug in the infusion bag. The administration time should not be less than 30 minutes. If the vial and / or intravenous infusion bag are refrigerated, they must be brought to room temperature before use; avoid mixing with other drugs; avoid intravenous bolus injection.

[0162] During administration, each subject should receive a single, constant-rate infusion of the corresponding drug. The infusion device must be equipped with an online filter of 0.2–5.0 μm. Depending on the local injection site and the subject's response, the investigator may adjust the infusion rate, temporarily discontinue the drug, or stop the study drug administration as needed, and record the changes. Record the start and end times of administration. If the diluted antibody / placebo HZ33B9C5 cannot be used immediately, store it at 2°C–8°C for up to 8 hours, or at room temperature (20°C–25°C) for up to 6 hours (6 hours including the administration time), protecting it from light.

[0163] Example 3. Clinical Study

[0164] The aim of this clinical study is to conduct a Phase 1 clinical trial to evaluate the pharmacokinetic characteristics, tolerability, and safety of HZ33B9C5 in patients with primary Sjögren's syndrome.

[0165] This study was conducted in accordance with the scientific and ethical principles of the Declaration of Helsinki and Good Clinical Practice (GCP), as well as national regulations. Prior to the study, the clinical trial protocol, case report forms, and informed consent forms were all approved by the ethics committee. Informed consent and informed consent forms were used to protect the rights, safety, and health of the participants, while strict inclusion and exclusion criteria were established to ensure the ethical principles of the research.

[0166] Before any research-related procedures are carried out, the subject or their legal representative and the researcher must sign an informed consent form that has been approved by the IEC / IRB. The researcher will provide a copy of the signed informed consent form to the subject or their legal representative, and the original will be kept by the researcher.

[0167] 3.1 Research Objectives

[0168] -Main purpose:

[0169] To observe the safety and tolerability of HZ33B9C5 with multiple doses in pSS subjects.

[0170] -Secondary objective:

[0171] To evaluate the PK characteristics and immunogenicity of HZ33B9C5 after multiple doses in pSS subjects.

[0172] -Other purposes:

[0173] To evaluate the effectiveness of multiple doses of HZ33B9C5 in treating pSS.

[0174] 3.2 Study Endpoints

[0175] ●Main endpoint:

[0176] 1. Adverse events (AEs) and serious adverse events (SAEs) following multiple doses of HZ33B9C5 in pSS subjects.

[0177] ●Secondary endpoint:

[0178] 1. PK Parameters: Concentration data will be subject to descriptive statistics, including but not limited to the area under the plasma drug concentration-time curve (AUC) and the maximum serum concentration of drug (C). max The following parameters are also considered: clearance (CL), volume of distribution (V), and elimination half-life (t). 1 / 2 );

[0179] 2. Immunogenicity: The production of anti-drug antibodies (ADA).

[0180] ●Other destinations:

[0181] 1. EULAR Sjögren's syndrome disease activity index (EULAR) of pSS subjects Sjögren's syndrome disease activity index (ESSDAI) and EULAR Sjögren's syndrome patient self-report index (EULAR). The change in the Syndrome Patient Reported Index (ESSPRI) score from baseline at week 16 and its change over time;

[0182] 2. Changes in serum antinuclear antibodies (ANA), immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin A (IgA), and rheumatoid factor (RF) titers in pSS subjects from baseline at week 16 and over time;

[0183] 3. Schirmer I test, total salivary gland flow rate (unstimulated, stimulated), and changes from baseline in pSS subjects at week 16;

[0184] 4. Changes in serum chemokine CXC motif ligand 13 (CXCL13) from baseline at week 16.

[0185] 3.3 Basis for Dosage Selection

[0186] The initial recommended dose for patients with Sjögren's syndrome (HZ33B9C5) is 7.5 mg / kg (Q4W), 15 mg / kg (Q4W), and 30 mg / kg (Q4W).

[0187] Basis for selecting multiple-dose administration in patients with Sjögren's syndrome:

[0188] Basis for selecting starting dose:

[0189] In a mouse KLH-induced T-cell-dependent antibody response model, a single dose of 3 mg / kg significantly inhibited the production of anti-KLH antibodies, with an average plasma concentration of 16.7 μg / mL at this dose. Based on human PK simulations, the average plasma concentration of HZ33B9C5 Q4W administered at 7.5 mg / kg in the first cycle was 59.7 μg / mL, which is greater than 16.7 μg / mL.

[0190] Basis for selecting the maximum tolerated dose:

[0191] Human pharmacokinetic simulations showed the steady-state C at 30 mg / kg Q4W. max Approximately 874.612 μg / mL, steady-state AUC ss The steady-state C2 after Q2W administration of approximately 223778 μg·h / mL in cynomolgus monkey toxicology NOAEL of 100 mg / kg. max The concentration was 3472 μg / mL, which is the steady-state concentration in the human body. max 3.9 times; steady-state AUC ss The concentration was 583433 μg·h / mL, which is the steady-state AUC in the human body. ss 2.6 times that.

[0192] 3.4 Research Design

[0193] 3.4.1 Treatment Plan

[0194] The antibody injection formulation and placebo prepared as in Example 2 were used in this study.

[0195] This study is a dose-escalation study of HZ33B9C5 in subjects with primary Sjögren's syndrome. It is a randomized, double-blind, dose-escalation clinical study exploring the safety and tolerability of multiple doses in pSS subjects.

[0196] This study was designed as a randomized, double-blind, multiple-dose, dose-escalation, placebo-controlled study. The study process included: a screening period (6 weeks before dosing), a dosing period (12 weeks), and a safety follow-up period (12 weeks).

[0197] The study was divided into three dosage groups and planned to enroll 30 pSS subjects. This study marks the first time this product has been administered to pSS subjects. Based on the results of the Phase I single-dose escalation trial in healthy individuals, the currently proposed escalation doses are as follows:

[0198] ●Dose Group 1: 7.5 mg / kg Q4W (N=10)

[0199] ●Dose Group 2: 15 mg / kg Q4W (N=10)

[0200] ●Third dose group: 30 mg / kg Q4W (N=10)

[0201] The treatment plan is shown in Table 1:

[0202] Table 1. Treatment Plan

[0203] The study design diagram is shown in Figure 1.

[0204] Eligible pSS subjects were admitted to the study ward before administration and randomly assigned in an 8:2 ratio to receive either HZ33B9C5 or placebo intravenously, once every four weeks for a total of four doses. The first and fourth doses were to be administered in the hospital, while the remaining doses required observation at the center for at least 60 minutes after administration. Subjects could leave after the observation period ended without adverse reactions. Based on the safety and tolerability data from the previous dose group, a decision was made on whether to escalate to the next dose or adjust the actual dose for the next dose group. If necessary, the dose escalation regimen would be adjusted based on a comprehensive assessment of safety, pharmacokinetic / pharmacodynamic (PK / PD) data.

[0205] All subjects in each dose group should be observed for at least 4 weeks after the first dose. The dose escalation study may continue if the safety assessment committee assesses that there is no risk to safety and tolerability when escalating to the next dose and agrees to explore the next dose group. If the dose escalation cessation criteria are met, the study for that dose group should be discontinued, and the need to explore other dosing methods should be assessed. Each dose group should be followed up safely for up to 12 weeks after the last dose in the last subject.

[0206] Similarly, the formulation of the present invention was administered to healthy individuals at doses of 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 7.5 mg / kg, 15 mg / kg, 25 mg / kg, and 35 mg / kg, respectively.

[0207] Dose escalation termination criteria: If any of the following criteria are met, the exploration of this dose and above doses shall be suspended.

[0208] ● Within the same dose group, any subject experienced a life-threatening SAE related to the study drug, or two subjects experienced a SAE related to the study drug.

[0209] ● Within the same dose group, ≥50% of subjects experienced anemia-related adverse events, defined as grade 2 or higher anemia in subjects with normal baseline or a decrease in Hb of ≥20% from baseline in subjects with abnormal baseline;

[0210] ● Within the same dose group, two subjects experienced grade ≥3 adverse events, which were considered to be related to the study drug;

[0211] ●Any subject experiences a thrombotic event related to the study drug;

[0212] ●Any subject experiences bleeding in an internal organ that is considered to be related to the study drug;

[0213] Once the dose escalation termination criteria are met, dosing should be discontinued for the remaining subjects in that dose group, and the exploration of doses above that dose group should be terminated. Dosing for the remaining subjects in that dose group may not continue unless the safety assessment committee, after analysis, deems the safety and tolerability of the dose exposure acceptable. Simultaneously, the safety assessment committee must assess whether to continue exploring the intermediate dose between this dose and the previous dose to determine the maximum tolerated dose.

[0214] Single subject intolerance criteria:

[0215] ● The subject experienced an adverse event of grade 3 or higher, and it was determined to be related to the study drug;

[0216] ●The subject experienced any grade of thromboembolic event or bleeding event in internal organs;

[0217] ● If a subject experiences an adverse event (AE) that the investigator deems relevant to the investigation drug, and after discussion with the sponsor, it is deemed necessary to discontinue the investigation at that dose, indicating that the subject is intolerant to that dose.

[0218] If a single subject is found to be intolerant, the remaining doses for that subject should be discontinued, and whether to resume dosing should be assessed by the Safety Data Committee.

[0219] 3.4.2 Previous and concomitant treatments

[0220] In this study, it is necessary to record the medications (including prescription or over-the-counter drugs, and traditional Chinese medicine) related to the primary disease that the subjects received in the past year, as well as any special treatments received since the subjects were diagnosed (such as: biologic therapy, hormone pulse therapy, human immunoglobulin therapy, plasma exchange, hemodialysis, etc.). Records should include: the date of administration (including start and end dates), dosage, and frequency.

[0221] Participants are permitted to receive appropriate treatments approved during the study, including but not limited to:

[0222] 1. Medications used to treat adverse events (AEs) with the consent of the investigator and the sponsor;

[0223] 2. Medications for treating the primary disease that can be used concurrently during the study period must exclude drugs prohibited by the exclusion criteria;

[0224] 3. Vitamins, calcium supplements, and oral contraceptives;

[0225] 4. Vaccines (excluding live vaccines) that have been approved for the prevention of infection or infection-related diseases.

[0226] Subjects are prohibited from receiving the following treatments during the study:

[0227] ●Study any medication other than the drug, unless it is a permitted concomitant treatment.

[0228] ● Exclude prohibited drugs from the criteria.

[0229] It is important for researchers to review every medication (prescription and over-the-counter) that subjects received before the start of the study and at each study visit.

[0230] ● During each visit, you must ask the subject about any new medications they are taking;

[0231] ● In order to reduce the risk of adverse drug interactions, all measures must be taken to limit the number of combined medications that are truly necessary;

[0232] ● Avoid taking hepatotoxic drugs (i.e., drugs that are warned of being hepatotoxic in the product information leaflet) during the course of treatment. Each potentially hepatotoxic drug can be reviewed by searching the website www.livertox.nih.gov.

[0233] 3.5 Study population

[0234] Selection criteria

[0235] All participants must meet the following criteria to be included in this study.

[0236] 1. Understand and sign the informed consent form;

[0237] 2. Age ≥ 18 years old, male or female;

[0238] 3. Body Mass Index (BMI) between 18.0 and 28.0 kg / m² 2 Between (including both ends);

[0239] 4. Meets the 2016 American College of Rheumatology (ACR) / European Congress of Rheumatology (EULAR) classification diagnostic criteria for Sjögren's syndrome;

[0240] 5. Positive for sicca syndrome A autoantibodies (SSA) and / or sicca syndrome B autoantibodies (SSB);

[0241] 6. Under unstimulated conditions, the total salivary gland flow rate is >0 ml / min;

[0242] 7. IgG > Upper Limit of Normal Value (ULN) or RF > ULN;

[0243] 8. The following medications (alone or in combination) may be used prior to enrollment and should remain stable throughout the study:

[0244] - Glucocorticoids, with a dose not exceeding 10 mg / day equivalent to prednisone, must be stable for at least 2 weeks before enrollment;

[0245] Hydroxychloroquine (maximum dose 0.4 g / day) should be stabilized for at least 4 weeks before enrollment;

[0246] - Methotrexate, the dose should not exceed 10 mg / week, and the patient should be stable for at least 8 weeks before enrollment;

[0247] -Total paeoniflorin, stabilized for at least 1 month before enrollment;

[0248] - Topical ophthalmic prescription medications (excluding artificial tears, gels, and lubricants) must be stable for at least 12 weeks prior to enrollment.

[0249] Exclusion criteria

[0250] Subjects who meet any of the following criteria are ineligible to participate in this clinical study:

[0251] 1. Individuals with allergies to any component of HZ33B9C5, those with allergic diseases, or those with an allergic constitution;

[0252] 2. Those who cannot tolerate frequent intravenous punctures;

[0253] 3. The diagnosis is secondary Sjögren's syndrome, or the subject's clinical symptoms (or abnormal laboratory tests) need to be explained by other connective tissue diseases (such as systemic lupus erythematosus, mixed connective tissue disease, etc.);

[0254] 4. Subjects who received the following medications prior to randomization:

[0255] a) Those who have used B-cell clearance therapy, such as CD20 monoclonal antibody, within 1 year prior to randomization (those who have previously used B-cell clearance therapy are not suitable for inclusion in this study unless there is evidence that peripheral blood B cells have recovered).

[0256] b) Those who have used cyclophosphamide, other biological agents such as belimumab, teltascept, etc. within the 6 months prior to randomization;

[0257] c) Individuals who received plasma exchange, intravenous human immunoglobulin therapy, or hormone pulse therapy within the previous 6 months;

[0258] d) Patients taking other immunosuppressants (other than those permitted under inclusion criterion 8), such as mycophenolate mofetil, cyclosporine A, tacrolimus, ellamod, leflunomide, etc., must have discontinued these medications for less than 8 weeks prior to randomization;

[0259] 5. Those who have participated in other drug clinical trials within the past 1 month, or whose time since the last clinical trial is less than 5 half-lives of the investigational drug (whichever is longer);

[0260] 6. An infection requiring systemic drug treatment has been present within the preceding 30 days;

[0261] 7. History of receiving live or attenuated vaccines within the month prior to randomization;

[0262] 8. Those who have donated blood or lost ≥400ml of blood within the past 3 months will be screened;

[0263] 9. Individuals who are positive for any of the following serological virological tests: human immunodeficiency virus antibody (HIV-Ab), rapid plasma reagin (RPR), hepatitis C virus antibody (HCV-Ab), hepatitis B virus (HBV) surface antigen, hepatitis B e antigen (HBeAg), or hepatitis B core antibody (HBcAb).

[0264] 10. There is clinical or imaging evidence that the subject has active tuberculosis, or there is evidence that the subject is in the latent period of tuberculosis, such as: those who are positive for the T-SPOT test, or those who have been in a closed space for several days with a confirmed active tuberculosis patient.

[0265] 11. Individuals with a positive fecal occult blood test at screening. If the researchers determine that the result is not caused by gastrointestinal bleeding, a repeat test can be conducted during the screening period. If gastrointestinal bleeding is confirmed to be absent, inclusion in this study can be considered.

[0266] 12. The following clinically significant diseases are present:

[0267] a) Individuals with a history of chronic congestive heart failure and NYHA class IV heart function; or those with an echocardiographic ejection fraction of less than 50%.

[0268] b) Those who have had a history of myocardial infarction, acute coronary syndrome, or pulmonary embolism; or those who have undergone coronary revascularization.

[0269] c) Those with severe arrhythmias requiring Class Ia or III antiarrhythmic drug therapy; those with sick sinus syndrome, type II or III atrioventricular block, and who have not yet had a pacemaker implanted.

[0270] d) Individuals whose electrocardiogram during the screening period shows a QTcB interval ≥450ms, or who have a history of QTc interval prolongation;

[0271] e) Those with a history of peptic ulcers or gastrointestinal bleeding;

[0272] f) Other serious, uncontrolled diseases (including but not limited to) cardiovascular diseases, neuromuscular diseases, blood diseases, respiratory diseases, digestive diseases, metabolic diseases, neurological or mental diseases, etc.

[0273] 13. During screening, subjects with the following abnormal laboratory test results:

[0274] a) Individuals whose urinalysis shows proteinuria (++) or higher and who are deemed unsuitable for enrollment by the researchers;

[0275] b) White blood cell count (WBC) <2.0 × 10⁻⁶ 9 / L, or absolute neutrophil count <1.0×10 9 / L, hemoglobin (Hb) <90g / L, platelet count (PLT) <75×10 9 / L;

[0276] c) Alanine aminotransferase (ALT) or aspartate aminotransferase (AST) ≥ 2 times the upper limit of normal;

[0277] d) Serum total bilirubin ≥ 1.5 times the upper limit of normal;

[0278] e)eGFR<90ml / min / 1.73m 2 Subjects with eGFR ≤ 60 < 90 ml / min / 1.73 mcg were generally excluded from this study. 2 Subjects deemed suitable for inclusion in this study by the researchers (e.g., those with decreased renal function related to the primary disease) may be included in this study.

[0279] 14. Patients with concurrent pulmonary interstitial fibrosis, or those requiring combined anti-fibrotic drug therapy, or those with abnormal lung function who are deemed unsuitable for enrollment by the investigator;

[0280] 15. Medications that cause dry mouth / dry eyes must be used during the research process;

[0281] 16. Pregnant or breastfeeding women;

[0282] 17. History of concurrent malignant tumors;

[0283] 18. Subjects who are unwilling to use contraception as required by the protocol, and who are women (or men) of reproductive age who plan to become pregnant (or donate sperm) during the study and within 6 months after the last dose, and who are unwilling to take reliable contraceptive measures.

[0284] 19. Long-term use of estrogen replacement therapy prior to screening or other conditions deemed by the investigator as having a high risk of hypercoagulability, such as poor glycemic control or nephrotic syndrome;

[0285] 20. Individuals who have been taking anticoagulants continuously or intermittently prior to screening, or who have been assessed by the investigator as having a bleeding risk.

[0286] 21. Underwent major surgical treatment within 8 weeks prior to screening, or plans to undergo surgical treatment during the study period;

[0287] 22. Individuals with a history of drug abuse, drug dependence, or a positive drug screening result within the 12 months prior to screening;

[0288] 23. Individuals with a history of neuropsychiatric disorders or other individuals deemed unsuitable to participate in this clinical trial by the researchers.

[0289] 3.6 Effectiveness Assessment

[0290] 3.6.1 Related research, thematic studies and functional imaging

[0291] Lacrimal gland function research

[0292] In this study, the Schirmer I test was used to assess the lacrimal gland function of the subjects. The Schirmer I test was conducted without anesthesia. Schirmer test strips (5mm×35mm or 5mm×30mm) were inserted into the conjunctival sac at the junction of the outer and middle 1 / 3 of the lower eyelid with the tip folded inward. The length of the test strip soaked with tear fluid within 5 minutes was measured and recorded as the result.

[0293] Salivary gland function research

[0294] This study used total salivary gland flow rate under both unstimulated and stimulated conditions to assess salivary gland function. The assessment was performed at a fixed time of day whenever possible. For unstimulated salivary flow rate measurement, subjects were prohibited from eating, drinking alcohol, or smoking for 60 minutes prior to saliva collection, and were also prohibited from taking secretagogues or foods that promote secretion on the day of collection. Saliva was collected from the subject over a 5-minute period, and the collection time for each subject at each visit point should be as consistent as possible. For stimulated salivary flow rate measurement, citric acid (or a centrally provided stimulant) was applied to the saliva before collection, and the time was measured for 5 minutes. The volume of saliva collected (in milliliters) divided by the collection time gave the subject's stimulated salivary flow rate.

[0295] 3.6.2 Indicators related to efficacy evaluation

[0296] This study will explore the preliminary efficacy of pSS in subjects.

[0297] 3.6.2.1 ESSDAI (EULAR Sjögren's Disease Activity Index) Syndrome Disease Activity Index (ESSDAI) score

[0298] The ESSDAI score is the EULAR Disease Activity Index for Sjögren's syndrome, which assesses damage to 12 systems throughout the body associated with pSS. Each indicator is scored according to its disease activity level multiplied by its weight. The final ESSDAI score is the sum of the scores for all affected systems. An ESSDAI < 5 indicates low disease activity, 5 ≤ ESSDAI ≤ 13 indicates moderate disease activity, and an ESSDAI ≥ 14 indicates high disease activity.

[0299] In the registration clinical studies developing the pSS indication, the change in ESSDAI score from baseline was used as the primary efficacy endpoint for assessment.

[0300] 3.6.2.2 ESSPRI (EULAR Self-Report Index for Sjögren's Syndrome) Syndrome Patient Reported Index (ESSPRI) score

[0301] The EULAR-developed ESSPRI score consists of three patient-reported symptoms: dryness, fatigue, and limb pain. It uses a visual analog scale (VAS), with each symptom scored individually, ranging from 0 to 10 points based on symptom severity, from asymptomatic to most severe. The final ESSPRI score is the mean of the three scores.

[0302] This score is entirely based on patient-submitted questionnaires and complements ESSDAI well, making it suitable for clinical assessment.

[0303] 3.7 Security Assessment

[0304] Laboratory tests:

[0305] This includes complete blood count, blood biochemistry, urinalysis, coagulation tests, viral serology, thyroid function tests, serological tests, hematological tests, and stool routine tests.

[0306] Clinical examination

[0307] Physical examination

[0308] A complete physical examination includes: general condition (including height and weight), skin and mucous membranes, superficial lymph nodes, head and neck (including ears, eyes, nose, and thyroid), chest (including heart and lung auscultation and percussion), abdomen, musculoskeletal system (including spine and limbs), genitals / anus (optional), and neurological assessment.

[0309] A routine physical examination includes: general condition, skin and mucous membranes, and auscultation of the heart and lungs.

[0310] vital signs

[0311] Vital signs include body temperature, pulse, respiratory rate, and blood pressure.

[0312] Lead electrocardiogram

[0313] Analyze resting 12-lead ECG in a local laboratory.

[0314] A 12-lead ECG should be performed on the subject after they have rested in a supine position for at least 5 minutes. All 12-lead ECGs should be recorded while the subject is resting in a supine position. Further ECGs will be performed if clinically necessary, such as in the event of a cardiac-related adverse event. The investigator will complete the ECG assessment on the day of the examination and record the results on the ECG. The same assessment method should be used throughout the study.

[0315] Researchers should evaluate all ECGs according to the category of clinically significant abnormality / non-clinically significant abnormality. If the abnormality is clinically significant, the researcher should record it as an adverse event (AE) on the electronic case report form (eCRF).

[0316] 3.8 Immunogenicity Analysis

[0317] Immunogenic specimens will be collected according to the immunogenicity sampling schedule (Table 2). Testing will be conducted in the central laboratory. 2 mL of whole blood will be collected using anticoagulated vacuum blood collection tubes, plasma will be separated, aliquoted, and cryopreserved for ADA and / or NAb analysis of HZ33B9C5. If necessary, drug concentration in the immunogenicity samples may be tested. Samples from all subjects who received HZ33B9C5 will be tested for immunogenicity.

[0318] The entire analytical method is based on the principle of electrochemiluminescence and mainly consists of the following steps: First, acid is used to dissociate the human plasma sample, rendering ADA in a free state. After neutralization with alkali, it is captured by HZ33B9C5 bound to the ELISA plate. Second, acidification is performed again to dissociate the ADA bound to the ELISA plate, which is then coated onto a blank MSD plate. After blocking, Ruthenylated-HZ33B9C5 is added and incubated to form the "ADA-Ruthenylated-HZ33B9C5" complex. Third, a reading buffer containing tripropylamine is added, and an applied voltage generates an electrochemiluminescence signal. Only HZ33B9C5 antibodies bound to Ruthenylated-HZ33B9C5 will produce a signal.

[0319] 3.9 Pharmacokinetic Analysis

[0320] PK sampling will be conducted according to the PK sampling schedule (Table 2), and the tests will be performed in the central laboratory. 2 ml of whole blood will be collected using anticoagulated vacuum blood collection tubes, plasma will be separated, aliquoted, and cryopreserved for HZ33B9C5 PK analysis. PK samples will be used to detect the plasma drug concentration of HZ33B9C5, and may be used to detect immunogenicity if necessary.

[0321] The PK assay is based on the principle of enzyme-linked immunosorbent assay (ELISA). Human CD40 ligand is used as the coating protein to bind to the ELISA plate to capture HZ33B9C5 in the sample. Then, Mouse Anti-Human IgG Fc Antibody HRP assay reagent is added to form a target-HZ33B9C5-assay reagent complex. Finally, TMB solution is added for color development. The absorbance measured after color development is directly proportional to the HZ33B9C5 concentration in the sample.

[0322] 3.10 Other endpoint indicators

[0323] This study will examine the serum CXCL13 levels in subjects. CXCL13, also known as a B lymphocyte chemokine, selectively attracts B cells (mostly central cells differentiated after antigen stimulation, which highly express CXCR5), promoting the differentiation of antigen-activated B cells into memory B cells or plasma cells in the germinal center. Serum CXCL13 expression reflects the activity level of the germinal center, indicating the level of B cell differentiation activity.

[0324] Blood samples were collected from the subjects during follow-up for serum CXCL13 analysis.

[0325] With the approval of the ethics committee, blood samples will be collected from the subjects according to the visit form for serum CXCL13 analysis.

[0326] The analytical method is based on the sandwich principle and employs electrochemiluminescence (ECL) technology. First, a biotin-labeled anti-CXCL13 antibody is coated onto an SA-MSD plate. Then, the sample is added to the coated MSD plate and incubated. The analyte (CXCL13) in the sample is captured and immobilized on the MSD plate. Subsequently, a Ru-labeled specific anti-CXCL13 antibody is added, binding the analyte to form a sandwich structure. The ECL signal is directly proportional to the concentration of CXCL13.

[0327] 3.11 Safety Reporting and Adverse Event Management

[0328] Definition of Adverse Events (AEs)

[0329] An adverse event (AE) is defined as any adverse medical event that occurs to a clinical trial participant after they have signed an informed consent form, regardless of whether it is causally related to the investigational drug. AEs include, but are not limited to, the following:

[0330] ● The worsening of existing (pre-clinical) medical conditions / diseases (including worsening of symptoms, signs, and abnormal laboratory test results);

[0331] ● Any new adverse medical condition (including symptoms, signs, and newly diagnosed diseases);

[0332] ● Abnormal laboratory test values ​​or results that are clinically significant.

[0333] Adverse events of particular concern

[0334] Adverse events of particular concern (AESIs) are adverse events that require close monitoring to enhance understanding of the investigational drug's safety. AESIs can be non-serious events. All AESIs must be reported in accordance with SAE requirements and timelines.

[0335] The adverse events that require special attention in this study are:

[0336] ● Hypersensitivity reactions caused by use of the investigational drug;

[0337] ● Grade 2 or higher anemia related to the research drug;

[0338] ● Grade 3 or higher infection related to the investigational drug;

[0339] ● Thromboembolic events of any grade;

[0340] ● New positive fecal occult blood test during the study period (i.e., after administration of the study drug) (excluding positive OB results caused by hemorrhoids and anal bleeding);

[0341] ●Pregnancy events.

[0342] Definition of Serious Adverse Events (SAE)

[0343] A serious adverse event is defined as an adverse event that meets at least one of the following criteria:

[0344] ●Causes death;

[0345] ●Life-threatening (The definition of "life-threatening" refers to an AE that poses a risk of death to the subject when it occurs, but does not include AEs that may cause death if the event is aggravated).

[0346] ●Hospitalization or extended hospital stay is required, excluding the following situations:

[0347] √Rehabilitation institutions;

[0348] √ Sanatorium;

[0349] √ Routine emergency room admission;

[0350] √Same-day surgery (e.g., outpatient / same-day / non-bedridden surgery);

[0351] √ Hospitalization or prolonged hospitalization unrelated to the worsening of an adverse event (AE) is not itself an SAE. Examples include: hospitalization due to a pre-existing condition without the occurrence of new adverse events or exacerbation of the pre-existing condition (e.g., to check for persistent laboratory abnormalities from before the trial); hospitalization for management reasons (e.g., annual routine checkups); hospitalization as required by the trial protocol during the clinical trial (e.g., following the trial protocol); elective hospitalization unrelated to the worsening of an adverse event (e.g., elective surgery); pre-planned treatments or surgeries should be documented throughout the trial protocol and / or the subject's baseline data; and hospitalization solely due to the use of blood products.

[0352] ●Causes permanent or severe disability / loss of ability (severely interferes with the ability to live a normal life);

[0353] ●Causes congenital abnormalities / birth defects (in the offspring of subjects who used the product).

[0354] ● Other significant medical events: Although they do not lead to death, are not life-threatening, or do not require hospitalization, if, based on appropriate medical judgment, the event may endanger the subject and may require medical or surgical intervention to prevent one of the above outcomes, such events may also be considered serious adverse events.

[0355] Assessment of the severity of adverse events

[0356] Researchers will assess the severity of the AE based on the five-level criteria established by NCI CTCAE v5.0.

[0357] For adverse event terms not included in NCI CTCAE v5.0, they shall be classified according to the following CTCAE classification principles:

[0358] ● Level 1 Mild: Asymptomatic or with mild signs; only clinical or diagnostic observation required; no medical intervention needed.

[0359] ● Grade 2 Moderate; requires minimal, local or non-invasive treatment; age-appropriate daily living functions (e.g., cooking, shopping, using the phone, and managing finances) are limited.

[0360] ● Grade 3 serious or clinically significant but not immediately life-threatening; hospitalization or prolonged hospitalization; disability; limited ability to perform daily living activities (e.g., bathing, dressing, eating, toileting and taking medication), but not bedridden.

[0361] ● Level 4: Life-threatening consequences; requires emergency treatment.

[0362] ●Deaths related to Level 5 AE

[0363] Determining the causal relationship between adverse events and investigational drugs

[0364] For adverse events in clinical trials, qualified researchers are required to provide an assessment of the causal relationship between the investigational drug and the adverse event.

[0365] This clinical trial uses "relevant" and "unrelevant" to record the correlation between adverse events and the study drug:

[0366] ●Relevant: This generally refers to situations where the timing of the AE (Adverse Event) and the product administration is reasonable, and the AE is more likely to be explained by the investigational drug than by other causes.

[0367] ●Irrelevant: This generally refers to the occurrence of an adverse event, the timing of the adverse event being poorly managed, or other reasons that can better explain the adverse event than the research drug.

[0368] Recording of adverse events

[0369] AEs or SAEs should be recorded using medical terminology / concepts. Colloquialisms and abbreviations should be avoided. All AEs (including SAEs) should be recorded in the eCRF adverse event table.

[0370] Timeframe for adverse event collection

[0371] Adverse events are identified by asking subjects non-leading questions.

[0372] From the signing of the informed consent form until the subject completes the study (completion of follow-up or withdrawal from the study), all adverse events, including serious adverse events, will be collected, whether observed by the investigator or reported by the subject.

[0373] AESI is collected from the moment a subject first uses the study drug until the completion of the study (either upon completion of follow-up or withdrawal from the study).

[0374] Follow-up of adverse events

[0375] Adverse events should be followed up until recovery to baseline or grade 0-1, or until the investigator deems follow-up unnecessary for a reasonable reason (e.g., irreversible or improved). If an adverse event is irreversible, a reasonable explanation must be recorded in the eCRF. Regardless of whether it is related to the investigational drug, the recovery status and date of the subject's AE or SAE should be recorded in the eCRF and medical record.

[0376] Content of adverse event records

[0377] Researchers must fully document any adverse event, including diagnosis (or, if no diagnosis, symptoms, signs, and laboratory abnormalities), start and end dates and times (if applicable), severity and changes of the adverse event, whether it was a serious adverse event, measures taken with the study drug, treatment given due to the AE and the outcome of the event, and the relationship between the adverse event and the study drug.

[0378] For serious adverse events (SAEs), investigators should also provide the date the AE meets the SAE criteria, the date the investigator became aware of the SAE, the basis for classifying the AE as an SAE, the date of hospitalization, the date of discharge, the possible cause of death, the date of death, whether an autopsy was performed, a causal assessment of the study process, a causal assessment of other drugs, and other possible causes of the SAE. Investigators should also provide the basis for their relevance assessment and a description of the SAE. The SAE description should also include the subject's ID, age, sex, height, and weight; the indications for the investigational drug treatment and the disease stage, as well as relevant general conditions; the clinical course of the SAE, including its occurrence, development, outcome, and results; laboratory test results related to the SAE (the date, units, and normal ranges must be provided); past medical history related to the SAE, comorbidities and their occurrence and duration; medication history related to the SAE, comorbid drugs and their initiation, duration, and dosage; and detailed information on the initiation, duration, and dosage of the investigational drug treatment.

[0379] SAE, rapid pregnancy report

[0380] (1) SAE Report

[0381] The reporting period for SAEs begins from the date of signing informed consent until the subject completes the study (either by completing follow-up or withdrawing from the study). If a subject experiences an SAE, the investigator must immediately report it to the sponsor's pharmacovigilance department within 24 hours of becoming aware of the SAE, following the methods and requirements specified by the sponsor.

[0382] For deaths and life-threatening serious adverse events, researchers should urgently follow up on missing information and provide a complete SAE report.

[0383] If an SAE determination occurs outside the aforementioned timeframe and is related to the investigational drug, it should also be reported to the sponsor.

[0384] (2)Pregnancy

[0385] To ensure the safety of the participants, all fertile participants in this clinical study must use effective contraception.

[0386] If a female subject exposed to the study drug becomes pregnant during a clinical study, the investigator should report the pregnancy to the sponsor within 24 hours of becoming aware of it, in accordance with the sponsor's instructions and requirements.

[0387] During the clinical trial, male participants exposed to the study drug may continue the trial if their partners become pregnant. Researchers should report the pregnancy to the sponsor within 24 hours of discovery, following the methods and requirements specified by the sponsor.

[0388] Researchers should continuously monitor subjects who become pregnant and follow up on pregnancy outcomes until 8 weeks after delivery, and report the results to the sponsor.

[0389] Pregnancy outcomes such as stillbirth, spontaneous abortion, fetal malformation (any congenital abnormality / birth defect), and medically induced abortion are considered SAEs and must be reported according to the SAE procedure and time limits.

[0390] If a subject develops an SAE during pregnancy, it must be reported in accordance with the SAE reporting procedure.

[0391] 3.12 Statistical Analysis

[0392] 3.12.1 Sample Size

[0393] This is a Phase I exploratory trial, and no statistical assumptions are made regarding the sample size estimate. A total of 30 pSS subjects are planned for enrollment.

[0394] 3.12.2 Analyzing the dataset

[0395] Safety Set (SS): Subjects who have given informed consent and received the investigational drug at least once. The SS will be used for analysis of demographic and other baseline characteristics, drug exposure, and safety indicators.

[0396] Full Analysis Set (FAS): Subjects who meet the inclusion and exclusion criteria, have been randomized, and have received the study drug at least once. The FAS will be used for efficacy analysis.

[0397] Pharmacokinetic Concentration Set (PKCS): This includes all subjects who have received the study drug at least once and have valid post-dose detection data for at least one component concentration.

[0398] Pharmacokinetic Parameters Set (PKPS): This includes all randomized subjects who have received the study drug at least once and have at least one valid pharmacokinetic parameter data.

[0399] Immunogenicity analysis dataset: includes all randomized subjects who have received the study drug at least once and have at least one post-dose immunogenicity assessment data.

[0400] 3.12.3 General Methods of Statistical Analysis

[0401] This study will primarily use descriptive statistics and will generally not include inter-group comparisons. For continuous variables, the number of cases, mean, standard deviation, median, Q1, Q3, minimum, and maximum values ​​will be calculated. For PK parameters, the geometric mean and coefficient of variation will also be provided. For categorical variables, the number of cases and percentage will be calculated.

[0402] 3.12.4 Demographic and Baseline Characteristics

[0403] Descriptive statistics include the demographic characteristics (sex, age) and baseline information of the subjects: height, weight, body mass index, medical history, etc.

[0404] In the safety analysis set, descriptive statistics of demographic data and baseline characteristics are summarized by treatment group.

[0405] The following continuous variables are summarized as follows: number of cases, mean, standard deviation, median, Q1 and Q3, minimum and maximum values:

[0406] ● Age (years) = (Date of informed consent – ​​Date of birth + 1) / 365.25, take the smallest integer;

[0407] ● Height (cm)

[0408] ●Weight(kg)

[0409] ● Body Mass Index (kg / m²) 2 )

[0410] The following categorical variables summarize the number and percentage of participants:

[0411] ●Gender: Male, Female

[0412] ●Ethnicity: Han, others

[0413] The demographic data and baseline characteristics of the subjects were listed.

[0414] 3.12.5 Pharmacokinetic Analysis

[0415] Descriptive statistical analyses were performed on the p-kJ parameters in each dose group, including but not limited to T. max C max Area Under Curve (AUC), Volume of Distribution (Vd), Elimination Half-Life (t) 1 / 2 ), clearance (CL). See separate pharmacokinetic analysis report for details.

[0416] List the blood drug concentrations of the subjects.

[0417] 3.12.6 Validity Analysis

[0418] For continuous efficacy endpoints, the mean ± standard deviation, maximum value, minimum value, median, first and third quantiles were calculated by summing the data according to the dosing stage and dosage group.

[0419] For the therapeutic endpoint indicators of different types, the frequency and ratio within each group were calculated according to the dosing stage and dosage group, and the 95% CI within each group was calculated using the Clopper-Pearson method.

[0420] ESSDAI rating

[0421] Definition of therapeutic endpoint:

[0422] The ESSDAI score is the EULAR Disease Activity Index for Sjögren's syndrome, which assesses damage to 12 systems throughout the body associated with pSS. Each indicator is scored according to its disease activity level multiplied by its weight. The final ESSDAI score is the sum of the scores for all affected systems. An ESSDAI < 5 indicates low disease activity, 5 ≤ ESSDAI ≤ 13 indicates moderate disease activity, and an ESSDAI ≥ 14 indicates high disease activity.

[0423] The theoretical range for ESSDAI is 0 to 123, and the final score is calculated as follows:

[0424] Final score = the sum of the scores of all 12 systems

[0425] System score = Activity level × System weight

[0426] A higher ESSDAI score indicates a more severe disease.

[0427] The changes in ESSDAI scores since baseline and from baseline will be analyzed using a repeated measures mixture model (MMRM). Treatment, visits, and the interaction between treatment and visits are used as fixed effects, baseline ESSDAI scores as covariates, and subjects as random effects. Restricted maximum likelihood (REML) method is used for parameter estimation, calculated using the Newton-Raphson algorithm, with the Kenward-Roger approximation used for the denominator degrees of freedom. Unstructured correlation matrices are used to model internal errors among subjects. If the model does not converge, internal errors are modeled sequentially using TOEPH, CSH, and TOEP correlation matrices until convergence. Missing data are not imputed. Curves (mean ± standard error) will be plotted for each treatment group, showing the changes in measurements since baseline over time.

[0428] ESSPRI rating

[0429] The EULAR-developed ESSPRI score consists of three patient-reported symptoms: dryness, fatigue, and limb pain. It uses a visual analog scale (VAS), with each symptom scored individually, ranging from 0 to 10 points based on symptom severity, from asymptomatic to most severe. The final ESSPRI score is the mean of the three scores.

[0430] The statistical analysis method for the ESSPRI score is the same as that for the ESSDAI score. The total score and each score are analyzed separately. Curves (mean ± standard error) of the total score and each score measurement, as well as the changes from baseline over time, will also be plotted for each treatment group.

[0431] Salivary gland function research

[0432] This study used total salivary gland flow rate under both unstimulated and stimulated conditions to assess the salivary gland function of the subjects.

[0433] The statistical analysis method for total salivary gland flow rate values ​​(unstimulated and stimulated) is the same as that for the ESSDAI score. Curves (mean ± standard error) of the changes in measurement values ​​from baseline over time will also be plotted for each treatment group.

[0434] Lacrimal gland function research

[0435] In this study, the Schirmer I test was used to assess the lacrimal gland function of the subjects. The average results for both the right and left eyes were obtained for each test; if only one eye's results were available, that result was used. The statistical analysis of the Schirmer I test results was performed using the same methods as the ESSDAI score, and curves showing the mean and changes from baseline over time for each treatment group were also plotted.

[0436] 3.12.7 Security Analysis

[0437] Drug exposure

[0438] The assessment of drug exposure includes the total exposure to the study drug and the relative dose intensity.

[0439] The total exposure and relative dose intensity of the study drugs were summarized by drug group, and the following statistics were given: number of cases, mean, standard deviation, median, Q1, Q3, minimum and maximum values.

[0440] The relative dose intensity is calculated as: (Actual dosage / Planned dosage) * 100%.

[0441] List the medication use of the subjects.

[0442] Adverse events

[0443] Categorized by the date the event occurred:

[0444] ○ Adverse events prior to medication: Adverse events that occurred before the signing of informed consent and prior to the first study use of the drug.

[0445] ○ Treatment Emergent Adverse Event (TEAE): Adverse events that occur or worsen during the period from the start of the first study medication to the last visit.

[0446] If the start date of an adverse event is completely or partially missing, fill it in according to the rules in Appendix 16.1.9 SAP Section 5.3 before marking the adverse event before or during medication.

[0447] For each medication group, all adverse events (AEs), treatment-related AEs (TEAEs), grade 2 or higher and grade 3 TEAEs, TEAEs related to the investigation drug, grade 2 or higher and grade 3 TRAEs, TEAEs leading to suspension of the investigation drug, TEAEs leading to suspension of the investigation drug and related to the investigation drug, TEAEs resulting in death, SAEs and SAEs related to the investigation drug, and adverse events of special concern during treatment (AESIs) were summarized, and their number and percentage were calculated.

[0448] Adverse events are summarized into the following categories according to the SOC, PT, and CTCAE classifications in MedDRA coding:

[0449] ●TEAE

[0450] ●Level 2 TEAE

[0451] ●Level 3 or above TEAE

[0452] ●TEAE related to investigational drugs

[0453] ●Level 2 or higher TEAE related to the investigational drug

[0454] ● ≥ Grade 3 TEAE related to the investigational drug

[0455] ●SAE

[0456] ●AESI

[0457] ●TEAE leading to medication discontinuation

[0458] ●TEAEs that lead to drug discontinuation

[0459] ●TRAE leading to medication discontinuation

[0460] ●TRAE leading to drug discontinuation

[0461] The above AE categories are summarized and sorted according to the total occurrence rate of AEs in the corresponding category.

[0462] All adverse events, treatment-associated adverse events (TEAEs) related to the investigational drug, TEAEs leading to discontinuation of the investigational drug, adverse events leading to the death of the subject, and SAEs will be listed (including at least the AE start date, end date, severity, relationship to the drug, measures taken, and outcome).

[0463] Laboratory tests

[0464] Cross-tabulations were compiled for each laboratory test, showing the clinical significance of normal and abnormal results before and after baseline, categorized by medication group. Based on the criteria for potentially clinically significant abnormality (PCSA), the worst-case results of complete blood count and blood biochemistry after baseline, which were potentially clinically significant, were compiled.

[0465] List the laboratory test results, and list the pregnancy test, abdominal ultrasound and / or viral serological test results during the trial separately.

[0466] Other security indicators

[0467] vital signs

[0468] Based on the criteria for Potential Clinically Significant Abnormalities (PCSA), post-baseline vital signs results that may have clinical significance were compiled.

[0469] List the results of the vital signs examination.

[0470] electrocardiogram

[0471] The ECG parameters to be analyzed include heart rate (HR), RR interval, QRS interval, QT interval, QTcF, and PR interval.

[0472] The results of 12-lead electrocardiograms and their relative baseline changes will be summarized at each evaluation time point. Post-baseline electrocardiogram results that may have clinical significance will be summarized according to PCSA criteria.

[0473] List the electrocardiogram (ECG) test results.

[0474] Physical examination

[0475] The subjects whose physical examinations showed abnormal changes relative to baseline were described in a list format.

[0476] 3.12.8 Analysis of other endpoint indicators

[0477] Serum antinuclear antibodies

[0478] A cross-tabulation of serum antinuclear antibody levels before and after baseline, and whether the abnormality was clinically significant, was compiled according to the treatment group.

[0479] Serum immunoglobulins IgG, IgM, IgA, and rheumatoid factor

[0480] To assess the effects of HZ33B9C5 on the changes in serum IgG, IgM, IgA, and RF over time, an MMRM model will be used for analysis. Curves (mean ± standard error) will be plotted for each treatment group showing the changes in measurements from baseline and the percentage change from baseline over time.

[0481] Erythrocyte sedimentation rate (ESR) and serum CXCL13 levels

[0482] The statistical analysis methods for ESR and CXCL13 are the same as those for the ESSDAI score. Curves will also be plotted for the mean, change from baseline, and percentage change from baseline over time for each treatment group. If applicable, a curve will be plotted for the median (interquartile) value of CXCL13 over time.

[0483] Immunogenicity analysis

[0484] The positive rate of anti-drug antibody (ADA) and the immunogenicity test results at each assessment time point were summarized according to the HZ33B9C5 dose group.

[0485] The list describes cases where subjects who received intravenous HZ33B9C5 developed anti-drug antibodies.

[0486] 3.13 Research Data and Results

[0487] As of July 25, 2025 (database locked), a total of 30 subjects were enrolled and administered the study drug in this study. Enrolled subjects were randomly assigned to the HZ33B9C5 7.5 mg / kg group (N=8), HZ33B9C5 15 mg / kg group (N=8), HZ33B9C5 30 mg / kg group (N=8), or placebo group (N=6).

[0488] The median ages (ranges) of the HZ33B9C5-7.5 mg / kg group, HZ33B9C5-15 mg / kg group, HZ33B9C5-30 mg / kg group, and placebo group were 50.0 (24–62) years, 38.5 (25–70) years, 32.5 (21–67) years, and 47.0 (28–61) years, respectively; the median BMI (range) was 21.55 (18.0–28.0) kg / m², respectively. 2 22.55 (18.8~25.6) kg / m 2 21.50 (18.0~24.4) kg / m 219.80 (18.2~27.9) kg / m 2 All participants were female; all participants were of Han ethnicity.

[0489] Twenty-four subjects had received at least one dose of HZ33B9C5 and were included in the safety analysis set (SS), with eight subjects in each of the 7.5 mg / kg, 15 mg / kg, and 30 mg / kg dose groups. Six subjects had received at least one dose of placebo and were included in the safety analysis set (SS).

[0490] Based on the SS population, the disease course was largely consistent among subjects in the HZ33B9C5 dose groups (7.5 mg / kg, 15 mg / kg, and 30 mg / kg) and the placebo group. The mean (standard deviation) of baseline ESSDAI scores in the HZ33B9C5 total group and the placebo group were 5.4 (4.80) and 5.0 (4.00), respectively, and the mean (standard deviation) of baseline ESSPRI scores were 4.0 (1.78) and 3.4 (1.93), respectively. The majority of subjects (96.7%) had hematologic impairment, a minority (10.0%) had arthritis, a minority (6.7%) had renal impairment, and a minority (3.3%) had interstitial lung disease.

[0491] Based on the SS population, the median total exposure (range) during the study period was 1575.60 mg (1111.2–2400.0) in the 7.5 mg / kg, 15 mg / kg, and 30 mg / kg dose groups and 2713.20 mg (1519.2–8755.2) in the placebo group.

[0492] Pharmacokinetic results:

[0493] In subjects with Sjögren's syndrome, dose-exposure to HZ33B9C5 increased nearly proportionally within a dose range of 7.5–30 mg / kg, exhibiting linear pharmacokinetic characteristics.

[0494] Following single and multiple intravenous infusions of 7.5–30 mg / kg HZ33B9C5 in subjects with Sjögren's syndrome, the geometric mean half-life ranged from 16 to 25.2 days, supporting a dosing regimen of once every 4 weeks.

[0495] The geometric mean of the drug exposure accumulation ratio calculated based on AUC after multiple (4) administrations ranged from 1.61 to 1.87. The PK characteristics of multiple (4) intravenous infusions were consistent with those of a single administration.

[0496] Validity results:

[0497] At week 16, the mean changes from baseline in ESSDAI scores for the 7.5 mg / kg, 15 mg / kg, 30 mg / kg, and placebo groups were -0.29 ± 1.799, -0.50 ± 4.375, -1.50 ± 3.625, and 0.40 ± 1.517 (mean ± standard deviation), respectively. Compared with the placebo group, the mean least-squares estimates of the mean differences in ESSDAI scores from baseline for the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were -0.61 (95% CI: -3.901, 2.687; P = 0.7076), -0.91 (-4.161, 2.335; P = 0.5677), and -1.61 (-4.857, 1.637; P = 0.3166), respectively.

[0498] At week 16, the mean changes from baseline in the ESSPRI total score for the 7.5 mg / kg, 15 mg / kg, 30 mg / kg, and placebo groups were -1.03 ± 1.119, -1.50 ± 1.466, and -0.89 ± 1.791, respectively (mean ± standard deviation). Compared with the placebo group, the mean least-squares estimates of the changes from baseline in the ESSPRI total score for the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were -1.47 (95% CI: -2.838, -0.099; P = 0.0366), -2.19 (-3.531, -0.859; P = 0.0023), and -1.55 (-2.886, -0.212; P = 0.0249), respectively.

[0499] At week 16, the mean changes from baseline in the Schirmer I trial results for the 7.5 mg / kg, 15 mg / kg, 30 mg / kg and placebo groups were -0.71 ± 7.399, -4.31 ± 4.877, -0.50 ± 8.120 and 3.20 ± 7.571 mm (mean ± standard deviation), respectively. Compared with the placebo group, the mean least-squares estimates of the changes from baseline in the Schirmer I trial for the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were -4.09 mm (95% CI: -11.872, 3.701; P = 0.2912), -7.44 mm (-15.115, 0.227; P = 0.0567), and -3.58 mm (-11.250, 4.094; P = 0.3467), respectively.

[0500] At week 16, the mean changes from baseline in whole salivary gland flow rate (unstimulated) in the 7.5 mg / kg, 15 mg / kg, 30 mg / kg and placebo groups were -0.01 ± 0.073, 0.10 ± 0.134, 0.01 ± 0.089 and -0.03 ± 0.055 (mean ± standard deviation) mL / min, respectively. Compared with the placebo group, the mean least-squares estimates of the changes in whole salivary gland flow rate (without stimulation) from baseline in the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were 0.03 mL / min (95% CI: -0.091, 0.145; P = 0.6433), 0.11 mL / min (-0.007, 0.226; P = 0.0641), and 0.02 mL / min (-0.093, 0.140; P = 0.6850), respectively.

[0501] At week 16, the mean changes from baseline in whole salivary gland flow rate (stimulation) in the 7.5 mg / kg, 15 mg / kg, 30 mg / kg and placebo groups were 0.99 ± 0.713, 0.62 ± 0.596, 0.96 ± 1.041 and 0.60 ± 0.480 (mean ± standard deviation) mL / min, respectively. Compared with the placebo group, the mean least-squares estimates of the changes in total salivary gland flow rate (stimulation) from baseline in the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were 0.25 mL / min (95% CI: -0.080, 0.573; P = 0.1321), -0.05 mL / min (-0.360, 0.266; P = 0.7594), and -0.07 mL / min (-0.386, 0.246; P = 0.6521), respectively.

[0502] At week 16, all HZ33B9C5 dose groups showed varying degrees of decrease in IgG, IgA, IgM, rheumatoid factor, chemokine 13, and erythrocyte sedimentation rate compared to baseline.

[0503] Safety results:

[0504] All levels of adverse events

[0505] Based on the SS population, 19 subjects (79.2%) in the HZ33B9C5 total group experienced 55 treatment-induced adverse events (TEAEs) during the study period; in the placebo group, 6 subjects (100%) reported 23 adverse events. Classified by PT level, the most frequent TEAE in both the HZ33B9C5 total group and the placebo group was upper respiratory tract infection, with an incidence of 50.0% (12 reported events) in the HZ33B9C5 total group and 33.3% (2 reported events) in the placebo group.

[0506] The incidence of grade ≥2 TEAEs was 62.5% in the HZ33B9C5 total group and 66.7% in the placebo group. The most common grade ≥2 TEAEs in both the HZ33B9C5 total group and the placebo group were upper respiratory tract infections, with an incidence of 41.7% in the HZ33B9C5 total group (10 events reported) and 33.3% in the placebo group (2 events reported).

[0507] Drug-related adverse events

[0508] Based on the SS population, 12 subjects (50.0%) in the HZ33B9C5 total group experienced at least one treatment-related adverse event (TRAE) during the study period, while 4 subjects (66.7%) in the placebo group experienced at least one TRAE. Classified by PT level, the most common TRAE in both the HZ33B9C5 total group and the placebo group was upper respiratory tract infection, with an incidence of 33.3% (6 reported events) in the HZ33B9C5 total group and 33.3% (2 reported events) in the placebo group.

[0509] The incidence of grade ≥2 TRAEs was 41.7% in the HZ33B9C5 combined group and 50.0% in the placebo group. Classified by PT level, the most common grade ≥2 TRAEs in both the HZ33B9C5 combined group and the placebo group were upper respiratory tract infections, with an incidence of 25.0% (6 reported events) in the HZ33B9C5 combined group and 33.3% (2 reported events) in the placebo group. No subjects experienced grade ≥3 TRAEs.

[0510] During the study, two subjects experienced TEAEs that led to the discontinuation of the study drug. According to the PT classification, both TEAEs were due to abnormal liver function, and the researchers determined that they were related to the study drug.

[0511] During the study, one subject experienced an adverse event that led to permanent discontinuation of HZ33B9C5. According to PT classification, this treatment-associated adverse event (TEAE) was myositis, which the investigators determined was unrelated to the study drug.

[0512] During the study, no subjects experienced serious adverse events (SAEs), adverse events of special concern during treatment (AESIs), or treatment-related adverse events (TEAEs) that resulted in death.

[0513] Physical examination, vital signs, complete blood count, blood biochemistry, coagulation, urinalysis, and electrocardiogram parameters remained generally stable across all groups during the study period.

[0514] Immunogenicity results:

[0515] The incidence of ADA positivity in all pSS subjects who received intravenous HZ33B9C5 was 4.2% (1 / 24). No immunogenicity-related adverse reactions, such as infusion reactions or hypersensitivity reactions, occurred during the observation period of this study. Therefore, after multiple intravenous infusions of HZ33B9C5 within the dose range of 7.5–30 mg / kg, immunogenicity has essentially no impact on safety.

[0516] 3.14 Discussion and Conclusion

[0517] discuss

[0518] This was a randomized, double-blind, single-center, multiple-dose, dose-escalation, placebo-controlled study designed to evaluate the safety, tolerability, pharmacokinetic characteristics, and therapeutic potential of HZ33B9C5 in pSS subjects with multiple doses. The study included a screening period (6 weeks before dosing), a dosing period (12 weeks), and a safety follow-up period (12 weeks). The dose range of HZ33B9C5 was 7.5 mg / kg–30 mg / kg, administered intravenously at a frequency of Q4W.

[0519] The primary endpoint of this study was the safety and tolerability of HZ33B9C5. During the observation period, the overall incidence and severity of treatment-associated adverse events (TEAEs) were comparable among subjects receiving multiple doses of HZ33B9C5 and placebo. Only one subject receiving HZ33B9C5 at 30 mg / kg experienced a grade 3 TEAE, which was determined to be unrelated to the study drug; all other TEAEs were grade CTCAEs 1-2. The incidence of treatment-associated adverse events (TRAEs) was also similar in the HZ33B9C5 and placebo groups, both being grade CTCAEs 1-2. One subject in each of the HZ33B9C5 7.5 mg / kg and placebo groups experienced a TRAE leading to discontinuation of the study drug. No subjects experienced SAEs, AESIs, or TEAEs resulting in death. In conclusion, in pSS subjects, HZ33B9C5 demonstrated good safety and tolerability within the dose range of 7.5 mg / kg–30 mg / kg.

[0520] This study preliminarily explored the efficacy of HZ33B9C5 in treating pSS using other endpoints. The ESSDAI score assessed the objective systemic disease activity in pSS patients. In this study, the proportion of patients with improved ESSDAI scores from baseline was higher in all HZ33B9C5 dose groups than in the placebo group, suggesting a potential improvement in ESSDAI scores.

[0521] The ESSPRI score assessed the subjective disease activity of patients with pSS. Compared with the placebo group, the mean least-squares estimate of the change in total ESSPRI score from baseline at week 16 was -1.47 (P = 0.0366), -2.19 (P = 0.0023), and -1.55 (P = 0.0249) in the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups, respectively. In another phase II clinical trial of the anti-CD40L antibody Dazodalibep, compared with the placebo group, the mean least-squares estimate of the change in total ESSPRI score from baseline at week 24 after treatment with Dazodalibep 1500 mg Q2W was -1.27 (P = 0.0002) (St Clair EW, Baer AN, Ng WF, et al. CD40 ligand antagonist dazodalibep in disease:arandomized, double-blinded, placebo-controlled, phase 2 trial. Nat Med. 2024; 30(6):1583-1592. doi:10.1038 / s41591-024-03009-3). Therefore, it can be seen that HZ33B9C5 administered Q4W in the dose range of 7.5 mg / kg-30 mg / kg resulted in better improvement of ESSPRI scores than Dazodalibep.

[0522] In addition to the primary endpoints for assessing pSS disease activity, ESSDAI and ESSPRI scores, this study also evaluated other endpoints correlated with efficacy. Assessments of the Schirmer I test and total salivary gland flow rate (unstimulated / unstimulated) showed that none of the HZ33B9C5 dose groups significantly improved lacrimal and salivary gland function compared to the placebo group, which may be related to the irreversible nature of lacrimal and salivary gland dysfunction in the subjects. Regarding serum biomarkers, although HZ33B9C5 did not significantly improve serum IgM, CXCL13, and ESR levels in pSS subjects, serum IgG, IgA, and RF levels in all HZ33B9C5 dose groups were significantly lower than in the placebo group at week 16. Compared with the placebo group, the mean least-squares estimated differences in serum IgG from baseline at week 16 in the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were -17.54% (P = 0.0017), -19.18% (P = 0.0009), and -17.66% (P = 0.0016), respectively. Compared with the placebo group, the mean least-squares estimated differences in serum RF from baseline at week 16 in the HZ33B9C5 7.5 mg / kg, 15 mg / kg, and 30 mg / kg groups were -39.79% (P = 0.0118), -57.18% (P = 0.0006), and -57.53% (P = 0.0005), respectively. Based on the CRESS standard (Arends S, de Wolff L, van Nimwegen JF, et al. Composite of Relevant Endpoints for Syndrome (CRESS): development and validation of a novel outcome measure. Lancet Rheumatol. 2021; 3(8):e553-e562. doi:10.1016 / S2665-9913(21)00122-3). In pSS patients, a decrease of ≥10% in IgG is clinically significant, while a decrease of ≥25% in RF is clinically significant. Therefore, it can be considered that HZ33B9C5 has clinical significance in improving important serum markers in pSS subjects, and also reflects the effective inhibition of the ability of B cells in pSS subjects to produce antibodies.

[0523] In terms of pharmacokinetics, in subjects with Sjögren's syndrome, HZ33B9C5 showed a near-proportional increase in dose-exposure within the dose range of 7.5–30 mg / kg, exhibiting linear PK characteristics. The PK characteristics after multiple (4) doses were consistent with those after a single dose. The half-life after single or multiple doses ranged from approximately 16 to 25.2 days. Combined with the aforementioned maintenance of clinical efficacy, this supports a Q4W or even longer dosing interval for HZ33B9C5 in pSS subjects. Regarding immunogenicity, the proportion of ADA-positive subjects among all pSS subjects receiving intravenous HZ33B9C5 was 4.2% (1 / 24), with only one subject experiencing treatment-induced persistent ADA positivity. This suggests that HZ33B9C5 has low immunogenicity and high safety and stability in pSS subjects.

[0524] in conclusion:

[0525] In Chinese subjects with primary Sjögren's syndrome, multiple intravenous injections of HZ33B9C5 at doses ranging from 7.5 mg / kg to 30 mg / kg showed good overall safety and tolerability.

[0526] In Chinese subjects with primary Sjögren's syndrome, repeated intravenous injections of HZ33B9C5 resulted in a decrease in the subjects' ESSPRI total score and serum markers, suggesting that HZ33B9C5 has the effect of reducing disease activity in primary Sjögren's syndrome.

[0527] Based on the observed safety, tolerability, and disease efficacy of HZ33B9C5, further development of HZ33B9C5 in patients with primary Sjögren's syndrome is supported.

[0528] The foregoing describes exemplary embodiments of the present invention. Those skilled in the art should understand that these disclosures are merely exemplary, and various other substitutions, adaptations, and modifications can be made within the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments listed herein.

[0529] Sequence information:

Claims

1. A method for preventing or treating an autoimmune disease in a patient, the method comprising administering an anti-CD40L antibody or an antigen-binding fragment thereof to a patient in need according to a dosing regimen comprising administering one or more doses of the anti-CD40L antibody or an antigen-binding fragment thereof to the patient in need, wherein the dose is 0.1 mg / kg to 40 mg / kg, for example 0.3 mg / kg to 35 mg / kg, 1 mg / kg to 35 mg / kg, 3 mg / kg to 35 mg / kg, 5 mg / kg to 35 mg / kg, 7.5 mg / kg to 35 mg / kg. g, 7.5 mg / kg-30 mg / kg or 15 mg / kg-35 mg / kg, wherein the anti-CD40L antibody or its antigen-binding fragment comprises a first heavy chain complementarity-determining region (HCDR1), a second heavy chain complementarity-determining region (HCDR2), and a third heavy chain complementarity-determining region (HCDR3), as well as a first light chain complementarity-determining region (LCDR1), a second light chain complementarity-determining region (LCDR2), and a third light chain complementarity-determining region (LCDR3), wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 respectively comprise or consist of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, respectively.

2. The method of claim 1, wherein the anti-CD40L antibody or its antigen-binding fragment comprises a heavy chain variable region (VH), wherein the heavy chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence, or comprises an amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19, or is composed of said amino acid sequence; or comprises a light chain variable region (VL), wherein the light chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:10 or the nucleic acid sequence shown in SEQ ID NO:20, or is composed of said amino acid sequence, or comprises an amino acid sequence encoded by ... The amino acid sequence encoded by the amino acid sequence shown in NO:10 or the nucleic acid sequence shown in SEQ ID NO:20, or composed of the amino acid sequence shown.

3. The method according to claim 1 or 2, wherein the anti-CD40L antibody or its antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:4 or the nucleic acid sequence shown in SEQ ID NO:19 or is composed of said amino acid sequence, and the light chain variable region comprises an amino acid sequence encoded by the amino acid sequence shown in SEQ ID NO:10 or the nucleic acid sequence shown in SEQ ID NO:20 or is composed of said amino acid sequence.

4. The method according to any one of claims 1-3, wherein the anti-CD40L antibody or its antigen-binding fragment further comprises an Fc region, for example, wherein the Fc region is or is derived from human IgG Fc, for example, is or is derived from human IgG1 Fc, is or is derived from human IgG2 Fc, is or is derived from human IgG3 Fc or is derived from human IgG4 Fc; preferably, the Fc region comprises a mutation that reduces or eliminates effector function, and / or eliminates the binding affinity to FcγR while retaining the binding affinity to FcRn, preferably, the mutation is an L234A / L235A and P329 deletion mutation, preferably, the Fc region (i) Contains or is composed of the amino acid sequence shown in SEQ ID NO: 13 or 14; (ii) Contains an amino acid sequence having at least 90% identity with the amino acid sequence shown in SEQ ID NO:13, for example, 95%, 96%, 97%, 99% or higher identity; or (iii) Contains an amino acid sequence that has at least 90% identity with the amino acid sequence shown in SEQ ID NO:14, such as 95%, 96%, 97%, 99% or higher, and contains L234A / L235A and P329 deletion mutations.

5. The method according to any one of claims 1-4, wherein the anti-CD40L antibody or its antigen-binding fragment further comprises a heavy chain constant region, said heavy chain constant region being or derived from a constant region of IgG1, IgG2, IgG3 or IgG4, preferably, said heavy chain constant region (i) Contains or is composed of the amino acid sequence shown in SEQ ID NO:5 or 16; (ii) comprising an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:16; or (iii) Contains an amino acid sequence that has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequence shown in SEQ ID NO:5, and has L234A / L235A and P329 deletion mutations; and / or The anti-CD40L antibody or its antigen-binding fragment further comprises a light chain constant region, which is or is derived from the lambda or Kappa light chain constant region. Preferably, the light chain constant region comprises or is composed of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:11; or comprises or is composed of the amino acid sequence shown in SEQ ID NO:

11.

6. The method according to any one of claims 1-5, wherein the anti-CD40L antibody or its antigen-binding fragment heavy chain, wherein the heavy chain (i) comprising or consisting of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO:6; or (ii) Contains or is composed of the amino acid sequence shown in SEQ ID NO:6; or It contains a light chain, wherein the light chain (i) comprising or consisting of an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 12; or (ii) Contains or consists of the amino acid sequence shown in SEQ ID NO:

12.

7. The method according to any one of claims 1-6, wherein the anti-CD40L antibody or its antigen-binding fragment comprises a heavy chain and a light chain, wherein the heavy chain comprises or is composed of the amino acid sequence shown in SEQ ID NO:6, and the light chain comprises or is composed of the amino acid sequence shown in SEQ ID NO:

12.

8. The method according to any one of claims 1-7, wherein the anti-CD40L antibody is a humanized antibody or a chimeric antibody.

9. The method according to any one of claims 1-8, wherein the antigen-binding fragment of the anti-CD40L antibody is selected from the following antibody fragments: Fv, Fab, Fab', Fab'-SH, F(ab')2, dAb (domain antibody), linear antibody, single-chain antibody (e.g., scFv); single-domain antibody (sdAb) such as VHH, bivalent antibody or fragment thereof, or camelid antibody or diabody antibody.

10. The method according to any one of claims 1-9, wherein the autoimmune disease is Sjögren's syndrome, such as primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

11. The method according to any one of claims 1-9, wherein the autoimmune disease is systemic lupus erythematosus.

12. The method according to any one of claims 1-11, wherein, The dosing regimen includes the anti-CD40L antibody or its antigen-binding fragment at 2-week, 4-week, or 6-week intervals.

13. The method according to any one of claims 1-12, wherein the administration regimen comprises at least four administrations, for example at least four consecutive administrations, preferably, each administration of the same dose.

14. The method according to any one of claims 1-13, wherein, The dosage of the dosing regimen is approximately 1-35 mg / kg, preferably 7.5-30 mg / kg, for example approximately 1 mg / kg, 3 mg / kg, 5 mg / kg, 7.5 mg / kg, 15 mg / kg, 25 mg / kg, 30 mg / kg or 35 mg / kg, preferably the same dosage for each administration.

15. The method according to any one of claims 1-14, wherein, The administration method is, for example, subcutaneous injection or intravenous injection, preferably intravenous infusion.

16. The method according to any one of claims 1-15, wherein, The dosing regimen involves administering the same dose of anti-CD40L antibody or its antigen-binding fragment to patients in need at 4-week intervals, with each dose being approximately 7.5 mg / kg to 30 mg / kg.

17. The method according to any one of claims 1-15, wherein, The dosing regimen administers the same dose of anti-CD40L antibody or its antigen-binding fragment to patients in need at 4-week intervals, with each dose being approximately 7.5 mg / kg, approximately 15 mg / kg, or approximately 30 mg / kg.

18. The method according to any one of claims 1-17, wherein, The anti-CD40L antibody or its antigen-binding fragment is formulated into a liquid preparation or a lyophilized preparation, for example, the anti-CD40L antibody or its antigen-binding fragment is formulated into a lyophilized powder.

19. The method according to any one of claims 1-18, wherein the anti-CD40L antibody or its antigen-binding fragment is contained in a pharmaceutical formulation, the pharmaceutical formulation further comprising histidine, histidine hydrochloride, sucrose, mannitol and polysorbate 80, preferably, the formulation being formulated as follows: 50-100 mg / mL of the CD40L antibody or its antigen-binding fragment, 10 mM histidine (composed of 0.12 mg / mL histidine and 1.94 mg / mL histidine hydrochloride), 50.00 mg / mL sucrose, 20.00 mg / mL mannitol, and 0.3 mg / mL polysorbate 80.

20. A unit dosage form comprising about 100 mg to 1000 mg, such as about 180 mg to 550 mg, 200 mg to 540 mg, 300 mg to 600 mg, 300 mg to 720 mg, 500 mg to 1000 mg, or 500 mg to 800 mg, such as 180 mg, 300 mg, 540 mg, or 600 mg, of an anti-CD40L antibody or an antigen-binding fragment thereof, said unit dosage form for the prevention or treatment of autoimmune diseases by the method according to any one of claims 1-19, preferably, said anti-CD40L antibody or antigen-binding fragment thereof is formulated as a lyophilized powder.

21. A medicine box comprising: (1) one or more unit dosage forms of claim 18; (2) a packaging insert printed with instructions for the use of an anti-CD40L antibody or its antigen-binding fragment in an individual for the prevention or treatment of an autoimmune disease by any one of claims 1-19; Optionally, the kit is in the form of a drug dosage unit, and the anti-CD40L antibody or its antigen-binding fragment is contained in a single-dose dosing unit, thereby providing a single-dose dosing unit according to the dosing regimen or drug administration interval; Optionally, a single-dose delivery unit comprising the anti-CD40L antibody or its antigen-binding fragment may contain approximately 0.1 mg / kg to 40 mg / kg, such as 0.3 mg / kg to 35 mg / kg, 1 mg / kg to 35 mg / kg, 3 mg / kg to 35 mg / kg, 5 mg / kg to 35 mg / kg, 7.5 mg / kg to 35 mg / kg, 7.5 mg / kg to 30 mg / kg, 15 mg / kg to 35 mg / kg, 15 mg / kg to 30 mg / kg, or 25 mg / kg to 35 mg / kg. mg / kg, for example, about 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg or 40 mg / kg of the said antiCD40L antibody or its antigen-binding fragment; Optionally, the kit comprises four single-dose delivery units, each containing the same dose of the anti-CD40L antibody or its antigen-binding fragment, wherein the dose is approximately 0.1 mg / kg, 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 17.5 mg / kg, 20 mg / kg, 22.5 mg / kg, 25 mg / kg, 27.5 mg / kg, 30 mg / kg, 32.5 mg / kg, 35 mg / kg, 37.5 mg / kg, or 40 mg / kg; optionally, the kit comprises four single-dose delivery units, each containing the same dose, wherein the dose is approximately 7.5 mg / kg to 30 mg / kg, for example 7.5 mg / kg, 15 mg / kg, or 30 mg / kg.

22. Use of an anti-CD40L antibody or its antigen-binding fragment in the preparation of a medicament for the prevention or treatment of autoimmune diseases, wherein the anti-CD40L antibody or its antigen-binding fragment or the medicament is administered according to the dosing regimen as described in any one of claims 1-19, and the anti-CD40L antibody or its antigen-binding fragment is as defined in any one of claims 1-9.

23. Use of the unit dosage form of claim 20 or the cassette of claim 21 in the preparation of a medicament for the prevention or treatment of autoimmune diseases.

24. The unit dosage form of claim 20, the kit of claim 21, or the use of claim 22 or 23, wherein the autoimmune disease is Sjögren's syndrome, such as primary Sjögren's syndrome or secondary Sjögren's syndrome, such as Sjögren's syndrome secondary to systemic lupus erythematosus.

25. The unit dosage form of claim 20, the kit of claim 21, or the use of claim 22 or 23, wherein the autoimmune disease is systemic lupus erythematosus.

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