Methods of treatment

Abstract

Provided are methods for treating certain diseases or disorders characterized by IL-17 and / or BAFF-mediated inflammation in a human subject in need thereof, comprising systemically administering to the subject a therapeutically effective amount of a bispecific antibody, or a variant thereof, such as a bispecific antibody comprising two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1, or a variant thereof, and the amino acid sequence of the second polypeptide is SEQ ID NO:2, or a variant thereof.

Description

Attorney Docket No. ZURAP002WO METHODS OF TREATMENT

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 636,375, filed April 19, 2024; U.S. Provisional Application No. 63 / 692,300, filed September 9, 2024; and U.S. Provisional Application No. 63 / 739,966, filed December 30, 2024, each of which is incorporated herein by reference in its entirety. SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted herewith as part of the specification and is hereby incorporated by reference in its entirety. Said .xml copy, created on April 3, 2025 is named ZURAP002WO.xml, and is 9,084 bytes in size.

[0003] “Systemic sclerosis” (SSc) or “scleroderma” is a complex and heterogeneous disease with skin and tissue fibrosis, vascular alterations, and autoantibodies against various cellular antigens being amongst its principal features. The clinical manifestations of systemic sclerosis can range from limited skin involvement to severe internal organ dysfunction. Internal visceral organ pathology is a major factor contributing to the morbidity of this disease, with the kidneys, esophagus, heart, and lungs being the most frequently involved.

[0004] The United States Food and Drug Administration (FDA) has approved two treatments for SSc with interstitial lung disease (SSc-ILD): the multi-tyrosine kinase inhibitor nintedanib, and the interleukin-6 receptor blocker tocilizumab. However, these treatments are indicated only to slow the rate of decline in pulmonary function in patients with SSc-ILD (OFEV® (nintedanib) package insert; ACTEMRA® (tocilizumab) package insert). Further, no approved treatment is indicated for any other manifestation of SSc, including skin disease (which is a major cause of morbidity) or digital ulceration and hand contractures that severely impact activities of daily living. A need therefore exists for additional SSc therapies, including differentiated treatments for SSc-ILD and treatments across organ systems.

[0005] Injection site reactions (such as erythema, pain, edema, and pruritis) have been observed following administration of certain antibody therapies, including formulations containing citrate. A need exists for formulations that reduce injection site reactions.

[0006] There are significant unmet needs for safe, effective, and tolerable treatments for the systemic sclerosis and related disorders. The present disclosure satisfies these needs and provides related advantages as well.Attorney Docket No. ZURAP002WO SUMMARY

[0007] The bispecific antibodies described herein have been engineered to target T-cell and B-cell pathways, but have not been evaluated in patients with systemic sclerosis (SSc). Described herein are studies to determine and evaluate doses and dosing regimens that generate sufficient target engagement in the heterogeneous tissues impacted by SSc. Also described herein are formulations to minimize injection site reactions, which may be particularly useful given the impact of SSc on the skin for many patients.

[0008] Provided are methods for treating certain diseases or disorders characterized by IL-17 and / or BAFF-mediated inflammation in a human subject in need thereof, comprising systemically administering to the subject a therapeutically effective amount of a bispecific antibody that specifically binds to human IL-17 and / or BAFF or a variant of such bispecific antibody.

[0009] In some embodiments, the bispecific antibody comprises two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1, or a variant thereof, and the amino acid sequence of the second polypeptide is SEQ ID NO:2, or a variant thereof. In some embodiments, the bispecific antibody comprises two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1 and the amino acid sequence of the second polypeptide is SEQ ID NO:2.

[0010] In some embodiments, the bispecific antibody is tibulizumab, or a variant thereof. In some embodiments, the bispecific antibody is tibulizumab.

[0011] These and other aspects of the invention disclosed herein will be set forth in greater detail as the patent disclosure proceeds. BRIEF DESCRIPTION OF THE FIGURES

[0012] Figure 1 shows model-predicted percentage change from baseline of free unbound BAFF (upper panel) and IL-17 (lower panel) concentration versus time following multiple tibulizumab administrations.

[0013] Figure 2 shows the revised CRISS endpoint in SSc assessment.

[0014] Figure 3 shows the model-predicted blood tibulizumab pharmacokinetics, total & free target time course following 24-week subcutaneous administration of 150 mg Q4W with an additional dose at Week 2.

[0015] Figure 4 shows the model-predicted skin / lung tibulizumab pharmacokinetics, total and free target time course following 24-week subcutaneous administration of 150 mg Q4W with an additional Dose at Week 2.Attorney Docket No. ZURAP002WO

[0016] Figure 5 shows the model-predicted blood tibulizumab pharmacokinetics, total & free target time course following 24-week subcutaneous administration of 300 mg Q4W with an additional dose at Week 2.

[0017] Figure 6 shows the model-predicted skin / lung tibulizumab pharmacokinetics, total & free target time course following 24-week subcutaneous administration of 300 mg Q4W with an additional dose at Week 2. DETAILED DESCRIPTION

[0018] Described herein is the first study to evaluate the effectiveness of tibulizumab in patients with SSc, including measures of disease activity and disease progression in the skin and lung.

[0019] Provided are methods of treating systemic sclerosis in a human subject in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody, wherein the bispecific antibody comprises two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1 and the amino acid sequence of the second polypeptide is SEQ ID NO:2.

[0020] Also provided are methods of treating systemic sclerosis in a human subject in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody against interleukin 17 (IL-17) and human B cell activating factor (BAFF).

[0021] Also provided are methods of treating systemic sclerosis in a human subject in need thereof, comprising administering an anti-human BAFF immunoglobulin G4 (IgG4) antibody fused with an anti-human IL-17 single-chain variable fragment (scFv) at the C-terminus of each heavy chain.

[0022] In some embodiments, the bispecific antibody is administered subcutaneously.

[0023] In some embodiments, the antibody or antigen-binding fragment thereof is administered once per week, once every other week, once every two weeks, once every three weeks, once every four weeks, once every month, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every two months, once every nine weeks, once every ten weeks, once every eleven weeks or once every twelve weeks. In some embodiments, the bispecific antibody is administered once every two weeks (Q2W). In some embodiments, the bispecific antibody is administered once every four weeks (Q4W).

[0024] In some embodiments, the bispecific antibody is administered in an amount of between about 100 mg and about 500 mg, such as between about 200 mg and about 400 mg, such as about 300 mg. In some embodiments, the bispecific antibody is administered in an amount of about 100, about 110, about 120, about 125, about 130, about 140, about 150,Attorney Docket No. ZURAP002WO about 160, about 170, about 175, about 180, about 190, about 200, about 210, about 220, about 225, about 230, about 240, about 250, about 260, about 270, about 275, about 280, about 290, or about 300 mg. In some embodiments, the bispecific antibody is administered in an amount of about 100, about 120, about 125, about 150, about 175, about 200, about 225, about 240, about 250, about 275, about 300, about 350, about 400, about 450, or about 500 mg.

[0025] In some embodiments, the bispecific antibody is administered by subcutaneous injections totaling about 120 mg every 2 weeks. In some embodiments, the subcutaneous injections are in a single injector. In some embodiments, the subcutaneous injections are in two injectors. In some embodiments, the injector is an autoinjector.

[0026] In some embodiments, the bispecific antibody is administered by subcutaneous injections totaling about 240 mg every month. In some embodiments, the bispecific antibody is administered by subcutaneous injections totaling about 300 mg every month. In some embodiments, the bispecific antibody is administered by subcutaneous injections totaling about 240 mg every four weeks. In some embodiments, the bispecific antibody is administered by subcutaneous injections totaling about 300 mg every four weeks. In some embodiments, the subcutaneous injections are in a single injector. In some embodiments, the subcutaneous injections are in two injectors. In some embodiments, the injector is an autoinjector. In some embodiments, two subcutaneous injections of 120 mg bispecific antibody each are administered together every four weeks (Q4W). In some embodiments, one subcutaneous injection of about 120 mg bispecific antibody is administered every two weeks (Q2W). In some embodiments, two subcutaneous injections of 150 mg bispecific antibody each are administered together every four weeks (Q4W). In some embodiments, one subcutaneous injection of about 150 mg bispecific antibody is administered every two weeks (Q2W). In some embodiments, two subcutaneous injections of 150 mg bispecific antibody each are administered two weeks following the first administration.

[0027] In some embodiments, the bispecific antibody is administered 300 mg Q4W. In some embodiments, an additional administration of 300 mg is provided between the first and second administration of 300 mg Q4W.

[0028] In some embodiments, the bispecific antibody is administered 300 Q2W for a first time period, and 300 mg Q4W for a second time period. In some embodiments, administration of the bispecific antibody occurs at the following time points: time zero, week 2, week 4, and monthly thereafter. In some embodiments, loading doses of the bispecific antibody are administered during a first time period. In some embodiments, a lower doseAttorney Docket No. ZURAP002WO and / or less frequent doses are administered during a second time period. In some embodiments, the difference between loading doses and subsequent doses is the frequency of dosing. In some embodiments, a maintenance dose is administered less frequently than loading doses.

[0029] In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 300 mg every 2 weeks.

[0030] In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 300 mg every 4 weeks. In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 300 mg every month.

[0031] In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 150 mg every 2 weeks.

[0032] In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 150 mg every 4 weeks. In some embodiments, the bispecific antibody is administered by subcutaneous injections of about 150 mg every month.

[0033] In some embodiments, the bispecific antibody is administered every four weeks with an additional dose administered between the first and second dose. In some embodiments, an additional dose of the bispecific antibody is administered between the first and second dose of a Q4W dosing regimen. In some embodiments, the additional dose is administered about two weeks after the first dose. In some embodiments, the additional dose is administered from about 10 days to about 18 days, such as from about 11 days to about 17 days, such as from about 12 days to about 16 days, such as from about 13 days to about 15 days, such as 14 days after the first dose. In some embodiments, the bispecific antibody is administered more frequently in the first month than in the second month. In some embodiments, an additional dose of the bispecific antibody is administered to expedite achievement of steady state.

[0034] In some embodiments, the bispecific antibody is administered in an amount of about 150 mg.

[0035] In some embodiments, the bispecific antibody is administered in an amount of about 240 mg.

[0036] In some embodiments, the bispecific antibody is administered in an amount of about 300 mg.

[0037] In some embodiments, the bispecific antibody is administered once per week, once every other week, once every two weeks, once every three weeks, once every four weeks, or once every month.Attorney Docket No. ZURAP002WO

[0038] In some embodiments, the bispecific antibody is administered in an amount of about 150 mg once every four weeks.

[0039] In some embodiments, the bispecific antibody is administered as tibulizumab 150 mg every four weeks.

[0040] In some embodiments, the method further comprises an additional administration of 150 mg tibulizumab between the first and second administration of 150 mg every four weeks.

[0041] In some embodiments, the bispecific antibody is administered in an amount of about 240 mg once every four weeks.

[0042] In some embodiments, the bispecific antibody is administered as tibulizumab 240 mg every four weeks.

[0043] In some embodiments, the method further comprises an additional administration of 240 mg tibulizumab between the first and second administration of 240 mg every four weeks.

[0044] In some embodiments, the bispecific antibody is administered in an amount of about 300 mg once every four weeks.

[0045] In some embodiments, the bispecific antibody is administered as tibulizumab 300 mg every four weeks.

[0046] In some embodiments, the method further comprises an additional administration of 300 mg tibulizumab between the first and second administration of 300 mg every four weeks.

[0047] In some embodiments, the bispecific antibody is administered according to a multiple variable dose regimen.

[0048] In some embodiments, the treatment is to be carried out with a multiple- variable dose comprising a first induction dose of a bispecific antibody to a subject in need thereof such that a threshold level of the bispecific antibody is achieved within an induction phase; and at least one treatment dose of the bispecific antibody for subsequent administration within a treatment phase, such that treatment occurs.

[0049] In some embodiments, the multiple variable dose regimen comprises administering at least one loading dose and administering a treatment dose which is different than the loading dose. In some embodiments, the multiple variable dose regimen comprises a loading dose and a treatment dose that is lower than the loading dose.

[0050] In some embodiments, a time interval between the loading dose and a first treatment dose is at least one week, at least two weeks, at least three weeks, at least four weeks, at least one month, at least five weeks, at least six weeks, at least seven weeks, at least eight weeks, at least two months, at least nine weeks, at least ten weeks, at least eleven weeks, or at least twelve weeks.Attorney Docket No. ZURAP002WO

[0051] In some embodiments, a biomarker is evaluated in conjunction with the administration of tibulizumab. In some embodiments, the biomarker is selected from one or more of the following: BAFF, IL-17, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), periostin (POSTN), collagen 1A1 (COL1A1), C-X-C motif chemokine ligand 1 (CXCL1), interleukin 11 (IL-11), advanced glycosylation end-product specific receptor (Ager), C-X-C motif chemokine receptor 4 (CXCR4), toll like receptor 2 (TLR2), collagen 1A2 (COL1A2), fibronectin 1 (FN1), tissue inhibitor of metalloproteinase 1 (TIMP1), connective tissue growth factor (CTGF), acetyl-CoA acetyltransferase 2 (ACTA2), transforming growth factor beta 1 (TGFβ1), vascular endothelial growth factor A (VEGFA), platelet-derived growth factor (PDGF), Nod-, LRR- and pyrin domain-containing protein 3 (NLRP3), C-C motif chemokine ligand 2 (CCL2), interleukin 8 (IL-8), chemokine ligand 3 (CXCL3), chemokine ligand 4 (CXCL4), C-C motif chemokine ligand 18 (CCL18), matrix metalloproteinase 7 (MMP7), matrix metalloproteinase 12 (MMP12), and surfactant protein D (SP-D). In some embodiments, the biomarker is CXCL1. In some embodiments, the biomarker is immunoglobulin G (IgG). In some embodiments, the biomarker is immunoglobulin 1 (IgG1). In some embodiments, the biomarker is BAFF. In some embodiments, the biomarker is IL- 17.

[0052] In some embodiments, the biomarker is a gene. In some embodiments, the biomarker is a transcript. In some embodiments, the biomarker is a protein. In some embodiments, an individual administered tibulizumab is evaluated for a reduction in one or more biomarkers. In some embodiments, an individual administered tibulizumab is evaluated for an increase in one or more biomarkers. In some embodiments, a biomarker is upregulated following administration of tibulizumab. In some embodiments, a biomarker is downregulated following administration of tibulizumab. Systemic Sclerosis

[0053] In some embodiments, the systemic sclerosis is limited systemic sclerosis (CREST syndrome). In some embodiments, the systemic sclerosis is generalized systemic sclerosis. In some embodiments, the systemic sclerosis systemic sclerosis sine scleroderma. In some embodiments, the systemic sclerosis is diffuse cutaneous systemic sclerosis. In some embodiments, the systemic sclerosis is early diffuse cutaneous systemic sclerosis. In some embodiments, the systemic sclerosis is localized systemic sclerosis. In some embodiments, the systemic sclerosis is morphea. In some embodiments, the systemic sclerosis is linearAttorney Docket No. ZURAP002WO systemic sclerosis. In some embodiments, the disorder or disorder is systemic sclerosis with interstitial lung disease.

[0054] In some embodiments, the subject has a symptom chosen from Raynaud syndrome, polyarthralgia, dysphagia, heartburn, and swelling, skin tightening and / or contractures of the fingers.

[0055] In some embodiments, the symptom is chosen from fibrosis, skin thickening, cutaneous pruritus, whitening of hands on exposure to cold, pain in affected digits, difficulty in swallowing foods or liquids, nausea, vomiting, weight loss, abdominal cramps, blotting diarrhea, fecal incontinence, shortness of breath on exertion, palpitations without characteristic pain in thoracic cavity, nonproductive cough, atypical chest pain, fatigue, dyspnea, hypertension, joint pain, limitation of movement, joint swelling, muscle pain, inflammatory myopathy or weakness.

[0056] In some embodiments, the subject is classified according to the American College of Rheumatology criteria for the classification of systemic scleroderma as having proximal diffuse (truncal) sclerosis (skin tightness, thickening, and non-pitting induration).

[0057] In some embodiments, the subject is classified according to the American College of Rheumatology criteria for the classification of systemic scleroderma as having (1) sclerodactyly (only fingers and / or toes), (2) digital pitting scars or loss of substance of the digital finger pads (pulp loss), and / or (3) bilateral basilar pulmonary fibrosis. In some embodiments, the subject fulfills classification criteria for SSC according to ACR and European Alliance of Association for Rheumatology (EULAR) 2013 criteria. In some embodiments, diffuse cutaneous SSc is defined as an mRSS greater than zero over at least one skin area proximal to the elbows and / or knees, in addition to acral fibrosis.

[0058] In some embodiments, prior to administration, the subject has a modified Rodnan skin score (mRSS) score of 10-29.

[0059] In some embodiments, a clinical response is defined by whether or not a subject achieves an improvement in the subject’s modified Rodnan skin score (mRSS) score, which is a measure of skin thickness. In some embodiments, mRSS scoring is as follows: • mRSS = 0 is “normal skin” where the examiner appreciates fine wrinkles but no skin thickness is present. • mRSS = 1 is defined as definite but “mild” skin thickness where the examiner can easily make skin folds between 2 fingers; fine wrinkles are acceptable.Attorney Docket No. ZURAP002WO • mRSS = 2 is defined as “moderate” skin thickness with difficulty in making skin folds and no wrinkles. • mRSS = 3 is defined as “severe” skin thickness with inability to make skin folds between 2 examining fingers.

[0060] In some embodiments, the subject’s mRSS score is improved after administration of the bispecific antibody.

[0061] In some embodiments, the subject’s scleroderma health assessment questionnaire disability index (SHAQ-DI) is improved after administration of the bispecific antibody.

[0062] In some embodiments, the subject’s visual analog scale (VAS) score for Raynaud's phenomenon, pain, ulcers, and / or breathing is improved after administration of the bispecific antibody.

[0063] In some embodiments, a visual analog scale (VAS) score is measured.

[0064] In some embodiments, a VAS score is improved compared to administration with a formulation containing citrate.

[0065] In some embodiments, the number of new upper limb digital ulcers is measured. In some embodiments, upper limb digital ulcers are evaluated using the World Scleroderma Foundation definition for SSc skin ulcers in clinical trials.

[0066] In some embodiments, a modified Rodnan skin score (mRSS) is measured. In some embodiments, high-resolution computed tomography (HRCT) is performed. In some embodiments, quantitative high-resolution computed tomography (qHRCT) is performed. In some embodiments, forced vital capacity (FVC) is measured. In some embodiments, forced expiratory volume (FEV) is measured. In some embodiments, forced expiratory volume in one second (FEV1) is measured. In some embodiments, diffusing capacity for carbon monoxide (DLCO) is measured. In some embodiments, the ratio of forced vital capacity to diffusing capacity (FVC / DLCO) is measured. In some embodiments, a spirometry measurement is taken. In some embodiments, clinician global assessment (CGA) is performed. In some embodiments, scleroderma skin patient reported outcome (SSPRO) is performed. In some embodiments, patient global assessment (PGA or PtGA) is performed. In some embodiments, health assessment questionnaire disability index (HAQ-DI) is performed. In some embodiments, scleroderma health assessment questionnaire disability index (SHAQ- DI) is performed. In some embodiments, the subject’s Composite Response Index in Systemic Sclerosis (CRISS) is measured. In some embodiments, the subject’s revised Composite Response Index in Systemic Sclerosis (rCRISS) is measured. In someAttorney Docket No. ZURAP002WO embodiments, the subject’s revised CRISS-25 is measured. In some embodiments, the subject’s revised CRISS-25 Step 1 worsening is measured. In some embodiments, a subject is assessed using the methodology in Figure 2.

[0067] In some embodiments, mRSS is improved following administration. In some embodiments, HRCT is improved following administration. In some embodiments, qHRCT is improved following administration. In some embodiments, FVC is improved following administration. In some embodiments, the subject’s DLCO is improved following administration. In some embodiments, the subject’s FVC / DLCO is improved following administration. In some embodiments, FEV is improved following administration. In some embodiments, FEV1 is improved following administration. In some embodiments, CGA is improved following administration. In some embodiments, SSPRO is improved. In some embodiments, PGA or PtGA is improved following administration. In some embodiments, HAQ-DI is improved following administration. In some embodiments, SHAQ-DI is improved following administration. In some embodiments, CRISS is improved following administration.

[0068] In some embodiments, the CRISS is the American College of Rheumatology Composite Response Index in Systemic Sclerosis (ACR-CRISS). In some embodiments, the CRISS is a revised CRISS (rCRISS). In some embodiments, the CRISS is CRISS25. In some embodiments, the CRISS is ACR-CRISS25. In some embodiments, the CRISS is a Step 1 worsening in revised CRISS-25. In some embodiments, the CRISS is as outlined in Figure 2.

[0069] In some embodiments, administration of the bispecific antibody prevents or delays new onset of left ventricular failure requiring treatment. In some embodiments, administration of the bispecific antibody prevents or delays new onset of pulmonary arterial hypertension (PAH) requiring treatment. In some embodiments, administration of the bispecific antibody prevents or delays gastrointestinal dysmotility requiring enteral or parenteral nutrition. In some embodiments, administration of the bispecific antibody prevents or delays digital ischemia with gangrene, amputation, or hospitalization requiring treatment.

[0070] In some embodiments, mRSS has been determined for a subject prior to administration. In some embodiments, mRSS is measured as described in Khanna, D. et al., Journal of Scleroderma and Related Disorders, doi:10.5301 / jsrd.5000231 or Ferreli, C., et al. Clinical Reviews in Allergy & Immunology, doi:10.1007 / s12016-017-8625-4. In some embodiments, mRSS assesses skin thickness by evaluating about 17 body sites (e.g., face, chest, abdomen, arms, and legs). In some embodiments, each site is scored from 0 to 3. InAttorney Docket No. ZURAP002WO some embodiments, the total mRSS ranges from 0 to 51, with higher scores indicating greater skin involvement.

[0071] In some embodiments, the subject has had systemic sclerosis for less than about five years. In some embodiments, the subject has had systemic sclerosis for less than about two years. In some embodiments, the subject has had systemic sclerosis for less than about one year. In some embodiments, the duration of systemic sclerosis is defined as the time since first non-Raynaud’s phenomenon symptom or sign attributed to systemic sclerosis. In some embodiments, the subject has had systemic sclerosis (defined as the first non-Raynaud phenomenon symptom or sign attributed to systemic sclerosis) for less than about five years.

[0072] In some embodiments, mRSS is 10-35 and the subject has had systemic sclerosis for less than about two years. In some embodiments, mRSS is 12-35 and the subject has had systemic sclerosis for less than about two years. In some embodiments, mRSS is greater than or equal to 15 and less than or equal to 45. In some embodiments, mRSS is greater than or equal to 15 and less than or equal to 45 and the subject has had systemic sclerosis for less than about two years. In some embodiments, mRSS is about 15-45. In some embodiments, mRSS is about 15-45 and the subject has had systemic sclerosis for about two to about five years. In some embodiments, mRSS is about 20-45. In some embodiments, mRSS is about 20-45 and the subject has had systemic sclerosis for about two to about five years. In some embodiments, mRSS is about 15-45 and the subject has had systemic sclerosis for less than about two years.

[0073] In some embodiments, the subject is RNA polymerase III negative and has had systemic sclerosis for about two years to about five years. In some embodiments, (i) mRSS is greater than or equal to 20 and less than or equal to 45; (ii) the subject has had systemic sclerosis for greater than or equal to 2 years and less than or equal to 5 years; and (iii) the subject (a) is either RNA polymerase III negative or (b) has evidence of disease progression defined as screening mRSS increased by greater than or equal to 3 units compared with the last assessment within the previous 6 months or involvement of two new mRSS body areas compared with the last assessment withing the previous 6 months. In some embodiments, the subject has evidence of recent progression and has had systemic sclerosis for about two years to about five years. In some embodiments, about a number of years is that number of years (for example, about two years is two years).

[0074] In some embodiments, treating SSc is reducing at least one symptom of the disease. In some embodiments, treating SSc is reducing symptoms in more than one organ system in the patient. In some embodiments, treating SSc is reducing symptoms in the lung and skin.Attorney Docket No. ZURAP002WO In some embodiments, treating SSc is slowing the rate of decline in the disease. For example, in some embodiments, treating SSc is slowing the rate of decline in SSc-ILD.

[0075] In some embodiments, anti-Scl-70 antibodies are measured. In some embodiments, patients are screened for anti-Scl-70 antibodies. In some embodiments, patients positive for anti-Scl70 antibodies are selected for treatment with tibulizumab.

[0076] In some embodiments, an autoantibody is measured. In some embodiments, anti- centromere antibody is measured. In some embodiments, patients are screened for anti- centromere antibody (ACA). In some embodiments, patients negative for anti-centromere antibody are selected for treatment with tibulizumab. In some embodiments, patients positive for anti-centromere antibody are not selected for treatment with tibulizumab. In some embodiments, a patient positive for anti-centromere antibody is selected for treatment with tibulizumab if they are also positive for RNA polymerase III. In some embodiments, a patient positive for anti-centromere antibody is selected for treatment with tibulizumab if they are also positive for anti-topoisomerase I. In some embodiments, tibulizumab administration is discontinued in patients positive for anti-centromere antibody.

[0077] In some embodiments, acute phase reactants are measured. In some embodiments, patients are screened for acute phase reactants. In some embodiments, patients with elevated acute phase reactants are selected for treatment with tibulizumab. In some embodiments, patients with normal acute phase reactants are not selected for treatment with tibulizumab.

[0078] In some embodiments, a subject has received background therapy. In some embodiments, a subject has received mycophenolate mofetil (MMF). In some embodiments, a subject has been receiving MMF for at least six months prior to administration of tibulizumab. In some embodiments, a subject has not initiated an antifibrotic agent within 6 months of administration of tibulizumab. In some embodiments, a subject has not initiated an antifibrotic agent within one month of administration of tibulizumab. In some embodiments, the antifibrotic agent is selected from nintedanib, pirfenidone, D-penicillamine, or another kinase inhibitor (e.g., nilotinib, imatinib, or dasatinib). The Bispecific Antibody

[0079] A bispecific antibody is understood to mean an immunoglobulin molecule comprising four antigen binding sites, which binds two different antigens with specificity for each antigen in the mAb-scFV format. The bispecific antibody is capable of binding each antigen alone or each antigen simultaneously.Attorney Docket No. ZURAP002WO

[0080] The bispecific antibody described herein comprises two first polypeptides and two second polypeptides. In some embodiments, the first polypeptide forms an inter-chain disulfide bond with each of the second polypeptides, and the first polypeptide forms two inter-chain disulfide bonds with the other first polypeptide, and each of the first polypeptides forms several intra-chain disulfide bonds. In some embodiments, the relationship of the polypeptides and the disulfide bonds are shown in the following schematic:

[0081] In some embodiments, the amino acid sequence of the first polypeptide is: QVQLQQWGAG LLKPSETLSL TCAVYGGSFS GYYWSWIRQP PGKGLEWIGE INHSGSTNYN PSLKSRVTIS VDTSKNQFSL KLSSVTAADT AVYYCARGYY DILTGYYYYF DYWGQGTLVT VSSASTKGPS VFPLAPCSRS TSESTAALGC LVKDYFPEPV TVSWNSGALT SGVHTFPAVL QSSGLYSLSS VVTVPSSSLG TKTYTCNVDH KPSNTKVDKR VESKYGPPCP PCPAPEFLGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS QEDPEVQFNW YVDGVEVHNA KTKPREEQFN STYRVVSVLT VLHQDWLNGK EYKCKVSNKG LPSSIEKTIS KAKGQPREPQ VYTLPPSQEE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SRLTVDKSRW QEGNVFSCSV MHEALHNHYT QKSLSLSPGG GGSGGGGTGG GGSQVQLVQS GAEVKKPGSS VKVSCKASGY KFTDYHIHWV RQAPGQCLEW MGVINPTYGT TDYNQRFKGR VTITADESTS TAYMELSSLR SEDTAVYYCA RYDYFTGTGV YWGQGTLVTV SSGGGGSGGG GSGGGGSGGG GSDIVMTQTP LSLSVTPGQP ASISCRSSRS LVHSRGETYL HWYLQKPGQS PQLLIYKVSN RFIGVPDRFS GSGSGTDFTL KISRVEAEDV GVYYCSQSTHLPFTFGCGTK LEIK (SEQ ID NO: 1).Attorney Docket No. ZURAP002WO

[0082] In some embodiments, the amino acid sequence of the second polypeptide is: EIVLTQSPAT LSLSPGERAT LSCRASQSVS RYLAWYQQKP GQAPRLLIYD 50 ASNRATGI PA RFSGSGSGTD STLTI SSLEP EDFAVYYCQQ RSNWPRTFGQ 100 GTKVE I KRTV AAPSVFI FPP SDEQLKSGTA SVVCLLNNFY PREAKVQWKV 150 DNALQSGNSQ ESVTEQDSKD STYSLSNTLT LSKADYEKHK VYACEVTHQG 200 LSSPVTKSFN RGEC 214 (SEQ ID NO:2).

[0083] In some embodiments, the inter-chain disulfide bond of each of the first polypeptides and each of the second polypeptides forms between cysteine residue 137 of SEQ ID NO: 1 and cysteine residue 214 of SEQ ID NO:2. In some embodiments, the first polypeptide forms two inter-chain disulfide bonds with the other first polypeptide. In some embodiments, the first inter-chain disulfide bond forms between cysteine residue 229 of the first polypeptide of SEQ ID NO: 1 and cysteine residue 229 of the other first polypeptide of SEQ ID NO: 1. In some embodiments, the second inter-chain disulfide bond forms between cysteine residue 232 of the first polypeptide of SEQ ID NO: 1 and cysteine residue 232 of the other first polypeptide of SEQ ID NO: 1.

[0084] In some embodiments, within the scFV, an engineered intra-chain disulfide bond is formed between cysteine residue 507 of SEQ ID NO: 1 and cysteine residue 707 of SEQ ID NO: 1. Also, in some embodiments, an intra-chain disulfide bond is formed between cysteine residue 625 of SEQ ID NO: 1 and cysteine residue 695 of SEQ ID NO: 1. In some embodiments, within the mAb, intra-chain disulfide bonds that normally occur in an IgG4 antibody are formed between cysteine residue 22 of SEQ ID NO: 1 and cysteine residue 95 of SEQ ID NO: 1, between cysteine residue 150 of SEQ ID NO: 1 and cysteine residue 206 of SEQ ID NO: 1, between cysteine residue 264 of SEQ ID NO: 1 and cysteine residue 324 of SEQ ID NO: 1, between cysteine residue 370 of SEQ ID NO: 1 and cysteine residue 428 of SEQ ID NO: 1, between cysteine residue 485 of SEQ ID NO: 1 and cysteine residue 559 of SEQ ID NO: 1, between cysteine residue 23 of SEQ ID NO:2 and cysteine residue 88 of SEQ ID NO:2, and between cysteine residue 134 of SEQ ID NO:2 and cysteine residue 194 of SEQ ID NO:2.

[0085] In some embodiments, the first polypeptide comprises a first heavy chain variable region (HCVR1), a heavy chain constant region (CH), a second heavy chain variable region (HCVR2) and a second light chain variable region (LCVR2). In some embodiments, the second polypeptide comprises a first light chain variable region (LCVRl) and a light chain constant region (CL). The HCVR and LCVR regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed withAttorney Docket No. ZURAP002WO framework regions (FR). In some embodiments, each HCVR and LCVR is composed of three CDRs and four FRs, arranged from amino- terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0086] The 3 CDRs of HCVR1 can be referred to as CDRHl-1, CDRHl-2, and CDRHl-3 and the 3 CDRs of HCVR2 are referred to as CDRH2-1, CDRH2-2, and CDRH2-3 and the 3 CDRs of LCVRl are referred to as CDRLl-1, CDRLl-2 and CDRLl-3 and the 3 CDRs of LCVR2 are referred to as CDRL2-1, CDRL2-2 and CDRL2-3.

[0087] In some embodiments, the CH is fused to HCVR2 by an amino acid linker (LI). HCVR2 is fused to LCVR2 by an amino acid linker (L2).

[0088] In some embodiments, the relationship of the various regions and linkers is as follows:

[0089] Significant problems associated with chemical and physical stability were encountered when constructing a bispecific antibody in the mAb-scFv format with the anti-Attorney Docket No. ZURAP002WO IL-17 binding portion in the scFv configuration. Chemical modifications were made in the CDRL2-1 and CDRH2-2 portions of the bispecific antibody that improved physical stability and reduced concentration-dependent aggregation. Extensive protein stability and solubility studies identified chemically unstable residues in CDRL2-1 and CDRH2-2. These labile residues were replaced with charge neutral amino acids using targeted libraries constructed by codon depletion. Additionally, the electrostatic surface of the bispecific antibody was calculated and charged patches were identified. Disruptions of these charged patches in the scFv led to a decrease in protein self-association. However, a mutation was identified in the CDRH2-1 portion of the bispecific antibody that rebalanced the surface electrostatic distribution, and improved physical stability and solubility at high concentrations. None of the above issues were encountered in the parental single antibodies. These problems were encountered only in the context of constructing a bispecific antibody in the mAb-scFv format, suggesting that the local environment around the mutated areas of the single antibody differed in the context of a bispecific antibody.

[0090] Further chemical modifications were made to stabilize the HCVR2 / LCVR2 interface in the IL-17 portion of the bispecific antibody, and to reduce bispecific antibody aggregation. Studies conducted to determine the aggregation showed that the observed protein self- association was not driven by conformational instability of the individual HCVR2 or LCVR2 domains, but rather by the opening or breathing of the HCVR2 / LCVR2 interface, leading to intermolecular protein interactions. Thus, various intra-chain disulfide bonds were introduced into the HCVR2 / LCVR2 interface of the IL- 17 portion of the bispecific antibody. One such intra-chain disulfide bond occurs in each of the first polypeptides between cysteine residue 507 of SEQ ID NO: 1 and cysteine residue 707 of SEQ ID NO: 1. This disulfide bond covalently connects the HCVR2 / LCVR2 interface in the IL-17 portion of the bispecific antibody, which stabilizes the HCVR2 / LCVR2 interface and reduces intermolecular protein interactions that can lead to physical instability and unfavorable formulation limitations. Out of the nine different disulfide bonds tested, 8 of which expressed functional protein, the magnitude of affinity loss ranged from about 2 to about 35-fold. The intra-chain disulfide bond in each of the first polypeptides between cysteine residue 507 of SEQ ID NO: 1 and cysteine residue 707 of SEQ ID NO: 1 best stabilized the HCVR2 / LCVR2 interface while maintaining optimal binding affinity for IL-17.

[0091] In addition, studies indicated that linker length for LI affected binding kinetics. Kinetic analysis (by surface plasmon resonance) showed that a 10 amino acid linker caused a 2-fold slower Konrate compared to 15 amino acid and 20 amino acid linkers. Thus, in someAttorney Docket No. ZURAP002WO embodiments, a linker length of > 10 amino acids is used. In some embodiments, the linker length is between 15 amino acids and 20 amino acids. In some embodiments, the linker length is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids. In some embodiments, a linker length of 15 is used.

[0092] The bispecific antibodies described herein bind both human BAFF and human IL-17. The bispecific antibodies described herein can inhibit IL-17 in the presence and absence of BAFF in vitro. The bispecific antibodies described herein can inhibit both soluble and membrane-bound BAFF in the presence or absence of IL-17 in vitro. The bispecific antibodies described herein are further characterized as having a binding affinity for human BAFF in the range of 150 pM to 1 pM and human IL-17 in the range of 50 pM to 1 pM. The bispecific antibodies have a binding affinity for human IL-17A / F heterodimer of about 90 pM.

[0093] The bispecific antibodies effectively neutralize soluble as well as membrane- bound BAFF and this neutralization is not affected by the presence of saturating amounts of human IL-17. The bispecific antibodies effectively neutralize human IL-17 and this neutralization is not affected by the presence of saturating amounts of human BAFF.

[0094] In some embodiments, the bispecific antibody is tibulizumab. In some embodiments, the bispecific antibody comprises components of tabalumab combined with components of ixekizumab.

[0095] Expression vectors capable of directing expression of genes to which they are operably linked are well known in the art. Expression vectors can encode a signal peptide that facilitates secretion of the polypeptide(s) from a host cell. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide. The first polypeptide and the second polypeptide may be expressed independently from different promoters to which they are operably linked in one vector or, alternatively, the first polypeptide and the second polypeptide may be expressed independently from different promoters to which they are operably linked in two vectors - one expressing the first polypeptide and one expressing the second polypeptide.

[0096] A host cell includes cells stably or transiently transfected, transformed, transduced or infected with one or more expression vectors expressing a first polypeptide, a second polypeptide or both a first polypeptide and a second polypeptide described herein.

[0097] Creation and isolation of host cell lines producing a bispecific antibody described herein can be accomplished using standard techniques known in the art. In some embodiments, mammalian cells are used as the host cells for expression of bispecificAttorney Docket No. ZURAP002WO antibodies. Particular mammalian cells are HEK 293, NSO, DG-44, and CHO. In some embodiments, the bispecific antibodies are secreted into the medium in which the host cells are cultured, from which the bispecific antibodies can be recovered or purified.

[0098] It is well known in the art that mammalian expression of antibodies results in glycosylation. Typically, glycosylation occurs in the Fc region of the antibody at a highly conserved N-glycosylation site. N-glycans typically attach to asparagine. In some embodiments, each of the first polypeptides is glycosylated at asparagine residue 300 of SEQ ID NO: l.

[0099] A particular DNA polynucleotide sequence encoding the first polypeptide having an amino acid sequence of SEQ ID NO: 1 is: CAGGTGCAACTACAGCAGTGGGGCGCAGGACTGTTGAAGCCTTCGGAGACCCTG TCCCTC ACCTGCGCTGTCTATGGTGGGTCCTTCAGTGGTTACTACTGGAGCTGGATTCGCC AGCCC CCAGGGAAGGGGCTGGAGTGGATTGGGGAAATCAATCATAGTGGAAGCACCAAC TACAAC CCGTCCCTCAAGAGTCGAGTCACCATATCAGTAGACACGTCCAAGAACCAGTTCT CCCTG AAACTGAGCTCTGTGACCGCCGCGGACACGGCTGTGTATTACTGTGCGAGAGGG TATTAC GATATTTTGACTGGTTATTATTACTACTTTGACTACTGGGGCCAGGGAACCCTGG TCACC GTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCGCTAGCGCCCTGCTCCA GGAGC ACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAAC CGGTG ACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTG TCCTA CAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCT TGGGC ACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC AAGAGA GTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGG GGGGAAttorney Docket No. ZURAP002WO CCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGA CCCCT GAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTC AACTGG TACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCA GTTCAAC AGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG GCAAG GAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACC ATCTCC AAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAG GAGGAG ATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGC GACATC GCCGTGGAGTGGGAAAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCC TCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA GGTGG CAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT ACACA CAGAAGAGCCTCTCCCTGTCTCCTGGAGGCGGAGGATCCGGGGGAGGGGGTACC GGAGGA GGGGGCTCGCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGG TCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGTTACAAGTTCACTGACTACCATA TTCATTGGGTG CGACAGGCCCCTGGACAATGCCTTGAGTGGATGGGAGTAATTAATCCTACTTATG GTACT ACTGACTACAATCAGCGGTTCAAAGGCCGTGTCACCATTACCGCGGACGAATCC ACGAGC ACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTAC TGTGCG AGATATGATTACTTTACTGGGACGGGTGTGTACTGGGGCCAAGGAACCCTGGTCA CCGTC TCCTCAGGTGGCGGAGGATCTGGTGGAGGTGGCTCAGGAGGTGGCGGAAGCGGCAttorney Docket No. ZURAP002WO GGAGGT GGAAGTGATATTGTGATGACTCAGACTCCACTCTCCCTGTCCGTCACCCCTGGAC AGCCG GCCTCCATCTCCTGCAGATCTAGTAGGAGCCTTGTACACAGTCGTGGAGAAACCT ATTTA CATTGGTATCTGCAGAAGCCAGGCCAATCTCCACAGCTCCTAATTTATAAAGTTT CCAAC CGGTTTATTGGGGTCCCAGACAGATTCAGCGGCAGTGGGTCAGGCACAGATTTCA CACTG AAAATCAGCAGGGTGGAGGCCGAAGATGTTGGGGTTTATTACTGCTCTCAAAGT ACACAT CTTCCATTCACGTTTGGCTGCGGGACCAAGCTGGAGATCAAA SEQ ID NO:3.

[0100] A particular DNA polynucleotide sequence encoding the second polypeptide having an amino acid sequence of SEQ ID NO:2 is: GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAG CCACC CTCTCCTGCAGGGCCAGTCAGAGTGTTAGCCGCTACTTAGCCTGGTACCAGCAGA AACCT GGCCAGGCTCCCAGGCTCCTCATCTATGATGCATCCAACAGGGCCACTGGCATCC CAGCC AGGTTCAGTGGCAGTGGGTCTGGGACAGACTCCACTCTCACCATCAGCAGCCTAG AGCCT GAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCGGACGTTCG GCCAA GGGACCAAGGTGGAAATCAAACGAACTGTGGCGGCGCCATCTGTCTTCATCTTCC CGCCA TCTGATGAGCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACT TCTAT CCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAAC TCCCAG GAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAACACC CTGACG CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCAT CAGGGCAttorney Docket No. ZURAP002WO CTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC SEQ ID NO:4.

[0101] Medium, into which a bispecific antibody has been secreted, may be purified by conventional techniques. For example, the medium may be applied to and eluted from a Protein A or G column using conventional methods. Soluble aggregate and multimers may be effectively removed by common techniques, including size exclusion, hydrophobic interaction, ion exchange, or hydroxyapatite chromatography. The product may be immediately frozen, for example at -70°C, or may be lyophilized.

[0102] There may be a need to reduce the level of misfolded bispecific antibody present in the medium. Misfolded bispecific antibody is also known as diabody. Misfolding results when one or more disulfide bonds form incorrectly, either inter or intra chains. The misfolded bispecific antibody may be purified by conventional techniques. For example, the medium containing the misfolded bispecific antibody may be applied to and eluted from strong cation exchange resin. For example, SP-Sepharose HP strong cation exchange resin is used to purify correctly folded bispecific antibody from diabody. The pH of the medium containing the diabody is adjusted to pH 8 using 1M Tris Base. The medium is loaded onto an SP-Sepharose HP column, washed with 2 column volumes of 20mM Tris, pH 8 and eluted with 20mM Tris, 100mM NaCl, pH8 over 30 column volumes (0-70 mM NaCl). The collected pools can be assessed for high molecular weight versus main peak. A typical result is an improvement from 10% diabody to 1% diabody with 71% recovery.

[0103] In another example, Poros HS 50 strong cation exchange resin is used to purify correctly folded bispecific antibody from diabody. The pH of the medium containing the diabody is adjusted to pH 8 using 1M Tris Base. The medium is loaded onto an SP- Sepharose HP column and eluted with 20mM Tris, 100mM NaCl, pH8 over 15 column volumes (15-50 mM NaCl). The collected pools can be assessed for high molecular weight versus main peak. A typical result is an improvement from 10% diabody to 1% diabody with 57% recovery.

[0104] The bispecific antibody of the example of expression and demonstration of properties described herein comprises two first polypeptides having amino acid sequences of SEQ ID NO: l and two second polypeptides having amino acid sequences of SEQ ID NO:2 wherein each of the first polypeptides forms an inter-chain disulfide bond with each of the second polypeptides between cysteine residue 137 of SEQ ID NO: l and cysteine residue 214 of SEQ ID NO:2, and the first polypeptide forms two inter-chain disulfide bonds with the other first polypeptide between cysteine residue 229 first polypeptide of SEQ ID NO: 1 andAttorney Docket No. ZURAP002WO cysteine residue 229 of the other first polypeptide of SEQ ID NO: 1 and between cysteine residue 232 first polypeptide of SEQ ID NO: 1 and cysteine residue 232 of the other first polypeptide of SEQ ID NO: 1, and each of the first polypeptides forms an intra-chain disulfide bond between cysteine residue 22 and cysteine residue 95 of SEQ IDNO: l, between cysteine residue 150 of SEQ ID NO: l and cysteine residue 206 of SEQ ID NO: 1, between cysteine residue 264 of SEQ ID NO: 1 and cysteine residue 324 of SEQ ID NO: 1, between cysteine residue 370 of SEQ ID NO: 1 and cysteine residue 428 of SEQ ID NO: l, between cysteine residue 485 of SEQ ID NO: l and cysteine residue 559 of SEQ ID NO: l, between cysteine residue 507 of SEQ ID NO: l and cysteine residue 707 of SEQ ID NO: 1, and between cysteine residue 625 of SEQ ID NO: 1 and cysteine residue 695 of SEQ ID NO: l, and each of the second polypeptides forms an intra-chain disulfide bond between cysteine residue 23 of SEQ ID NO:2 and cysteine residue 88 of SEQ ID NO:2, and between cysteine residue 134 of SEQ ID NO:2 and cysteine residue 194 of SEQ ID NO:2, and wherein each of the first polypeptides is glycosylated at asparagine residue 300 of SEQ ID NO: l. The ratio of correctly folded bispecific antibody to misfolded diabody is on the order to 90: 10. Variants

[0105] A “variant” antibody, refers herein to a molecule which differs in amino acid sequence from a “parent” antibody amino acid sequence by virtue of addition, deletion and / or substitution of one or more amino acid residue(s) relative to the parent antibody sequence. In some embodiments, the variant antibody comprises at least one amino acid (e.g., from one to about ten, and such as 2, 3, 4, 5, 6, 7 or 8) addition, deletion and / or substitution in the CDR regions of the parent antibody. Identity or homology with respect to the variant antibody sequence is defined herein as the percentage of amino acid residues in the variant antibody sequence that are identical with the parent antibody residues after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. The variant antibody retains the ability to bind the antigen, or in some embodiments, the epitope, to which the parent antibody binds. In some embodiments, the variant antibody has at least one property or bioactivity that is superior to that of the parent antibody. For example, in some embodiments, the variant antibody has stronger binding affinity, slower off-rate, lower IC50 or enhanced ability to inhibit an antigen bioactivity than the parent antibody. In some embodiments, the variant antibody displays at least about 2-fold, such as at least about 5-fold, 10-fold or 20-fold enhancement in a property or bioactivity when compared to the parent antibody. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%,Attorney Docket No. ZURAP002WO 95%, 97%, 98%, or 99% amino acid sequence identity with the parent antibody. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with the parent antibody heavy or light chain variable domain sequences. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with the parent antibody heavy chain variable domain sequence. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with the parent antibody light chain variable domain sequence. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with SEQ ID NO: 1. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with SEQ ID NO: 2. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with SEQ ID NO: 3. In some embodiments, the variant antibody has at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity with SEQ ID NO: 4. In some embodiments, the sequence identity is calculated without reference to the cysteine residues, i.e., the sequence of the variant antibody can be varied except for cysteines required for maintaining the conformation of the bispecific antibody.

[0106] In some embodiments, the “parent” antibody herein is one which is encoded by an amino acid sequence used for the preparation of a variant antibody. The parent antibody may have framework sequence of murine origin, but in some embodiments, the framework sequence is entirely or substantially of human origin. The parent antibody may be a murine, chimeric, humanized or human antibody.

[0107] A murine monoclonal antibody or a human antibody (produced e.g., in a transgenic mouse) raised against IL-17 and / or BAFF may be a parent antibody. A parent antibody may be further altered to create a chimeric or humanized form of the antibody or other variant form of the antibody using methods available in the art, e.g., PCR mutagenesis. Such chimeric, humanized, or otherwise variant antibodies, may serve as parent antibodies for further variation or mutagenesis. Parent antibodies may be mutagenized, e.g., within the CDR domain(s) to create variant antibodies that may be screened for presence of a property of interest, e.g., binding affinity (lower KD), IC50, specificity, preferential binding, etc. In some embodiments, the property of interest in the variant antibody is an improvement over that property in the parent antibody. In some embodiments, an amino acid substitution variant antibody has at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residue(s) of the parent antibodyAttorney Docket No. ZURAP002WO molecule removed and a different residue inserted in its place. The site of greatest interest for substitutional mutagenesis is one or more CDR regions, but FR alterations are also contemplated. Conservative amino acid substitutions are typically used; although, for more substantial changes, non-conservative amino acid changes may be introduced and the resulting antibodies screened for the property of interest.

[0108] A convenient way for generating substitution variants of a parent antibody is affinity maturation using phage display. Briefly, a polynucleotide molecule encoding a parent antibody is mutated within one or more CDR regions to generate all possible amino acid substitutions at each amino acid residue at which a substitution is desired. The antibody variants thus generated are displayed in a monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage- displayed variant antibodies are then screened for their biological activity (e.g., binding affinity, specificity, IC50). In order to identify candidate CDR region sites for modification, alanine scanning mutagens can be performed to identify CDR region residues contributing significantly to antigen binding.

[0109] Alternatively, or in addition, it may be beneficial to analyze a crystal structure of the antigen-antibody complex to identify contact points between the antibody and IL-17 or BAFF. Such contact residues and neighboring residues are candidates for substitution according to the techniques elaborated herein or known in the art. Alternatively, or in addition, random mutagenesis or point mutagenesis may be performed, on one or more polynucleotide molecules encoding at least one CDR. The mutagenesis may be performed, at one or more positions, either while the CDR is operably linked to the framework region within the variable region or while the CDR is independent of other variable region sequence and then the altered CDR returned to the variable region using recombinant DNA technology. Once such variant antibodies are generated the panel of variants is subjected to screening for a property or activity of interest and antibodies with superior properties in one or more relevant assays may be selected for further development.

[0110] Any cysteine residue not involved in maintaining the proper conformation of the antibody may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability (particularly where the antibody is an antibody fragment such as an Fv fragment).

[0111] Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. In some embodiments, altering is deleting one or more carbohydrateAttorney Docket No. ZURAP002WO moieties found in the antibody, and / or adding one or more glycosylation sites that are not present in the parent antibody. Glycosylation of antibodies is typically either N-linked or O- linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X- threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine’s side chain. Thus the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N- aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.

[0112] Addition of glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).

[0113] In some embodiments, the parent antibody is the polypeptide having SEQ ID NO: 1. In some embodiments, the parent antibody is the polypeptide having SEQ ID NO: 2. In some embodiments, the parent antibody is the bispecific antibody comprising two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1 and the amino acid sequence of the second polypeptide is SEQ ID NO:2. Pharmaceutical Compositions

[0114] A bispecific antibody described herein can be incorporated into a pharmaceutical composition suitable for administration to a patient. Such pharmaceutical compositions are designed to be appropriate for the selected mode of administration, and pharmaceutically acceptable diluents, carrier, and / or excipients such as dispersing agents, buffers, surfactants, preservatives, solubilizing agents, isotonicity agents, stabilizing agents and the like are used as appropriate. Said compositions can be designed in accordance with conventional techniques disclosed in, e.g., Remington, The Science and Practice of Pharmacy, 19thEdition, Gennaro, Ed., Mack Publishing Co., Easton, PA 1995 which provides a compendium of formulation techniques as are generally known to practitioners. Suitable carriers for pharmaceutical compositions include any material which, when combined with a bispecific antibody described herein, retains the molecule's activity and is non-reactive with the patient'sAttorney Docket No. ZURAP002WO immune system. A pharmaceutical composition described herein comprises a bispecific antibody and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0115] A pharmaceutical composition comprising a bispecific antibody described herein can be administered to a patient at risk for or exhibiting diseases or disorders as described herein using standard administration techniques.

[0116] A pharmaceutical composition described herein contains an effective amount of a bispecific antibody described herein. An effective amount refers to an amount necessary (at dosages and for periods of time and for the means of administration) to achieve the desired therapeutic result. An effective amount of the bispecific antibody may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual. An effective amount is also one in which any toxic or detrimental effect of the bispecific antibody, are outweighed by the therapeutically beneficial effects.

[0117] In some embodiments, the pharmaceutical composition comprises tibulizumab.

[0118] In some embodiments, the pharmaceutical composition is formulated for injection.

[0119] In some embodiments, the pharmaceutical composition is formulated for subcutaneous injection.

[0120] In some embodiments, the pharmaceutical composition is formulated for intravenous injection.

[0121] In some embodiments, the pharmaceutical composition is in a liquid formulation.

[0122] In some embodiments, the pharmaceutical composition comprises a buffer. In some embodiments, the pharmaceutical composition comprises a buffering agent. In some embodiments, the pharmaceutical composition comprises buffer system. In some embodiments, the pharmaceutical composition comprises a stabilizer. In some embodiments, the pharmaceutical composition comprises a tonicity agent. In some embodiments, the pharmaceutical composition comprises a surfactant. In some embodiments, the pharmaceutical composition comprises water for injection.

[0123] In some embodiments, the pharmaceutical composition does not comprise citrate. In some embodiments, the pharmaceutical composition does not comprise sodium citrate dihydrate. In some embodiments, hypersensitivity reactions are evaluated following administration. In some embodiments, injection site reactions are evaluated following administration. In some embodiments, the injection site reaction is selected from pain, a nodule, erythema or redness, and induration or swelling. In some embodiments, the injection site reaction is mild (Grade 1), moderate (Grade 2), severe (Grade 3), or potentially life-Attorney Docket No. ZURAP002WO threatening (Grade 4). In some embodiments, the risk of injection site reaction is reduced compared to the administration of ixekizumab with citrate. In some embodiments, the buffer does not comprise citrate.

[0124] In some embodiments, the pharmaceutical composition comprises histidine. In some embodiments, the buffer is histidine.

[0125] In some embodiments, the pharmaceutical composition is lyophilized. In some embodiments, the pharmaceutical composition has not been lyophilized.

[0126] In some embodiments, the pharmaceutical composition comprises sucrose. In some embodiments, the stabilizer is sucrose. In some embodiments, the tonicity agent is sucrose.

[0127] In some embodiments, the pharmaceutical composition comprises polysorbate. In some embodiments, the solubilizer is polysorbate. In some embodiments, the polysorbate is polysorbate 80 (PS80). In some embodiments, the concentration of polysorbate is about 0.01, 0.02, 0.025, 0.03, 0.04, or 0.05% of the pharmaceutical composition.

[0128] In some embodiments, the pharmaceutical composition has a pH of about 5-6. In some embodiments, the pH is about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0. In some embodiments, the pH is about 5.4. In some embodiments, the pH is about 5.5. In some embodiments, the pH is about 5.6.

[0129] In some embodiments, the pharmaceutical composition is isotonic. In some embodiments, the pharmaceutical composition is isotonic with patient tissue impacted by disease. In some embodiments, the pharmaceutical composition is isotonic with the injection site for a patient with systemic sclerosis. In some embodiments, the pharmaceutical composition has a pH close to that of human skin. In some embodiments, the pharmaceutical composition has a pH close to that of patient skin impacted by systemic sclerosis.

[0130] In some embodiments, the isoelectric point of the pharmaceutical composition is about 8-9 under denaturing conditions. In some embodiments, the isoelectric point of the pharmaceutical composition is about 8.5 under denaturing conditions. In some embodiments, the isoelectric point of the pharmaceutical composition is about 8.6 under denaturing conditions. In some embodiments, the isoelectric point of the pharmaceutical composition is about 8.7 under denaturing conditions. In some embodiments, the isoelectric point of the pharmaceutical composition is about 8.8 under denaturing conditions.

[0131] In some embodiments, the concentration of tibulizumab in the pharmaceutical composition is about 25-100 mg / ml. In some embodiments, the concentration of tibulizumab in the pharmaceutical composition is about 45-75 mg / ml. In some embodiments, the concentration of tibulizumab in the pharmaceutical composition is about 45, 50, 55, 60,Attorney Docket No. ZURAP002WO 65, 70, 75, 80, 85, 90, 95, or 100 mg / ml. In some embodiments, the concentration of tibulizumab in the pharmaceutical composition is about 75 mg / ml. In some embodiments, the concentration of tibulizumab in the pharmaceutical composition is about 100 mg / ml.

[0132] In some embodiments, the pharmaceutical composition comprises tibulizumab, a buffer, a stabilizer or tonicity agent, and a solubilizer.

[0133] In some embodiments, the pharmaceutical composition is a subcutaneous injection in a liquid formulation comprising tibulizumab, histidine, sucrose, and polysorbate.

[0134] In some embodiments, the pharmaceutical composition comprises tibulizumab and has a pH of about 5.5.

[0135] In some embodiments, the pharmaceutical composition comprises about 75 mg / mL tibulizumab and has a pH of about 5.5.

[0136] In some embodiments, the pharmaceutical composition comprises about 75 mg / mL tibulizumab and histidine.

[0137] In some embodiments, the pharmaceutical composition has a pH of about 5.5 and comprises histidine.

[0138] In some embodiments, the pharmaceutical composition has a pH of about 5.5 and does not comprise citrate.

[0139] In some embodiments, the pharmaceutical composition has a pH of about 5.5 and comprises histidine.

[0140] In some embodiments, the pharmaceutical composition has a pH of about 5.5 and does not comprise citrate.

[0141] In some embodiments, the pharmaceutical composition comprises about 75 mg / mL tibulizumab and does not comprise citrate.

[0142] In some embodiments, the pharmaceutical composition comprises about 75 mg / mL tibulizumab; about 20 mM histidine; about 270 mM sucrose; and polysorbate.

[0143] In some embodiments, the pharmaceutical composition comprises about 75 mg / mL tibulizumab; about 20 mM histidine; about 270 mM sucrose; and polysorbate 80; and has a pH of about 5.5.

[0144] In some embodiments, the polysorbate is polysorbate 80.

[0145] In some embodiments, the pharmaceutical composition has a viscosity of about 10- 20 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 15-20 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 15 centiPoise. In some embodiments,Attorney Docket No. ZURAP002WO the pharmaceutical composition has a viscosity of about 16 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 17 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 18 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 19 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of about 20 centiPoise. In some embodiments, the pharmaceutical composition has a viscosity of less than about 30, 25, or 20 centiPoise.

[0146] In some embodiments, the pharmaceutical composition is stored at about 0 to about 10 degrees Celsius. In some embodiments, the pharmaceutical composition is stored at about 2 to about 8 degrees Celsius. In some embodiments, the pharmaceutical composition is stored at about 5 degrees Celsius. In some embodiments, the pharmaceutical composition is stored at about 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 degrees Celsius. In some embodiments, the pharmaceutical composition is allowed to come to room temperature prior to administration. In some embodiments, about 2, 3, 4, 5, or 6 mL of pharmaceutical composition is delivered per administration. In some embodiments, about 2 mL of pharmaceutical composition is delivered per administration. In some embodiments, about 4 mL of pharmaceutical composition is delivered per administration. In some embodiments, the 2, 3, 4, 5, or 6 mL per administration is delivered in more than one injection.

[0147] In some embodiments, the pharmaceutical composition is delivered in one injection per administration. In some embodiments, the pharmaceutical composition is delivered in two injections per administration. In some embodiments, the pharmaceutical composition is delivered in three injections per administration.

[0148] In some embodiments, the pharmaceutical composition is delivered in three or less injections per administration. In some embodiments, the pharmaceutical composition is delivered in two or less injections per administration. In some embodiments, the pharmaceutical composition is delivered in one injection per administration.

[0149] In some embodiments, 100 mg tibulizumab is in each of two vials for an administration. In some embodiments, 100 mg tibulizumab is in each of three vials for an administration. In some embodiments, 200 mg tibulizumab is in a single vial for an administration. In some embodiments, 120 mg tibulizumab is in each of two vials for an administration. In some embodiments, 240 mg tibulizumab is in a single vial for an administration. In some embodiments, 125 mg tibulizumab is in each of two vials for an administration. In some embodiments, 250 mg tibulizumab is in a single vial for an administration. In some embodiments, 150 mg tibulizumab is in each of two vials for anAttorney Docket No. ZURAP002WO administration. In some embodiments, 300 mg tibulizumab is in a single vial for an administration. In some embodiments, each vial contains about 1, 1.25, 1.5, 1.75, or 2 mL of pharmaceutical composition. In some embodiments, each vial contains 1.5 mL of pharmaceutical composition. In some embodiments, each vial contains 2.0 mL of pharmaceutical composition. In some embodiments, each vial contains about 100, 125, 150, 175, or 200 mg of bispecific antibody. In some embodiments, each vial contains about 125 mg of bispecific antibody. In some embodiments, each vial contains about 150 mg of bispecific antibody.

[0150] In some embodiments, the pharmaceutical composition is in an autoinjector. In some embodiments, the pharmaceutical composition is administered by autoinjector.

[0151] As used in the present specification, the following words and phrases are generally intended to have the meanings as set forth herein.

[0152] PREVENT, PREVENTING, OR PREVENTION: As used herein, the term “prevent,” “preventing”, or “prevention,” such as prevention of a particular disorder or the occurrence or onset of one or more symptoms associated with the particular disorder and does not necessarily mean the complete prevention of the disorder. For example, the term “prevent,” “preventing” and “prevention” means the administration of therapy on a prophylactic or preventative basis to an individual who may ultimately manifest at least one symptom of a disease or condition but who has not yet done so. Such individuals can be identified on the basis of risk factors that are known to correlate with the subsequent occurrence of the disease. Alternatively, prevention therapy can be administered without prior identification of a risk factor, as a prophylactic measure. Delaying the onset of at least one symptom can also be considered prevention or prophylaxis.

[0153] TREAT, TREATING, OR TREATMENT: As used herein, the term “treat,” “treating”, or “treatment” means the administration of therapy to an individual who already manifests at least one symptom of a disease or condition or who has previously manifested at least one symptom of a disease or condition. For example, “treating” can include alleviating, abating or ameliorating a disease or condition symptoms, preventing additional symptoms, ameliorating the underlying metabolic causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition. For example, the term “treating” in reference to a disorder means a reduction in severity of one or more symptoms associated with that particular disorder. Therefore, treating a disorder does notAttorney Docket No. ZURAP002WO necessarily mean a reduction in severity of all symptoms associated with a disorder and does not necessarily mean a complete reduction in the severity of one or more symptoms associated with a disorder.

[0154] TOLERATE: As used herein, an individual is said to “tolerate” a dose of a compound if administration of that dose to that individual does not result in an unacceptable adverse event or an unacceptable combination of adverse events. One of skill in the art will appreciate that tolerance is a subjective measure and that what may be tolerable to one individual may not be tolerable to a different individual. For example, one individual may not be able to tolerate headache, whereas a second individual may find headache tolerable but is not able to tolerate vomiting, whereas for a third individual, either headache alone or vomiting alone is tolerable, but the individual is not able to tolerate the combination of headache and vomiting, even if the severity of each is less than when experienced alone.

[0155] INTOLERANCE: As used herein, “intolerance” means significant toxicities and / or tolerability issues that led to a reduction in dose or discontinuation of the medication. “Intolerance” can be replaced herein with the term “unable to tolerate.”

[0156] ADVERSE EVENT: As used herein, an “adverse event” is an untoward medical occurrence that is associated with treatment with the active agent.

[0157] IN NEED OF TREATMENT and IN NEED THEREOF: As used herein, “in need of treatment” and “in need thereof” when referring to treatment are used interchangeably to mean a judgment made by a caregiver (e.g., physician, nurse, nurse practitioner, etc.) that an individual requires or will benefit from treatment. This judgment is made based on a variety of factors that are in the realm of a caregiver’s expertise, but that includes the knowledge that the individual is ill, or will become ill, as the result of a disease, condition or disorder that is treatable by the compounds described herein. Accordingly, the compounds described herein can be used in a protective or preventive manner; or compounds described herein can be used to alleviate, inhibit or ameliorate the disease, condition or disorder.

[0158] INDIVIDUAL: As used herein, “individual” means any human. In some embodiments, a human individual is referred to a “subject” or “patient.” In some embodiments, the patient is a human with a disease, disorder or condition that would benefit from a decreased level of BAFF and / or IL-17 or decreased bioactivity of BAFF and / or IL- 17.

[0159] DOSE: As used herein, “dose” means a quantity of a bispecific antibody described herein, given to the individual for treating or preventing the disease or disorder at one specific time.Attorney Docket No. ZURAP002WO

[0160] THERAPEUTICALLY EFFECTIVE AMOUNT: As used herein, “therapeutically effective amount” of an agent, compound, drug, composition or combination is an amount which is nontoxic and effective for producing some desired therapeutic effect upon administration to a subject or patient (e.g., a human subject or patient). The precise therapeutically effective amount for a subject may depend upon, e.g., the subject’s size and health, the nature and extent of the condition, the therapeutics or combination of therapeutics selected for administration, and other variables known to those of skill in the art. The effective amount for a given situation is determined by routine experimentation and is within the judgment of the clinician. In some embodiments, the therapeutically effective amount is the standard dose.

[0161] CLINICAL RESPONSE: As used herein, a "clinical response" refers to an indicator of therapeutic effectiveness of an agent.

[0162] PHARMACEUTICAL COMPOSITION: As used herein, “pharmaceutical composition” means a composition comprising at least one active ingredient, such as a bispecific antibody described herein, whereby the composition is amenable to investigation for a specified, efficacious outcome. Those of ordinary skill in the art will understand and appreciate the techniques appropriate for determining whether an active ingredient has a desired efficacious outcome based upon the needs of the artisan.

[0163] When an integer is used in a method disclosed herein, the term “about” can be inserted before the integer.

[0164] Throughout this specification, unless the context requires otherwise, the word “comprise”, or variations such as “comprises” or “comprising” will be understood to imply the inclusion of a stated step or element or integer or group of steps or elements or integers but not the exclusion of any other step or element or integer or group of elements or integers.

[0165] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps, or group of compositions of matter shall be taken to encompass one and a plurality (i.e., one or more) of those steps, compositions of matter, groups of steps, or groups of compositions of matter.

[0166] Each embodiment described herein is to be applied mutatis mutandis to each and every other embodiment unless specifically stated otherwise.

[0167] Those skilled in the art will appreciate that the invention(s) described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention(s) includes all such variations and modifications. The invention(s) also includes all the steps, features, compositions and compounds referred to orAttorney Docket No. ZURAP002WO indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features unless specifically stated otherwise.

[0168] The present invention(s) is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of exemplification only. Functionally equivalent products, compositions, and methods are clearly within the scope of the invention(s), as described herein.

[0169] It is appreciated that certain features of the invention(s), which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the invention(s), which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination. For example, a method that recites prescribing and / or administering a bispecific antibody described herein can be separated into two methods; one method reciting prescribing a bispecific antibody described herein and the other method reciting administering a bispecific antibody described herein. In addition, for example, a method that recites prescribing a bispecific antibody described herein and a separate method of the invention reciting administering a bispecific antibody described herein can be combined into a single method reciting prescribing and / or administering a bispecific antibody described herein.

[0170] Further embodiments include the embodiments disclosed in the following Examples, which is not to be construed as limiting in any way.Attorney Docket No. ZURAP002WO EXAMPLES Example 1: Bispecific Antibodies

[0171] The bispecific antibody can be expressed and purified essentially as follows. A glutamine synthetase (GS) expression vector containing the DNA of SEQ ID NO:3 (encoding the first polypeptide having amino acid sequence of SEQ ID NO: 1) and SEQ ID NO:4 (encoding the light chain amino acid sequence of SEQ ID NO:2) is used to transfect the Chinese hamster cell line, CHOK1SV (Lonza Biologies PLC, Slough, United Kingdom) by electroporation. The expression vector encodes an SV Early (Simian Virus 40E) promoter and the gene for GS. Expression of GS allows for the biochemical synthesis of glutamine, an amino acid required by the CHOK1SV cells. Post-transfection, cells undergo bulk selection with 50μΜ L-methionine sulfoximine (MSX). The inhibition of GS by MSX is utilized to increase the stringency of selection. Cells with integration of the expression vector cDNA into transcriptionally active regions of the host cell genome can be selected against CHOK1 SV wild type cells, which express an endogenous level of GS. Transfected pools are plated at low density to allow for close-to-clonal outgrowth of stable expressing cells. The masterwells are screened for bispecific antibody expression and then scaled up in serum-free, suspension cultures to be used for production. Clarified medium, into which the bispecific antibody has been secreted, is applied to a Protein A affinity column that has been equilibrated with a compatible buffer, such as phosphate buffered saline (pH 7.4). The column is washed to remove nonspecific binding components. The bound bispecific antibody is eluted, for example, by pH gradient (such as 0.1 M sodium phosphate buffer pH 6.8 to 0.1 M sodium citrate buffer pH 2.5). Bispecific antibody fractions are detected, such as by SDS-PAGE or analytical size-exclusion, and then are pooled. Soluble aggregate and multimers may be effectively removed by common techniques, including size exclusion, hydrophobic interaction, ion exchange, or hydroxyapatite chromatography. The bispecific antibody may be concentrated and / or sterile filtered using common techniques. The purity of the bispecific antibody after these chromatography steps is greater than 98%. The

[0172] Binding affinity and binding stoichiometry of the bispecific antibody to human IL-17 and human BAFF is determined using a surface plasmon resonance assay on a Biacore 2000 instrument primed with HBS-EP+ (GE Healthcare, 10 mM Hepes pH7.4 + 150 mM NaCl + 3 mM EDTA + 0.05% surfactant P20) running buffer and analysis temperature set at 25 °C. A CM5 chip containing immobilized protein A (generated using standard NHS-EDC amine coupling) on all four flow cells (Fc) is used to employ a capture methodology. Antibody samples are prepared at 10 mcg / mL by dilution into running buffer. Human IL-17 or humanAttorney Docket No. ZURAP002WO BAFF are prepared at final concentrations of 20.0, 10.0, 5.0, 2.5, 1.25 and 0 (blank) nM by dilution into running buffer. Each analysis cycle consists of (1) capturing antibody samples on separate flow cells (Fc2, Fc3, and Fc4), (2) injection of 250 mcL (300-sec) of human IL- 17 or human BAFF over all Fc at 50 mcL / min, (3) return to buffer flow for 20 min to monitor dissociation phase, (4) regeneration of chip surfaces with a 5 mcL (30-sec) injection of glycine, pH1.5, (5) equilibration of chip surfaces with a 10 mcL (60-sec) injection of HBS- EP+. Data are processed using standard double-referencing and fit to a 1 : 1 binding model using Biacore 2000 Evaluation software, version 4.1, to determine the association rate (kon, M'Y1units), dissociation rate (kof¾ s"1units), and Rmax (RU units). The equilibrium dissociation constant (¾) is calculated as from the relationship ¾ = k0ff / kon, and is in molar units. Table 1 : Binding affinity to human IL-17 and human BAFF by the bispecific antibody.

[0173] These results demonstrate that the bispecific antibody described herein binds human IL-17 and human BAFF.

[0174] BIAcore 2000 instrument is used to determine whether human IL- 17 and human BAFF can bind to the bispecific antibody simultaneously. Except as noted, all reagents and materials are purchased from BIAcore AB (Upsala, Sweden). All measurements are performed at 25°C. HBS-EP+ buffer (150 mM sodium chloride, 3 mM EDTA, 0.05 % (w / v) surfactant P-20, and 10 mM HEPES, pH7.4) is used as the running buffer and sample buffer. Protein A is immobilized on flow cells 1 and 2 of a CM4 sensor chip using an amine coupling kit. The bispecific antibody is first captured on flow cell 2, followed by injection of human IL-17 at 20 nM for 5 min to saturate IL-17 binding site. After binding of IL-17, human BAFF at 20 nM is then injected for 5 min. and additional binding signal is observed. Chip surface is then regenerated using l0 mM Glycine pH 1.5. The same process is repeated except with a different order of human IL-17 and human BAFF. The stoichiometry is calculated to ensure complete saturation of human IL-17 or human BAFF to the bispecific antibody. The stoichiometry of human IL-17 to the bispecific antibody is typically at -1.3 based on a kinetic binding experiment. Similarly, the stoichiometry of human BAFF to the bispecific antibody isAttorney Docket No. ZURAP002WO typically at ~ 1.0 based on a kinetic binding experiment. Control BAFF Ab is 4A5-3.1.1-B4 of US7,317,089.

[0175] Control IL-17 Ab is Fab 126 of US7,838,638. Table 2: Simultaneous binding of human IL-17 and human BAFF to the bispecific antibody.

[0176] These results demonstrate that the bispecific antibody described herein can bind human IL-17 and human BAFF simultaneously as shown by the increase in response units (Δ RU) from the two ligands binding to the bispecific antibody.

[0177] HT-29 cells are human colorectal adenocarcinoma epithelial cells that naturally express the IL-17 receptor. Incubation of HT29 cells with human IL-17 results in the production of CXCL1, which can be measured using a commercially available ELISA.

[0178] A dose range of the bispecific antibody from 41200 to 2.64 pM is evaluated (final concentration based on monomeric MW of bispecific antibody (=100 kDa)). Each test concentration of bispecific antibody is then added (50 mcl) to wells containing recombinant IL-17 (final IL-17 concentration in the well is 3.75 nM (based on monomeric MW of IL-17 (=16 kDa)). Testing is carried out in triplicate wells per treatment. Assay medium is used for "medium alone" and "IL-17 alone" controls. An IL-17 neutralizing antibody (Fab 126 of US7, 838,638) is used as positive control in the assay. Plates containing IL-17 and antibody mixtures are incubated for 60 to 90 minutes at 37°C, 95% relative humidity, 5% C02 in the inner wells of tissue-culture treated 96 well plates. In a variation of this assay, a saturating concentration of human BAFF is added (1.25 nM final concentration based on monomeric MW of BAFF (=20 kDa)), with the goal to determine if bispecific antibody would still be able to neutralize IL-17 when simultaneously bound to BAFF. HT-29 cells are routinely cultured in assay medium (McCoy's 5A containing10% FBS, penicillinG (0.2 U / mL) and streptomycin (0.2 mcg / mL)). On the day of the assay, the cells are rinsed with HBSS and detached from the culture flasks with trypsin + EDTA. The trypsin is inactivated with assay medium. HT-29 cells are then centrifuged at 500Xg for 5 minutes at RT. The cell pellet isAttorney Docket No. ZURAP002WO resuspended in assay medium. Cell density is measured with a hemocytometer, and 20,000 HT-29 cells (in 100 mcl) are added to the 96-well plates containing the antibody / IL-17 mixture. Two hundred mcl of PBS is added to each of the unused edge wells (without cells) to reduce edge effects resulting from evaporation. The 96-well plates are placed in a tissue culture incubator (37°C, 95% relative humidity, 5%C02) for approximately 48 hours.

[0179] At the end of the assay, the plates are centrifuged (500Xg for 5 minutes at RT), and the cell culture medium is transferred to polypropylene 96-well plates, which are sealed and frozen at -80°C. On the day of measuring CXCLl by ELISA, the plates are thawed at RT. CXCLl levels in medium (either undiluted, or diluted 1 : 3) are measured with a CXCLl sandwich ELISA (R&D Systems DuoSet #DY275), as per the manufacturer's instructions, using the following buffers and modifications: IX ELISA wash buffer from BioFX Labs (from 10X, #WSHW- 1000-01); sample and standard volume of 50 mcL per well; substrate from BioFX Labs (1 component HRP substrate, #TMBW- 1000-01); a stop solution from BioFX Labs (#LSTP- 1000-01; 100 mcl per well). At the end of the ELISA reactions, plates were read at 450 nm on a microplate reader (Molecular Devices SpectraMax 190). Data are collected as % of maximum amount of CXCLl produced (with IL-17 alone being 100%). The concentration where 50% of the IL-17-induced response is inhibited (IC50) by either bispecific antibody or the positive control is calculated using a 4 parameter sigmoidal fit of the data (GraphPad Prism).

[0180] The results demonstrate that the bispecific antibody described herein inhibited IL-17- induced secretion of CXCLl by HT-29 cells in a concentration-dependent manner. The inhibition is comparable to that observed with the positive control antibody (with an IC50 for bispecific antibody of 2.00 + 0.21 nM versus 1.86 + 0.22 nM for the positive control antibody (average of 3 independent experiments + SEM)), whereas the negative control antibody did not inhibit proliferation. Moreover, a similar inhibition is observed in the presence of a saturating amount of BAFF, with an IC50 for bispecific antibody of 1.57 + 0.45 nM versus 1.41 + 0.50 nM for the positive control antibody(average of 3 independent experiments + SEM). The bispecific antibody described herein effectively neutralizes IL-17 and that this neutralization is not affected by the presence of saturating amounts of BAFF.

[0181] Tl 165.17 is a murine plasmacytoma cell line that is dependent on external factors (IL-l beta or BAFF) for survival and growth. These cells naturally express the receptor for BAFF and their response to human BAFF is measured by monitoring proliferation.

[0182] A dose range of the bispecific antibody from 1 nM to 4.1 pM (final concentration based on monomeric MW of bispecific antibody (=100 kDa)) is evaluated for the ability toAttorney Docket No. ZURAP002WO neutralize soluble BAFF (final soluble BAFF concentration in assay is 150 pM based on monomeric MW of BAFF (=20 kDa)). Various concentrations of bispecific antibody are incubated with soluble BAFF for 30-60 minutes at 37°C in the inner wells of a flat bottom 96 well tissue culture plate in a total volume of 50 mcl. A BAFF neutralizing antibody (4A5- 3.1.1-B4 of US7, 317,089) is used as positive control in the assay. Each condition is tested in triplicate. In a variation of this assay, the ability of bispecific antibody to neutralize membrane BAFF is tested. Various concentrations of bispecific antibody are incubated with the membrane fraction of HEK293 cells expressing a non- cleavable form of BAFF (achieved by mutating the furin cleavage site in BAFF, resulting in permanent expression of BAFF on the cell membrane). In another variation of this assay, a saturating concentration of human IL-17 is added (15.6 nM final concentration, based on monomeric MW of IL-17 (=16 kDa)), with the goal to determine if bispecific antibody would still be able to neutralize either soluble or membrane BAFF when simultaneously bound to IL-17.

[0183] Tl 165 cells are routinely cultured in assay medium (RPMI1640 containing 10% FBS, HEPES, L-Glutamine, l mM Sodium Pyruvate, 5 x 10"5M 2-mercaptoethanol, IX Antibiotic-Antimycotic) supplemented with 2 ng / mL recombinant human IL-lbeta. On the day of the assay cells are washed 3 times with assay medium and resuspended to lxlO5cells / mL in assay medium. Fifty mcl of the cell suspension is added to the 96 well plate, containing the mixture of antibody and BAFF. One hundred mcl of assay medium is added to each of the unused edge wells (without cells) to reduce edge effects resulting from evaporation. Plates are placed in a tissue culture incubator (37°C, 95% relative humidity, 5%C02) for approximately 44 hours. At the end of the assay 20 mcl of Promega Cell Titer 96 Aqueous One Solution is added to each well and incubated for 1 to 4 hours at 37°C. Plates are read at 490 nm on a microplate reader (Molecular Devices SpectraMax 190). Data are collected as % inhibition, using wells without BAFF as the minimum and wells with 150 pM BAFF as the maximum responses. The concentration where 50% of the BAFF-induced response is inhibited (IC50) by either bispecific antibody or the positive control is calculated using a 4 parameter sigmoidal fit of the data (SigmaPlot).

[0184] The results demonstrate that the bispecific antibody inhibits soluble BAFF- induced proliferation of Tl 165 cells in a concentration-dependent manner. This inhibition is comparable to that observed with the positive control antibody (with an IC50 for bispecific antibody of 0.064 + 0.021 pM versus 0.071 + 0.002 pM for the positive control antibody (average of 2 independent experiments + SEM)), whereas the negative control antibody did not inhibit proliferation. Moreover, a similar inhibition is observed in the presence of aAttorney Docket No. ZURAP002WO saturating amount of IL-17, with an IC50 for bispecific antibody of 0.060 + 0.014 pM versus 0.073 + 0.012 pM for the positive control antibody (average of 2 independent experiments + SEM). The bispecific antibody effectively neutralizes BAFF and this neutralization is not affected by the presence of saturating amounts of IL- 17.

[0185] The ability of bispecific antibody to inhibit proliferation of Tl 165 cells induced by membrane-bound BAFF is also demonstrated. The bispecific antibody described herein effectively inhibits proliferation induced by membrane-bound BAFF similarly to the positive control BAFF antibody (4A5-3.1.1-B4 of US7,317,089). Moreover, similar inhibition is observed in the presence of a saturating amount of IL-17.

[0186] [Injection of human IL-17 leads to a rapid and transient increase in mouse CXCL1 in the circulation. Regular female C57B16 mice (n=8 per group) are injected SC with either bispecific antibody (66 meg / mouse), or a positive control anti-IL-17 antibody (Fab 126 of US7, 838,638, 50 meg / mouse) or negative control antibody (huIgG4, 50 meg / mouse). Two days later, mice receive a single IP injection of human IL-17 (3 meg / mouse) and 2 hours later serum is collected and stored at -80°C until analysis. The concentration of CXCL1 is determined by ELISA. Microtiter plates are coated with an antibody capturing human Fc (Jackson ImmunoResearch 109-005-098, 1 mcg / mL) and incubated overnight at 4°C. Plates are washed, blocked with casein, and 100 mcl of serum (1 : 1000 dilution) is added. Plates are incubated for 2h at RT, washed and an HRP-labeled detection antibody (anti-human IgG, Jackson ImmunoResearch 709-035-149) is added. Plates are incubated for l hour at RT, washed and developed using TMB substrate and read using a plate reader. The concentration is calculated based on appropriate standard curves. IL-17-induced levels of CXCL1 after exposure to bispecific antibody.

[0187] These data confirms that human IL-17 results in an increase in serum CXCL1 levels. However, in the presence of the bispecific antibody these results demonstrate that the IL-17- induced increase of CXCL1 is reduced (P<0.01, ANOVA) relative to animals that receive the negative control antibody. The reduction in CXCL1 with bispecific antibody is comparable to that observed with the positive control anti-IL-17 antibody. Equivalent exposure to eitherAttorney Docket No. ZURAP002WO bispecific antibody, the positive and negative control antibodies within each group is confirmed by quantitative ELISA. Thus, bispecific antibody described herein effectively neutralizes biological effects induced by human IL-17 in the mouse. P value determination is compared to negative control / IL-17 group.

[0188] Mice that carry a transgene encoding soluble human BAFF have an abnormally high number of B lymphocytes in the spleen.

[0189] Mice transgenic for human BAFF (n=5 per group) are injected IP with either a single dose of bispecific antibody (660 meg / mouse), or a positive control anti-BAFF antibody (4A5- 3.1.1-B4 US7,317,089, 500 meg / mouse) or negative control antibody (huIgG4, 500 meg / mouse). Eight days later, serum and spleens are collected. A single cell suspension of spleen cells is prepared and the total number of leukocytes is determined after lysing the red blood cells. The relative percentage of B lymphocytes is determined using the cell surface marker B220 by flow cytometry. The total number of B cells per spleen is calculated by multiplying the percentage of B220 positive cells by the total number of lymphocytes in the spleen. Microtiter plates are coated with an antibody capturing human Fc (Jackson ImmunoResearch 109-005-098, 1 mcg / mL) and incubated overnight at 4°C. Plates are washed, blocked with casein, and 100 mcl of serum (1 :5000 dilution) is added. Plates are incubated for 2h at RT, washed and an HRP-labeled detection antibody (anti-human IgG, Jackson ImmunoResearch 709-035-149) is added. Plates are incubated for l hour at RT, washed and developed using TMB substrate and read using a plate reader. The concentration is calculated based on appropriate standard curves. B cell numbers in the spleen of mice transgenic for human BAFF after exposure of bispecific antibody.

[0190] These results demonstrate that the number of B cells in the spleens of mice transgenic for human BAFF is reduced (p<0.0001, ANOVA) by a single administration of bispecific antibody. This normalization of B cell numbers is equivalent to that observed with the positive control BAFF antibody. Equivalent exposure to either bispecific antibody, the positive and negative control antibodies within each group is confirmed by quantitativeAttorney Docket No. ZURAP002WO ELISA. Thus, bispecific antibody described herein effectively neutralizes biological effects induced by human BAFF in the mouse. P value determination is compared to negative control group.

[0191] The bispecific antibody is formulated in PBS at pH 7.4. The bispecific antibody is concentrated from 1-2 mg / mL to a concentration ranging from 52 mg / mL to 58 mg / mL using Amicon concentrators. Concentrated samples are stored at 25°C over a period of 4 weeks. Samples are analyzed for percent high molecular weight (%HMW) with size exclusion chromatography (SEC) at initial concentration, 1 day, 1 week, and 4 weeks incubations. SEC is performed on an Agilent 1100 system using a TSK G3000SW-XL (Tosoh Bioscience) column. PBS + 0.35M NaCl, pH 7.4 is used as the mobile phase running at 0.5 mL / min for 35 minutes. A volume of l uL of the concentrated antibody is injected into the column and the detection is measured at 280nm. Chromatograms are analyzed using ChemStation and % high molecular weight (HMW) is calculated using the ratio of AUC of the peaks eluted before the monomer peak to total AUC. Samples stored at 25°C at different time points are analyzed for %HMW and the results are summarized in Table 3. Table 3. Summary of % high molecular weight species measured by SE-HPLC.

[0192] Preliminary studies with a starting bispecific antibody comprising a BAFF antibody of WO2003016468 and an IL-17 antibody of WO2007070750 demonstrated that after concentration to only 6 mg / mL, a 25% increase in % HMW species was detected by SE- HPLC after 3 weeks storage at 4°C in PBS, and at 30 mg / mL the increase in % HMW species was 15% after just 2 days storage at 4°C in PBS. These results demonstrate that the bispecific antibody described herein has much improved properties, including decreased aggregation and increased physical stability, over the starting bispecific antibody. Example 2: Dose Selection for Tibulizumab A combined PK dataset from individuals administered tibulizumab or placebo (including 24 patients with Sjögren's syndrome, 33 patients with rheumatoid arthritis, and 8 healthyAttorney Docket No. ZURAP002WO participants) was evaluated to predict BAFF and IL-17 response through population PK modeling. Free BAFF concentration and free IL-17 concentration relative to baseline were predicted using a pharmacokinetic / pharmacodynamic PK / PD model based on assayed free drug, total soluble BAFF, and total soluble IL-17. Results are shown in Table 4 and Figure 1. Table 4. Model-predicted maximum change from baseline of free unbound BAFF and IL-17 concentration following the last dose of tibulizumab administrationCI = confidence interval; Q2W = once every two weeks; Q4W = once every four weeks

[0193] The pharmacokinetics of tibulizumab have not been measured clinically in skin or lungs. A PK / PD model with a theoretical skin / lung compartment was developed to provide comparative predictions for diseases involving those tissues. Total and free target BAFF and IL-17 were simulated for 150 mg and 300 mg Q4W dosing regimens administered subcutaneously for 24 weeks (with an additional dose of 150 mg and 300 mg at Week 2, respectively).

[0194] Predicted median trough changes from baseline (5th / 95thpercentile) were calculated for free BAFF and IL-17 for peripheral blood and skin / lung at weeks 16 and 24. Results are shown in Table 5, Figure 3 (predicted blood PK for the 150 mg regimen), Figure 4 (predicted skin / lung PK for the 150 mg regimen), Figure 5 (predicted blood PK for the 300 mg regimen), and Figure 6 (predicted skin / lung PK for the 300 mg regimen).

[0195] Steady state was reached by week 12 for both dosing regimens. For the 150 mg regimen, the lower predicted concentration of tibulizumab in skin and lung tissue relative to peripheral blood resulted in lower target engagement (Table 2), with median steady state trough target engagement ~ 75% for BAFF and ~79% for IL-17. In addition, variability for skin and lung was higher than in peripheral blood.

[0196] For the 300 mg regimen, the lower predicted concentration of tibulizumab in skin and lung tissue relative to peripheral blood again resulted in lower target engagement (TableAttorney Docket No. ZURAP002WO 5), with median steady state trough target engagement between 83 and 87% for both BAFF and IL-17. Variability for skin and lung was also higher than in blood, but was reduced relative to the 150 mg regimen. Table 5. Model-predicted change from baseline of free BAFF and IL-17 trough concentration in peripheral blood and skin / lung following the last dose of tibulizumab administrationCFB = change from baseline; Q4W = once every 4 weeks

[0197] The efficacy and safety of tibulizumab in systemic sclerosis patients will be investigated in a phase 2 study with 300 mg Q4W (with additional 300 mg dose at Week 2). This dosage regimen was selected for the study based on factors including predicted target engagement in lung and skin (particularly with respect to trough concentrations), predicted safety risk and margin, and feasibility for administration. The additional dose at Week^2 will be included with the aim of facilitating more rapid achievement of steady state. Example 3: Open Label Clinical Trial in Patients with Systemic Sclerosis

[0198] A phase 2 open label study in patients with systemic sclerosis will be conducted. There will be assessments performed at Screening (-14 days to Study Hour 0), Baseline (Study Visit 1 / Study Hour 0), Week 12 (Study Visit 2), and Week 24 (Study Visit 3). Safety will be monitored throughout the Treatment Period. The study will evaluate about twentyAttorney Docket No. ZURAP002WO patients with SSc having a mRSS score of 10-29 and presenting the first symptoms of sclerosis other than Raynaud’s phenomenon within 60 months of enrollment.

[0199] The objectives of the study may include the determination of: • safety of the bispecific antibody in patients with SSc as measured by treatment emergent adverse events (TEAE) and standard laboratory assessments. • whether administration of the bispecific antibody improves the patient’s mRSS score. • whether administration of the bispecific antibody improves the patient’s scleroderma health assessment questionnaire disability index. • whether administration of the bispecific antibody improves the patient’s visual analog scale (VAS) score for Raynaud's phenomenon, pain, ulcers, and / or breathing. • whether administration of the bispecific antibody improves the patient’s ratio of forced vital capacity to diffusion capacity (FVC / DLCO). Example 4: Randomized Controlled Clinical Trial in Patients with Systemic Sclerosis

[0200] A Phase 2 randomized placebo-controlled study will be conducted to assess efficacy, safety, and tolerability of tibulizumab in patients with systemic sclerosis.

[0201] A high-resolution computed tomography (HRCT) scan will be performed to assess SSc-ILD status at screening. A minimum number of participants will be enrolled with visually confirmed scleroderma lung disease (with moderate or more severe extent, defined as QILD ≥10% lung involvement as determined by HRCT). On Day^1, participants will be randomized 1:1 to receive 240^mg^tibulizumab or placebo, administered Q4W, with stratification by SSc-ILD per screening HRCT and anti-Scl-70 antibody status (positive / negative). Participants assigned to active treatment will receive 240^mg (administered as 2 x 1.6^mL subcutaneous injection)(75 mg / mL) tibulizumab Q4W. Participants assigned to placebo will receive matching injections. Study visits will occur every 4^weeks throughout a 24-week randomized treatment period. Safety follow-up visits will occur thereafter.

[0202] Safety assessments will include adverse events, vital signs, clinical laboratory findings, ECGs, physical examinations, and concomitant medications. Efficacy assessments will include modified Rodnan skin score (mRSS), lung assessments (HRCT and forced vital capacity (FVC)), revised Composite Response Index in Systemic Sclerosis (CRISS), digital ulcers, and patient reported outcomes (PROs) from baseline to the end of the study.Attorney Docket No. ZURAP002WO

[0203] For mRSS, scoring will be conducted by palpation of skin on multiple areas of the body using a 0-3 scale. Total score will range from 0 (no thickening) to 51 (severe thickening in all examined areas). For HRCT, standardized thin-section (<1.25 mm) reduced-dose (80- 100 mAs) volumetric non-contrast HRCT at end inspiration will be performed at screening, and again at Week 24 for participants with lung involvement. Whole lung and lobe images will be analyzed utilizing computer-aided diagnostic scores for the extent of lung involvement based on QLF (fibrotic reticulation patterns) and QILD (the sum of QLF, as well as quantitative ground glass (QGG) and quantitative honeycomb QHC). Spirometry assessment of FVC (absolute and % predicted) will be conducted per the 2005 ATS / ERS guidelines.

[0204] For ACR-CRISS responder assessment, participants will be assessed for new internal organ involvement or worsening, defined as: • New scleroderma renal crisis^ • New decline in percent predicted FVC ≥15% in established^interstitial lung disease or new percent predicted FVC below 80%^ • New onset of left ventricular failure requiring treatment^ • New onset of pulmonary arterial hypertension requiring treatment^ • Gastrointestinal dysmotility requiring enteral or parenteral^nutrition^ • Digital ischemia with gangrene, amputation, or hospitalization^requiring treatment

[0205] Participants who do not meet the above criteria will be evaluated for responder status, defined as: • improvement in ≥2 components: ≥5% improvement in % predicted FVC, and / or ≥25%^improvement in any of the following:^mRSS, HAQ-DI,^Patient Global Assessment (PtGA), Clinician Global Assessment (CGA); and • worsening in ≤1 component: ≥5% worsening in % predicted FVC, and / or ≥25%^worsening in any of the following:^mRSS, HAQ-DI,^PtGA, CGA

[0206] Pharmacodynamic assessments will include changes in biomarkers of response and serum proteins at Baseline and Week 4, 8, and 24. Pharmacokinetic assessments will include blood samples for trough plasma concentrations of tibulizumab.Attorney Docket No. ZURAP002WO Example 5: Randomized Controlled Clinical Trial in Patients with Systemic Sclerosis

[0207] A phase 2 study will be conducted with a randomized, double-blind, placebo- controlled period (Period 1) followed by an open-label extension (OLE) period (Period 2) to assess the efficacy, safety, and tolerability of tibulizumab in systemic sclerosis.

[0208] In Period 1, initial dose administration will occur on Day 1. Participants will be randomly assigned to receive either 300 mg tibulizumab or placebo, with stratification by SSc-ILD (yes / no), anti-RNA polymerase III antibody status (positive / negative), and disease duration (<2 years or ≥2 years). Of approximately 80 participants dosed in the study, approximately 60 participants will be randomized with visually confirmed SSc-ILD. Following an initial dose on Day 1, study treatment will be administered at Week 2 and Week 4, and Q4W thereafter through Week 20. A final double-blind treatment period visit for Period 1 will occur at Week 24. During Period 2, participants will receive 300 mg tibulizumab Q4W at study visits occurring from Week 24 through Week 48, with a final visit on Week 52. A final safety follow-up visit will occur 12 weeks after the final dose.

[0209] During Period 1, participants will receive randomized treatment on Day 1, Week 2, and Week 4, and Q4W from Week 4 through Week 20. During Period 2, participants will receive 300 mg tibulizumab Q4W at visits from Week 24 through Week 48.

[0210] Efficacy assessments will include modified Rodnan skin score (mRSS), lung assessments (high-resolution computed tomography (HRCT) and forced vital capacity (FVC)), diffusing capacity of the lungs for carbon monoxide (DLCO), revised-CRISS-25, digital ulcer (DU) assessments, patient reported outcomes (patient global assessment (PtGA), health assessment questionnaire disability index (HAQ-DI), and scleroderma skin patient reported outcome (SSPRO)), and clinician global assessment (CGA). Additional assessments may include changes in QILD and QLF with HRCT; the number of upper limb digital ulcers; levels of anti-centromere antibodies (ACA), anti-topoisomerase I antibodies (ATA), and anti- RNA-polymerase III antibodies; changes in gene expression in skin; and assessment of DNA samples for gene polymorphisms correlated with clinical response.

[0211] Assessments in mRSS score may include improvement or worsening by ≥5 units and ≥25% from baseline. Assessments in FVC and DLCO may include ≥10% relative decline in % predicted FVC, or ≥5% and <10% relative decline in % predicted FVC and ≥15% relative decline in % predicted DLCO. Immunogenicity assessments will include the measurement of anti-drug antibodies against tibulizumab. Pharmacokinetic assessments will include blood samples for the measurement of plasma concentrations of tibulizumab. PharmacodynamicAttorney Docket No. ZURAP002WO assessments will include blood samples and skin biopsies for the measurement of biomarkers. Genetic assessments will evaluate possible genetic variations that may influence the clinical response to tibulizumab when administered to patients with SSc. Safety assessments will include adverse events, vital signs, clinical laboratory sampling, ECGs, physical examinations, Columbia-Suicide Severity Rating Scale (C-SSRS), and concomitant medications.

[0212] Tibulizumab will be supplied as 75 mg / mL (150 mg / 2 mL per vial) and administered via subcutaneous injection. During the randomization treatment period, participants will receive 300 mg (administered as 2 × 2.0 mL subcutaneous injections) of tibulizumab or placebo through Week 20. Participants who complete the randomization treatment period and continue to the OLE period will receive 300 mg (administered as 2 × 2.0 mL subcutaneous injections) of tibulizumab through Week 48.

[0213] Although the disclosure has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the disclosure. The various embodiments described above can be combined to provide further embodiments. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and / or listed in the Application Data Sheet are incorporated herein by reference, in their entirety. Aspects of the embodiments can be modified, if necessary to employ concepts of the various patents, applications and publications to provide yet further embodiments.

[0214] These and other changes can be made to the embodiments in light of the above- detailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

Claims

Attorney Docket No. ZURAP002WO What is claimed in:

1. A method of treating systemic sclerosis in a human subject in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody against interleukin 17 (IL-17) and human B cell activating factor (BAFF).

2. The method of claim 1, wherein the bispecific antibody comprises two first polypeptides and two second polypeptides.

3. The method of claim 2, wherein the first polypeptide comprises a first heavy chain variable region (HCVR1), a heavy chain constant region (CH), a second heavy chain variable region (HCVR2) and a second light chain variable region (LCVR2).

4. The method of claim 3, wherein HCVR1 comprises CDRHl-1, CDRHl-2, and CDRHl- 3; HCVR2 comprises CDRH2-1, CDRH2-2, and CDRH2-3; and / or LCVR2 comprises CDRL2-1, CDRL2-2 and CDRL2-3.

5. The method of claim 2, wherein the second polypeptide comprises a first light chain variable region (LCVRl) and a light chain constant region (CL).

6. The method of claim 5, wherein LCVRl comprises CDRLl-1, CDRLl-2 and CDRLl-3.

7. A method of treating systemic sclerosis in a human subject in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody, or a variant thereof, wherein the bispecific antibody comprises two first polypeptides and two second polypeptides wherein the amino acid sequence of the first polypeptide is SEQ ID NO: 1, or a variant thereof, and the amino acid sequence of the second polypeptide is SEQ ID NO:2, or a variant thereof.

8. The method of any one of the preceding claims, wherein the systemic sclerosis is limited systemic sclerosis (CREST syndrome).

9. The method of any one claims 1 to 7, wherein the systemic sclerosis is generalized systemic sclerosis.

10. The method of any one claims 1 to 7, wherein the systemic sclerosis is systemic sclerosis sine scleroderma.

11. The method of any one claims 1 to 7, wherein the subject has interstitial lung disease.

12. The method of any one claims 1 to 7, wherein the systemic sclerosis is systemic sclerosis with interstitial lung disease (SSc-ILD).

13. The method of any one claims 1 to 7, wherein the subject does not have SSc-ILD.Attorney Docket No. ZURAP002WO 14. The method of any one of the preceding claims, wherein the subject has a symptom chosen from Raynaud syndrome, polyarthralgia, dysphagia, heartburn, swelling, skin tightening and / or contractures of the fingers.

15. The method of any one of the preceding claims, wherein the subject has a symptom chosen from fibrosis, skin thickening, cutaneous pruritus, whitening of hands on exposure to cold, pain in affected digits, difficulty in swallowing foods or liquids, nausea, vomiting, weight loss, abdominal cramps, bloating, diarrhea, fecal incontinence, shortness of breath on exertion, palpitations without characteristic pain in thoracic cavity, nonproductive cough, atypical chest pain, fatigue, dyspnea, hypertension, joint pain, limitation of movement, joint swelling, muscle pain, inflammatory myopathy or weakness.

16. The method of any one of the preceding claims, wherein the subject is classified according to the American College of Rheumatology criteria for the classification of systemic scleroderma as having proximal diffuse (truncal) sclerosis (skin tightness, thickening, and non-pitting induration).

17. The method of any one of the preceding claims, wherein the subject is classified according to the American College of Rheumatology criteria for the classification of systemic scleroderma as having (1) sclerodactyly (only fingers and / or toes), (2) digital pitting scars or loss of substance of the digital finger pads (pulp loss), and / or (3) bilateral basilar pulmonary fibrosis.

18. The method of any one of the preceding claims, wherein the subject has diffuse cutaneous systemic sclerosis.

19. The method of any one of the preceding claims, wherein prior to administration, the subject has a modified Rodnan skin score (mRSS) score of 10-29 or 15-45.

20. The method of any one of the preceding claims, wherein the subject has had systemic sclerosis for about 5 years or less.

21. The method of any one of the preceding claims, wherein the subject’s mRSS score is improved after administration of the bispecific antibody.

22. The method of any one of the preceding claims, wherein the subject’s Composite Response Index in Systemic Sclerosis (CRISS) is improved after administration of the bispecific antibody.

23. The method of any one of the preceding claims, wherein the subject’s forced vital capacity (FVC) is improved after administration of the bispecific antibody.Attorney Docket No. ZURAP002WO 24. The method of any one of the preceding claims, wherein the subject’s scleroderma health assessment questionnaire disability index (SHAQ-DI) is improved after administration of the bispecific antibody.

25. The method of any one of the preceding claims, wherein the subject’s visual analog scale (VAS) score for Raynaud's phenomenon, pain, ulcers, and / or breathing is improved after administration of the bispecific antibody.

26. The method of any one of the preceding claims, wherein the subject’s ratio of forced vital capacity to diffusion capacity (FVC / DLCO) is improved after administration of the bispecific antibody.

27. The method of any one of the preceding claims, further comprising measuring a modified Rodnan skin score (mRSS) for the subject.

28. The method of any one of the preceding claims, further comprising measuring Composite Response Index in Systemic Sclerosis (CRISS) for the subject.

29. The method of any one of the preceding claims, further comprising performing high- resolution computed tomography (HRCT) for the subject.

30. The method of any one of the preceding claims, further comprising measuring forced vital capacity (FVC) for the subject.

31. The method of any one of the preceding claims, further comprising performing a clinical global assessment (CGA) for the subject.

32. The method of any one of the preceding claims, further comprising performing patient global assessment (PGA) for the subject.

33. The method of any one of the preceding claims, further comprising performing a health assessment questionnaire disability index (HAQ-DI) for the subject.

34. The method of any one of the preceding claims, wherein there is an increase in CRISS, FVC, or HRCT in the subject following administration of the bispecific antibody.

35. The method of any one of the preceding claims, wherein there is a reduction in mRSS, CGA, PGA, HAQ-DI in the subject following administration of the bispecific antibody.

36. The method of any of the preceding claims, wherein the bispecific antibody is administered in an amount of about 100, about 110, about 120, about 125, about 130, about 140, about 150, about 160, about 170, about 175, about 180, about 190, about 200, about 210, about 220, about 225, about 230, about 240, about 250, about 260, about 270, about 275, about 280, about 290, about 300, about 310, about 320, about 325, about 330, about 340, about 350, about 360, about 370, about 375, about 380, about 390, or about 400 mg.Attorney Docket No. ZURAP002WO 37. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 150 mg.

38. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 240 mg.

39. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 300 mg.

40. The method of any one of the preceding claims, wherein the bispecific antibody is administered once per week, once every other week, once every two weeks, once every three weeks, once every four weeks, or once every month.

41. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 150 mg once every four weeks.

42. The method of claim 36, wherein the bispecific antibody is administered as tibulizumab 150 mg every four weeks.

43. The method of claim 42, further comprising an additional administration of 150 mg tibulizumab between the first and second administration of 150 mg every four weeks.

44. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 240 mg once every four weeks.

45. The method of claim 36, wherein the bispecific antibody is administered as tibulizumab 240 mg every four weeks.

46. The method of claim 45, further comprising administering an additional 240 mg tibulizumab between the first and second administration of 240 mg every four weeks.

47. The method of claim 36, wherein the bispecific antibody is administered in an amount of about 300 mg once every four weeks.

48. The method of claim 36, wherein the bispecific antibody is administered as tibulizumab 300 mg every four weeks.

49. The method of claim 48, further comprising administering an additional 300 mg tibulizumab between the first and second administration of 300 mg every four weeks.

50. The method of any one of the preceding claims, wherein the bispecific antibody is administered as a subcutaneous injection.

51. The method of claim 50, wherein the subcutaneous injection is in a liquid formulation comprising 75 mg / mL tibulizumab; 20 mM histidine; 270 mM sucrose; and polysorbate.

52. The method of claim 51, wherein the liquid formulation has a pH of about 5.5.Attorney Docket No. ZURAP002WO 53. The method of any one of the preceding claims, wherein the bispecific antibody is administered by autoinjector.

54. The method of any one of claims 1 to 49, wherein the bispecific antibody is administered as an intravenous injection.

55. The method of any one of the preceding claims, further comprising evaluating a biomarker.

56. A liquid pharmaceutical composition comprising: tibulizumab; a buffer; a stabilizer or tonicity agent; and a solubilizer.

57. A liquid pharmaceutical composition comprising: tibulizumab; histidine; sucrose; and polysorbate.

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