CD40L-specific Tn3-derived scaffolds for the treatment and prevention of Sjogren's syndrome

Abstract

Compositions and methods comprising a CD40L-specific Tn3 scaffold are provided, as are methods for preventing and treating Sjögren's syndrome. Sjögren's syndrome (SS) is a systemic autoimmune disease characterized by chronic lymphocytic inflammation of exocrine glands, primarily the salivary and lacrimal glands, leading to loss of function and manifesting as excessive dryness. In addition, extraglandular manifestations have been described as multisystemic involvement affecting the musculoskeletal, pulmonary, renal, neurological, dermatological, gastrointestinal, hematological, hepatobiliary, or vascular systems, while fatigue is a particularly prominent comorbidity.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 597,860, filed November 10, 2023, and U.S. Provisional Patent Application No. 63 / 479,830, filed January 13, 2023, which are hereby incorporated by reference in their entireties for all purposes.

[0002] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (HOPA_065_02WO_SeqList_ST26.xml; size: 24,509 bytes; and creation date: January 12, 2024) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates to compositions comprising a Tn3 scaffold and methods of use thereof in the treatment and prevention of Sjogren's syndrome. Methods of use thereof in the treatment of rheumatoid arthritis are also provided. [Background technology]

[0004] Sjögren's syndrome (SS) is a systemic autoimmune disease characterized by chronic lymphocytic inflammation of the exocrine glands, primarily the salivary and lacrimal glands, leading to loss of function and manifesting as excessive dryness. In addition, extraglandular manifestations affect the musculoskeletal, pulmonary, renal, neurological, and dermatological systems. It has been described as a multisystem disease affecting the gastrointestinal, hematological, hepatobiliary, or vascular systems, while fatigue is a particularly prominent comorbidity. The joints, lungs, skin, and peripheral nerves are the organ systems most frequently affected, while cytopenia, hypocomplementemia, and cryoglobulinemia at the time of diagnosis are strongly associated with high systemic activity.

[0005] The subjective aspects of SS, including patient perceptions of dryness, musculoskeletal pain, and fatigue, can be debilitating and have been shown to have a substantial negative impact on quality of life (QoL). Dryness and fatigue are major factors contributing to reduced QoL. QoL is also influenced by the psychological and emotional challenges and social disruption associated with dependence on close relatives for daily living and difficulties at work as well as in other occupations. The majority of SS patients have exocrine insufficiency and do not meet the European League Against Rheumatism (EULAR) Sjoegren's Syndrome Patient-Reported Index (SJOR) score, which is considered the cut-off point for "unsatisfactory symptom state." Patients with SS who have severe symptoms, as defined by an ESSPRI score of ≥ 5, but low systemic disease activity (EULAR Sjoegren's Syndrome Disease Activity Index (ESSDAI) score < 5), have been excluded from recent clinical trials despite their substantial disease burden and overall unacceptable health status.

[0006] Approximately 15–20% of patients with SS have systemic manifestations beyond the commonly affected exocrine glands (salivary and ocular glands). Moderate to severe systemic activity in SS patients can be defined by an ESSDAI score of ≥ 5. These manifestations most commonly include arthritis, pulmonary disease, renal disease, vasculitis, neuropathy, and autonomic nervous system dysfunction, which are associated with glandular involvement. Moderate to severe disease activity is not only debilitating, but can also lead to dysfunction of one or more affected organs, which is associated with an increased risk of mortality, or hematologic morbidity, including thrombocytopenia and lymphoma. No biologic or disease-modifying antirheumatic drug (DMARD) has been shown to significantly reduce systemic disease activity in SS. Currently, there are no approved immunomodulatory agents or evidence-based treatment guidelines available for the treatment of extraglandular manifestations of SS. Therefore, the standard of care for extraglandular manifestations varies widely and is based on local practice, expert opinion, and the personal experience of the treating physician.

[0007] New treatments aimed at reducing the substantial disease burden and overall unacceptable health status are needed. Summary of the Invention [Means for solving the problem]

[0008] Described herein are methods for treating Sjogren's syndrome (SS) in a subject in need of such treatment. forMethods are provided that include administering to a subject a Tn3 scaffold that includes a CD40L-specific monomeric subunit that binds to CD40L, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop SEQ ID NO: 16, and wherein a subject in need thereof experiences an increase in a subset of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to the subject's baseline level or the level of an untreated subject. have In embodiments, at least one CD40L-specific monomeric subunit comprises seven β-strands designated A, B, C, D, E, F, and G. wherein beta strand A comprises SEQ ID NO:5, beta strand B comprises SEQ ID NO:6, beta strand C comprises SEQ ID NO:17, beta strand D comprises SEQ ID NO:18, beta strand E comprises SEQ ID NO:19, beta strand F comprises SEQ ID NO:20, and beta strand G comprises SEQ ID NO:21. In embodiments, the subject has a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≧5. In embodiments, the ESSDAI score is assessed based on the ESSDAI domains consisting of skin, kidney, joint, muscle, hematology, gland, constitution, lymphadenopathy, and biology. In embodiments, the subject has (a) an ESSDAI score <5, (b) an EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score >5, and (c) a stimulated total salivary flow rate >0.1 mL / min. haveIn embodiments, the Tn3 scaffold is administered as an induction dose and then as maintenance doses. In embodiments, the induction dose comprises administering the Tn3 scaffold once every two weeks for at least three doses. In embodiments, the maintenance dose comprises administering the Tn3 scaffold once every four weeks for at least four doses. In embodiments, the time between the final induction dose and the first maintenance dose is about four weeks. In embodiments, the Tn3 scaffold is administered once every four weeks, once every two months, once every three months, once every four months, or once every six months. In embodiments, the Tn3 scaffold is administered over at least four doses. In embodiments, the Tn3 scaffold is administered over at least five doses. In embodiments, the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, dermally, systemically, topically, transdermally, or by inhalation. In embodiments, the Tn3 scaffold is administered intravenously. In embodiments, the Tn3 scaffold comprises two CD40L-specific monomer subunits linked in tandem. In embodiments, the two CD40L-specific monomer subunits each comprise SEQ ID NO: 3. In embodiments, the CD40L-specific monomer subunits are linked by a linker. In embodiments, at least one CD40L-specific monomer subunit is directly fused to polyethylene glycol (PEG). I'm In embodiments, at least one CD40L-specific monomer subunit is fused to polyethylene glycol (PEG) via a linker. I'm In embodiments, the linker comprises a peptide linker. In embodiments, the linker comprises SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10. In embodiments, at least one CD40L-specific monomer subunit is fused to albumin. I'mIn embodiments, the albumin is human serum albumin (HSA). In embodiments, the HSA is a mutant HSA comprising SEQ ID NO: 4. In embodiments, the Tn3 scaffold comprising a CD40L-specific monomer subunit is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg. In embodiments, the dose is about 1500 mg. In embodiments, the dose is about 2500 mg. In embodiments, the dose is about 3000 mg.

[0009] Provided are methods of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising administering to the subject a dose of about 1500 mg, about 2500 mg, or about 3000 mg of a Tn3 scaffold, wherein the Tn3 scaffold comprises SEQ ID NO: 1, and wherein the subject in need thereof exhibits an increase in B cells of a subset selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to an otherwise healthy subject. have In embodiments, the Tn3 scaffold is administered as an induction dose, followed by one or more maintenance doses. In embodiments, the induction dose and the maintenance dose are the same amount. In embodiments, the induction dose and the maintenance dose are different amounts. In embodiments, at least one dose is 3000 mg. In embodiments, the induction dose and at least one maintenance dose are administered about one month apart, about two months apart, The administration is about 3 months apart, about 4 months apart, or about 6 months apart. In embodiments, the induction dose comprises administering the Tn3 scaffold about every 2 weeks for at least 3 doses, and the maintenance dose comprises administering the Tn3 scaffold once every 4 weeks for at least 4 doses. In embodiments, the Tn3 scaffold comprises SEQ ID NO: 1. In embodiments, the administration is effective to reduce ESSDAI score when compared to an otherwise similar subject receiving a placebo control. In embodiments, the reduction is at least about a 1-point, 2-point, 3-point, 4-point, or 5-point reduction. In embodiments, the reduction is at least about a 6-point reduction. In embodiments, the administration is effective to reduce B-cell levels when compared to the subject's baseline level or the level in an untreated subject. In embodiments, the administration is effective to reduce serum CXCL13 levels when compared to the subject's baseline level or the level in an untreated subject. In embodiments, the administration is effective to reduce rheumatoid factor (RF) autoantibodies when compared to the subject's baseline level or the level in an untreated subject. In embodiments, the reduction is at least 14 days after administration. to It is detected.

[0010]

[0010] Provided is a method of treating an autoimmune disease in a subject in need thereof, comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L when biomarker levels are increased in the subject's sample compared to healthy baseline levels, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16. In embodiments, the biomarker is expressed on the surface of a cell. In embodiments, the biomarker is an antibody. In embodiments, the biomarker is secreted by a cell. In embodiments, the biomarker is selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, CD11c high atypical memory cells, CXCL13, RF autoantibodies, and any combination thereof. In embodiments, the biomarker is a protein. In embodiments, the protein biomarker is selected from the group consisting of TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, CD80, and any combination thereof. In embodiments, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjogren's syndrome, dermatomyositis, myositis, sclerosis, diabetes, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, rheumatic carditis, psoriasis, arthritis, inflammation, rheumatoid arthritis, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonopathy. In embodiments, the arthritis is rheumatoid arthritis.In embodiments, administration is continued until biomarker levels return to a healthy baseline. In embodiments, biomarker levels are analyzed periodically. In embodiments, the Tn3 scaffold comprises SEQ ID NO: 1. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0011] Provided is a method of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, and the DE loop comprises SEQ ID NO: 14. wherein the EF loop comprises SEQ ID NO: 15 and the FG loop SEQ ID NO: 16, and treatment of a subject with a Tn3 scaffold results in a decrease in the level of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells when compared to the subject's baseline level or the level in an untreated subject. In embodiments, the Tn3 scaffold comprises SEQ ID NO: 1. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg. In embodiments, B cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold.

[0012] Provided are methods of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16, and wherein treating the subject with the Tn3 scaffold reduces or eliminates the subject's baseline or untreated CD40L-specific monomelic subunits. When compared to the levels in a control, the level of T cells selected from the group consisting of Ki67+ / CD3+, Ki67+ / CD4+ cells, Ki67+ / CD3+ / CD4+ cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCRT7+ cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCR7- cells, Ki67+ / CD3+ / CD4+ / CXCR5+ / ICOS+ cells, Ki67+ / CD3+ / CD8+ cells, Ki67+ / CD3+ / CD8+ / CD45RA- / CCR7+ cells, Ki67+ / CD3+ / CD8+ / CXCR5+ cells, and CXCR5+ / ICOS+ / CD3+ / CD4+ cells is reduced. In embodiments, the Tn3 scaffold comprises SEQ ID NO: 1. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg. In embodiments, T cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold. [Brief explanation of the drawings]

[0013] [Figure 1]An exemplary study flow diagram is shown, including adults with moderate-to-severe (moderate-to-high) systemic disease activity, as defined by an ESSDAI score of ≥ 5, and the presence of anti-Ro autoantibodies and / or rheumatoid factor (RF). Seventy-four subjects were randomized (1:1) to receive three doses of intravenous VIB4920 1500 mg or placebo every two weeks, followed by four additional doses every four weeks (Q4W) (Stage I). Starting on Day 169, subjects randomized to VIB4920 received five doses of placebo Q4W, and subjects randomized to placebo received five doses of VIB4920 Q4W (Stage II). Randomization was stratified by EULAR Sjoegren's Syndrome Disease Activity Index (ESSDAI) score at screening (< 10 vs. ≥ 10). Δ = difference; D = days; EoS = end of study; EP = endpoint; ESSDAI = European League Against Rheumatism Sjoegren's Syndrome Disease Activity Index; ESSPRI = EULAR Sjoegren's Syndrome Patient Reported Index; IA = interim analysis; mos. = months; PE = primary endpoint. [Figure 2] Heat maps depicting the effects of dazodarivep on disease activity score, tender and swollen joint counts, and blood biomarkers of T cell and B cell costimulation in patients with moderate to severe active rheumatoid arthritis (top); CXCL13 and rheumatoid factor (middle); and post-switch memory B cells, plasmablasts, memory B progenitor cells, T cells, and Tfh cells (bottom). DAZ, dazodarivep; FC, fold change; DAS28-CRP, Disease Activity Score 28 with C-reactive protein; SDAI, Simple Disease Activity Index; CDAI, Clinical Disease Activity Index; post-switch memory B cells; Tfh cells, T follicular helper cells. [Figure 3A]Figures 3A-3G show that subjects in both cohorts 1 and 2 who received VIB4920 exhibited decreased levels of CXCL13 (Figure 3A), rheumatoid factor (Figure 3B), post-switch memory B cells (Figure 3C), plasmablasts (Figure 3D), memory progenitor B cells (Figure 3E), T cells (Figure 3F), and T follicular helper cells (Figure 3G). Data were analyzed using the Mann-Whitney U test for fold change relative to baseline values; *p<0.05; **p<0.01, ***p<0.001; DAZ, dazodaribep (VIB4920); DAZ-PBO, subjects who received DAZ in Stage I and PBO in Stage II; PBO, placebo; FC, fold change; PBO-DAZ, subjects who received PBO in Stage I and DAZ in Stage II; RF subset only, subjects who were RF positive at baseline were included. [Figure 3B-C] Same as above. [Figure 3D-E] Same as above. [Figure 3F-G] Same as above. [Figure 4] 1 is a heat map showing that subjects in both Cohorts 1 and 2 who received VIB4920 exhibited various reductions in T and B cell biomarkers. [Figure 5] Heat maps are shown showing the effect of dazodaribep (DAZ) versus placebo (PBO) on Population 1 (Pop1) and Population 2 (Pop2) at baseline, 15 days, and 169 days post-dose when compared to a healthy, treatment-naive cohort. Levels of 29 biomarkers were quantified as FC relative to baseline. FC = fold change. [Figure 6A-B]Figures 6A to 6DDD show the results of PDCD1 (Figures 6A to 6B), TNFRSF8 (Figures 6C to 6D), CD22 (Figures 6E to 6F), IL12B (Figures 6G to 6H), TNFRSF9 (Figures 6I to 6J), CXCL13 (Figures 6K to 6L), TNFRSF4 (Figures 6M to 6N), IL2RA (Figures 6O to 6P), TIGIT (Figures 6Q to 6R), IL2A / IL2B (Figures 6S to 6T), CCL19 (Figures 6U to 6V), NOS3 (Figures 6W to 6X), CD80 (Figures 6Y to 6Z), CD72 (Figures 6AA to 6BB), FCAMR (Figures 6CC to 6DD), and TNFRS. Graphs of biomarker levels are shown for F6B (Figures 6EE-6FF), CD79B (Figures 6GG-6HH), CXCL9 (Figures 6II-6JJ), CXCL11 (Figures 6KK-6LL), ADGRE1 (Figures 6MM-6NN), SLAMF7 (Figures 6OO-6PP), CD27 (Figures 6QQ-6RR), LAG3 (Figures 6SS-6TT), CXCL10 (Figures 6UU-6VV), ADA2 (Figures 6WW-6XX), IL10 (Figures 6YY-6ZZ), CCL21 (Figures 6AAA-6BBB), and TNFRSF13B (Figures 6CCC-6DDD). Levels are shown as absolute normalized protein expression (NPX) values ​​and fold-change levels from baseline. Data points were taken on days 1, 15, 169, and 309 post-dose. *=p ≤ 0.05; **=p ≤ 0.01; ***=p ≤ 0.001. [Figure 6C-D] Same as above. [Figure 6E-F] Same as above. [Figure 6G-H] Same as above. [Figure 6I-J] Same as above. [Figure 6K-L] Same as above. [Figure 6M-N] Same as above. [Figure 6O-P] Same as above. [Figure 6Q-R] Same as above. [Figure 6S-T] Same as above. [Figure 6U-V] Same as above. [Figure 6W-X] Same as above. [Figure 6Y-Z] Same as above. [Figures 6AA-BB] Same as above. [Figure 6CC-DD] Same as above. [Figure 6EE-FF] Same as above. [Figure 6GG-HH] Same as above. [Figure 6II-JJ] Same as above. [Figure 6KK-LL] Same as above. [Figure 6MM-NN] Same as above. [Figure 6OO-PP] Same as above. [Figure 6QQ-RR] Same as above. [Figure 6SS-TT] Same as above. [Figure 6UU-VV] Same as above. [Figure 6WW-XX] Same as above. [Figure 6YY-ZZ] Same as above. [Figure 6AAA-BBB] Same as above. [Figure 6CCC-DDD] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0014] Provided herein is a cluster of differentiation (CD) 40 ligand (CD40L)-human tenascin-C protein third fibronectin type III (Fn3) protein domain (Tn3) scaffold, a fusion protein, and methods for its use in SS. In some embodiments, the provided compositions and methods are utilized to treat glandular and extraglandular manifestations in SS patients with moderate to severe systemic disease activity, and to treat subjective symptoms of dryness, fatigue, and pain in SS patients with exocrine dysfunction. Cluster of differentiation (CD40) activation has been shown to be critical for germinal center formation, immunoglobulin (Ig) class switching, and cytokine expression, such as interferon-α, tumor necrosis factor-α, and interleukin-6, all of which have been previously implicated in the pathophysiology of SS. Dysregulation of CD40 / CD40L has been observed in both circulating cells and epithelial salivary gland cells in SS patients. These observations suggest that inhibition of the CD40L / CD40 pathway may be beneficial in SS.

[0015] The following description includes information that may be useful in understanding the present disclosure. This is not an admission that any of the information provided herein is prior art or relevant to the claimed disclosure, or that any publication specifically or implicitly referenced is prior art.

[0016] definition Although the following terms are believed to be well understood by those of ordinary skill in the art, the following definitions are provided to facilitate description of the subject matter disclosed herein.

[0017] All technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art, unless otherwise defined below. References to techniques used herein are intended to refer to techniques as commonly understood in the art, including variations on and / or equivalent technique alternatives that would be apparent to one of ordinary skill in the art.

[0018] As used herein, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0019] The terms "about" or "approximately," when placed immediately before a numerical value, refer to a range (e.g., ±10% of that value). For example, unless the context of this disclosure indicates otherwise or contradicts such an interpretation, "about 50" can mean 45 to 55, "about 25,000" can mean 22,500 to 27,500, etc. For example, in a list of numerical values ​​such as "about 49, about 50, about 55, ...," "about 50" refers to a range extending to less than half of one or more intervals between the preceding and following values, e.g., a range greater than 49.5 and less than 52.5. Furthermore, the phrases "less than about (a value)" or "greater than about (a value)" should be understood in light of the definition of the term "about" provided herein. Similarly, the term "about," when placed before a series of numerical values ​​or range of values ​​(e.g., "about 10, 20, 30" or "about 10 to 30"), refers to every value in that series or to each of the endpoints of that range.

[0020] As used herein, the term "subject" refers to any individual, e.g., a human or non-human mammal, for whom diagnosis, prognosis, or therapy is desired. The term "subject" can refer to a human or non-human mammal suffering from, susceptible to, or suspected of suffering from a disease. The terms "subject" and "patient" are used interchangeably herein. In embodiments, the subject is a mammal. Mammals include primates, e.g., humans, monkeys, chimpanzees, and apes, as well as non-primates, e.g., laboratory animals (rabbits and rodents, e.g., guinea pigs, rats, or mice), and domestic animals, including domestic pets and farm animals (e.g., cats, dogs, pigs, cows, sheep, goats, horses, rabbits), as well as non-domestic animals, such as wildlife, birds, reptiles, fish, and the like. Typically, the subject is a human subject.

[0021] As used herein, the term "subject in need thereof" includes a subject who may or will benefit from the methods described herein. Subjects in need of treatment include, without limitation, subjects already with a condition or disorder, subjects prone to having a condition or disorder, subjects suspected of having a condition or disorder, and subjects in whom a condition or disorder is to be prevented, ameliorated, or reversed.

[0022] As used herein, "treating" or "treating" refers to the management and care of a subject for the purpose of combating a disease, condition, or disorder, and includes alleviating the symptoms or complications of a disease, condition, or disorder, or eliminating the disease, condition, or disorder through administration of a Tn3 scaffold used in the methods described herein. Thus, the term "treating" or "treating" refers to both therapeutic and prophylactic or preventative treatments, the goal of which is to prevent, slow (reduce), or ameliorate the progression of a disease (e.g., SS). Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of the disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, improvement or palliation of the disease state, and reversal of the disease (whether partial or complete). The term "treating" can also include treatment of cells in vitro or in animal models.

[0023] As used herein, "fused" refers to at least two polypeptides that have been recombinantly joined together. As used herein, "conjugated" refers to the formation of a bond between two components by a chemical reaction. The bond may be covalent or non-covalent. Typically, the two components that are conjugated to each other are chemically linked by a covalent bond.

[0024] When referring to nucleic acid or protein sequences, the term "identity" is used to refer to the similarity between two sequences. Unless otherwise indicated, the percent identity reported herein is determined using the BLAST algorithm, available on the world wide web at the following address: blast.ncbi.nlm.nih.gov / Blast.cgi, with default parameters. It is determined using a meter.

[0025] Described herein is the use of a Tn3 scaffold containing a CD40L-specific monomeric subunit to treat SS. for A method is described.

[0026] In one embodiment, the Tn3 scaffold is used in a method for treating SS. SS is a systemic autoimmune disease characterized by chronic lymphocytic inflammation of exocrine glands, primarily the salivary and lacrimal glands, leading to loss of function and manifesting as excessive dryness. Extraglandular manifestations have been described as multisystemic involvement affecting the musculoskeletal, pulmonary, renal, neurological, dermatological, gastrointestinal, hematological, hepatobiliary, or vascular systems, while fatigue is a particularly prominent comorbidity. SS may also be seen in association with other autoimmune diseases. In one embodiment, the method involves treating a patient with SS with moderate to severe systemic disease activity by administering the Tn3 scaffold. In one embodiment, moderate to severe systemic disease activity is defined as a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≥ 5. In embodiments, the method comprises treating a patient with SS with moderate to severe (and in some cases severe) subjective symptoms by administration of a Tn3 scaffold, in embodiments, moderate to severe subjective symptoms are defined by a EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score of ≥ 5, and mild systemic disease activity as defined by an ESSDAI score < 5 with residual stimulated salivary flow.

[0027] In embodiments, the method includes treating SS in a subject in need thereof by administering a Tn3 scaffold comprising a CD40L-specific monomeric subunit. In embodiments, the Tn3 scaffold binds to CD40L. In embodiments, the monomeric subunit of the Tn3 scaffold comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, where the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16. In embodiments, the Tn3 scaffold is VIB4920. The CD40 receptor is a member of the TNF receptor family expressed on the plasma membrane of antigen-stimulated B cells, macrophages, and dendritic cells. The CD40 receptor functions to provide costimulatory signals to antigen-bound B cells. The cognate ligand for CD40 is CD40L (also known as CD154), which is expressed on the plasma membrane of T cells and other cell types, including platelets.

[0028] Tn3 scaffold Provided herein are compositions that bind to CD40L. In embodiments, the compositions provided comprise a CD40L antagonist. In embodiments, provided herein are compositions comprising a Tn3 scaffold (e.g., a "Tn3 scaffold") comprising a CD40L-specific monomer subunit. In embodiments, provided herein are compositions comprising a Tn3 scaffold comprising two CD40L-specific monomer subunits.

[0029] In embodiments, provided compositions may comprise an amino acid sequence as described in PCT / US2012 / 059477 and PCT / US2019 / 052997, which are incorporated by reference herein in their entireties. In embodiments, provided compositions may comprise an amino acid sequence as set forth in SEQ ID NO: 1 (referred to herein as VIB4920). VIB4920 comprises a bivalent CD40L-specific Tn3 protein fused to a protein.

[0030] In embodiments, the CD40L monomer subunit of the Tn3 scaffold comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG. In embodiments, the Tn3 scaffold comprises a single CD40L-specific monomer subunit. In embodiments, the Tn3 scaffold comprises two CD40L-specific monomer subunits. In embodiments, the two CD40L-specific monomer subunits are linked in tandem. In embodiments, the two CD40L-specific monomer subunits are linked by a linker. In embodiments, the linker comprises a peptide linker, which may be a flexible peptide linker. In embodiments, the peptide linker is a (G m X) n wherein X is serine (S), alanine (A), glycine (G), Leu (L), isoleucine (I), or valine (V); m and n are integer values; m is 1, 2, 3, or 4; and n is 1, 2, 3, 4, 5, 6, or 7.

[0031] In embodiments, the Tn3 scaffold comprises a linker comprising a functional moiety. In embodiments, the functional moiety is an immunoglobulin or a fragment thereof. In embodiments, the immunoglobulin or fragment thereof comprises an Fc domain. In embodiments, the Fc domain is incapable of inducing at least one FcγR-mediated effector function (e.g., is Fc-deficient). In embodiments, the at least one FcγR-mediated effector function is antibody-dependent cellular cytotoxicity (ADCC).

[0032] In embodiments, the Tn3 scaffold comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises or consists of SEQ ID NO: 11, the BC loop comprises or consists of SEQ ID NO: 12, the CD loop comprises or consists of SEQ ID NO: 13, the DE loop comprises or consists of SEQ ID NO: 14, the EF loop comprises or consists of SEQ ID NO: 15, and the FG loop comprises or consists of SEQ ID NO: 16. In embodiments, the Tn3 scaffold comprises or consists of SEQ ID NO: 1 (also known as VIB4920). In embodiments, beta strand A comprises or consists of SEQ ID NO:5, beta strand B comprises or consists of SEQ ID NO:6, beta strand C comprises or consists of SEQ ID NO:17, beta strand D comprises or consists of SEQ ID NO:18, beta strand E comprises or consists of SEQ ID NO:19, beta strand F comprises or consists of SEQ ID NO:20, and beta strand G comprises or consists of SEQ ID NO:21.

[0033] In embodiments, one or more CD40L-specific Tn3 monomers have a β-strand A that comprises or consists of IEV (SEQ ID NO:5), RLDAPSQIEV (SEQ ID NO:23), or SQIEV (SEQ ID NO:24). In embodiments, a Tn3 scaffold may comprise one or more CD40L-specific Tn3 monomers with the same or different β-strand A sequences. For example, a first CD40L-specific Tn3 monomer β-strand A may comprise or consist of IEV (SEQ ID NO:5), and a second CD40L-specific Tn3 monomer β-strand A may comprise or consist of RLDAPSQIEV (SEQ ID NO:23) or SQIEV (SEQ ID NO:24).

[0034] The Tn3 scaffold may have the amino acid sequence as set forth in SEQ ID NO: 1 and described above, or it may have one or more amino acid residue changes compared to the amino acid sequence as set forth in SEQ ID NO: 1. For example, if there is an amino acid sequence change in the scaffold compared to that set forth in SEQ ID NO: 1, the change may be in one of the linkers. The Tn3 scaffold may include a Gly15 linker separating the two CD40L-specific monomers and a Gly10 linker separating the CD40L-specific monomer from the HSA sequence. Both of these linkers Or one of them may be modified, (G m X) n where X is serine (S), alanine (A), glycine (G), Leu (L), isoleucine (I), or valine (V); m and n are integer values; m is 1, 2, 3, or 4; and n is 1, 2, 3, 4, 5, 6, or 7. For example, one or both linkers may be modified to have an amino acid sequence comprising one of GGGGSGGGGS (SEQ ID NO: 7), GGGGSGGGGSGGGGS (SEQ ID NO: 8), GGGGGGGGGG (SEQ ID NO: 9), or GGGGGGGGGGGGGG (SEQ ID NO: 10). If the Tn3 scaffold has an amino acid sequence different from the amino acid sequence as provided in SEQ ID NO: 1, it may be due to one or more changes in the HSA amino acid sequence fused to the two CD40L-specific monomers. The HSA fused to two CD40L-specific monomers may be modified compared to the HSA fused to two CD40L-specific Tn3 monomers, except for at least one amino acid substitution at a position selected from the group consisting of 407, 415, 463, 500, 506, 508, 509, 511, 512, 515, 516, 521, 523, 524, 526, 535, 550, 557, 573, 574, and 580, numbered based on their positions in full-length mature HSA; wherein the at least one amino acid substitution does not include a lysine (K) to glutamic acid (E) at position 573.

[0035] Exemplary sequences of Tn3 scaffolds are shown in Table 1. In embodiments, the Tn3 scaffold comprises at least about or at most about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or up to about 100% identity to any one of SEQ ID NOs: 1-25 shown in Table 1. In embodiments, any one of the sequences from Table 1 may be modified. In embodiments, the modification comprises one or more truncations, deletions, insertions, and combinations thereof. Modifications can be made to any of the residues provided in Table 1, and to any number of residues from Table 1. In embodiments, the modification can comprise 1 to 3, 1 to 5, 1 to 10, 5 to 20, 1 to 3, 1 to 5, 1 to 10, 1 to 20, 3 to 8, 3 to 10, 3 to 15, 5 to 8, 5 to 10, or 5 to 20 residues. In embodiments, modifications can be made to up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, or 450 residues.

[0036] [Table 1-1]

[0037] [Table 1-2]

[0038] [Table 1-3]

[0039] [Table 1-4]

[0040] When a Tn3 scaffold has an amino acid sequence change compared to that set forth in SEQ ID NO: 1, the change may be in the amino acid sequence of one or both of the CD40L-specific Tn3 monomers, for example, such that one or both CD40L-specific Tn3 monomers have the amino acid sequence as set forth in SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 22, and SEQ ID NO: 25, so long as it does not adversely affect the in vivo efficacy of the scaffold. In embodiments, the first one or two amino acid residues (SQ) at the N-terminus may be absent and / or substituted with alternative amino acid residues. In embodiments, the Tn3 scaffold comprises a monomer subunit comprising SEQ ID NO: 22, SEQ ID NO: 25, or both SEQ ID NO: 22 and SEQ ID NO: 25.

[0041] In embodiments, the Tn3 scaffold comprises at least one CD40L-specific monomeric subunit linked to a heterologous moiety. In embodiments, the heterologous moiety is selected from the group consisting of a protein, peptide, protein domain, linker, drug, toxin, cytotoxic agent, imaging agent, radionuclide, radioactive compound, organic polymer, inorganic polymer, polyethylene glycol (PEG), biotin, albumin, HSA FcRn binding moiety, antibody or fragment thereof, single-chain antibody, domain antibody, albumin binding domain, enzyme, ligand, receptor, binding peptide, non-FnIII scaffold, epitope tag, recombinant polypeptide polymer, cytokine, and combinations of two or more of the foregoing moieties. In embodiments, the heterologous moiety is albumin, and the albumin comprises human serum albumin. In embodiments, the heterologous moiety is an antibody. In embodiments, the antibody is selected from the group consisting of an Fc domain of an antibody, an antibody fragment, and a single-chain antibody.

[0042] In embodiments, the heterologous moiety is an antibody. In embodiments, the antibody is selected from the group consisting of an Fc domain of an antibody, an antibody fragment, and a single chain antibody.

[0043] In embodiments, the heterologous moiety is an imaging agent, such as a radionuclide or biotin. In embodiments, the heterologous moiety is a drug, such as a cytotoxic agent or a radioactive compound.

[0044] In embodiments, the heterologous moiety comprises PEG. In embodiments, the Tn3 scaffold is fused to PEG directly or via a linker. I'm or at least one conjugated In embodiments, the CD40L-specific monomer subunits are fused, conjugated, or linked to PEG via a linker. In embodiments, the Tn3 scaffold is fused to PEG directly or via a linker. I'm The CD40L-specific monomeric subunit comprises at least one (eg, two) CD40L-specific monomeric subunit that is either or conjugated to the CD40L-specific monomeric subunit.

[0045] In embodiments, the heterologous moiety comprises albumin. In embodiments, the Tn3 scaffold is fused to albumin directly or via a linker. I'm The albumin comprises at least one CD40L-specific monomer subunit attached or conjugated to it. In embodiments, the albumin is HSA. In embodiments, the HSA is a mutant HSA. In embodiments, the amino acid sequence of the mutant HSA is SEQ ID NO: 4. In embodiments, the mutant HSA has at least one improved property compared to native HSA or a native HSA fragment. In embodiments, the amino acid sequence of the mutant HSA is SEQ ID NO: 4 or a sequence having at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% identity to SEQ ID NO: 4. In embodiments, the improved property is an altered plasma half-life compared to the plasma half-life of native HSA or a native HSA fragment. In embodiments, the altered plasma half-life is a longer plasma half-life compared to the plasma half-life of native HSA or a native HSA fragment. In embodiments, the altered plasma half-life is a plasma half-life that is shorter compared to the plasma half-life of native HSA or a fragment of native HSA.

[0046] Dose setting In embodiments, any of the compositions comprising the Tn3 scaffold of the present disclosure can be administered in any form. In embodiments, the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, cutaneously, systemically, topically, transdermally, or by inhalation. In embodiments, the Tn3 scaffold is administered intravenously. In embodiments, the Tn3 scaffold is administered by intravenous infusion.

[0047] The Tn3 scaffold of the present disclosure can be administered at any dose. In embodiments, the Tn3 scaffold is administered at a dose of about 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1450 mg, 1500 mg, 1550 mg, 1600 mg, 1650 mg, 1700 mg, 1750 mg, 1800 mg, 1900 mg, 2000 mg, 2100 mg, 2200 mg, 2300 mg, 2400 mg, 2500 mg, 2600 mg, 2700 mg, 2800 mg, 2900 mg, 3000 mg, 3100 mg, 3200 mg, 3300 mg, 3400 mg, 350 mg, 3600 mg, 3700 mg, 3800 mg, 3900 mg, 4000 mg, 4100 mg, 4200 mg, 4300 mg, 4400 mg, 450 mg, 4600 mg, 4700 mg, 4800 mg, 4900 mg, 5000 mg, 5100 mg, 5200 mg, 5300 mg, 5400 mg, 550 mg, 5600 mg, 5700 mg, 5800 mg, 5900 mg, 6000 mg, 6100 mg, 6200 mg, 6300 mg, 6400 mg, 650 mg, 6600 mg, mg, 1850mg, 1900mg, 1950mg, 2000mg, 2050mg, 2100mg, 2150mg, 2200mg, 2250mg, 2300mg, 2350 mg, 2400mg, 2450mg, 2500mg, 2550mg, 2600mg, 2650mg, 2700mg, 2750mg, 2800mg, 2850mg, 2900m g, 2950mg, 3000mg, 3050mg, 3100mg, 3150mg, 3200mg, 3250mg, 3300mg, 3350mg, 3400mg, 3450m g, 3500mg, 3550mg, 3600mg, 3650mg, 3700mg, 3750mg, 3800mg, 3850mg, 3900mg, 3950mg, 4000mg , 4050 mg, 4100 mg, 4150 mg, 4200 mg, 4250 mg, 4300 mg, 4350 mg, 4400 mg, 4450 mg, 4500 mg, 4550 mg, 4600 mg, 4650 mg, 4700 mg, 4750 mg, 4800 mg, 4850 mg, 4900 mg, 4950 mg, or about 5000 mg. Any of the foregoing dosages can be effective dosages for methods including treatment, reduction, or elimination.

[0048] In embodiments, the Tn3 scaffold is administered at a dose of between about 800-5000 mg, 900-4900 mg, 1000-4800 mg, 1100-4700 mg, 1200-4600 mg, or 1300-4500 mg. In embodiments, the Tn3 scaffold is administered at a dose of between about 1300 mg, 1350 mg, 1400 mg, 1450 mg, 1500 mg, 1550 mg, 1600 mg, g, 1650mg, 1700mg, 1750mg, 1800mg, 1850mg, 1900mg, 1950mg, 2000mg, 2050mg, 2100mg, 2150mg, 2200mg, 2250mg, 2300mg, 2350mg, 2400mg, 2450mg, 2500mg, 2550mg, 2600mg, 2650mg, 2700mg, 2750mg, 2800mg, 2850mg, 2900mg, 2950mg, 3000mg, 3050mg, 3100mg, 315 In an embodiment, the Tn3 scaffold is administered at a dose selected from the group consisting of 0 mg, 3200 mg, 3250 mg, 3300 mg, 3350 mg, 3400 mg, 3450 mg, 3500 mg, 3550 mg, 3600 mg, 3650 mg, 3700 mg, 3750 mg, 3800 mg, 3850 mg, 3900 mg, 3950 mg, 4000 mg, 4050 mg, 4100 mg, 4150 mg, 4200 mg, 4250 mg, 4300 mg, 4350 mg, 4400 mg, 4450 mg, and 4500 mg. In an embodiment, the Tn3 scaffold is administered at a dose selected from the group consisting of 1500 mg and 3000 mg. In an embodiment, the Tn3 scaffold is administered at a dose of about 1500 mg. In embodiments, the Tn3 scaffold is administered at a dose of about 3000 mg.

[0049] Dosage frequency In embodiments, the Tn3 scaffold of the present disclosure is administered according to a schedule that produces optimal results.In embodiments, the Tn3 scaffold is administered to a subject in need thereof about once a week, about twice a week, about every two weeks, about once a month, about every four weeks, about every two months, about every three months, about every 12 weeks, about every 15 weeks, about every 16 weeks, about every four months, about every five months, about every six months, or every six months.Any number of administrations can be provided to a subject in need thereof.

[0050] The Tn3 scaffold of the present disclosure can be administered in a total of about 1-10, 10-50, 50-75, 75-100, 100-200, 200-300 doses, or over the course of the subject's lifetime. In embodiments, the Tn3 scaffold is administered in about 1, 2, 3, 4, 5, 6, 7, 8, 9 doses, or up to about 10 doses. In embodiments, the Tn3 scaffold is administered in a total of at least about or at most about 2, 3, 4, or 5 doses.

[0051] In embodiments, a subject is administered an effective dose about every 1, 2, 3, 4, 5, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, or 5 years after initiation of treatment, or for the lifetime of the subject. In embodiments, a subject receives an effective dose on days 1, 15, 29, and 57 after initiation of treatment. In embodiments, a subject receives 1500 mg to 3000 mg of a Tn3 scaffold on days 1, 15, 29, and 57 after initiation of treatment. In embodiments, a subject receives about 1500 mg to 3000 mg of a Tn3 scaffold every 2 weeks for about three administrations, and then every 4 weeks thereafter. In embodiments, a subject is administered about 1500 mg of Tn3 scaffold every two weeks for about three administrations, and then every four weeks thereafter. In embodiments, a subject is administered about 3000 mg of Tn3 scaffold at weeks 1, 4, and 12, and then every 12 weeks thereafter. In embodiments, a subject in need thereof is administered 1500 mg of Tn3 scaffold on days 1, 15, 29, and 57 after initiation of treatment. In embodiments, a subject in need thereof is administered 1500 mg of Tn3 scaffold on days 1 and 57 after initiation of treatment. In embodiments, a subject in need thereof is administered 3000 mg of Tn3 scaffold on days 1, 15, 29, and 57 after initiation of treatment. In embodiments, a subject in need thereof is administered 3000 mg of Tn3 scaffold on days 1 and 57 after initiation of treatment, and then every six months thereafter as needed. In embodiments, a subject is administered an initial dose of about 1500-3000 mg of a Tn3 scaffold of the present disclosure administered every two weeks for at least two, at least three, or more administrations, and then every four weeks thereafter. The dose is administered every two weeks for at least two, at least three, or more administrations, and then every four weeks thereafter. In embodiments, the subject is administered an initial dose of about 3000 mg of the Tn3 scaffold of the present disclosure for at least two, at least three, or more administrations every two weeks, and then every 12 weeks thereafter. In embodiments, the subject is administered 1500 mg of the Tn3 scaffold of the present disclosure every four weeks, with a loading dose of 1500 mg of the Tn3 scaffold administered in week two. In embodiments, the subject is administered 3000 mg of the Tn3 scaffold of the present disclosure every 12 weeks, with a loading dose of 3000 mg of the Tn3 scaffold administered in week four. In embodiments, the subject is administered 1500 mg of the Tn3 scaffold of the present disclosure every four weeks, with a loading dose of 1500 mg of the Tn3 scaffold administered in week two. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 1 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 22 and / or SEQ ID NO: 25 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 1 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 1500 mg of a Tn3 scaffold comprising SEQ ID NO: 22 or SEQ ID NO: 25 every 4 weeks, wherein a loading dose of 1500 mg of Tn3 scaffold is administered in week 2. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 22 or SEQ ID NO: 25 every 12 weeks, where a loading dose of 3000 mg of Tn3 scaffold is administered in week 4.

[0052] Therapeutic or preventative methods of the present disclosure include administering a Tn3 scaffold of the present disclosure to a subject in need thereof. In embodiments, the treatment method includes administering an effective amount of a Tn3 scaffold to a subject in need thereof for about three doses (±2 doses) every two weeks, followed by administration of a Tn3 scaffold every four weeks thereafter. In embodiments, the treatment method includes administering an effective amount of a Tn3 scaffold to a subject in need thereof at weeks 1, 4, and 12, followed by administration of a Tn3 scaffold every 12 weeks thereafter. In embodiments, the treatment method includes administering about 1400 mg to 1600 mg of a Tn3 scaffold to a subject in need thereof for about three doses (±2 doses) every two weeks, followed by administration of a Tn3 scaffold every four weeks thereafter. In embodiments, the treatment method includes administering about 2000 mg to 4000 mg of a Tn3 scaffold to a subject in need thereof at weeks 1, 4, and 12, followed by administration of a Tn3 scaffold every 12 weeks thereafter. In embodiments, the method of treatment comprises administering about 1500 mg of Tn3 scaffold to a subject in need thereof at about three doses (±2 doses) every two weeks, followed by administration of the Tn3 scaffold every four weeks thereafter. In embodiments, the method of treatment comprises administering about 3000 mg of Tn3 scaffold to a subject in need thereof at weeks 1, 4, and 12, followed by administration of the Tn3 scaffold every 12 weeks thereafter. In embodiments, administration occurs every four weeks and continues thereafter for up to about 1, 2, 3, 4, or 5 years. In embodiments, administration of the Tn3 scaffold continues until the subject dies. Any of the foregoing doses can be delayed by about 0-5 days, 0-4 days, 0-3 days, 0-2 days, or about 1 day.

[0053] In embodiments, a subject receives an effective dose of Tn3 scaffold on days 1, 15 (± 1 day), 29 (± 3 days), and 57 (± 3 days) after initiation of treatment. In embodiments, a subject in need thereof is administered 1500 mg of Tn3 scaffold on days 1, 15 (± 1 day), 29 (± 3 days), and 57 (± 3 days) after initiation of treatment, and then every 6 months thereafter as needed. In embodiments, a subject in need thereof is administered 1500 mg of Tn3 scaffold on days 1, 15 (± 1 day), and 29 (± 3 days) after initiation of treatment. In embodiments, a subject in need thereof is administered 1500 mg of Tn3 scaffold on days 1 and 57 (± 3 days) after initiation of treatment. In embodiments, a subject in need thereof is administered 3000 mg of Tn3 scaffold on days 1, 15 (± 1 day), 29 (± 3 days), and 57 (± 3 days) after initiation of treatment, and then every 6 months thereafter as needed. In embodiments, a subject in need thereof is administered 3000 mg of Tn3 scaffold on days 1, 15 (± 1 day), and 29 (± 3 days) after initiation of treatment. In embodiments, a subject in need thereof is administered 3000 mg of Tn3 scaffold on days 1 and 57 (± 3 days) after initiation of treatment, and then every 6 months thereafter as needed.

[0054] In embodiments, a subject in need thereof is administered a dose of the Tn3 scaffold on days 1, 15 (± 1 day), 29 (± 3 days), 57 (± 3 days), 85 (± 3 days), 113 (± 3 days), and 141 (± 3 days). In embodiments, a subject in need thereof is administered an effective dose of the Tn3 scaffold on days 169 (± 3 days), 197 (± 3 days), 225 (± 3 days), 253 (± 3 days), and 281 (± 3 days).

[0055] In embodiments, a subject in need thereof is administered an effective dose of the Tn3 scaffold once every 2-4 weeks. In embodiments, a subject in need thereof is administered an effective dose of the Tn3 scaffold once every 2 weeks, every 4 weeks, or every 12 weeks. In embodiments, a subject in need thereof is administered 1500 mg of the Tn3 scaffold once every 2 weeks for at least 3 doses, once every 4 weeks for at least 4 doses, once every 4 weeks for at least 5 doses, or a combination thereof. In embodiments, a subject in need thereof is administered 3000 mg of the Tn3 scaffold once every 2 weeks for at least 3 doses, once every 4 weeks for at least 4 doses, once every 4 weeks for at least 5 doses, or a combination thereof. In embodiments, a subject in need thereof is administered an effective dose of the Tn3 scaffold as an induction dose, followed by maintenance doses. In embodiments, 3000 mg of the Tn3 scaffold is administered every 3 months. In embodiments, 3000 mg of the Tn3 scaffold is administered every 12 weeks.

[0056] In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every two weeks for at least two doses, and then about once a month. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every two weeks for at least three doses, and then about once a month. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every two weeks for at least three doses, and then every four weeks. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once a month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered at a dose of about 3000 mg about once a month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered in two or more doses. In embodiments, a subject is administered 1500 mg of the Tn3 scaffold of the present disclosure every 4 weeks, with a loading dose of 1500 mg of the Tn3 scaffold administered in week 2. In embodiments, a subject is administered 3000 mg of the Tn3 scaffold of the present disclosure every 12 weeks, with a loading dose of 3000 mg of the Tn3 scaffold administered in week 4. In embodiments, a subject is administered 1500 mg of a Tn3 scaffold comprising SEQ ID NO: 1 every 4 weeks, with a loading dose of 1500 mg of the Tn3 scaffold administered in week 2. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 1 every 12 weeks, with a loading dose of 3000 mg of the Tn3 scaffold administered in week 4. In embodiments, a subject is administered 1500 mg of a Tn3 scaffold comprising SEQ ID NO: 22 or SEQ ID NO: 25 every 4 weeks, with a loading dose of 1500 mg of Tn3 scaffold administered on week 2. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 22 or SEQ ID NO: 25 every 12 weeks, with a loading dose of 3000 mg of Tn3 scaffold administered on week 4.

[0057] In embodiments, a subject is administered 1500 mg of a Tn3 scaffold of the present disclosure every 4 weeks, where a loading dose of 1500 mg of the Tn3 scaffold is administered in week 2. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO: 1 every 12 weeks, where a loading dose of 1500 mg of the Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO:22 and / or SEQ ID NO:25 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO:1 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4. In embodiments, a subject is administered 1500 mg of a Tn3 scaffold comprising SEQ ID NO:22 or SEQ ID NO:25 every 4 weeks, wherein a loading dose of 1500 mg of Tn3 scaffold is administered in week 2. In embodiments, a subject is administered 3000 mg of a Tn3 scaffold comprising SEQ ID NO:22 or SEQ ID NO:25 every 12 weeks, wherein a loading dose of 3000 mg of Tn3 scaffold is administered in week 4.

[0058] Treatment method Methods for treating autoimmune diseases are provided. The compositions of the present disclosure may be used to treat any autoimmune disease. In some autoimmune diseases, the CD40 / CD40L pathway is activated in various cell types, including T cells, B cells, antigen-presenting cells, and activated epithelial cells. In embodiments, the compositions of the present disclosure can be used in autoimmune diseases in which the CD40 / CD40L pathway is activated in various cell types, including T cells, B cells, antigen-presenting cells, and activated epithelial cells. In embodiments, the Tn3 scaffold of the present disclosure blocks costimulatory signals between T cells and CD40-expressing B cells, resulting in the destruction of germinal center development, pathogenic B cells, plasma cells, and autoantibodies, hallmarks of autoimmune diseases.

[0059] In embodiments, the autoimmune disease is systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjogren's syndrome, dermatomyositis, myositis, sclerosis, diabetes, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, rheumatic carditis, psoriasis, arthritis (e.g., rheumatoid arthritis; RA), inflammation, rheumatoid arthritis, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, type 1 interferonopathy, and any combination thereof. In embodiments, the autoimmune disease or other disease or condition comprises an increased level of a T cell or B cell biomarker of the present disclosure.

[0060] Rheumatoid arthritis In embodiments herein, a method relates to treating RA. In embodiments, the method includes administering a Tn3 scaffold of the present disclosure. In embodiments, the Tn3 scaffold is administered to a subject in need thereof for the treatment of RA using any of the dosing schedules disclosed herein. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every two weeks for at least two doses, and then about once monthly. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every two weeks for at least three doses, and then about once monthly. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg about once every month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered at a dose of about 3000 mg about once every month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered in two or more doses.

[0061] In an embodiment of the method disclosed herein, the subject in need thereof has been administered one or more standard care therapies for the treatment of RA before administering the Tn3 scaffold. In an embodiment, the provided method is described in PCT / US2022 / 077192, which is incorporated herein by reference in its entirety.

[0062] Sjögren's syndrome In embodiments herein, methods relate to treating SS. In embodiments, the methods comprise administering a Tn3 scaffold of the present disclosure. In embodiments, the Tn3 scaffold is The Tn3 scaffold is administered to a subject in need thereof for the treatment of SS using any of the dosing schedules disclosed herein. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg once every two weeks for at least two doses, and then about once a month. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg once every two weeks for at least three doses, and then about once a month. In embodiments, the Tn3 scaffold is administered at a dose of about 1500 mg once every month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered at a dose of about 3000 mg once every month, about once every two months, or about once every three months. In embodiments, the Tn3 scaffold is administered in two or more doses.

[0063] In embodiments, the methods involve treating a patient with SS. In embodiments, the methods involve treating a patient with SS with a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of about 1, about 2, about 3, about 5, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or more. In embodiments, the methods involve treating a patient with SS with a EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score of about 1, about 2, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, or more.

[0064] In embodiments, the method comprises treating a patient in need thereof. In embodiments, the method comprises treating a patient with SS. In embodiments, the method comprises treating a patient with SS with moderate systemic disease activity. In embodiments, the method comprises treating a patient with SS with high systemic disease activity. In embodiments, the method comprises treating a patient with SS with moderate to high systemic disease activity. In embodiments, the method comprises treating a patient with SS with moderate to high systemic disease activity by administering any dose of a Tn3 scaffold disclosed herein on any schedule. In embodiments, moderate to high systemic disease activity may be defined by an ESSDAI score of ≥ 5. In embodiments, the method comprises treating a patient with SS with moderate to high (or severe) subjective symptoms by administering a Tn3 scaffold. In embodiments, moderate to severe (or severe) subjective symptoms can be defined by a EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score of ≧5, and mild systemic disease activity can be defined by an ESSDAI score of <5 with residual stimulated salivary flow. In embodiments, the Tn3 scaffold is administered at a dose of 1500 mg. In embodiments, the Tn3 scaffold is administered at a dose of 3000 mg.

[0065] Also provided are diagnostic methods that help identify subjects who may benefit from the administration of the compositions of the present disclosure. In embodiments, the methods include determining whether a subject's sample contains a biomarker of the present disclosure that provides information about treatment. For example, the methods may include determining whether a subject in need has Ki67+CD27+ memory cells, CD27 high CD38 In some embodiments, the method may include determining whether a subject in need thereof is positive for one or more markers or cells selected from the group consisting of CXCL13, RF antibodies, Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, and administering a composition of the present disclosure if a positive result is obtained. In some embodiments, the method may include determining whether a subject in need thereof is positive for one or more markers or cells selected from the group consisting of CXCL13, RF antibodies, Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, and administering a composition of the present disclosure if a positive result is obtained. In some embodiments, a positive result is determined by ... In embodiments, a positive result includes an increase in the number of one or more cells compared to an otherwise identical healthy subject. In embodiments, the method includes periodically determining whether a subject in need is positive for one or more cells, and administering treatment as needed, thereby returning the level of one or more cells to the subject's baseline value. A baseline level may refer to the level of a biomarker before administration of a composition of the present disclosure and thus can be associated with a disease. In embodiments, a baseline level may refer to the level of a biomarker after administration of a composition of the present disclosure when the subject has or does not have a disease set as a control (e.g., a healthy baseline).

[0066] evaluation In embodiments, the subject is evaluated. The evaluation can be performed at any time before, during, or after administration of the Tn3 scaffold. The evaluation can be performed at any time before, during, or after administration of the Tn3 scaffold. In embodiments, the evaluation is performed before administration. In embodiments, the evaluation is performed during administration. In embodiments, the evaluation is performed after administration.

[0067] Any of the assessments referred to below can be performed at any time. In some embodiments, the subject is assessed every minute, every hour, every day, every week, every month, or every year. In some embodiments, assessments are completed twice daily, every other week, every other month, or every six months. In embodiments, the ratings are -10, -9, -8, -7, -6, -5, -4, -3, -2, -1, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187 45, 150, 155, 160, 165, 169, 170, 180, 190, 197, 200, 205, 210, 215, 220, 225, 230, 235, 240, 250, 252, 253, 255, 260, 265, 270, 280, 281, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316 , 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, or until about day 365 ± 7 days after treatment.

[0068] As discussed above, in embodiments, subjects undergo periodic assessment to determine the level of cells expressing a biomarker (or level of a biomarker) of the present disclosure. In embodiments, the level of cells expressing a biomarker of the present disclosure correlates with disease activity. For example, T cells and / or B cells may be costimulated in a diseased state, causing upregulation of cells expressing a biomarker of the present disclosure, thus leading to an increase in disease in the subject. Periodic assessment of a subject may allow for management or reduction of disease through monitoring and appropriate intervention. Periodic assessment may include daily, weekly, monthly, or yearly monitoring. In embodiments, subjects are assessed 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times per year. In embodiments, subjects are assessed 1-3, 2-4, 3-4, or 4-5 times per year. 5, 4-6, or 5-7 times. In embodiments, subjects are assessed annually, every 2 years, every 3 years, every 4 years, or every 5 years.

[0069] In embodiments, treatment of SS may be characterized by a reduction in clinical symptoms of the disease or disorder, or a reduction in inflammation, or a reduction in biomarkers of the disease or disorder by at least about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about 100% compared to levels prior to treatment with the Tn3 scaffold. The reduction in any of these symptoms, inflammation, or biomarkers can be at least about a 10%, 15%, 20%, 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about a 100% reduction in symptoms, inflammation, or biomarkers compared to their levels before treatment with the Tn3 scaffold was initiated. The reduction can be such that SS is characterized as being in remission.

[0070] In embodiments, administration of a composition of the present disclosure is effective to reduce the level of a biomarker, specific cell type, or level of cells expressing a biomarker described herein. In embodiments, the biomarker is reduced by at least or at most about 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject. In embodiments, the level of B cells is reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold after administration of a composition of the present disclosure.

[0071] In embodiments, the level of Ki67+CD27+ memory cells is reduced by at least or at most about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject. In embodiments, the level of CD27 high CD38 high plasmablasts is reduced by at least or at most about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject. In embodiments, the level of CDl lc high atypical memory cells is reduced by at least or by at most about 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject. In embodiments, the level of plasmablasts is reduced by at least or by at most about 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject. In embodiments, the level of CXCL13 is reduced by at least or at most about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject.In embodiments, the level of RF antibodies is reduced by at least or at most about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or up to about 100% compared to the subject's baseline level or the level in an otherwise comparable untreated subject.

[0072] In embodiments, the efficacy of treatment is assessed by the European League Against Rheumatism (ELR) standard. EULAR Sjoegren's Syndrome Disease Activity Index (ESSDAI), EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI), 36-Item Short Form Survey Version 2 (SF-36v2), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-Fatigue), visual analog scale (VAS) oral / ocular, Patient-Reported The Outcomes Measurement Information System® (PROMIS-29 Profile v2.1), Ocular Surface Disease Index (OSDI), Patient's Global Impression of Change (PGIC), Patient's Global Impression of Severity (PGIS), stimulated salivary flow measurement, Schirmer test, 28-joint assessment, Physician's Global Impression of Severity, Clinical EULAR Sjoegren's Syndrome Disease Activity Index (ClinESSDAI), and combinations thereof may be used to determine efficacy of treatment. In embodiments, efficacy of treatment may be measured using assessments that are patient-reported outcome (PRO) instruments.

[0073] Systemic disease activity and patient-reported outcomes ESSDAI In some embodiments, the assessment includes the ESSDAI, a systemic disease activity index that includes definitions of disease activity by organ (see Seror R, Ravaud P, Bowman SJ, Baron G, Tzioufas A, Theander E, et al.; EULAR Sjoegren's Task Force. EULAR Sjoegren's syndrome disease activity index: development of a consensus systemic disease activity index for primary Sjoegren's syndrome. Ann Rheum Dis. 2010;69(6):1103-9, which is incorporated herein by reference). The ESSDAI grades disease activity in 12 domains: skin, respiratory (lung), kidney, joint, muscle, peripheral nervous system, central nervous system, hematology, gland, constitution, lymphadenopathy, and biology. Using a physician's global assessment of activity as the gold standard, weights for each domain were determined using multiple regression modeling.

[0074] Each domain is assigned a weight ranging from 1 (biological domain) to 6 (muscle domain), and each domain has three or four activity levels ranging from 0 (no activity) to 3 (high activity).

[0075] In embodiments, the following domains may be scored but do not contribute to the minimum ESSDAI score of 5 required for inclusion in the studies of the present disclosure: peripheral nervous system, central nervous system, and lung.

[0076] The theoretical range of values ​​for the ESSDAI is 0 to 123 points, and the final score is calculated as follows: Final score = sum of scores from all 12 domains Domain score = activity level x domain weight Low disease activity was defined as an ESSDAI score <5, moderate disease activity as 5 points ≤ ESSDAI ≤ 13 points, and high disease activity as ESSDAI ≥ 14 points.

[0077] In embodiments, a change from baseline in ESSDAI is determined. In embodiments, the baseline ESSDAI score is reduced by at least about 1, 2, 3, 4, or 5 points after treatment with a composition provided herein. In embodiments, the baseline ESSDAI score is reduced by 3 points after treatment with a composition provided herein. In embodiments, the baseline ESSDAI score is reduced by 4 points after treatment with a composition provided herein. In embodiments, the post-treatment ESSDAI score is ≦2, ≦5, or ≦13 points. Administration of a Tn3 scaffold of the present disclosure can be effective in reducing the ESSDAI score by at least about or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 18, or 20 points when compared to the ESSDAI baseline score before administration. Administration of a Tn3 scaffold of the present disclosure can be effective in reducing the ESSDAI score by at least about or at most about 1-5, 3-5, 5-8, 5-10, or 10-15 points when compared to the ESSDAI baseline score before administration. In embodiments, administration of a Tn3 scaffold of the present disclosure is effective in reducing the ESSDAI score by at least about 6 points. In embodiments, administration of a Tn3 scaffold of the present disclosure is effective in reducing the ESSDAI score by at least about 6.3 points.

[0078] The ESSDAI score may refer to an absolute value or a relative value. For example, a relative value may include the difference between a subject treated with a Tn3 scaffold of the present disclosure and a subject treated with a placebo control. In embodiments, the ESSDAI score refers to an absolute value, e.g., a decrease in ESSDAI compared to a baseline level or a level determined before treatment. In embodiments, the ESSDAI score refers to a relative value. A relative value may refer to the difference between the ESSDAI scores of a Tn3 scaffold-treated subject and a control-treated subject. In embodiments, the relative value is the least squares mean difference between a Tn3 scaffold-treated subject and a control-treated subject.

[0079] In embodiments, a subject administered a Tn3 scaffold of the present disclosure achieves a 3, 4, 5, 6, 7, 8, 9, or 10 point reduction in their ESSDAI when compared to the ESSDAI before administration (e.g., baseline level). In embodiments, a subject administered a Tn3 scaffold of the present disclosure achieves a 6.4 point reduction in their ESSDAI when compared to the ESSDAI before administration (e.g., baseline level).

[0080] In embodiments, ESSDAI assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 9 The ESSDAI assessment can be performed at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the ESSDAI assessment for the Tn3 scaffold can be performed at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, the ESSDAI assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0081] ClinESSDAI In some embodiments, the assessment includes the Clinical EULAR Sjoegren's Syndrome Disease Activity Index (ClinESSDAI). The ClinESSDAI is a validated Sjoegren's Syndrome disease activity index based on the ESSDAI, but excluding the biological domain and assigning different weights to each domain. The ClinESSDAI was developed in an attempt to reduce potential correlations between B-cell biomarkers measured by the ESSDAI biological domain and clinical activity measures. The theoretical range of values ​​for the ClinESSDAI is 0-135 points. Similar to the ESSDAI, low activity is defined as <5 points, moderate activity as 5 points ≤ ClinESSDAI ≤ 13 points, and high activity as ≥ 14 points. The ClinESSDAI has been validated and shown to correlate well with the ESSDAI, and is considered a useful tool for detecting changes independent of the biological effects of drugs. In some embodiments, the change from baseline in the ClinESSDAI is determined. In embodiments, the baseline ClinESSDAI score is reduced by at least about 1, 2, 3, 4, or 5 points after treatment with a composition provided herein. In embodiments, the post-treatment ClinESSDAI score is ≦2, ≦5, or ≦13 points.

[0082] In embodiments, ClinESSDAI assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, The assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the ClinESSDAI assessment of the Tn3 scaffold can be assessed at about 85±3 days, 169±3 days, and 309±7 days after the start of treatment. In embodiments, the ClinESSDAI assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0083] ESSPRI In embodiments, the assessment comprises the ESSPRI (see Seror R, Ravaud P, Mariette X, Bootsma H, Theander E, Hansen A, et al. EULAR Sjoegren's Task Force. EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI): development of a consensus patient index for primary Sjoegren's syndrome. Ann Rheum Dis. 2011;70(6):968-72, which is incorporated herein by reference). The ESSPRI is a self-assessment tool developed in a multicenter, international cohort of 230 patients. The ESSPRI uses a 0-10 point numeric analog scale (ranging from 0 [no symptoms] to 10 [maximum conceivable severity]) for the assessment of each of three domains: dryness, fatigue, and pain (joint and / or muscle). The domains are weighted equally, and the mean of the scores across the three domains represents the final score. Each question defines a recall period, such as "the last two weeks." In embodiments, subjects in Cohort #2 achieve an ESSPRI response, defined as a ≥ 1-point or 15% reduction from baseline in the ESSPRI score in the absence of early discontinuation of Tn3 scaffold and in the absence of rescue therapy. In embodiments, the baseline ESSPRI score is approximately 3%, 5%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 340%, 350%, 360%, 370%, 380%, 390%, 400%, 410%, 420%, 430%, 440%, 450%, 460%, 470%, 480%, 490%, 500%, 510%, 520%, 530%, 540%, 550%, 560%, 570%, 580%, 590%, 610%, 620%, 630%, 640%, 650%, 660%, 670%, 680%, 690%, 700%, 710%, 720%, 730%, 740%, 750%, 760%, 770%, 780%, 790%, 800%, 810%, 820%, 830%, 840%, 850%, 860%, A reduction of 0%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to about 100% is achieved. In embodiments, the post-treatment ESSPRI score is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9 points. In embodiments, the post-treatment ESSPRI score is reduced by at least about 1, 2, 3, 4, 5, 6, 7, 8, or 9 points when compared to the pre-treatment ESSPRI score.

[0084] In embodiments, ESSPRI assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The evaluation may be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, ESSPRI evaluations of the Tn3 scaffold may be evaluated at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, ESSPRI evaluations may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0085] Functional Assessment of Chronic Illness Therapy (FACIT)-Fatigue In embodiments, the assessment includes the FACIT-Fatigue Scale. The FACIT-Fatigue Scale is a 13-item subject-administered questionnaire used to assess the impact of fatigue. The FACIT-Fatigue Scale has a recall period of approximately 7 days. Responses on the FACIT-Fatigue Scale range from 0 (not at all) to 4 (very much). To calculate the overall score, negatively worded items were reversed by subtracting the response from "4." The final score is the sum of the responses and ranges from 0 to 52. Higher scores indicate better quality of life (QoL). In embodiments, the change from baseline in the FACIT-Fatigue Scale can be determined. In embodiments, methods in which a Tn3 scaffold is administered to a subject result in a higher FACIT-Fatigue Scale when compared to comparable methods in which the subject is not administered a Tn3 scaffold.

[0086] In embodiments, the FACIT-Fatigue scale ratings for the Tn3 scaffold are measured at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 9 The FACIT-Fatigue scale assessments of the Tn3 scaffold can be assessed at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, the FACIT-Fatigue scale assessments of the Tn3 scaffold can be assessed at any time before, during, or after administration of the Tn3 scaffold.

[0087] Ocular Surface Disease Index (OSDI (Copyright)) In some embodiments, the assessment may include the OSDI. The OSDI is a valid and reliable instrument for assessing the effect on vision-related function and dry eye disease severity (normal, mild, moderate, and severe). The recall period for the OSDI is one week. The OSDI assessment consists of 12 questions, which a physician may ask the subject, who may circle the number that best describes each question. Responses to each question range from 0 (never) to 4 (always). The OSDI score is calculated as (total score of answered questions) / (number of answered questions) x 25, ranging from 0 to 100, with higher scores indicating more severe disability. In some embodiments, the change from baseline in the OSDI is determined.

[0088] In embodiments, OSDI assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 weeks The OSDI may be assessed at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, OSDI assessments of the Tn3 scaffold may be assessed at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, OSDI assessments may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0089] Patient Global Impression of Severity (PGIS) In some embodiments, the assessment involves completing the PGIS. The PGIS is a single item designed to capture the subject's perception of overall symptom severity over the past week with a 5-point categorical scale (none, mild, moderate, severe, or very severe) response. In some embodiments, the PGIS score is reduced after administration of the Tn3 scaffold of the present disclosure. The reduction can be from about 1, 2, 3, 4 points, or up to about 5 points.

[0090] In embodiments, PGIS assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The assessment may be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, PGIS assessment of the Tn3 scaffold may be assessed at about day 1, 29±3 days, 57±3 days, 85±3 days, 113±3 days, 141±3 days, 169±3 days, 197±3 days, 225±3 days, 253±3 days, 281±3 days, 309±7 days, and 365±7 days after initiation of treatment. In embodiments, PGIS assessment may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0091] Physician's overall impression of severity In some embodiments, the assessment may include a Physician's Global Impression of Severity (MDGIS). The Physician's Global Impression of Severity may include an overall assessment of SS disease severity or the severity of an autoimmune disease of the present disclosure. The PGIS may be scored with responses on a 5-point categorical scale (none, mild, moderate, severe, or very severe). In some embodiments, the Physician's Global Impression of Severity may include an overall assessment of SS disease severity or the severity of an autoimmune disease of the present disclosure. The PGIS may be scored with responses on a 5-point categorical scale (none, mild, moderate, severe, or very severe). The change from baseline in global impression is determined. In embodiments, the physician's global impression of severity score is reduced after administration of the Tn3 scaffold of the present disclosure. The reduction can be from about 1, 2, 3, 4 points, or up to about 5 points.

[0092] In embodiments, the Tn3 Scaffold Physician Global Impression of Severity assessment is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, The Tn3 scaffold may be evaluated at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the Physician's Global Impression of Severity assessment of the Tn3 scaffold may be evaluated at about days 1, 15±1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, the MDGIS assessment may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0093] composite indicator In embodiments, the evaluation includes determining a composite index, which may be defined as either an ESSDAI-3 point improvement (i.e., a 3 point reduction) or an ESSPRI-1 point improvement (i.e., a ≥ 1 point or 15% reduction from baseline in the ESSPRI score) plus no worsening from baseline in the Physician's Global Impression of Severity.

[0094] In embodiments, the composite index assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the composite endpoint assessment of the Tn3 scaffold can be assessed at about 85±3 days, 169±3 days, and 309±7 days after the start of treatment. In embodiments, the composite endpoint assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0095] 28 joint evaluation In some embodiments, the evaluation may include a 28-joint count. The 28-joint count may evaluate the following joints for tenderness and swelling: left and right shoulder joints, elbow joints, wrist joints, metacarpophalangeal (MCP) 1, MCP2, MCP3, MCP4, MCP5, proximal interphalangeal joint (PIP) 1, PIP2, PIP3, PIP4, PIP5 joints of the upper limbs, and left and right knee joints of the lower limbs. The presence of synovitis may be determined for each of the 28 joints. At the start of the 28-joint count (before the evaluation of tenderness and swelling), the subject may be asked whether they have experienced or are experiencing pain in any of the 28 joints. In some embodiments, the evaluation may include a tender joint count (TJC). In some embodiments, the evaluation may include a swollen joint count (SJC).

[0096] In embodiments, 28 joint count assessments of the Tn3 scaffold are performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks after initiation of treatment. , 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, 28 joint count assessments of the Tn3 scaffold can be assessed at about day 1, day 29±3, day 57±3, day 85±3, day 113±3, day 141±3, day 169±3, day 197±3, day 225±3, day 253±3, day 281±3, day 309±7, and day 365±7 after initiation of treatment. In embodiments, 28 joint count assessments can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0097] Salivary and lacrimal gland function Total stimulated saliva flow rate In some embodiments, the assessment involves determining stimulated total saliva flow. Stimulated total saliva flow can be measured to objectively assess changes in salivary gland function. Subjects receiving standard treatment for dry mouth at screening must discontinue use of pilocarpine or cevimeline for at least 12 hours and artificial saliva for at least 3 hours prior to saliva collection. Subjects must refrain from eating or drinking for at least 90 minutes prior to saliva collection. To minimize diurnal variation, all saliva collection attempts should be performed at the same time of day for all subsequent assessments.

[0098] In an exemplary stimulated total saliva flow rate measurement, a roll of parafilm approximately 5 x 5 cm square is presented to the subject to chew like a chewing stick at a rate of approximately 60 chews per minute. The subject must sit upright, keep their eyes open, tilt their head slightly forward, and begin chewing the parafilm. After 60 seconds, they must expectorate all collected saliva into an additional (unweighed) Falcon tube while still holding the parafilm in their mouth. This initial collection familiarizes the subject with the procedure. Three rounds of saliva collection are performed, each after 20 seconds of chewing, into a pre-weighed Falcon tube. These rounds are performed consecutively, with the timing stopped during saliva collection and restarted immediately after saliva is collected into the pre-weighed Falcon tube, for a total stimulation time of 60 seconds. If the subject is unable to complete the 60-second collection due to incapacity, the total collection time must be recorded. The total stimulated saliva collection volume is assessed by subtracting the final tube weight from the pre-collection weight. In embodiments, the change from baseline in stimulated total saliva flow rate is determined. In embodiments, subjects in Cohort #1 have a post-treatment unstimulated total saliva flow rate of >0.1 ml / min.

[0099] In embodiments, stimulated salivary flow assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, The stimulated salivary flow rate of the Tn3 scaffold can be evaluated at about 1 day, 85±3 days, 169±3 days, 253±3 days, 309±7 days, and 365±7 days after treatment initiation. Assessment of salivary slowness can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0100] Visual analog scale (VAS) oral / ocular In embodiments, the assessment involves determining a (VAS) Oral / Ocular. These two instruments assess changes in the severity of oral and ocular dryness using a continuous 100mm VAS ranging from 0mm (best) to 100mm (worst). Respondents are asked to place a line perpendicular to the VAS at the point representing the intensity of their symptoms. The Oral VAS may be rated for the question, "Usually, how dry does your mouth feel?" (0mm: not dry at all; 100mm: very dry). The Ocular VAS may be rated for the question, "Usually, how dry do your eyes feel?" (0mm: not dry at all; 100mm: very dry). In embodiments, the baseline (VAS) Oral / Ocular score is reduced by about 3%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or up to about 100% when compared to the subject's VAS score prior to administration of the Tn3 scaffold.

[0101] In embodiments, the VAS ratings of the Tn3 scaffold are measured at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The VAS may be assessed at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, VAS assessments of the Tn3 scaffold may be assessed at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, VAS assessments may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0102] Schirmer test In some embodiments, a Schirmer test can be performed without topical anesthesia. The Schirmer test measures lacrimal gland function. The Schirmer test can measure tear flow using a calibrated test strip, which is non-toxic filter paper. One end of the test strip is placed in the lower eyelid. Both eyes should be measured simultaneously. After placement, the subject is asked to gently close their eyes, and after 5 minutes, the test strip is removed from the eyelid and the degree to which each strip is wet is recorded. In some embodiments, the change from baseline in the Schirmer test can be determined. In some embodiments, the post-treatment score is >5mm / 5 minutes for at least one eye.

[0103] In embodiments, Schirmer test assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 9 The evaluation can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, Schirmer test evaluation of the Tn3 scaffold can be evaluated at about day 1, 85±3, 169±3, 253±3, 309±7, and 365±7 days after initiation of treatment. In embodiments, Schirmer test evaluation can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0104] Subjective symptoms Patient Global Impression of Change (PGIC) In embodiments, the assessment includes PGIC. PGIC is the progression of symptoms from the start of study medication. This is a single item designed to capture the subject's perception of overall change in severity.The change in severity is captured using a 5-point scale (much better, slightly better, no change, slightly worse, or much worse).In this embodiment, the change from baseline in PGIC is determined.

[0105] In embodiments, PGIC assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, PGIC assessment of the Tn3 scaffold can be assessed at about 15±1 day, 29±3 day, 57±3 day, 85±3 day, 113±3 day, 141±3 day, 169±3 day, 197±3 day, 225±3 day, 253±3 day, 281±3 day, 309±7 day, and 365±7 day after initiation of treatment. In embodiments, PGIC assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0106] 36-Item Short Form Survey Version 2 (SF36v2) physical and mental component scores In embodiments, the assessment includes an SF-36v2 profile. The SF-36v2 (immediate recall) is a 36-item global health assessment that captures information on eight health domains: physical functioning, role functioning (physical), bodily pain, general well-being, vitality, social functioning, role functioning (mental), and mental health. The SF-36v2 provides scores for each domain and two psychometric total scores: a physical component score and a mental component score. The recall period for the immediate version is one week (i.e., "last week"). In embodiments, the change from baseline in the SF36v2 physical component score and mental component score is determined.

[0107] In embodiments, the 36v2 profile assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks The SF36v2 profile assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the 36v2 profile assessment of the Tn3 scaffold can be assessed at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, the SF36v2 assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0108] Patient-Reported Outcomes Measurement Information System® (PROMIS-29 Profile) In embodiments, the assessment includes a PROMIS-29 profile. The 2016 WHO 2016-10-19 (2016-10-19) assesses seven domains (depression, anxiety, physical function, pain interference, fatigue, sleep disturbance, and ability to participate in social roles and activities) with four questions for each domain. The domains are assessed for the past seven days, excluding physical function, which has no time limit. In some embodiments, treated subjects show improved PROMIS-29 scores compared to otherwise comparable subjects who do not receive the Tn3 scaffold.

[0109] In embodiments, the PROMIS-29 profile assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 The Tn3 scaffold may be evaluated at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the PROMIS-29 profile assessment of the Tn3 scaffold may be evaluated at about days 1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, the PROMIS-29 profile assessment may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0110] Pharmacokinetic (PK) analysis In embodiments, the methods provided herein may include determining the concentration of Tn3 scaffold after administration in a subject in need thereof. In embodiments, the method includes a pharmacokinetic assessment. In embodiments, the sample is a blood sample or a plasma sample, or a combination of both. In embodiments, suitable assays for measuring pharmacokinetics may include electrochemiluminescence (ECL) assays, bead-based assays, cell-based assays, and combinations thereof. In embodiments, the sample may include plasma, and the plasma may be analyzed to determine the highest observed concentration (C) for the Tn3 scaffold concentration. max ), area under the concentration-time curve (AUC), CL, and terminal elimination half-life (t 1 / 2 ) is evaluated by measuring

[0111] In embodiments, PK assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 weeks The PK assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, PK assessment of the Tn3 scaffold can be assessed at about days 1, 15±1, 29±3, 57±3, 85±3, 113±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, PK assessment can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0112] immunogenicity In embodiments, the evaluation includes determining the level of immunogenicity, if any, of the Tn3 scaffold. Immunogenicity includes determining the presence of anti-drug antibodies (ADA) against the Tn3 scaffold. The presence of ADA can be determined using a plasma sample from a subject administered the Tn3 scaffold. In embodiments, no ADA is detected after administration of the Tn3 scaffold. In embodiments, In an otherwise equivalent method, ADA levels are reduced when compared to an otherwise equivalent method in which the subject is not administered a Tn3 scaffold; for example, the reduction can be about 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% when compared to ADA levels in an otherwise equivalent method in which the subject is not administered a Tn3 scaffold.

[0113] In embodiments, the immunogenicity of the Tn3 scaffold is assessed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 The assessment may be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the assessment of immunogenicity of the Tn3 scaffold may be assessed at about day 1, day 29±3, day 85±3, day 169±3, day 197±3, day 253±3, day 309±7, and day 365±7 after initiation of treatment. In embodiments, the assessment of immunogenicity may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0114] Pharmacodynamic and biomarker analysis In embodiments, the method includes a pharmacodynamic assessment. In embodiments, the assessment includes biomarkers: T cell subsets, B cell subsets, SS-A, SS-B, and IgG and IgM. In embodiments, serum and plasma may be collected to measure changes in exploratory biomarkers of disease activity, such as pro-inflammatory cytokine and CXCL13 levels. In embodiments, biomarkers related to B cell subsets, RF autoantibodies, and CXCL13 are analyzed. In embodiments, erythrocyte sedimentation rate (ESR) is analyzed. In embodiments, biomarker analysis may include determining changes in T and B cell subsets and salivary proteins by methods such as flow cytometry or mass spectrometry (e.g., LC-MS), as well as changes in the fecal microbiome. Exemplary B and T cell subsets include, but are not limited to, plasmablasts (e.g., CD27 in CD19+ cells), plasmablasts (e.g., CD27 in CD19+ cells), and plasmacytoids (e.g., CD27 in CD19+ cells). br / CD38br / IgD- subset), memory B precursor cells (e.g., CD11c br subset), T follicular helper (Tfh) cells (e.g., the CXCR5+ / ICOS+ subset of CD3+ / CD4+ cells), post-switch memory B cells (e.g., CD27 br These include proliferation of T cells (e.g., the Ki67+ subset of CD3+ cells) and / or proliferation of total T cells (e.g., the Ki67+ subset of CD3+ cells).

[0115] In embodiments, samples may be assessed for protein biomarkers. Exemplary protein biomarkers include, but are not limited to, TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, CXCL13, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, CD80, or any combination thereof. In embodiments, the protein biomarker is TNFRSF13B. In embodiments, the protein biomarker is BAFF. In embodiments, the protein biomarker is CXCL10. In embodiments, the protein biomarker is CXCL12. In embodiments, the protein biomarker is IL6. In some embodiments, the protein biomarker is CCL21. In some embodiments, the protein biomarker is CXCL9. In some embodiments, the protein biomarker is IL12B. In some embodiments, the protein biomarker is TNFRSF4. In some embodiments, the protein biomarker is LAMP3. In some embodiments, the protein biomarker is CD22. In some embodiments, the protein biomarker is CD79B. In some embodiments, the protein biomarker is SLAMF7. In some embodiments, the protein biomarker is IL10. In some embodiments, the protein biomarker is TNFRSF8. In some embodiments, the protein biomarker is CXCL11. In some embodiments, the protein biomarker is CCL19. In some embodiments, the protein biomarker is CXCL13. In some embodiments, the protein biomarker is TNFRSF9. In some embodiments, the protein biomarker is TNFRSF6B. In some embodiments, the protein biomarker is NOS3. In some embodiments, the protein biomarker is CD72. In embodiments, the protein biomarker is ADA2. In embodiments, the protein biomarker is IL2RA. In embodiments, the protein biomarker is CD27. In embodiments, the protein biomarker is PDCD1. In embodiments, the protein biomarker is IL12A. In embodiments, the protein biomarker is IL12B. In embodiments, the protein biomarker is LAG3. In embodiments, the protein biomarker is ADGRE1. In embodiments, the protein biomarker is TIGIT. In embodiments, the protein biomarker is FCAMR. In embodiments, the protein biomarker is CD80.

[0116] In embodiments, the sample to be assessed for protein biomarkers is obtained from a subject. In embodiments, the subject has symptoms of or has been diagnosed with a disease or disorder. In embodiments, the subject has symptoms of or has been diagnosed with an autoimmune disease of the present disclosure (e.g., Sjogren's syndrome, rheumatoid arthritis, etc.). In embodiments, the subject has symptoms of or has been diagnosed with Sjogren's syndrome. In embodiments, the subject has symptoms of or has been diagnosed with rheumatoid arthritis.

[0117] In embodiments, the level of a protein biomarker in a sample obtained from a subject who has symptoms of or has been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) is increased or decreased compared to a healthy subject. In embodiments, the level of a protein biomarker in a subject who has symptoms of or has been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) is increased or decreased after administration of a compound or composition of the present disclosure compared to a comparable subject who has not been administered the compound or composition.

[0118] In some embodiments, the level of a protein biomarker in a sample obtained from a subject who has symptoms of or has been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) is increased compared to a healthy subject. In some embodiments, the level of a protein biomarker in a subject who has symptoms of or has been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) is increased after administration of a compound or composition of the present disclosure, compared to a comparable subject who has not been administered the compound or composition. Exemplary protein biomarkers that are increased include, but are not limited to, TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, CXCL13, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, Examples of the increased protein biomarker include PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, CD80, or any combination thereof. In embodiments, the increased protein biomarker is TNFRSF13B. In embodiments, the increased protein biomarker is BAFF. In embodiments, the increased protein biomarker is CXCL10. In embodiments, the increased protein biomarker is CXCL12. In embodiments, the increased protein biomarker is IL6. In embodiments, the increased protein biomarker is CCL21. In embodiments, the increased protein biomarker is CXCL9. In embodiments, the increased protein biomarker is IL12B. In embodiments, the increased protein biomarker is TNFRSF4. In embodiments, the increased protein biomarker is LAMP3. In embodiments, the increased protein biomarker is CD22. In embodiments, the increased protein biomarker is CD79B. In embodiments, the increased protein biomarker is SLAMF7. In embodiments, the increased protein biomarker is IL10. In embodiments, the increased protein biomarker is TNFRSF8. In embodiments, the increased protein biomarker is CXCL11. In embodiments, the increased protein biomarker is CCL19. In embodiments, the increased protein biomarker is CXCL13. In embodiments, the increased protein biomarker is TNFRSF9. In embodiments, the increased protein biomarker is TNFRSF6B. In embodiments, the increased protein biomarker is NOS3. In embodiments, the increased protein biomarker is CD72. In embodiments, the increased protein biomarker is ADA2. In embodiments, the increased protein biomarker is IL2RA. In embodiments, the increased protein biomarker is CD27. In embodiments, the increased protein biomarker is PDCD1. In embodiments, the increased protein biomarker is IL12A.In embodiments, the increased protein biomarker is IL12B. In embodiments, the increased protein biomarker is LAG3. In embodiments, the increased protein biomarker is ADGRE1. In embodiments, the increased protein biomarker is TIGIT. In embodiments, the increased protein biomarker is FCAMR. In embodiments, the increased protein biomarker is CD80.

[0119] In embodiments, the level of a protein biomarker is decreased in a sample obtained from a subject having symptoms of or having been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) compared to a healthy subject. In embodiments, the level of a protein biomarker is decreased following administration of a compound or composition of the present disclosure in a subject having symptoms of or having been diagnosed with an autoimmune disease (e.g., Sjogren's syndrome, rheumatoid arthritis) compared to a comparable subject not administered the compound or composition. Exemplary protein biomarkers that are decreased include, but are not limited to, TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, CXCL13, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, CD80, or any combination thereof. In an embodiment, the protein biomarker that is decreased is TNFRSF13B. In an embodiment, the protein biomarker that is decreased is BAFF. In an embodiment, the protein biomarker that is decreased is CXCL10. In an embodiment, the protein biomarker that is decreased is CXCL12. In embodiments, the protein biomarker that is decreased is IL6. In embodiments, the protein biomarker that is decreased is CCL21. In embodiments, the protein biomarker that is decreased is CXCL9. In embodiments, the protein biomarker that is decreased is IL6. In embodiments, the protein biomarker is IL12B. In embodiments, the protein biomarker that is decreased is TNFRSF4. In embodiments, the protein biomarker that is decreased is LAMP3. In embodiments, the protein biomarker that is decreased is CD22. In embodiments, the protein biomarker that is decreased is CD79B. In embodiments, the protein biomarker that is decreased is SLAMF7. In embodiments, the protein biomarker that is decreased is IL10. In embodiments, the protein biomarker that is decreased is TNFRSF8. In embodiments, the protein biomarker that is decreased is CXCL11. In embodiments, the protein biomarker that is decreased is CCL19. In embodiments, the protein biomarker that is decreased is CXCL13. In embodiments, the protein biomarker that is decreased is TNFRSF9. In embodiments, the protein biomarker that is decreased is TNFRSF6B. In embodiments, the protein biomarker that is decreased is NOS3. In embodiments, the protein biomarker that is decreased is CD72. In embodiments, the protein biomarker that is decreased is ADA2. In embodiments, the protein biomarker that is decreased is IL2RA. In embodiments, the protein biomarker that is decreased is CD27. In embodiments, the protein biomarker that is decreased is PDCD1. In embodiments, the protein biomarker that is decreased is IL12A. In embodiments, the protein biomarker that is decreased is IL12B. In embodiments, the protein biomarker that is decreased is LAG3. In embodiments, the protein biomarker that is decreased is ADGRE1. In embodiments, the protein biomarker that is decreased is TIGIT. In embodiments, the protein biomarker that is decreased is FCAMR. In embodiments, the protein biomarker that is decreased is CD80.

[0120] In some embodiments, cell markers are determined using flow cytometry. Generally, positive staining can be classified as dim, moderate, bright, or variable. Weak positive staining may be slightly increased compared to the negative control. Moderate staining may be at least one logarithmic decade brighter than the negative control. Strong positive staining may be at least two logarithmic decades brighter than the negative control. In some embodiments, the terms bright and high positive may be used interchangeably herein.

[0121] In embodiments, B cell subsets are determined. In embodiments, the determined B cell subsets are selected from the group consisting of transitional, naive, plasma, memory, and any combination of these B cell subsets. In embodiments, the B cell subset comprises transitional cells. In embodiments, the B cell subset comprises naive cells. In embodiments, the B cell subset comprises plasma cells. In embodiments, the B cell subset comprises memory cells. In embodiments, the determined B cell subsets are plasma and memory cells. In embodiments, the B cell subsets comprise Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells. In embodiments, the B cell subsets comprise the Ki67+ subset of CD19+ cells. In embodiments, the B cell subsets comprise the CD86+ subset of CD19+ cells. In embodiments, the B cell subset comprises a CD86+ subset of CD27+ / IgD- / CD19+ cells. In embodiments, the B cell subset comprises a Ki67+ subset of CD27+ / IgD- / CD19+ cells. In embodiments, the B cell subset comprises a CD27+ subset of CD19+ cells. br / IgD- / CD38 br In embodiments, the B cell subset comprises a Ki67+ subset of CD27+ / IgD- / CD19+ cells. In embodiments, the B cell subset comprises a CD11c subset of CD19+ cells. br In embodiments, the B cell subset comprises CD11cbr / Contains the CD86+ subset of CD19+ cells.

[0122] In embodiments, the B cell subset comprises transitional cells comprising any combination of biomarkers selected from the group consisting of CD19+, CD20+, CD27-, CD10+, CD24 high, CD38 high, IgD+, CD5+, and BR3+. In embodiments, the B cell subset comprises mature naive cells comprising any combination of biomarkers selected from the group consisting of CD19+, CD20+, CD27, CD24 int, CD38 int, CD21+, CD10-, IgD+, and IgM-. In embodiments, the B cell subset comprises germinal center cells comprising any combination of biomarkers selected from the group consisting of CD19+, CD10+, CD20+, CD27+, CD38+, FAS+, and IgDBR3+. In embodiments, the B cell subset comprises memory cells comprising any combination of biomarkers selected from the group consisting of CD19+, CD20+, CD27+, and CD38. In embodiments, the B cell subset comprises plasmablasts comprising any combination of biomarkers selected from the group consisting of CD19+, CD20, CD27+, CD38high, IgD, and CD138-. In embodiments, the B cell subset comprises plasma cells comprising any combination of biomarkers selected from the group consisting of CD19+, CD20-, IgD, CD27high, CD38high, CD24lo, CD138high, and TACI+. In embodiments, the B cells are identified based on the presence or absence of any combination of markers selected from the group including CD10, CD11c, CD19, CD20, CD21, CD24, CD27, CD38, CD138, Ki67, TACI+, IgM, or IgD.

[0123] In embodiments, T cell subsets are determined. In embodiments, CD4+ subsets, such as Th1, Th2, Th9, Th17, Th22, Treg, or Tfh, or a combination thereof, are determined. In embodiments, a CD4+ Th1 subset is determined. In embodiments, a CD4+ Th2 subset is determined. In embodiments, a CD4+ Th9 subset is determined. In embodiments, a CD4+ Th17 subset is determined. In embodiments, a CD4+ Th22 subset is determined. In embodiments, a CD4+ Treg subset is determined. In embodiments, a CD4+ Tfh subset is determined. In embodiments, CD8+ memory and effector subsets (CD25, CD45RO, CD45RA, CCR-7, L-selectin [CD62L], and / or intracellular markers (FOXP3) are determined. In embodiments, naive CD4+ T cells comprising any combination of markers selected from the group consisting of CD4+, CD45RA+, CD45RO-, and CCR7+ are determined. In embodiments, Th1 cells comprising any combination of biomarkers selected from the group consisting of CD4+, CXCR3+, CCR5+, and IL12Rβ2+ are determined. In embodiments, Th2 cells comprising any combination of biomarkers selected from the group consisting of CD4+, CCR4+, and IL12Rβ2- are determined. In embodiments, Th2 cells comprising any combination of biomarkers selected from the group consisting of CD4+, CCR3+, and CCR6+ are determined. In some embodiments, Th9 cells are determined comprising any combination of biomarkers selected from the group consisting of: CD4+, CCR6+, CCR4+, and NK1.1+. In some embodiments, Th17 cells are determined comprising any combination of biomarkers selected from the group consisting of: CD4+, CCR10+, CCR4+, and CCR6+. In some embodiments, Th22 cells are determined comprising any combination of biomarkers selected from the group consisting of: CD4+, CD127+, CD25+, and CTLA-4+. In some embodiments, Tfh cells are determined comprising any combination of biomarkers selected from the group consisting of: CD3+, CD4+, CXCR5+, CD40L+, and ICOS+.In embodiments, γδ T cells comprising any combination of biomarkers selected from the group consisting of TCRγ / δ+ and IL23R+ are determined. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ cells. In embodiments, the T cell subset comprises Ki67+ cells of CD3+ cells. In embodiments, the T cell subset comprises Ki67+ cells of CD3+ / CD4+ cells. In embodiments, the T cell subset is CD3+ / CD4+ / CD45RA- / CCRT. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD4+ / CD45RA- / CCR7- cells. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD4+ / CXCR5+ / ICOS+ cells. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD8+ cells. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD8+ / CD45RA- / CCR7+ cells. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD8+ / CD45RA / CCR7- cells. In embodiments, the T cell subset comprises a Ki67+ subset of CD3+ / CD8+ / CXCR5+ cells. In embodiments, the Tfh subset comprises a CXCR5+ / ICOS+ subset of CD3+ / CD4+ cells. In some embodiments, NKT cells are determined that contain any combination of biomarkers selected from the group consisting of TCR Vα24+ and TCR Vβ11+. In some embodiments, naive CD8+ T cells are determined that contain any combination of biomarkers selected from the group consisting of CD8+, CD45RA+, CD45RO-, CCR7+, and CD28+. In some embodiments, cytotoxic CD8+ T cells are determined that contain any combination of biomarkers selected from the group consisting of CD8+ and CCR7-.

[0124] In embodiments, the plasmablast subset is CD27c br / CD38 br / IgD-containing.

[0125] In embodiments, dendritic cells (DCs) are determined. In embodiments, the DCs are plasmacytoid DCs (pDCs). In embodiments, the DCs are circulating pDCs. In embodiments, DC subsets are determined with respect to one or more markers. In embodiments, the marker is LAMP3.

[0126] In embodiments, whole blood samples may be collected for assessment of changes in number, activation state, and frequency of major leukocyte populations, including B and T lymphocytes, using flow cytometry.

[0127] In embodiments, administration of the Tn3 scaffold of the present disclosure reduces the levels of certain B cell and plasmablast / plasma cell populations. In embodiments, administration of the Tn3 scaffold reduces the levels of T cell and / or other immune cell populations.

[0128] In embodiments, the Tn3 scaffold may provide at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in biomarker levels in subjects administered the Tn3 scaffold when compared to subjects at baseline or subjects not treated with the Tn3 scaffold. In embodiments, the Tn3 scaffold may provide at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in one or more protein biomarker levels in subjects administered the Tn3 scaffold when compared to subjects at baseline or subjects not treated with the Tn3 scaffold. In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in plasmablast subsets of the present disclosure in subjects administered the Tn3 scaffold when compared to subjects at baseline or subjects not treated with the Tn3 scaffold.

[0129] In embodiments, the Tn3 scaffold reduces the expression of at least about 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% of one or more B cell subsets of the present disclosure in subjects administered the Tn3 scaffold compared to subjects at baseline or compared to subjects not treated with the Tn3 scaffold. In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in one or more post-switch memory B cell subsets of the present disclosure in a subject administered the Tn3 scaffold when compared to the subject at baseline or a subject not treated with the Tn3 scaffold. In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in one or more memory progenitor B cell subsets of the present disclosure in a subject administered the Tn3 scaffold when compared to the subject at baseline or a subject not treated with the Tn3 scaffold.

[0130] In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% decrease in one or more T cell subsets of the present disclosure in a subject administered the Tn3 scaffold when compared to the subject at baseline or a subject not treated with the Tn3 scaffold. In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% increase in one or more Tfh subsets in a subject administered the Tn3 scaffold when compared to the subject at baseline or a subject not treated with the Tn3 scaffold.

[0131] In embodiments, the Tn3 scaffold may achieve at least about a 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to a 100% reduction in plasmablast subsets of the present disclosure in subjects administered the Tn3 scaffold when compared to subjects at baseline or subjects not treated with the Tn3 scaffold.

[0132] In embodiments, pharmacodynamic assessment of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, The evaluation can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the pharmacodynamic evaluation of the Tn3 scaffold can be evaluated at about day 1, day 15±1, day 29±3, day 57±3, day 85±3, day 113±3, day 141±3, day 169±3, day 197±3, day 225±3, day 253±3, day 281±3, day 309±7, and day 365±7 after the start of treatment. In embodiments, the pharmacodynamic evaluation can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0133] Fecal microbiome samples In some embodiments, the assessment may include changes in the fecal microbiome. In some embodiments, stool samples from whole stool may be collected, and 16S microarray and / or deep sequencing may be used to assess changes in the composition of the gut microbiome over time. In some embodiments, fecal samples are collected and tested by providing a self-collection microbial DNA collection kit from feces before or during a clinic visit. In some embodiments, changes in the fecal microbiome from baseline are determined.

[0134] In embodiments, fecal microbiome assessment of Tn3 scaffolds is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks after initiation of treatment. , 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, fecal microbiome assessment of the Tn3 scaffold can be assessed at about day 1, day 169±3, and day 309±7 after initiation of treatment. In embodiments, assessment of the fecal microbiome can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0135] autoantibodies In embodiments, the evaluation includes determining the subject's autoantibody levels. Autoantibodies include those reactive to autoantigens. Exemplary autoantibodies include SS-A, SS-B, and IgG and / or IgM rheumatoid factor (RF) autoantibodies, and combinations thereof. In embodiments, serum is collected and assessed for the presence of anti-SSA (Ro), anti-SSB (La), antinuclear antibodies, and RF. Exemplary methods for determining autoantibody levels include EliA immunoassay, microarray, ELISA, or a combination thereof. In embodiments, no autoantibodies are detected after administration of the Tn3 scaffold. In embodiments, treatment includes a reduction of the subject's autoantibodies by about 20%, 30%, 40%, 45%, 50%, 60%, 75%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% when compared to the autoantibody levels in an otherwise equivalent subject in an otherwise equivalent procedure in which the Tn3 scaffold is not administered. In embodiments, changes from baseline in SS-A, SS-B, and IgG and / or IgM rheumatoid factor (RF) autoantibodies are determined. In embodiments, autoantibodies are detected at levels that are at most about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, or 50-fold higher than in an otherwise equivalent method in which the subject is not administered a Tn3 scaffold. In embodiments, administration of a Tn3 scaffold of the present disclosure is effective to reduce autoantibody levels in a subject by at least about, or at most about, 3% to 5%, 5% to 10%, 10% to 20%, or 5% to 25% compared to baseline levels prior to administration. In embodiments, administration of a Tn3 scaffold is effective to eliminate autoantibodies in a subject in need thereof.

[0136] In embodiments, assessment of Tn3 scaffold autoantibodies is at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 weeks The assessment can be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, the assessment of autoantibodies of the Tn3 scaffold can be assessed at about day 1, day 85±3, day 169±3, and day 253±3 after initiation of treatment. In embodiments, the assessment of autoantibodies can be performed at any time before, during, or after administration of the Tn3 scaffold.

[0137] Inflammatory markers In embodiments, the evaluation includes determining levels of inflammatory markers. Exemplary inflammatory markers include, but are not limited to, immunoglobulins (IgM, IgG, IgA), beta-2 microglobulin, C-reactive protein (CRP), CXCL13, C3, C4 and serum-free light chains, cryoglobulins, and serum and urine immunofixation, and combinations thereof. In embodiments, whole blood, plasma, serum, and urine are collected to assess inflammatory markers. In embodiments, changes compared to baseline are determined. In embodiments, changes from baseline in levels of inflammatory markers (immunoglobulins, beta-2 microglobulin, C-reactive protein [CRP], CXCL13, C3, C4, and serum-free light chains) are determined. In embodiments, suitable assays for assessing inflammation levels include ELISA, hs-CRP testing, CRP testing, Luminex, and combinations thereof. In embodiments, administration of a Tn3 scaffold of the present disclosure is effective to reduce a subject's inflammatory biomarker levels by at least about or at most about 20%, 30%, 40%, 45%, 50%, 60%, 75%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% when compared to autoantibody levels in an otherwise equivalent method in which the subject is not administered a Tn3 scaffold. In embodiments, administration of a Tn3 scaffold of the present disclosure is effective to reduce inflammatory biomarker levels in a subject by at least about or at most about 3% to 5%, 5% to 10%, 10% to 20%, or 5% to 25% when compared to baseline levels before administration.

[0138] In embodiments, assessment of inflammatory markers in the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, The Tn3 scaffold may be evaluated at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, evaluation of inflammatory markers of the Tn3 scaffold may be evaluated at about day 1, day 15±1, day 29±3, day 57±3, day 85±3, day 113±3, day 141±3, day 169±3, day 197±3, day 225±3, day 253±3, day 281±3, day 309±7, and day 365±7 after initiation of treatment. In embodiments, evaluation of inflammatory markers may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0139] RNA and DNA analysis In embodiments, the evaluation comprises quantifying the expression levels of genes associated with disease activity using RNA and / or DNA analysis. In embodiments, RNA testing is performed. In embodiments, RNA testing is performed to measure the expression levels of genes associated with disease activity, specific cell types including plasma cell gene signatures, T follicular helper gene signatures, and signal transduction including the CD40L / CD40 pathway.

[0140] In embodiments, blood RNA is used to measure expression levels of genes associated with disease activity, specific cell types including plasma cell gene signatures, T follicular helper gene signatures, and signal transduction including the CD40L / CD40 pathway. In embodiments, blood samples will be collected using the PAXgene Blood RNA system for stabilization of intracellular RNA in tubes after collection, transport, and storage of the blood and sealing, and subsequent isolation and purification of intracellular RNA from whole blood for microarray analysis and quantitative polymerase chain reaction.

[0141] In embodiments, DNA testing may be performed. Given that certain CD40 SNPs have been identified as susceptibility loci for certain disease subsets in SLE or Graves' disease, DNA may be isolated from blood at baseline for pharmacogenomic (single nucleotide polymorphism [SNP]) profiling of CD40 and other genes involved in the CD40 / CD40L axis. In embodiments, epigenetic testing, such as DNA methylation in immune-related genes, may be performed. Blood DNA is collected for genetic analysis. In embodiments, epigenomic profiling is assessed by methods including, but not limited to, ATACseq and DNA methylation sequencing.

[0142] In embodiments, expression levels of genes associated with disease activity using RNA and / or DNA analysis may be reduced by at least about 10%, 15%, 20%, 25%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or up to 100% when compared to an otherwise equivalent method in which the Tn3 scaffold is not administered to a subject in an otherwise equivalent method.

[0143] In embodiments, RNA and / or DNA analysis of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, The assessment may be at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, RNA and / or DNA analysis of the Tn3 scaffold may be assessed at about days 1, 15±1, 29±3, 57±3, 85±3, 141±3, 169±3, 197±3, 225±3, 253±3, 281±3, 309±7, and 365±7 after initiation of treatment. In embodiments, RNA and / or DNA assessment may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0144] Actigraphy In embodiments, the assessment may include measuring overall activity and / or sleep patterns. In embodiments, the assessment may include actigraphy. In embodiments, actigraphy is collected throughout the test to measure overall activity and sleep patterns, including but not limited to, sleep duration and / or fragmentation. In embodiments, the activity monitoring device is an ActiGraph, a wearable wristwatch-type device. In embodiments, changes in overall activity and sleep patterns during the test may be determined.

[0145] In embodiments, actigraphy analysis of the Tn3 scaffold is performed at about 0 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, The actigraphy may be assessed at 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, or at least about 53 weeks. In embodiments, actigraphy analysis of the Tn3 scaffold may be assessed at about 365±7 days after initiation of treatment. In embodiments, actigraphy assessment may be performed at any time before, during, or after administration of the Tn3 scaffold.

[0146] Duration of response (duration of clinical response) In embodiments, the evaluation comprises determining the duration of response of the Tn3 scaffold. In embodiments, the evaluation comprises determining the duration of response of the Tn3 scaffold by quantifying the time to initiation of salvage therapy. In embodiments, administration of the Tn3 scaffold reduces the initiation of salvage therapy in treated subjects compared to subjects not administered the Tn3 scaffold. In embodiments, administration of the Tn3 scaffold is effective to extend the time to initiation of salvage therapy by at least about 1 day, 6 days, 11 days, 16 days, 21 days, 26 days, 30 days, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months, 11 months, 1 year, 2 years, or up to about 5 years after administration.

[0147] Pharmaceutical Composition In an embodiment, a pharmaceutical composition is provided. The pharmaceutical composition can comprise the Tn3 scaffold of the present disclosure. In an embodiment, the pharmaceutical composition is part of a treatment regimen that comprises the Tn3 scaffold of the present disclosure and one or more additional therapeutic agents provided herein.

[0148] Many drugs can be administered orally as a liquid, capsule, tablet, or chewable tablet. The oral route is the most commonly used route because it is the most convenient and usually the safest and least expensive. However, the oral route has limitations due to the way drugs typically travel through the digestive tract. For orally administered drugs, absorption can begin in the mouth and stomach. However, most drugs are usually absorbed from the small intestine. Drugs pass through the intestinal wall and travel to the liver before being transported via the bloodstream to their target site. The intestinal wall and liver chemically modify (metabolize) many drugs, reducing the amount of drug that reaches the bloodstream. As a result, these drugs are often given in lower doses to produce the same effect when injected intravenously.

[0149] For the subcutaneous route, a needle is inserted into the fatty tissue just below the skin. After injection, the drug then migrates into the microvasculature (capillaries) and is carried away by the bloodstream. Alternatively, the drug reaches the bloodstream through lymphatic vessels. The intramuscular route is preferred over the subcutaneous route when a larger volume of drug product is needed. Because the muscle is located beneath the skin and fatty tissue, a longer needle is used. Drugs are usually injected into the muscles of the upper arm, thigh, or buttocks. How quickly a drug is absorbed into the bloodstream depends, in part, on the blood supply to the muscle: the thinner the blood supply, the longer it takes for the drug to be absorbed. For the intravenous route, a needle is inserted directly into a vein. A solution containing the drug may be given in a single dose or by continuous infusion. For infusion, the solution is moved by gravity (from a crushable plastic bag) or, more commonly, by an infusion pump through thin, flexible tubing and into a tube (catheter) inserted into a vein, usually in the forearm.

[0150] In embodiments, the pharmaceutical compositions provided herein are administered by infusion. The infusion can be administered over a period of time. For example, the infusion can administer the pharmaceutical agent over a period of about 5 minutes to about 10 hours. The infusion can be administered over a period of about 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, or up to about 10 hours. In embodiments, intravenous administration allows for rapid, well-controlled systemic delivery of a precise dose. Infusions are also used for irritating solutions that can cause pain and tissue damage if administered by subcutaneous or intramuscular injection. When administered intravenously, the drug is delivered to the bloodstream immediately and tends to take effect more quickly than when administered by any other route. Consequently, medical professionals monitor intravenous recipients more closely for signs that the drug is working or causing unwanted side effects. Additionally, the effects of drugs administered via this route tend to be short-lived. Therefore, some drugs must be administered by continuous infusion to maintain consistent efficacy. In some embodiments, infusion reactions may occur, including headache, nausea, somnolence, difficulty breathing, fever, muscle pain, rash, or other symptoms. Potential risks associated with administration of the Tn3 scaffold include infection, redness, swelling, pain, and hardening at the administration site. Before each intravenous infusion, subjects should be monitored to reduce the risk or severity of potential reactions. To reduce the risk of pulmonary embolism, patients may receive prophylaxis with IV methylprednisolone, oral diphenhydramine, and oral acetaminophen, or one or more equivalent drugs.

[0151] In some embodiments, the pharmaceutical agent is administered intrathecally. For the intrathecal route, a needle is inserted between two vertebrae in the lower spine and into the space surrounding the spinal cord. The drug is then injected into the spinal canal. A small amount of local anesthetic is often used to numb the injection site. This route is used when the drug needs to have a rapid or local effect on the brain, spinal cord, or the layers of tissue covering them (meninges), for example, to treat infections in these structures.

[0152] Drugs administered by inhalation through the mouth can be atomized into finer droplets than those administered via the nasal route, allowing the drug to pass through the throat (trachea) and enter the lungs. How deep into the lungs the drug penetrates depends on the size of the droplets. The finer the droplets, the deeper they penetrate, thereby increasing the amount of drug absorbed. Inside the lungs, the drug is absorbed into the bloodstream. Drugs applied to the skin are usually used for their local effect and are therefore most commonly used to treat superficial skin disorders such as psoriasis, eczema, skin infections (viral, bacterial, and fungal), pruritus, and xerosis. The drug is mixed with an inactive substance. Depending on the consistency of the inactive substance, the formulation can be an ointment, cream, lotion, solution, powder, or gel.

[0153] In embodiments, a treatment regimen including a pharmaceutical composition may be titrated according to a subject's body weight. For subjects determined to be obese (BMI>35), it may be necessary to utilize practical body weight. BMI is calculated by BMI=weight (kg) / [height (m)]2. Ideal body weight may be calculated as 50 kg + 2.3 x (inches over 60 inches) for men or 45.5 kg + 2.3 (inches over 60 inches) for women. For subjects who are more than 20% heavier than their ideal body weight, an adjusted body weight may be calculated. The adjusted body weight may be the sum of ideal body weight + (0.4 x (actual body weight - ideal body weight)). In embodiments, body surface area may be used to calculate dosage. Body surface area (BSA) may be calculated by BSA (m2) = V height (cm) x weight (kg) / 3600.

[0154] In some embodiments, the pharmaceutical composition can be administered by any route, either alone or together with a pharmaceutically acceptable carrier or excipient, and such administration can be carried out in both single and multiple doses. More specifically, the pharmaceutical composition can be combined with various pharmaceutically acceptable inert carriers in the form of tablets, capsules, lozenges, troches, hand candies, powders, sprays, aqueous suspensions, injectable solutions, elixirs, syrups, and the like. Such carriers include solid diluents or fillers, sterile aqueous media, and various non-toxic organic solvents. Furthermore, pharmaceutical preparations may be suitably sweetened and / or flavored with various agents of the type commonly used for such purposes. Exemplary carriers and excipients may include dextrose, sodium chloride (NaCl), sucrose, lactose, cellulose, xylitol, sorbitol, maltitol, gelatin, PEG, PVP, histidine / histidine hydrochloride, trehalose dihydrate, polysorbate 80, and any combination thereof. In embodiments, the excipients include histidine / histidine hydrochloride, NaCl, trehalose dihydrate, and polysorbate 80.

[0155] Numbered Embodiments Notwithstanding the appended claims, the following numbered embodiments also form part of this disclosure.

[0156] Embodiment Set 1 Embodiment 1. Treating Sjogren's syndrome (SS) in a subject in need thereof for 1. A method comprising: The method is Tn3 scaffold containing CD40L-specific monomeric subunits This includes administering fruitwherein the Tn3 scaffold binds to CD40L; and the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G and six loop regions designated AB, BC, CD, DE, EF, and FG, where the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop SEQ ID NO: 16, wherein the Tn3 scaffold comprising the CD40L-specific monomeric subunit is administered at a dose of about 1500 mg, about 2500 mg, about 3000 mg, about 75-1500 mg, or about 300-3000 mg.

[0157] Embodiment 2. The method of embodiment 1, wherein at least one CD40L-specific monomeric subunit comprises seven β strands designated A, B, C, D, E, F, and G, wherein β strand A comprises SEQ ID NO:5, β strand B comprises SEQ ID NO:6, β strand C comprises SEQ ID NO:17, β strand D comprises SEQ ID NO:18, β strand E comprises SEQ ID NO:19, β strand F comprises SEQ ID NO:20, and β strand G comprises SEQ ID NO:21.

[0158] Embodiment 3. The subject has a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≥ 5. have 3. The method of embodiment 1 or 2, wherein

[0159] Embodiment 4. The method of embodiment 3, wherein the ESSDAI score is assessed based on the ESSDAI domains consisting of skin, kidney, joint, muscle, hematology, gland, constitution, lymphadenopathy, and biology.

[0160] Embodiment 5. The subject has (a) an ESSDAI score of <5; (b) a EULAR Sjoegren's Syndrome Patient Reported Index score of <5; 3. The method of embodiment 1 or 2, wherein the patient has (a) an Essay Reporting Index (ESSPRI) score of > 5, and (b) a stimulated total saliva flow rate > 0.1 mL / min.

[0161] Embodiment 6. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered as an induction dose and then as a maintenance dose.

[0162] Embodiment 7. The method of embodiment 6, wherein the induction dose comprises administering the Tn3 scaffold once about every two weeks for at least three doses.

[0163] Embodiment 8. The method of embodiment 6, wherein the maintenance dose comprises administering the Tn3 scaffold once about every four weeks for at least four doses.

[0164] Embodiment 9. The method of embodiment 8, wherein the time between the final induction dose and the first maintenance dose is about 4 weeks.

[0165] Embodiment 10. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered about once every 4 weeks, about once every 2 months, about once every 3 months, about once every 4 months, or about once every 6 months.

[0166] Embodiment 11. The method of embodiment 10, wherein the Tn3 scaffold is administered over at least four doses.

[0167] Embodiment 12. The Tn3 scaffold is administered over at least 5 doses. The method described below.

[0168] Embodiment 13. The method of any one of embodiments 1-12, wherein the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, cutaneously, systemically, topically, transdermally, or by inhalation.

[0169] Embodiment 14 The method of embodiment 13, wherein the Tn3 scaffold is administered intravenously.

[0170] Embodiment 15. The method of any one of embodiments 1 to 13, wherein the Tn3 scaffold comprises two CD40L-specific monomeric subunits linked in tandem.

[0171] Embodiment 16. The method of embodiment 15, wherein the two CD40L-specific monomer subunits each comprise SEQ ID NO:3.

[0172] Embodiment 17 The method of any one of embodiments 1 to 16, wherein the CD40L-specific monomeric subunits are linked by a linker.

[0173] Embodiment 18. At least one CD40L-specific monomer subunit is directly fused to polyethylene glycol (PEG). I'm 18. The method of any one of embodiments 1 to 17, wherein the compound is or is conjugated to the compound.

[0174] Embodiment 19. At least one CD40L-specific monomeric subunit is fused to polyethylene glycol (PEG) via a linker. I'm 18. The method of any one of embodiments 1 to 17, wherein the compound is or is conjugated to the compound.

[0175] Embodiment 20. The method of embodiment 17, wherein the linker comprises a peptide linker.

[0176] Embodiment 21. The method of embodiment 20, wherein the linker comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0177] Embodiment 22. At least one CD40L-specific monomeric subunit is fused to albumin I'm 22. The method of embodiment 21, wherein the antibody is or is conjugated to the antibody.

[0178] Embodiment 23. The method of embodiment 22, wherein the albumin is human serum albumin (HSA).

[0179] Embodiment 24. The method of embodiment 23, wherein the HSA is a mutant HSA comprising SEQ ID NO:4.

[0180] Embodiment 25. A method of treating Sjögren's syndrome (SS) in a subject in need thereof, comprising: The method is administering to a subject a dose of about 1500 mg, about 2500 mg, or about 3000 mg of a Tn3 scaffold. fruit , wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0181] Embodiment 26. The method of embodiment 25, wherein the Tn3 scaffold is administered as an induction dose, followed by one or more maintenance doses.

[0182] Embodiment 27. The method of embodiment 26, wherein the induction dose and the maintenance dose are the same amount.

[0183] Embodiment 28 The method of embodiment 27, wherein the induction dose and the maintenance dose are different amounts.

[0184] Embodiment 29. The method of any one of embodiments 25 to 28, wherein at least one dose is 3000 mg.

[0185] Embodiment 30. The method of any one of embodiments 25-29, wherein the induction dose and at least one maintenance dose are administered about 1 month apart, about 2 months apart, about 3 months apart, about 4 months apart, or about 6 months apart.

[0186] Embodiment 31. The method of any one of embodiments 25-30, wherein the induction dose comprises administering the Tn3 scaffold approximately every two weeks for at least three doses, and the maintenance dose comprises administering the Tn3 scaffold once every four weeks for at least four doses.

[0187] Embodiment 32. The method of any one of embodiments 1 to 31, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0188] Embodiment 33. The method of any one of embodiments 1-32, wherein the administration is effective to reduce the ESSDAI score when compared to otherwise similar subjects receiving a placebo control.

[0189] Embodiment 34. The method of embodiment 33, wherein the decrease is at least about a 1-point, 2-point, 3-point, 4-point, or 5-point decrease.

[0190] Embodiment 35. The method of embodiment 33, wherein the reduction is at least about a 6-point reduction.

[0191] Embodiment Set 2 1. Treating Sjögren's syndrome (SS) in subjects in need of SS treatment for 1. A method comprising: The method is administering to a subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L. fruit wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16; and wherein a subject in need thereof is treated with an antibody to the antibody to the antibody, to induce an increase in a subset of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to the subject's baseline level or the level in an untreated subject. have ,method.

[0192] 2. The method of embodiment 1, wherein at least one CD40L-specific monomeric subunit comprises seven β strands designated A, B, C, D, E, F, and G, wherein β strand A comprises SEQ ID NO:5, β strand B comprises SEQ ID NO:6, β strand C comprises SEQ ID NO:17, β strand D comprises SEQ ID NO:18, β strand E comprises SEQ ID NO:19, β strand F comprises SEQ ID NO:20, and β strand G comprises SEQ ID NO:21.

[0193] 3. The subject has a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≥ 5. have 3. The method of embodiment 1 or 2, wherein

[0194] 4. As described in embodiment 3, wherein the ESSDAI score is assessed based on the ESSDAI domains consisting of skin, kidney, joint, muscle, hematology, gland, constitution, lymphadenopathy, and biology. How to do it.

[0195] 5. Subjects have (a) an ESSDAI score <5, (b) an EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score ≥5, and (c) a stimulated total salivary flow rate >0.1 mL / min. have 3. The method of embodiment 1 or 2, wherein

[0196] 6. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered as an induction dose and then as a maintenance dose.

[0197] 7. The method of embodiment 6, wherein the induction dose comprises administering the Tn3 scaffold once about every two weeks for at least three doses.

[0198] 8. The method of embodiment 6, wherein the maintenance dose comprises administering the Tn3 scaffold once about every four weeks for at least four doses.

[0199] 9. The method of embodiment 8, wherein the time between the final induction dose and the first maintenance dose is about 4 weeks.

[0200] 10. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered about once every 4 weeks, about once every 2 months, about once every 3 months, about once every 4 months, or about once every 6 months.

[0201] 11. The method of embodiment 10, wherein the Tn3 scaffold is administered in at least four doses.

[0202] 12. The method of embodiment 11, wherein the Tn3 scaffold is administered over at least 5 doses.

[0203] 13. The method of any one of embodiments 1-12, wherein the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, cutaneously, systemically, topically, transdermally, or by inhalation.

[0204] 14. The method of embodiment 13, wherein the Tn3 scaffold is administered intravenously.

[0205] 15. The method of any one of embodiments 1-14, wherein the Tn3 scaffold comprises two CD40L-specific monomeric subunits linked in tandem.

[0206] 16. The method of embodiment 15, wherein the two CD40L-specific monomer subunits each comprise SEQ ID NO:3.

[0207] 17. The method of any one of embodiments 1 to 16, wherein the CD40L-specific monomeric subunits are linked by a linker.

[0208] 18. At least one CD40L-specific monomeric subunit is directly fused to polyethylene glycol (PEG). I'm 18. The method of any one of embodiments 1 to 17, wherein the compound is or is conjugated to the compound.

[0209] 19. At least one CD40L-specific monomeric subunit is fused to polyethylene glycol (PEG) via a linker. I'm or conjugated, 18. The method according to any one of aspects 1 to 17.

[0210] 20. The method of embodiment 17, wherein the linker comprises a peptide linker.

[0211] 21. The method of embodiment 20, wherein the linker comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0212] 22. At least one CD40L-specific monomeric subunit is fused to albumin. I'm 22. The method of embodiment 21, wherein the antibody is or is conjugated to the antibody.

[0213] 23. The method of embodiment 22, wherein the albumin is human serum albumin (HSA).

[0214] 24. The method of embodiment 23, wherein the HSA is a mutant HSA comprising SEQ ID NO:4.

[0215] 25. The method of any one of embodiments 1-24, wherein the Tn3 scaffold comprising a CD40L-specific monomeric subunit is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0216] 26. The method of embodiment 25, wherein the dose is about 1500 mg.

[0217] 27. The method of embodiment 25, wherein the dose is about 2500 mg.

[0218] 28. The method of embodiment 25, wherein the dose is about 3000 mg.

[0219] 29. A method of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising: The method is administering to a subject a dose of about 1500 mg, about 2500 mg, or about 3000 mg of a Tn3 scaffold. fruit wherein the Tn3 scaffold comprises SEQ ID NO: 1, and wherein a subject in need thereof has an increase in B cells of a subset selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to an otherwise healthy subject. have ,method.

[0220] 30. The method of embodiment 29, wherein the Tn3 scaffold is administered as an induction dose, followed by one or more maintenance doses.

[0221] 31. The method of embodiment 30, wherein the induction dose and the maintenance dose are the same amount.

[0222] 32. The method of embodiment 30, wherein the induction dose and the maintenance dose are different amounts.

[0223] 33. The method of any one of embodiments 29-32, wherein at least one dose is 3000 mg.

[0224] 34. The method of any one of embodiments 29-33, wherein the induction dose and at least one maintenance dose are administered about 1 month apart, about 2 months apart, about 3 months apart, about 4 months apart, or about 6 months apart.

[0225] 35. The method of any one of embodiments 29-34, wherein the induction dose comprises administering the Tn3 scaffold approximately every two weeks for at least three doses, and the maintenance dose comprises administering the Tn3 scaffold once every four weeks for at least four doses.

[0226] 36. The method of any one of embodiments 1-35, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0227] 37. The method of any one of embodiments 1-36, wherein the administration is effective in reducing the ESSDAI score when compared to otherwise identical subjects receiving a placebo control.

[0228] 38. The method of embodiment 37, wherein the decrease is at least about a 1-point, 2-point, 3-point, 4-point, or 5-point decrease.

[0229] 39. The method of embodiment 38, wherein the reduction is at least about a 6-point reduction.

[0230] 40. The method of any one of embodiments 1-39, wherein the administration is effective to decrease B cell levels when compared to the subject's baseline level or the level in an untreated subject.

[0231] 41. The method of any one of embodiments 1-40, wherein the administration is effective to reduce serum CXCL13 levels when compared to the subject's baseline level or the level in an untreated subject.

[0232] 42. The method of any one of embodiments 1-41, wherein the administration is effective in reducing rheumatoid factor (RF) autoantibodies when compared to the subject's baseline level or the level in an untreated subject.

[0233] 43. The method of any one of embodiments 40-42, wherein the decrease is detected by at least 14 days after administration.

[0234] 44. Treating an autoimmune disease in a subject in need thereof for It is a method , the method and administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L when the biomarker level is increased in the subject's sample compared to a healthy baseline level. fruit, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16.

[0235] 45. The method of embodiment 44, wherein the biomarker is expressed on the surface of a cell.

[0236] 46. ​​The method of embodiment 44, wherein the biomarker is an antibody.

[0237] 47. The method of embodiment 44, wherein the biomarker is secreted by the cell.

[0238] 48. The method of any one of embodiments 44-47, wherein the biomarkers are selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, CD11c high atypical memory cells, CXCL13, RF autoantibodies, and any combination thereof.

[0239] 49. Autoimmune diseases include systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjogren's syndrome, dermatomyositis, myositis, sclerosis, diabetes, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, rheumatic carditis, psoriasis, arthritis, inflammation, and rheumatoid arthritis. The method according to any one of embodiments 44 to 48, wherein the disease is selected from the group consisting of gore, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonopathy.

[0240] 50. The method of any one of embodiments 44-49, wherein administration is continued until the level of the biomarker returns to a healthy baseline.

[0241] 51. The method according to any one of embodiments 44 to 50, wherein the level of the biomarker is analyzed periodically.

[0242] 52. The method of any one of embodiments 44-51, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0243] 53. The method of any one of embodiments 44-52, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0244] 54. Treating Sjögren's syndrome (SS) in a subject in need of SS treatment for 1. A method comprising: The method is administering to a subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L. fruit wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop SEQ ID NO: 16, and wherein treatment of a subject with the Tn3 scaffold reduces the level of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells when compared to the subject's baseline level or the level in an untreated subject.

[0245] 55. The method of embodiment 54, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0246] 56. The method of embodiment 54 or 55, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0247] 57. The method of any one of embodiments 54-56, wherein B cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold.

[0248] Embodiment Set 3 1. Treating Sjögren's syndrome (SS) in subjects in need of SS treatment for 1. A method comprising: The method is administering to a subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L. fruit wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16; and wherein a subject in need thereof is treated with an antibody to the antibody to the antibody, to induce an increase in a subset of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to the subject's baseline level or the level in an untreated subject. have ,method.

[0249] 2. The method of embodiment 1, wherein at least one CD40L-specific monomeric subunit comprises seven β strands designated A, B, C, D, E, F, and G, wherein β strand A comprises SEQ ID NO:5, β strand B comprises SEQ ID NO:6, β strand C comprises SEQ ID NO:17, β strand D comprises SEQ ID NO:18, β strand E comprises SEQ ID NO:19, β strand F comprises SEQ ID NO:20, and β strand G comprises SEQ ID NO:21.

[0250] 3. The subject has a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≥ 5. have 3. The method of embodiment 1 or 2, wherein

[0251] 4. The method of embodiment 3, wherein the ESSDAI score is assessed based on the ESSDAI domains consisting of skin, kidney, joint, muscle, hematology, gland, constitution, lymphadenopathy, and biology.

[0252] 5. The subject is (a) ESSDAI score < 5 points; (b) EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score ≥ 5 points, and (c) Total saliva flow rate during stimulation >0.1mL / min have 3. The method of embodiment 1 or 2, wherein

[0253] 6. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered as an induction dose and then as a maintenance dose.

[0254] 7. The method of embodiment 6, wherein the induction dose comprises administering the Tn3 scaffold once about every two weeks for at least three doses.

[0255] 8. The method of embodiment 6, wherein the maintenance dose comprises administering the Tn3 scaffold once about every four weeks for at least four doses.

[0256] 9. The method of embodiment 8, wherein the time between the final induction dose and the first maintenance dose is about 4 weeks.

[0257] 10. The method of any one of embodiments 1-5, wherein the Tn3 scaffold is administered about once every 4 weeks, about once every 2 months, about once every 3 months, about once every 4 months, or about once every 6 months.

[0258] 11. The method of embodiment 10, wherein the Tn3 scaffold is administered in at least four doses.

[0259] 12. The method of embodiment 11, wherein the Tn3 scaffold is administered over at least 5 doses.

[0260] 13. The method of any one of embodiments 1-12, wherein the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, cutaneously, systemically, topically, transdermally, or by inhalation.

[0261] 14. The method of embodiment 13, wherein the Tn3 scaffold is administered intravenously.

[0262] 15. The method of any one of embodiments 1-14, wherein the Tn3 scaffold comprises two CD40L-specific monomeric subunits linked in tandem.

[0263] 16. The method of embodiment 15, wherein the two CD40L-specific monomer subunits each comprise SEQ ID NO:3.

[0264] 17. The method of any one of embodiments 1 to 16, wherein the CD40L-specific monomeric subunits are linked by a linker.

[0265] 18. At least one CD40L-specific monomeric subunit is directly fused to polyethylene glycol (PEG). I'm 18. The method of any one of embodiments 1 to 17, wherein the compound is or is conjugated to the compound.

[0266] 19. At least one CD40L-specific monomeric subunit is fused to polyethylene glycol (PEG) via a linker. I'm 18. The method of any one of embodiments 1 to 17, wherein the compound is or is conjugated to the compound.

[0267] 20. The method of embodiment 17, wherein the linker comprises a peptide linker.

[0268] 21. The method of embodiment 20, wherein the linker comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0269] 22. At least one CD40L-specific monomeric subunit is fused to albumin. I'm 22. The method of embodiment 21, wherein the antibody is or is conjugated to the antibody.

[0270] 23. The method of embodiment 22, wherein the albumin is human serum albumin (HSA).

[0271] 24. The method of embodiment 23, wherein the HSA is a mutant HSA comprising SEQ ID NO:4.

[0272] 25. The method of any one of embodiments 1-24, wherein the Tn3 scaffold comprising a CD40L-specific monomeric subunit is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0273] 26. The method of embodiment 25, wherein the dose is about 1500 mg.

[0274] 27. The method of embodiment 25, wherein the dose is about 2500 mg.

[0275] 28. The method of embodiment 25, wherein the dose is about 3000 mg.

[0276] 29. A method of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising: The method is administering to a subject a dose of about 1500 mg, about 2500 mg, or about 3000 mg of a Tn3 scaffold. fruit, wherein the Tn3 scaffold comprises SEQ ID NO: 1, and the subject in need thereof originally 10. A method comprising the step of administering to a subject a therapeutically effective amount of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, when compared to a healthy subject.

[0277] 30. The method of embodiment 29, wherein the Tn3 scaffold is administered as an induction dose, followed by one or more maintenance doses.

[0278] 31. The method of embodiment 30, wherein the induction dose and the maintenance dose are the same amount.

[0279] 32. The method of embodiment 30, wherein the induction dose and the maintenance dose are different amounts.

[0280] 33. The method of any one of embodiments 29-32, wherein at least one dose is 3000 mg.

[0281] 34. The method of any one of embodiments 29-33, wherein the induction dose and at least one maintenance dose are administered about 1 month apart, about 2 months apart, about 3 months apart, about 4 months apart, or about 6 months apart.

[0282] 35. The method of any one of embodiments 29-34, wherein the induction dose comprises administering the Tn3 scaffold approximately every two weeks for at least three doses, and the maintenance dose comprises administering the Tn3 scaffold once every four weeks for at least four doses.

[0283] 36. The method of any one of embodiments 1-35, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0284] 37. The method of any one of embodiments 1-36, wherein the administration is effective in reducing the ESSDAI score when compared to otherwise identical subjects receiving a placebo control.

[0285] 38. The method of embodiment 37, wherein the decrease is at least about a 1-point, 2-point, 3-point, 4-point, or 5-point decrease.

[0286] 39. The method of embodiment 38, wherein the reduction is at least about a 6-point reduction.

[0287] 40. The method of any one of embodiments 1-39, wherein the administration is effective to decrease B cell levels when compared to the subject's baseline level or the level in an untreated subject.

[0288] 41. The method of any one of embodiments 1-40, wherein the administration is effective to reduce serum CXCL13 levels when compared to the subject's baseline level or the level in an untreated subject.

[0289] 42. The method of any one of embodiments 1-41, wherein the administration is effective in reducing rheumatoid factor (RF) autoantibodies when compared to the subject's baseline level or the level in an untreated subject.

[0290] 43. Reduction continues for at least 14 days after administration to 43. The method of any one of embodiments 40 to 42, wherein the marker is detected.

[0291] 44. Treating an autoimmune disease in a subject in need thereof for It is a method , the method and administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L when the biomarker level is increased in the subject's sample compared to a healthy baseline level. fruit , the monomeric subunits are composed of seven β-strands designated A, B, C, D, E, F, and G, and seven β-strands designated AB, BC, CD, DE, EF, and F. and six loop regions designated G, wherein the AB loop comprises SEQ ID NO:11, the BC loop comprises SEQ ID NO:12, the CD loop comprises SEQ ID NO:13, the DE loop comprises SEQ ID NO:14, the EF loop comprises SEQ ID NO:15, and the FG loop comprises SEQ ID NO:16.

[0292] 45. The method of embodiment 44, wherein the biomarker is expressed on the surface of a cell.

[0293] 46. ​​The method of embodiment 44, wherein the biomarker is an antibody.

[0294] 47. The method of embodiment 44, wherein the biomarker is secreted by the cell.

[0295] 48. The method of any one of embodiments 44-47, wherein the biomarkers are selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, CD11c high atypical memory cells, CXCL13, RF autoantibodies, and any combination thereof.

[0296] 49. The method of any one of embodiments 44-48, wherein the biomarkers are selected from the group consisting of TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, and CD80, and any combination thereof.

[0297] 50. The method of any one of embodiments 44-49, wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjogren's syndrome, dermatomyositis, myositis, sclerosis, diabetes, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, rheumatic carditis, psoriasis, arthritis, inflammation, rheumatoid arthritis, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonopathy.

[0298] 51. The method of embodiment 50, wherein the arthritis is rheumatoid arthritis.

[0299] 52. The method of any one of embodiments 44-51, wherein administration is continued until the level of the biomarker returns to a healthy baseline.

[0300] 53. The method according to any one of embodiments 44 to 52, wherein the level of the biomarker is analyzed periodically.

[0301] 54. The method of any one of embodiments 44-53, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0302] 55. The method of any one of embodiments 44-54, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0303] 56. Treating Sjögren's syndrome (SS) in a subject in need of SS treatment for 1. A method comprising: The method is administering to a subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L. fruit , the monomeric subunit comprises seven β-strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, where the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, and the DE loop comprises SEQ ID NO: No. 14, wherein the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16, and wherein treating a subject with the Tn3 scaffold reduces the level of B cells selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells when compared to the subject's baseline level or the level in an untreated subject.

[0304] 57. The method of embodiment 56, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0305] 58. The method of embodiment 56 or 57, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0306] 59. The method of any one of embodiments 56-58, wherein B cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold.

[0307] 60. Treat Sjögren's syndrome (SS) in a subject in need of SS treatment. for 1. A method comprising: The method administering to a subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L. fruit, wherein the monomeric subunit comprises seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, where the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16, and the Tn3 scaffold increases Ki67+ / CD3+, Ki67+ / CD4+ cells, Ki67+ / CD3+ / CD4+ cells, when compared to the subject's baseline levels or the levels of an untreated subject. cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCRT7+ cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCR7- cells, Ki67+ / CD3+ / CD4+ / CXCR5+ / ICOS+ cells, Ki67+ / CD3+ / CD8+ cells, Ki67+ / CD3+ / CD8+ / CD45RA- / CCR7+ cells, Ki67+ / CD3+ / CD8+ / CD45RA / CCR7- cells, and Ki67+ / CD3+ / CD8+ / CXCR5+ cells, and CXCR5+ / ICOS+ / CD3+ / CD4+ cells. of increase have The method comprises administering the compound to a subject.

[0308] 61. The method of embodiment 60, wherein the Tn3 scaffold comprises SEQ ID NO:1.

[0309] 62. The method of embodiment 60 or 61, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

[0310] 63. The method of any one of embodiments 60-62, wherein T cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold. [Example]

[0311] Example 1 - A Phase 2, Randomized, Double-Blind, Placebo-Controlled, Proof-of-Concept Study to Determine the Efficacy and Safety of VIB4920 in Subjects with Sjogren's Syndrome (SS) Study design This study was a randomized, double-blind, placebo-controlled, parallel-group study to evaluate the efficacy, safety, and tolerability of VIB4920, an anti-CD40L-Tn3 fusion protein, in adult subjects diagnosed with SS according to the 2016 American College of Rheumatology (ACR) / European League Against Rheumatism (EULAR) criteria. Two SS populations were enrolled in this study. Population #1 consisted of subjects with moderate to severe (moderate to high) systemic disease activity as defined by an ESSDAI score of ≥ 5. Population #2 consisted of subjects with mild systemic disease activity as defined by an ESSDAI score < 5, but with moderate to high (or severe) subjective symptoms and residual stimulated salivary flow as defined by an ESSPRI score of ≥ 5.

[0312] Within each cohort, eligible subjects were randomized to receive VIB4920 1500 mg IV or placebo once every 2 weeks (Q2W) for 3 doses, followed by 4 additional doses once every 4 weeks (Q4W) (Stage I). Starting on Day 169, subjects randomized to VIB4920 received 5 doses of placebo Q4W, and subjects randomized to placebo received 5 doses of VIB4920 Q4W (Stage II). Subjects who discontinued VIB4920 were ineligible for treatment in Stage II. All subjects were followed for at least 12 weeks after their final IP dose.

[0313] An exemplary study schematic is presented in Figure 1.

[0314] group Two SS populations distinguish between patients with predominantly glandular dysfunction and those with both glandular dysfunction and systemic manifestations. These two SS populations represent subsets with significant unmet need and require different endpoints for efficacy assessment. The ESSDAI is a validated instrument for assessing systemic disease activity. The ESSPRI uses a 0-10 point numeric analog scale and is a scale for the assessment of each of the three domains of SS symptoms: dryness, fatigue, and pain (joint and / or muscle).

[0315] Population #1 included SS patients with moderate to high systemic disease activity as defined by an ESSDAI score of ≥ 5. The following domains were scored but did not contribute to the minimum ESSDAI score of 5 required for inclusion: peripheral nervous system, central nervous system, and lung. Population #2 included SS patients with exocrine dysfunction and subjective symptoms as defined by an ESSPRI score of ≥ 5, considered the cutoff point for "unsatisfactory symptom status," but with low systemic disease activity (ESSDAI score < 5).

[0316] A complete physical examination, including weight and height, and 28 joint evaluations were performed. The classification of SS for both cohorts #1 and #2 was performed using the 2016 ACR / EULAR criteria and confirmed by a medically qualified individual. Patients who met the inclusion criteria were 1 , and do not have any of the conditions listed as exclusion criteria 2 The classification of SS was applied to anyone who had a score ≥ 4 when summing the weights from the following items shown in Table 2:

[0317] [Table 2]

[0318] Eligibility for ESSDAI scores for Group #1 was also confirmed by a medical licensed professional. Stimulated salivary flow, ESSDAI, and ESSPRI were performed at screening to determine study eligibility. Blood tests required for ESSDAI were also completed. In addition, an autoantibody panel including RF was collected.

[0319] Inclusion criteria for group #1 For inclusion in Cohort #1 of this study, each individual may have met the following criteria: 1. Adult, 18 years of age or older at the time of informed consent (in countries with different laws, the minimum age for adult participants may be higher than 18 years). 2. The patient was diagnosed with SS based on the 2016 ACR / EULAR classification criteria. 3. ESSDAI score ≥ 5 at screening have the following domains were scored but did not contribute to the minimum ESSDAI score of 5 required for inclusion: peripheral nervous system, central nervous system, and lung. 4. Positive anti-Ro autoantibodies and / or RF at screening as defined by standard central laboratory testing. 5. Written informed consent obtained from subject / legal representative prior to performing any protocol-related procedures, including screening assessments, and any authorizations required locally (e.g., Health Insurance Portability and Accountability Act in the United States, European Union Data Privacy Directive in Europe). 6. Sexually active women of childbearing potential with an unsterilized male partner should From the time you sign the Informed Consent Form (ICF), you must use highly effective contraception and agree to continue using such protection until the end of study follow-up; discontinuation of contraception after this point must be discussed with your physician. Highly effective contraception includes: Combined (estrogen and progestogen-containing) hormonal contraceptive methods with ovulation suppression: -Oral -In the vagina -Transdermal Progestogen-only hormonal contraception with ovulation suppression: -Oral -injection -Implantation Intrauterine device (IUD) Intrauterine hormone-releasing system (IUS) Bilateral tubal occlusion Partner who has had a vasectomy ·abstinence Abstinence will be considered highly effective only if it is the subject's preferred and usual lifestyle and if the subject agrees to abstain from heterosexual intercourse from the time of signing the ICF until the end of study follow-up. Periodic abstinence, rhythm methods, and pull-out methods are not acceptable methods of contraception. It is recommended that female partners (of childbearing potential) of male study participants must use a highly effective method of contraception other than a barrier method. - A woman of childbearing potential is defined as a woman who has not been surgically sterilized (sterilization includes bilateral tubal ligation, bilateral oophorectomy, or hysterectomy) or who is not postmenopausal (defined as the absence of menstruation for 12 months without another medical cause). - Vasectomy partners are a highly effective method of birth control, provided they are the only sexual partners of the female study participant of childbearing potential, and the vasectomy partners have been medically evaluated for the success of the procedure. 7. Unsterilized male subjects who are sexually active with a female partner of childbearing potential must use spermicide-coated condoms from day 1 through the end of the study. 8. Meet all of the following tuberculosis (TB) criteria: a. No history of latent or active TB prior to screening, except for latent TB with documented completion of appropriate treatment. b. No signs or symptoms suggestive of active TB by medical history or physical examination. c. Not have had recent (within 12 weeks of screening) close contact with someone with active TB (close contact is defined as 4 hours / week or more or living in the same household or a home frequently visited by someone with active TB). d. A negative interferon-gamma release assay (IGRA) test result for TB obtained within 12 weeks prior to randomization. Subjects with an indeterminate test result may be retested, but if the retest is also indeterminate, the subject will be excluded. e. Chest radiograph (obtained at any time during the screening period or within 12 weeks prior to signing of the ICF) has no evidence of current active TB or other infection, or clinically significant abnormalities (other than those attributable to SS) suggestive of previous active TB, malignancy, or an active process.

[0320] If an individual in cohort #1 met any of the following criteria, they were ineligible: 1. Patients with a history of definite deep vein thrombosis or arterial thromboembolism within 2 years at the time of ICF signing. 2. Risk factors for venous thromboembolism or arterial thrombosis (e.g., ≥12 weeks prior to screening) Immobilization or major surgery), prothrombotic states (including but not limited to congenital or hereditary antithrombin III deficiency, protein C, protein S, or a confirmed diagnosis of fulminant antiphospholipid syndrome). 3. Patients who require treatment with anticoagulants (clopidogrel, prasugrel, warfarin, low molecular weight heparin, etc.). Low-dose aspirin therapy (up to 325 mg / day) is permitted. 4. Concomitant polymyositis or dermatomyositis or systemic sclerosis. 5. Active malignancy or history of malignancy within the past 5 years, except in the following cases: a. Treated cervical intraepithelial neoplasia with apparently successful definitive therapy more than 12 months prior to screening; or b. Basal cell carcinoma of the skin after apparently curative therapy. 6. Subjects who are pregnant or breastfeeding, or planning to become pregnant during the duration of the study. 7. Subject has tested positive for or been treated for Hepatitis B, Hepatitis C, or HIV infection. For hepatitis B, a positive test for chronic hepatitis B infection at screening is defined as either (1) a positive hepatitis B surface antigen (HBsAg) or (2) a positive hepatitis B core antibody (anti-HBc). 8. Subjects with: a. History of two or more episodes of herpes zoster and / or opportunistic infections in the past 12 months, excluding oral candidiasis, vaginal candidiasis, and fungal skin infections. b. Active viral, bacterial, or other infection requiring systemic treatment at screening or randomization, or a history of 3 or more infections requiring IV antibiotics within 12 months prior to signing of the ICF. Health-related COVID-19 severity risk based on current understanding of COVID-19 epidemiological risk (recent exposure, high-risk residence) and risk factors for severe disease when making decisions regarding the individual subject's risk of participation. Subjects with active COVID-19 infection or disease or other serious infection, or who, in the investigator's judgment, may be at unacceptable risk of COVID-19 or its complications, must not be randomized. d. Documented positive SARS-CoV-2 test within 2 weeks prior to randomization. Subjects who test positive for SARS-CoV-2 may be rescreened at least 2 weeks after the positive test if asymptomatic and at least 3 weeks after symptomatic COVID-19 illness. 9. Subject has a history of severe allergy or reaction to any component of the VIB4920 formulation or any other biologic therapy. 10. Subject has any severe cardiovascular, respiratory, endocrine, gastrointestinal, hematological, neurological, psychiatric, or systemic disorder or any other condition that, in the opinion of the investigator, may place the subject at unacceptable risk of complications, interfere with the assessment of VIB4920, or confound the subject's safety or the interpretation of the study results. 11. Subjects who are unable or unwilling to comply with the requirements of the protocol (e.g., active drug or alcohol abuse). 12.Subjects who have received a live (attenuated) vaccine within 4 weeks prior to signing of the ICF. 13. Last dose of an experimental biologic (except those listed in item 14) or oral medication within 3 months or 5 half-lives prior to randomization. 14. Subject has previously received treatment with any biologic B-cell depleting therapy (e.g., rituximab, ocrelizumab, or ofatumumab) within 12 months prior to randomization or other B-cell targeted therapy (e.g., belimumab) within 3 months prior to randomization. 15. Treatment with injectable corticosteroids (including intra-articular) or oral prednisone or equivalent at doses greater than 10 mg / day within 6 weeks prior to randomization. Concomitant treatment with oral corticosteroids ≤ 10 mg / day of prednisone or equivalent is permitted, provided this dose is stable from ≥ 2 weeks prior to screening through randomization (Day 1) and is expected to remain stable for the duration of treatment. Asthma, chronic obstructive pulmonary disease Alternatively, inhaled or topical corticosteroids given for skin conditions are permitted provided the dose is expected to remain stable during the study. 16.Subject is treated with systemic corticosteroids for a total of more than 2 weeks for an indication other than SS, RA, and SLE within the 24 weeks prior to the screening visit. 17. Use of the following medications: Antimalarials (e.g., chloroquine, hydroxychloroquine, quinacrine) if started within 8 weeks prior to signing the ICF or during the screening period, or if the dose was changed. b. MTX if the dose is greater than 20 mg / week; or if there is any change or initiation of a new dose within 4 weeks prior to ICF signing and before randomization (Day 1), or if there is any change in route of administration. c. Azathioprine (AZA) if the dose is greater than 150 mg / day and there is any change or initiation of a new dose and any change in route of administration within 4 weeks prior to ICF signing and before randomization (Day 1). d. Leflunomide if the dose is greater than 20 mg / day; or if there is any change or initiation of a new dose within 4 weeks prior to ICF signing and before randomization (Day 1). e. Mycophenolate mofetil (MMF) if the dose is greater than 2 g / day; or if there is any change or initiation of a new dose within 4 weeks prior to ICF signing and before randomization (Day 1). f. Any other DMARD, immunosuppressant, or antiproliferative agent if the last dose was taken within: - 4 weeks prior to signing the ICF or Drug-specific 5 half-life elimination period (if longer than 4 weeks). g. Any medication that, in the opinion of the investigator, may interfere with the assessment of VIB4920 or the subject's safety or interpretation of the study results. h. Any increase in or initiation of a new dose of cevimeline or pilocarpine and cyclosporine eye drops (Restasis) within 2 weeks prior to ICF signing through randomization (Day 1). 18. Subject has previously received treatment with an anti-CD40L compound at any time prior to screening. 19.Subject has any of the following blood tests at screening: Aspartate aminotransferase (AST) > 2 × upper limit of normal (ULN) Alanine aminotransferase (ALT) > 2 × ULN Total bilirubin (TBL) > 2 × ULN Hemoglobin < 75g / L ·Neutrophil<1.0×109 / L ·Platelets<100×109 / L Prothrombin or partial thromboplastin time (PTT) > ULN

[0321] Inclusion criteria for group #2 For inclusion in Cohort #2 of this study, each individual met all of the following criteria: 1. Adult, 18 years of age or older at the time of informed consent (in countries with different laws, the minimum age for adult participants may be higher than 18 years). 2. The patient was diagnosed with SS based on the 2016 ACR / EULAR classification criteria. 3. ESSPRI score ≥ 5 at screening have do. 4. ESSDAI score <5 at screening have a point do. 5. Positive anti-Ro autoantibodies and / or RF at screening as defined by available standard testing methods from a central laboratory. 6. Residual salivary gland function as defined by stimulated total salivary flow rate >0.1 mL / min. 7. Written informed consent obtained from subject / legal representative prior to performing any protocol-related procedures, including screening assessments, and any authorizations required locally (e.g., Health Insurance Portability and Accountability Act in the United States, EU Data Privacy Directive in Europe). 8. Sexually active women of childbearing potential with an unsterilized male partner must use highly effective contraception from the time of ICF signing and agree to continue using such protection until the end of study follow-up; discontinuation of contraception after this point must be discussed with their physician. Highly effective contraception includes: Combined (estrogen and progestogen-containing) hormonal contraceptive methods with ovulation suppression: -Oral -In the vagina -Transdermal Progestogen-only hormonal contraception with ovulation suppression: -Oral -injection -Implantation Intrauterine device (IUD) Intrauterine hormone-releasing system (IUS) Bilateral tubal occlusion Partner who has had a vasectomy ·abstinence Abstinence will be considered highly effective only if it is the subject's preferred and usual lifestyle and if the subject agrees to abstain from heterosexual intercourse from the time of signing the ICF until the end of study follow-up. Periodic abstinence, rhythm methods, and pull-out methods are not acceptable methods of contraception. It is recommended that female partners (of childbearing potential) of male study participants must use a highly effective method of contraception other than a barrier method. - A woman of childbearing potential is defined as a woman who has not been surgically sterilized (sterilization includes bilateral tubal ligation, bilateral oophorectomy, or hysterectomy) or who is not postmenopausal (defined as the absence of menstruation for 12 months without another medical cause). - Vasectomy partners are a highly effective method of birth control, provided they are the only sexual partners of the female study participant of childbearing potential, and the vasectomy partners have been medically evaluated for the success of the procedure. 9. Male subjects who are sexually active with an unsterilized female partner of childbearing potential must use spermicide condoms from Day 1 through the end of the study. 10. Meet all of the following TB criteria: i. No history of latent or active TB prior to screening, except for latent TB with documented completion of appropriate treatment. j. No signs or symptoms suggestive of active TB by medical history or physical examination. k. Not have had recent (within 12 weeks of screening) close contact with someone with active TB (close contact is defined as 4 hours / week or more or living in the same household or a home frequently visited by someone with active TB). l. A negative IGRA test result for TB obtained within 12 weeks prior to randomization. Subjects with an indeterminate test result may be retested, but if the retest is also indeterminate, the subject will be excluded. m. Chest radiograph (obtained at any time during the screening period or within 12 weeks prior to signing the ICF) showing current active TB or other infection, or past active TB, malignant There is no evidence of a chronic tumor or clinically significant abnormalities (other than those attributable to SS) suggesting an active process.

[0322] Exclusion criteria for population #2 If an individual in group #2 met any of the following criteria, they were ineligible: 1. Patients with a history of definitive deep vein thrombosis or arterial thromboembolism within 2 years at the time of signing the ICF. 2. Patients with risk factors for venous thromboembolism or arterial thrombosis (e.g., immobilization or major surgery within 12 weeks prior to screening), prothrombotic states (including but not limited to congenital or hereditary antithrombin III deficiency, protein C, protein S, or a confirmed diagnosis of fulminant antiphospholipid syndrome). 3. Patients who require anticoagulant therapy (clopidogrel, prasugrel, warfarin, low molecular weight heparin, etc.). Low-dose aspirin therapy (up to 325 mg / day) is permitted. 4. Concomitant polymyositis or dermatomyositis or systemic sclerosis. 5. Active malignancy or history of malignancy, except in the following cases: n. Treated cervical intraepithelial neoplasia with apparently successful definitive therapy more than 12 months prior to screening; or o. Treated basal cell carcinoma of the skin after apparently successful definitive treatment 6. Subjects who are pregnant or breastfeeding, or planning to become pregnant during the duration of the study. 7. Subject has tested positive for or been treated for Hepatitis B, Hepatitis C, or HIV infection. For hepatitis B, a positive test for chronic hepatitis B infection at screening is defined as detection of either (1) hepatitis B surface antigen (HBsAg); or (2) hepatitis B core antibody (anti-HBc). 8. Subjects with: a. History of two or more episodes of herpes zoster and / or opportunistic infections in the past 12 months, excluding oral candidiasis, vaginal candidiasis, and fungal skin infections. b. Active viral, bacterial, or other infection requiring systemic treatment at screening or randomization, or a history of 3 or more infections requiring IV antibiotics within 12 months prior to signing of the ICF. Health-related COVID-19 severity risk based on current understanding of COVID-19 epidemiological risk (recent exposure, high-risk residence) and risk factors for severe disease when making decisions regarding the individual subject's risk of participation. Subjects with active COVID-19 infection or disease or other serious infection, or who, in the investigator's judgment, may be at unacceptable risk of COVID-19 or its complications, must not be randomized. d. Documented positive SARS-CoV-2 test within 2 weeks prior to randomization. Subjects who test positive for SARS-CoV-2 may be rescreened at least 2 weeks after the positive test if asymptomatic and at least 3 weeks after symptomatic COVID-19 illness. 9. Subject has a history of severe allergy or reaction to any component of the VIB4920 formulation or any other biologic therapy. 10. Subject has any severe cardiovascular, respiratory, endocrine, gastrointestinal, hematological, neurological, psychiatric, or systemic disorder or any other condition that, in the opinion of the investigator, may interfere with the assessment of VIB4920 or the subject's safety or interpretation of the study results. 11. Subjects who are unable or unwilling to comply with the requirements of the protocol (e.g., active drug or alcohol abuse). 12.Subjects who have received a live (attenuated) vaccine within 4 weeks prior to signing of the ICF. 13. Last dose of an experimental biologic (except those listed in item 14) or oral medication within 3 months or 5 half-lives prior to randomization. 14. Any biological B-cell depleting therapy (e.g., rituximab) within 12 months prior to randomization Subjects who have previously received treatment with rituximab (e.g., ocrelizumab, ofatumumab) or other B-cell targeted therapies (e.g., belimumab) within 3 months prior to randomization. 15. Use of the following medications: Antimalarials (e.g., chloroquine, hydroxychloroquine, quinacrine) if started within 8 weeks prior to signing the ICF or during the screening period, or if the dose was changed. b. Oral, intramuscular, IV, or intra-articular corticosteroids within 4 weeks prior to ICF signing until randomization (Day 1). c. MTX, AZA, leflunomide, other cDMARDs, or immunosuppressive or antiproliferative medications if the last dose was taken within the following time periods: - 4 weeks prior to signing the ICF or Drug-specific 5 half-life elimination period (if longer than 4 weeks). d. Any medication that, in the opinion of the investigator, may interfere with the assessment of VIB4920 or the subject's safety or interpretation of the study results. e. Any increase in or initiation of a new dose of regular nonsteroidal anti-inflammatory medication within 2 weeks prior to ICF signing until randomization (Day 1). f. Any increase in or initiation of a new dose of cevimeline or pilocarpine and cyclosporine eye drops (Restasis) within 2 weeks prior to ICF signing and up to randomization (Day 1). 16. Subject has previously received treatment with an anti-CD40L compound at any time prior to screening. 17.Subject has any of the following blood tests at screening: AST>2×ULN ALT>2×ULN TBL>2×ULN Hemoglobin < 75g / L ·Neutrophil<1.0×109 / L ·Platelets<100×109 / L Prothrombin or PTT > ULN

[0323] Safety-related screening assessments included adverse events (AEs), safety laboratory tests (blood chemistry, hematology, and urinalysis), coagulation parameters, chest X-ray, thyroid function tests, and, for women of childbearing potential, serum β-human chorionic gonadotropin (β-hCG) pregnancy test.

[0324] Safety evaluation During the active treatment period, safety assessments were performed according to the following assessments, as shown in Table 5: - Monitor and record all adverse events (AEs) (including AEs of special interest) and serious adverse events - Safety laboratory tests (blood chemistry, hematology, and urinalysis) -Concomitant medication - Vital signs, physical examination (complete or symptom-based depending on visit), and weight -ECG -Urine pregnancy test (for women of childbearing potential) -Coagulation tests

[0325] Adverse events (AEs) An AE is any untoward medical occurrence associated with the use of an intervention in humans, whether or not it is considered related to the intervention.

[0326] Serious Adverse Events (SAEs) An SAE is considered "serious" if it results in any of the following outcomes: -death - Life-threatening AE (An event is considered "life-threatening" if, in the opinion of either the investigator or sponsor, its occurrence places the patient or subject at immediate risk of death. This does not include AEs or suspected adverse reactions (SARs) that, if they occurred in a more severe form, could have caused death.) -Hospitalization or extension of existing hospitalization - Persistent or significant incapacity or substantial failure to perform normal life functions -Congenital anomalies / birth defects A significant medical event that is not fatal, life-threatening, or requires hospitalization may be considered serious if, based on sound medical judgment, it may endanger the patient or subject and may require medical or surgical intervention to prevent one of the outcomes listed in this definition. Examples of such a medical event include allergic bronchospasm requiring intensive care in an emergency room or at home, blood dyscrasia or seizures that do not result in hospitalization, or the development of drug addiction or drug abuse.

[0327] Causation or association The relationship of AEs and SAEs to VIB4920 was assessed. An event was considered "not related" to the use of VIB4920 if it met any of the following tests: a. The temporal relationship between administration of VIB4920 and the occurrence of the event is unreasonable (e.g., the event either occurred before administration of VIB4920 or occurred too long after its administration to be considered VIB4920-related). b. A causal relationship between VIB4920 and the event is biologically implausible (e.g., death as a passenger in a car accident). c. There is a clearly more plausible alternative explanation for the event (e.g., a typical adverse reaction [AR] to a concomitant medication and / or a typical disease-related event)

[0328] Individual AE / SAE reports were considered "related" to the use of VIB4920 if they did not meet the criteria for "unrelated." "Related" means that the event was considered "associated with the use of the drug," implying that there was a "reasonable possibility" that the event could have been caused by the product (i.e., there were facts, evidence, or arguments suggesting a possible cause).

[0329] Severity / Intensity Severity was assessed according to the following scale: Grade 1: An event of mild intensity that is usually transient and may require only minimal treatment or intervention. The event generally does not interfere with normal daily living activities. Grade 2: An event of moderate intensity that is usually alleviated by additional specific therapeutic intervention. The event interferes with normal daily living activities and causes discomfort, but does not pose a significant or lasting risk of harm to the subject. Grade 3: A severe event requiring intensive therapeutic intervention. The event interferes with normal daily living activities or has a significant impact on the subject's clinical status. Grade 4: An event associated with an imminent risk of death or associated with physical or mental disability that affects or limits the subject's ability to perform daily living activities (eating, walking, toileting, etc.), and / or its immediate sequelae. Grade 5: Death (loss of life) as a result of the event.

[0330] Adverse Events of Special Interest (AESI) Adverse Events of Special Interest (AESI) were assessed. AESIs are among the scientific and medical concerns specific to the understanding of VIB4920 that warrant close monitoring and the collection of additional information. AESI may be serious or non-serious.

[0331] The following AESIs were monitored: Thrombotic and embolic events Abnormal liver function (meeting the definition of Hy's Law (HL)) Anaphylaxis and clinically significant (grade 3 or greater) hypersensitivity reactions Severe infusion reactions (Common Terminology Criteria for Adverse Events (CTCAE) grade 3 or higher). Severity was assessed according to CTCAE v5. - Grade 1: Mild, transient reaction; infusion interruption not indicated; intervention not indicated. - Grade 2: Interruption of therapy or infusion indicated but prompt response to symptomatic treatment (e.g., antihistamines, NSAIDs, narcotics, intravenous fluids); prophylactic drug therapy within 24 hours indicated Grade 3: Prolonged (e.g., not responding promptly to symptomatic medication and / or brief interruption of infusion); initial improvement followed by recurrence of symptoms; clinical sequelae warrant hospitalization - Grade 4: Life-threatening consequences; urgent intervention is indicated Malignant neoplasms ·Immune complex disease ·Infectious diseases: - Clinically significant (Grade 3 or greater) - Opportunistic infections, including but not limited to reactivation of latent viral infections, invasive fungal infections, and TB

[0332] Table 3. Overview of the study's screening procedures. Completion of screening may require more than one visit.

[0333] [Table 3-1]

[0334] [Table 3-2]

[0335] Dosage Formulation The formulation of VIB4920 is shown in Table 4.

[0336] [Table 4]

[0337] VIB4920 was prepared at 100 mg / mL in 10 mM sodium phosphate buffer, 250 mM sucrose, 0.02% (weight / volume [w / v]) poloxamer 188, pH 7.4. VIB4920 is formulated in 100 mg of acetaminophen. Each vial has a nominal volume of 5.0 mL. VIB4920 is a sterile liquid drug product (500 mg VIB4920 per vial, nominal) intended for intravenous infusion after dilution with normal saline. VIB4920 should not be shaken and does not require special biohazard handling. It should be stored at 2°C to 8°C (36°F to 46°F) in a refrigerator with appropriate temperature monitoring. VIB4920 should not be frozen.

[0338] The placebo was 0.9% (w / v) normal saline provided by each center as a 250 mL prefilled IV bag.

[0339] treatment Both Cohorts #1 and #2 received the following treatment regimens: Arm 1 received 1500 mg VIB4920 as an intravenous infusion Q2W x 3, followed by four additional doses Q4W (Stage I). Starting on Day 169, subjects received placebo Q4W for five doses (Stage II). Arm 2 received placebo as an intravenous infusion Q2W x 3, followed by four additional doses Q4W (Stage I). Starting on Day 169, subjects received VIB4920 1500 mg Q4W for five doses (Stage II).

[0340] VIB4920 was infused using an IV infusion pump. During Stage I, each subject received the entire volume of VIB4920 solution in the IV bag over at least 90 minutes (approximately 2.8 mL / min). VIB4920 was infused through a low protein-binding 0.2 or 0.22 line filter. Vital signs were obtained before the start of each IP infusion. For Stage II, the infusion was administered over at least 60 minutes (approximately 4.2 mL / min).

[0341] For both Stage I (Days 1 and 15) and Stage II (Days 169 and 197), on the first two dosing days, vital signs were measured within 30 minutes before the start of the infusion, every 30 minutes (± 5 minutes) during the infusion, at the end of the infusion (+ 5 minutes), and then 1 and 2 hours (± 10 minutes) after the end of the infusion. On subsequent dosing days, vital signs were measured within 30 minutes before the start of the infusion, every 30 minutes (± 5 minutes), at the end of the infusion (+ 5 minutes), and 30 minutes (± 5 minutes) after the end of the infusion. At the end of these observation periods, subjects were discharged if stable.

[0342] Posttreatment evaluation After treatment, subjects in Cohort #1 were evaluated to determine one or more of the following: Change from baseline in ESSDAI at days 85, 169, and 309 Change from baseline in ESSPRI at days 85, 169, and 309 The proportion of subjects who achieved an ESSDAI (i.e., at least a 3-point reduction) and an ESSDAI[4] (i.e., at least a 4-point reduction) response, defined as a reduction of at least 3[4] points from baseline on the ESSDAI at days 85, 169, and 309, if they did not prematurely discontinue the study and did not receive rescue therapy. Change from baseline in Functional Assessment of Chronic Illness Therapy (FACIT) fatigue score at days 85, 169, and 309 Change from baseline in Ocular Surface Disease Index (OSDI (copyright)) at days 85, 169, and 309 Patient's Global Impression of Severity (PGIS) at days 85, 169, and 309 The safety and tolerability of multiple IV doses of VIB4920 as measured by the incidence of treatment-emergent adverse events (TEAEs), treatment-emergent serious adverse events (TESAEs), adverse events of special interest (AESIs), and laboratory, vital sign, and electrocardiogram (ECG) abnormalities. PK under investigation The proportion of subjects with a positive immunogenic response measured by anti-VIB4920 antibodies until study completion Change from baseline in total stimulated salivary flow rate Change from baseline in Schirmer test Changes from baseline in SS-A, SS-B, and IgG and IgM rheumatoid factor (RF) autoantibodies Patient's Global Impression of Change (GPI) at 169 days Impression of Change (PGIC) Change from baseline in physician's global impression of severity Change from baseline in levels of inflammatory markers (immunoglobulins, beta-2 microglobulin, C-reactive protein [CRP], CXCL13, C3, C4, and serum-free light chains) The proportion of subjects who achieved a composite response, defined as either an improvement in ESSDAI-3 or an improvement in ESSPRI-1 at days 85, 169, and 309, plus no worsening from baseline in Physician's Global Impression of Severity Changes from baseline in SF36 physical and mental component scores Visual Analogue Scale (VAS) dry mouth ·VAS eye dryness Patient-Reported Outcomes Measurement Information System® (PROMIS-29 Profile v2.1) Biomarkers including blood flow, cytometry for changes in T and B cell subsets, and salivary proteins · Blood gene expression. Changes in fecal microbiome (optional) Baseline and post-treatment DNA epigenetics (optional) Actigraphy: Changes in overall activity and sleep patterns during the study (optional) Change from baseline in the Clinical EULAR Sjoegren's Syndrome Disease Activity Index (ClinESSDAI) at days 85, 169, and 309 The proportion of subjects who achieved ClinESSDAI and ClinESSDAI response, defined as at least a 3[4]-point reduction from baseline on the ClinESSDAI at Days 85, 169, and 309, if they did not prematurely discontinue the study and did not receive rescue therapy, with the objectives of: 1. Determine the clinical efficacy of multiple doses of VIB4920 in glandular and extraglandular manifestations in SS patients with moderate to severe systemic disease activity 2. Determine the effect of VIB4920 on general activity and patient-reported outcomes in subjects with SS 3. Determine the safety and tolerability of multiple doses of VIB4920 in subjects with SS 4. Characterize the pharmacokinetics (PK) of VIB4920 in subjects with SS 5. Evaluate the immunogenicity of VIB4920 in subjects with SS 6. Determine the pharmacodynamics of VIB4920 on the CD40 / CD40L pathway and disease biomarkers in subjects with SS 7. Determine changes in T cell and B cell populations and inflammatory biomarkers and autoantibodies in subjects with SS upon treatment with VIB4920 8. Determine the effect of VIB4920 on systemic disease activity and salivary and lacrimal gland function in subjects with SS 9. Determine changes in major subjective symptoms related to SS

[0343] After treatment, subjects in Cohort #2 were evaluated to determine one or more of the following: Change from baseline in ESSPRI at days 85, 169, and 309 The proportion of subjects who achieved an ESSPRI response, defined as a ≥1-point or 15% reduction from baseline in the ESSPRI score at days 85, 169, and 309, if they did not prematurely discontinue the study and did not receive salvage therapy Change from baseline in FACIT-Fatigue at days 85, 169, and 309 Change from baseline in OSDI at days 85, 169, and 309 PGIS at days 85, 169, and 309 TEAEs, TESAEs, AESIs, and abnormalities in laboratory tests, vital signs, and ECG Safety and Tolerability of Multiple IV Doses of VIB4920 as Measured by Incidence of PK under investigation The proportion of subjects with a positive immunogenic response measured by anti-VIB4920 antibodies until study completion Change from baseline in ESSDAI Change from baseline in total stimulated salivary flow rate Change from baseline in Schirmer test Changes from baseline in SS-A, SS-B, and IgG and IgM RF autoantibodies. PGIC at 169 days. Change from baseline in physician's global impression of severity Changes from baseline in levels of inflammatory markers (immunoglobulins, beta-2 microglobulin, CRP, CXCL13, C3, C4, and serum-free light chains) The proportion of subjects who achieved a composite response, defined as either an improvement in ESSDAI-3 or an improvement in ESSPRI-1 at days 85, 169, and 309, plus no worsening from baseline in Physician's Global Impression of Severity Changes from baseline in SF36 physical and mental component scores ·VAS dry mouth ·VAS eye dryness PROMIS-29 Profile v2.1 Biomarkers including blood flow, cytometry for changes in T and B cell subsets, and salivary proteins · Blood gene expression. Changes in fecal microbiome (optional) Baseline and post-treatment DNA epigenetics (optional) Actigraphy: Changes in overall activity and sleep patterns during the study (optional) Change from baseline in ClinESSDAI at days 85, 169, and 309 These aim to: 1. Determine the clinical efficacy of multiple doses of VIB4920 on the major subjective symptoms of SS (dryness, fatigue, and pain) 2. Determine the effect of VIB4920 on patient-reported outcomes in subjects with SS 3. Determine the safety and tolerability of multiple doses of VIB4920 in subjects with SS 4. Characterize the PK of VIB4920 in SS subjects 5. Evaluate the immunogenicity of VIB4920 in subjects with SS 6. Determine the pharmacodynamics of VIB4920 on the CD40 / CD40L pathway and disease biomarkers in subjects with SS 7. Determine changes in T cell and B cell populations and inflammatory biomarkers and autoantibodies in subjects with SS upon treatment with VIB4920 8. Determine the effect of VIB4920 on systemic disease activity and salivary and lacrimal gland function in subjects with SS 9. Determine changes in major subjective symptoms related to SS

[0344] Clinical Laboratory After treatment, blood and urine samples were collected for clinical safety testing. The hematology panel included complete blood count, differential (including basophils, eosinophils, lymphocytes, monocytes, and neutrophils), hemoglobin, hematocrit, platelets, and white blood cell count.

[0345] Blood biochemistry was analyzed for the following: Creatinine, blood urea nitrogen, fasting blood glucose, total protein, creatine kinase and Solutes (including sodium, potassium, chloride, calcium, bicarbonate, and phosphorus) Liver profile: albumin, TBL, indirect bilirubin, AST, ALT, alkaline phosphatase (ALP), and gamma-glutamyltransferase

[0346] Coagulation parameters were assessed. Subjects were assessed for prothrombin time and PTT at screening and at all study visits during treatment and off-treatment periods (Tables 2 and 3, respectively).

[0347] Urinalysis included protein, glucose, blood, ketones, white blood cells, and pH by dipstick analysis. Microscopy (crystals, casts, white blood cells, red blood cells) was performed if any abnormalities were observed.

[0348] Testing for SARS-COV-2 was conducted.

[0349] Vital signs were measured, including systolic and diastolic blood pressure (mmHg), pulse rate (beats / min), respiratory rate (breaths / min), body temperature (°C), and weight (kg).

[0350] A 12-lead ECG was performed at screening, during treatment, and during the off-treatment period. Each ECG included ventricular heart rate and intervals (PR, QRS, QT, QTc).

[0351] All women of childbearing potential completed one or more serum β-hCG pregnancy tests during the screening period (before day 0) and urine pregnancy tests at clinic visits during the treatment and off-treatment periods.

[0352] Safety evaluation Adverse Events (AEs) and Serious Adverse Events (SAEs) The study assessment schedules for the active treatment and off-treatment periods are presented in Tables 5 and 6, respectively. Table 6 also provides a summary of all assessments that may have been performed at unscheduled visits during the study.

[0353] [Table 5-1]

[0354] [Table 5-2]

[0355] [Table 5-3]

[0356] [Table 5-4]

[0357] [Table 5-5]

[0358] [Table 6-1]

[0359] [Table 6-2]

[0360] Subjects with moderate to severe systemic disease activity (as defined by ESSDAI), results for "Population 1" The results of the analysis of the change from baseline in ESSDAI at Day 169 and the responders are provided in Tables 7 and 8, respectively. The data in Table 7 show that at Week 24, subjects treated with dazodaribep achieved a 6.3±0.6 point reduction in their ESSDAI score compared with subjects treated with placebo. The results show that treated subjects achieved a 4.1 ± 0.6 point reduction, resulting in a statistically significant least squares mean difference of 2.2 points (p=0.017). Dazodarivep achieved statistical significance for the primary endpoint in subjects with moderate to severe systemic disease activity as defined by the ESSDAI, representing an important milestone in the development of a treatment for Sjogren's syndrome, a disease for which there is no available FDA-approved treatment.

[0361] [Table 7]

[0362] In addition to the primary endpoint analysis described above, key secondary, exploratory, and post-hoc analyses demonstrated numerical improvements. These included measures of dryness, an important symptom for subjects living with Sjögren's syndrome because it affects chewing, swallowing, and dental health. Fatigue, as measured by the Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), and physical function, measured using the 36-Item Short Form Health Survey (SF-36), demonstrated numerical improvements, as did tender and swollen joint counts. Additionally, a post-hoc responder analysis of subjects who achieved a high level of improvement on the ESSDAI demonstrated superiority of dazodaribep over placebo. Additionally, dazodaribep was well tolerated in this study.

[0363] Furthermore, compared with the placebo group, greater numerical improvements were observed in the dazodaribep group for the EULAR Sjögren's Syndrome Patient-Reported Index score, Functional Assessment of Chronic Illness Therapy-Fatigue score, 36-Item Short-Term Health Survey physical component score, and stimulated salivary flow rate. Additionally, a post-hoc response analysis of subjects who achieved high levels of improvement (5 and 6 points) on the ESSDAI favored dazodaribep over placebo.

[0364] [Table 8]

[0365] Example 2 - Hypothetical Phase III Study to Determine the Efficacy and Safety of VIB4920 in Subjects with Sjogren's Syndrome (SS) A study will be conducted to determine the efficacy and safety of VIB4920 when administered to subjects with Sjogren's syndrome (SS). The study will determine the effect of VIB4920, using two different dosing regimens, compared with placebo on systemic symptom expression in SS patients with moderate to high systemic disease activity.

[0366] Study design The study will enroll two cohorts of Sjogren's disease patients: Cohort #1 will include SS patients with moderate to high systemic activity as defined by an ESSDAI score of ≥ 5. The following domains: peripheral nervous system, central nervous system, and lung will be scored but may not contribute to the minimum ESSDAI score of 5 required for inclusion. Cohort #2 will include patients with exocrine insufficiency and subjective symptoms as defined by an ESSPRI score of ≥ 5, which is considered the cut-off point for "unsatisfactory symptom state," but with low systemic disease activity. This will include patients with SS who have low risk of developing the disease (ESSDAI score <5).

[0367] A complete physical examination including weight, height, and 28 joint evaluation will be performed.

[0368] Dose Administration Schedule Subjects will be divided into three dosage regimens: Dose 1, 1500 mg VIB4920 every two weeks for three doses, then every four weeks thereafter; Dose 2, 3000 mg VIB4920 at weeks 1, 4, and 12, then every 12 weeks thereafter; and placebo. The expected total duration of participation in the study for each subject is 392±7 days. An exemplary dosing schedule is provided in Table 9.

[0369] [Table 9]

[0370] Example 3 - Blockade of CD40L with Dazodaribep (VIB4920) reduces blood biomarkers of T and B cell costimulation in subjects with systemic Sjogren's syndrome Objective: To determine the effect of VIB4920 (dazodaribep) on blood biomarkers of B and T cell costimulation in adult SS subjects with moderate to high systemic disease activity. In several autoimmune diseases, the CD40 / CD40L pathway is activated in various cell types, including T cells, B cells, antigen-presenting cells, and activated epithelial cells. Dazodaribep blocks costimulatory signals between T cells and CD40-expressing B cells, resulting in the development of germinal centers, the destruction of pathogenic B cells, plasma cells, and autoantibodies, which are hallmarks of some autoimmune diseases, such as Sjögren's syndrome (SS).

[0371] A study was conducted to determine the efficacy of VIB4920 when administered to subjects with Sjögren's syndrome (SS). The study assessed B cell subsets downstream of T cell stimulation: Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells. Serum CXCL13 concentrations, a chemokine involved in germinal center formation produced by activated follicular T cells, and rheumatoid factor (RF) autoantibodies were also assessed.

[0372] method This randomized, double-blind, placebo-controlled, crossover study evaluated dazodarivep therapy in adult SS subjects with moderate to severe systemic disease activity, as defined by an EULAR Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≥ 5. Subjects were randomized 1:1 to receive intravenous dazodarivep 1500 mg or placebo (PBO) Q2W x 3 doses, followed by four additional doses Q4W x 4 doses (Stage 1). Beginning on Day 169, subjects originally randomized to dazodarivep received PBO Q4W x 5 doses, and subjects originally randomized to PBO received dazodarivep Q4W x 5 doses and were then followed for 12 weeks (Stage 2). Throughout the study, B cell subsets downstream of T cell stimulation (Ki67+CD27+ memory, CD27highCD38high plasmablasts, and CD11chigh atypical memory cells) were assayed by FACS. Serum CXCL13 levels, a chemokine produced by activated follicular T cells essential for germinal center (GC) formation, and rheumatoid factor (RF) autoantibodies were also assayed.

[0373] result Concomitant with the dazodarivep-associated reduction in ESSDAI observed in Stage 1, significant and rapid decreases in Ki67+CD27+ memory B cells, plasmablasts, CD11c high memory B cells, CXCL13, and RF antibodies were observed in subjects receiving dazodarivep compared with PBO from Day 15 onward. Similar decreases in these biomarkers were observed in PBO-treated subjects transitioning to dazodarivep treatment in Stage 2. Although these biomarkers returned to baseline values ​​in dazodarivep-treated subjects transitioning to PBO in Stage 2, a sustained decrease in ESSDAI from baseline was observed in these subjects throughout the duration of Stage 2. In independent subgroups of patients with elevated levels of Ki67+CD27+ memory B cells or CD11c high memory B cells at baseline, a greater decrease in ESSDAI with dazodarivep compared with placebo was observed.

[0374] Table 10 shows the median fold change from baseline in the placebo-dazodaribep and dazodaribep-placebo groups.

[0375] [Table 10]

[0376] conclusion Blockade of CD40-CD40L with dazodaribep in patients with SS reduces systemic disease activity through inhibition of T-cell and B-cell costimulation, as evidenced by treatment-associated decreases in blood biomarkers downstream of these pathways.

[0377] Example 4 - In the MIDORA Phase 2 study in patients with moderately to severely active rheumatoid arthritis, blocking CD40L with dazodarib significantly improved disease activity and swollen and tender joint counts while reducing multiple T-cell and B-cell biomarkers the purpose To evaluate the effects of dazodaribep on T and B cells in the MIDORA phase 2 clinical trial in patients with rheumatoid arthritis.

[0378] method This biomarker study focused on a total of 47 adult patients with moderately to severely active rheumatoid arthritis (RA) who had an inadequate response to methotrexate, other conventional disease-modifying antirheumatic drugs, or tumor necrosis factor-alpha (TNFα) inhibitors. Patients in this biomarker study were randomized to receive 1500 mg or 3000 mg of dazodaribep intravenously or placebo. Disease Activity Score 28-C-reactive protein (DAS28-CRP), Patient Global Assessment (PGA), Simplified Disease Activity Index (SDAI), Clinical Disease Activity Index (CDAI), Physician Global Assessment of Disease Activity (MDGA), tender joint count (TJC), and swollen joint count (SJC) were assessed. Plasmablasts (CD19+ cells) expressed CD27 br / CD38 br / IgD- subset), memory B precursor cells (CD11c of CD19+ cells) br subset), T follicular helper cells (CXCR5+ / ICOS+ subset of CD3+ / CD4+ cells), post-switch memory B cells (CD27 br Immunophenotyping of B and T cell subsets, including proliferation of B / IgD- / CD19+ cells (Ki67+ subset of CD3+ cells) and total T cell proliferation (Ki67+ subset of CD3+ cells), was performed using flow cytometry on whole blood. Serum levels of CXCL13 and rheumatoid factor (RF) autoantibodies were assessed by immunoassay and nephelometry, respectively.

[0379] result DAZ significantly improved the DAS28-CRP, PGA, SDAI, CDAI, and MDGA scores, as well as the total number of tender and swollen joints, in RA patients (Figure 2, top). In addition, DAZ treatment reduced CXCL13 and RF levels in RA patients compared with placebo (Figure 2, center). Compared with placebo, DAZ treatment significantly reduced the CD27 expression of plasmablasts (CD19+ cells). br / CD38 br / IgD- subset), T follicular helper cells (CXCR5+ / ICOS+ subset of CD3+ / CD4+ cells), post-switch memory B cells (CD27 br A decrease in the number of proliferating memory B progenitor cells (Ki67+ subset of CD3+ cells) and total T cells (Ki67+ subset of CD3+ cells), as well as decreased serum levels of CXCL13 and RF, also led to an increase in the number of proliferating memory B progenitor cells (CD11c+ subset of CD19+ cells). br subset) occurred (Fig. 2, bottom).

[0380] conclusion DAZ-mediated CD40L blockade in patients with moderately to severely active RA demonstrated significant improvements in disease activity scores, including DAS28-CRP, Patient Global Assessment, SDAI, CDAI, and Physician Global Assessment, including tender and swollen joint counts. Additionally, DAZ reduced multiple CD40L-CD40 downstream biomarkers through disruption of T- and B-cell costimulation, downstream biomarkers of GC formation, activity, and autoantibody production. Collectively, these data demonstrate the broad impact of dazodaribep in patients with rheumatoid arthritis and support findings of clinical efficacy.

[0381] Example 5 - Biomarker evaluation A study was conducted to determine the efficacy of dazodaribep when administered to subjects with Sjögren's syndrome (SS). This study assessed the activity of markers downstream of CD40L-mediated T cell and B cell costimulation: CXCL13, Ki67 (proliferation), and CD11c. br , CD27 br CD38 br We focused on SS-A, SS-B, RF, and ANA.

[0382] result Results show that subjects treated with dazodaribep experienced significant and rapid reductions in multiple pathogenic biomarkers of CD40L-mediated B- and T-cell pathway costimulation and proliferation; and downstream biomarkers of germinal center formation, activity, and autoantibody production. As shown in Figures 3A-G and 4, many of the reductions in these biomarkers occurred as early as day 15 and were sustained through day 169 (primary endpoint). Indeed, treatment with dazodaribep significantly reduced This allows blocking the binding of CD40L to its cognate receptor, CD40, in patients with Sjogren's syndrome, a population with either moderate to severe systemic disease activity or high symptom burden.

[0383] Example 6. Dazodaribep (anti-CD40L) effectively reduces multiple proteins associated with B cell, T cell, and dendritic cell biomarkers, confirming immunophenotyping findings from a Phase 2 study in Sjogren's syndrome populations with either moderate to severe disease activity or high symptom burden. the purpose To determine the effect of dazodaribep on the serum proteome in a phase 2 clinical trial in patients with Sjögren's disease.

[0384] method Two distinct Sjögren's disease populations were enrolled in this study. Population 1 (Pop1) included 74 patients with moderate to severe systemic disease activity as defined by an EULAR Sjögren's Syndrome Disease Activity Index (ESSDAI) of ≥ 5. Population 2 (Pop2) consisted of 109 patients with unacceptable symptom burden and limited extraglandular systemic involvement as defined by an EULAR Sjögren's Syndrome Patient Reported Index (ESSPRI) of ≥ 5 and an ESSDAI of < 5. Eligible participants were randomized 1:1 to receive intravenous DAZ 1500 mg or placebo. The serum proteome was comprehensively profiled at baseline, day 15, and day 169 using the Olink® Explore 3072 panel. Protein profiles were analyzed using AUC-ROC analysis to identify proteins that distinguished Sjögren's disease patients from age-matched healthy controls (n = 50) at baseline. Mixed linear models were applied to examine proteins affected by dazodaribep treatment. Matrix factorization and pathway analysis were applied to identify protein networks within the dataset. The 29 protein biomarkers evaluated included TNFRSF13B, BAFF, CXCL10, CXCL12, CXCL13, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, and CD80.

[0385] result Unsupervised machine learning identified a core set of 29 proteins elevated in Sjögren's disease patients at baseline compared with healthy controls, but there were no significant differences between Pop1 and Pop2 at baseline for these biomarkers. Pathway assessment revealed that these 29 biomarkers were enriched for B cell, T cell, and dendritic cell activation (e.g., LAMP3), which were significantly reduced by DAZ treatment in both Sjögren's syndrome populations 1 and 2 (Figure 5). Protein profiles were analyzed using AUC-ROC analysis to identify proteins that distinguished Sjögren's disease subjects from healthy controls at baseline (Figure 5, Figure 6A-6D). For all 29 biomarkers, a Benjamini-Hochberg false discovery rate (FDR) <10% was performed to determine the significance of changes from baseline. Factor analysis followed by varimax rotation was further used to identify proteins that shared high loadings (>0.45) and correlations with a common vector. By day 169, nine biomarkers (PDCD11, TNFRSF8, CD22, IL12B, TNFRSF9, CXCL13, TNFRSF4, IL2RA, and TIGIT) were near or in the range of expression levels in healthy subjects It entered inside (Figures 6A to 6R).

[0386] conclusion Serum proteomic profiling revealed a core set of biomarkers elevated in patients with Sjögren's disease at baseline compared with healthy cohorts, which were associated with inflammatory and adaptive immune responses, particularly B cell, T cell, and dendritic cell activation, and were significantly reduced (FDR<10%) by dazodaribep treatment in Sjögren's disease Pop1 and Pop2 subjects. Importantly, these biomarkers were shown to be significantly reduced by dazodaribep.

[0387] Incorporation by Reference All references, articles, publications, patents, patent application publications, and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, mention of any reference, article, publication, patent, patent application publication, or patent application cited herein should not be construed as an admission or any form of suggestion that they constitute prior art or form part of the common general knowledge anywhere in the world.

Claims

1. 1. A method of treating Sjogren's syndrome (SS) in a subject in need of treatment, the method comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L, the monomeric subunit comprising seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16, and wherein the subject in need has a higher or lower number of Ki67+ CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c plasmablasts compared to the subject's baseline levels or levels in an untreated subject.

2. A method comprising increasing the number of B cells of a subset selected from the group consisting of high atypical memory cells.

2. The method of claim 1, wherein at least one CD40L-specific monomer subunit comprises seven beta strands designated A, B, C, D, E, F, and G, wherein beta strand A comprises SEQ ID NO: 5, beta strand B comprises SEQ ID NO: 6, beta strand C comprises SEQ ID NO: 17, beta strand D comprises SEQ ID NO: 18, beta strand E comprises SEQ ID NO: 19, beta strand F comprises SEQ ID NO: 20, and beta strand G comprises SEQ ID NO:

21.

3. 3. The method of claim 1 or 2, wherein the subject has a European League Against Rheumatism (EULAR) Sjoegren's Syndrome Disease Activity Index (ESSDAI) score of ≧5.

4. 4. The method of claim 3, wherein the ESSDAI score is assessed based on the ESSDAI domains consisting of skin, kidneys, joints, muscles, hematology, glands, constitution, lymphadenopathy, and biology.

5. The object is (a) ESSDAI score < 5 points; (b) EULAR Sjoegren's Syndrome Patient Reported Index (ESSPRI) score ≥ 5; and (c) Total saliva flow rate during stimulation >0.1 mL / min 3. The method of claim 1 or 2, comprising:

6. 6. The method of any one of claims 1 to 5, wherein the Tn3 scaffold is administered as an induction dose and then as a maintenance dose.

7. 7. The method of claim 6, wherein the induction dose comprises administering the Tn3 scaffold once about every two weeks for at least three doses.

8. 7. The method of claim 6, wherein the maintenance dose comprises administering the Tn3 scaffold once about every four weeks for at least four doses.

9. 9. The method of claim 8, wherein the time between the final induction dose and the first maintenance dose is about 4 weeks.

10. 6. The method of any one of claims 1-5, wherein the Tn3 scaffold is administered about once every 4 weeks, about once every 2 months, about once every 3 months, about once every 4 months, or about once every 6 months.

11. 11. The method of claim 10, wherein the Tn3 scaffold is administered over at least four doses.

12. 12. The method of claim 11, wherein the Tn3 scaffold is administered over at least 5 doses.

13. 13. The method of any one of claims 1 to 12, wherein the Tn3 scaffold is administered intravenously, subcutaneously, orally, intramuscularly, intrathecally, sublingually, rectally, vaginally, cutaneously, systemically, topically, transdermally, or by inhalation.

14. 14. The method of claim 13, wherein the Tn3 scaffold is administered intravenously.

15. The method of any one of claims 1 to 14, wherein the Tn3 scaffold comprises two CD40L-specific monomeric subunits linked in tandem.

16. The method of claim 15, wherein the two CD40L-specific monomer subunits each comprise SEQ ID NO:

3.

17. The method of any one of claims 1 to 16, wherein the CD40L-specific monomeric subunits are linked by a linker.

18. The method of any one of claims 1 to 17, wherein at least one CD40L-specific monomer subunit is directly fused or conjugated to polyethylene glycol (PEG).

19. The method of any one of claims 1 to 17, wherein at least one CD40L-specific monomer subunit is fused or conjugated to polyethylene glycol (PEG) via a linker.

20. 18. The method of claim 17, wherein the linker comprises a peptide linker.

21. 21. The method of claim 20, wherein the linker comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:

10.

22. 22. The method of claim 21, wherein at least one CD40L-specific monomeric subunit is fused or conjugated to albumin.

23. 23. The method of claim 22, wherein the albumin is human serum albumin (HSA).

24. 24. The method of claim 23, wherein the HSA is a mutant HSA comprising SEQ ID NO:

4.

25. 25. The method of any one of claims 1 to 24, wherein the Tn3 scaffold comprising the CD40L-specific monomeric subunit is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

26. 26. The method of claim 25, wherein the dose is about 1500 mg.

27. 26. The method of claim 25, wherein the dose is about 2500 mg.

28. 26. The method of claim 25, wherein the dose is about 3000 mg.

29. 1. A method of treating Sjogren's syndrome (SS) in a subject in need thereof, comprising:

10. The method of claim 1, wherein the subject in need thereof has an increase in a subset of B cells selected from the group consisting of Ki67+ CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c high atypical memory cells, relative to an otherwise healthy subject.

30. 30. The method of claim 29, wherein the Tn3 scaffold is administered as an induction dose followed by one or more maintenance doses.

31. 31. The method of claim 30, wherein the induction dose and the maintenance dose are the same amount.

32. 31. The method of claim 30, wherein the induction dose and the maintenance dose are different amounts.

33. 33. The method of any one of claims 29 to 32, wherein at least one dose is 3000 mg.

34. 34. The method of any one of claims 29-33, wherein the induction dose and at least one maintenance dose are administered about 1 month apart, about 2 months apart, about 3 months apart, about 4 months apart, or about 6 months apart.

35. 35. The method of any one of claims 29-34, wherein the induction dose comprises administering the Tn3 scaffold about every two weeks for at least three doses, and the maintenance dose comprises administering the Tn3 scaffold once about every four weeks for at least four doses.

36. 36. The method of any one of claims 1 to 35, wherein the Tn3 scaffold comprises SEQ ID NO:

1.

37. 37. The method of any one of claims 1-36, wherein said administration is effective in reducing the ESSDAI score when compared to an otherwise similar subject receiving a placebo control.

38. 38. The method of claim 37, wherein the decrease is at least about a 1 point, 2 point, 3 point, 4 point, or 5 point decrease.

39. 39. The method of claim 38, wherein the decrease is at least about a 6 point decrease.

40. 40. The method of any one of claims 1-39, wherein the administration is effective to decrease B cell levels when compared to the subject's baseline level or the level in an untreated subject.

41. 41. The method of any one of claims 1 to 40, wherein the administration is effective to reduce serum CXCL13 levels when compared to the subject's baseline level or the level in an untreated subject.

42. 42. The method of any one of claims 1-41, wherein the administration is effective to reduce rheumatoid factor (RF) autoantibodies when compared to the subject's baseline level or compared to levels in an untreated subject.

43. 43. The method of any one of claims 40 to 42, wherein the decrease is detected by at least 14 days after the administration.

44. A method for treating an autoimmune disease in a subject in need of such treatment, the method comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L when biomarker levels are increased in a sample from the subject compared to a healthy baseline level, the monomeric subunit comprising seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO:

16.

45. 45. The method of claim 44, wherein the biomarker is expressed on the surface of a cell.

46. 45. The method of claim 44, wherein the biomarker is an antibody.

47. 45. The method of claim 44, wherein the biomarker is secreted by a cell.

48. 48. The method of any one of claims 44 to 47, wherein the biomarker is selected from the group consisting of Ki67+CD27+ memory cells, CD27 high CD38 high plasmablasts, CD11c high atypical memory cells, CXCL13, RF autoantibodies, and any combination thereof.

49. 49. The method of any one of claims 44 to 48, wherein the biomarkers are selected from the group consisting of TNFRSF13B, BAFF, CXCL10, CXCL12, IL6, CCL21, CXCL9, IL12B, TNFRSF4, LAMP3, CD22, CD79B, SLAMF7, IL10, TNFRSF8, CXCL11, CCL19, TNFRSF9, TNFRSF6B, NOS3, CD72, ADA2, IL2RA, CD27, PDCD1, IL12A, IL12B, LAG3, ADGRE1, TIGIT, FCAMR, and CD80, and any combination thereof.

50. 50. The method of any one of claims 44 to 49, wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjogren's syndrome, dermatomyositis, myositis, sclerosis, diabetes, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, rheumatic carditis, psoriasis, arthritis, inflammation, rheumatoid arthritis, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonopathy.

51. 51. The method of claim 50, wherein the arthritis is rheumatoid arthritis.

52. 52. The method of any one of claims 44-51, wherein said administration is continued until the level of said biomarker returns to said healthy baseline.

53. 53. The method of any one of claims 44 to 52, wherein the levels of the biomarkers are analyzed periodically.

54. 54. The method of any one of claims 44 to 53, wherein the Tn3 scaffold comprises SEQ ID NO:

1.

55. 55. The method of any one of claims 44-54, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

56. 1. A method for treating Sjogren's syndrome (SS) in a subject in need thereof, the method comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L, the monomeric subunit comprising seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16; and wherein said treating the subject with the Tn3 scaffold increases Ki67+ CD27+ memory cells, CD27 high CD38 high plasmablasts, and CD11c plasmablasts compared to the subject's baseline levels or levels in an untreated subject. The method of claim 1, wherein the level of B cells selected from the group consisting of: high atypical memory cells is reduced.

57. 57. The method of claim 56, wherein the Tn3 scaffold comprises SEQ ID NO:

1.

58. 58. The method of claim 56 or 57, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

59. 59. The method of any one of claims 56-58, wherein the B cell levels are reduced by at least about 1-fold, 5-fold, 10-fold, 30-fold, 50-fold, 75-fold, 100-fold, 150-fold, or 200-fold.

60. 1. A method for treating Sjogren's syndrome (SS) in a subject in need of treatment, the method comprising administering to the subject a Tn3 scaffold comprising a CD40L-specific monomeric subunit that binds to CD40L, the monomeric subunit comprising seven beta strands designated A, B, C, D, E, F, and G, and six loop regions designated AB, BC, CD, DE, EF, and FG, wherein the AB loop comprises SEQ ID NO: 11, the BC loop comprises SEQ ID NO: 12, the CD loop comprises SEQ ID NO: 13, the DE loop comprises SEQ ID NO: 14, the EF loop comprises SEQ ID NO: 15, and the FG loop comprises SEQ ID NO: 16, and the Tn3 scaffold is administered to the subject at a baseline level or at a level in an untreated subject. When compared with the control group, Ki67+ / CD3+, Ki67+ / CD4+ cells, Ki67+ / CD3+ / CD4+ cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCRT7+ cells, Ki67+ / CD3+ / CD4+ / CD45RA- / CCR7- cells, Ki67+ / CD3+ / CD4+ / CXCR5+ / ICOS+ cells, and Ki67+ / CD3+ / CD8+ cells, Ki67+ / CD3+ / CD8+ / CD45RA- / CCR7+ cells, Ki67+ / CD3+ / CD8+ / CD45RA / CCR7- cells, and Ki67+ / CD3+ / CD8+ / CXCR5+ cells, and CXCR5+ / ICOS+ / CD3+ / CD4+ cells,

61. 61. The method of claim 60, wherein the Tn3 scaffold comprises SEQ ID NO:

1.

62. 62. The method of claim 60 or 61, wherein the Tn3 scaffold is administered at a dose of about 1500 mg, about 2500 mg, or about 3000 mg.

63. 63. The method of any one of claims 60-62, wherein the T cell levels are reduced by at least about 1 fold, 5 fold, 10 fold, 30 fold, 50 fold, 75 fold, 100 fold, 150 fold, or 200 fold.

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