GP130 antigen binding antibodies

Abstract

Disclosed are Gp130 antigen binding antibodies that inhibit IL-6, IL-11, OSM, LIF, CNTF, CT-1 mediated signals and bind to a cytokine binding region. Also disclosed are nucleic acids and expression vectors encoding the Gp130 antigen binding antibodies, compositions comprising the Gp130 antigen binding antibodies, humanized antibodies, and methods of treatment using the Gp130 antigen binding antibodies.

Description

[0001] This application claims priority to GB 2311050.5, filed July 19, 2023, the contents and elements of which are incorporated herein by reference for all purposes. Technical Field

[0002] This disclosure relates to the field of molecular biology, and more specifically to the field of antibody technology. This disclosure also relates to methods for medical treatment and prevention. Background Technology

[0003] Gp130 is a constituent protein of the receptors for a variety of cytokines, including IL-6, IL-11, OSM, LIF, CNTF, CT-1, CLC, IL-27, and IL-35, which are generally referred to collectively as IL-6 family cytokines. Certain IL-6 family cytokines (such as IL-6 and IL-11) are associated with the pathology of a variety of diseases / disorders characterized as inflammation and / or fibrosis (see [link to relevant documentation]). For example Rose-John, F1000Res. (2020) 9:F1000 Faculty Rev-1013, Tanaka et al. , Cold SpringHarb Perspect Biol. (2014) 6(10): a016295, Hirano et al. International Immunology(2021) 33(3): 127–148, Putoczki and Ernst, Immunotherapy (2015) 7(4): 441-453, Nguyen et al. Growth Factors (2019) 37(1-2):1-11, Cook and Schafer Annu. Rev. Med. (2020) 71:263-276 and Fung et al. , Cytokine (2022) 149:155750).

[0004] Monoclonal antibodies that bind to gp130 and antagonize gp130-mediated signaling have been previously described. Monoclonal mouse anti-human gp130 antibody B-R3 is described in... For example Garbers et al. , J Biol Chem. (2013) 288(6): 4346–4354. Garbers et al. The study disclosed B-R3 that binds to the cytokine binding module (CBM) of gp130.

[0005] WO 2019 / 126071 A1 describes gp130-binding antibodies mAb16692, mAb16680, mAb16673, mAb16646, mAb16636, and mAb16618, which are disclosed for inhibiting OSM, LIF, and CNTF-mediated signaling. WO 2019 / 126071 A1 does not describe the ability of these antibodies to inhibit signaling mediated by other IL-6 family cytokines. None of the antibodies showed binding to mouse gp130—see Tables 9 and 10 of WO 2019 / 126071 A1. A concentration of 1.66 x 10⁻⁶ is disclosed. -7 M to 2.91 x 10 -9 K within the range of M D Antibodies that bind to human gp130, as determined by surface plasmon resonance analysis using Biacore. Summary of the Invention

[0006] In a first aspect, this disclosure provides an optionally isolated antigen-binding molecule that binds to gp130, wherein the antigen-binding molecule inhibits IL-6-mediated signaling, IL-11-mediated signaling, OSM-mediated signaling, CNTF-mediated signaling, CT-1-mediated signaling, and LIF-mediated signaling.

[0007] In some implementations, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:89.

[0008] In some implementations, the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12.

[0009] In some implementations, the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence having at least 70% amino acid sequence identity with the VH sequence shown in column A of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VL sequences shown in column B of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C. The VH and VL sequences are selected from the same row of Table C.

[0010] In some implementations, the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:218; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:204; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:20; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:195; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (e) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:214; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (f) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:215; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28.

[0011] In some implementations, the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence that is at least 70% amino acid sequence identical to the VH sequence shown in column A of rows 89, 90, 80-88, 74-79 or 2 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VH sequence shown in column B of rows 89, 90, 80-88, 74-79 or 2 of Table C. The VH and VL sequences are selected from the same row of Table C.

[0012] In some embodiments, the antigen-binding molecule is a multispecific antigen-binding molecule, wherein the antigen-binding molecule further includes an antigen-binding domain that binds to antigens other than gp130.

[0013] This disclosure also provides a chimeric antigen receptor (CAR) comprising an antigen-binding molecule according to this disclosure.

[0014] This disclosure also provides one or more nucleic acids (optionally isolated) that encode an antigen-binding molecule or CAR according to this disclosure.

[0015] This disclosure also provides an expression vector or multiple expression vectors comprising one or more nucleic acids according to this disclosure.

[0016] This disclosure also provides a cell comprising an antigen-binding molecule according to this disclosure, a CAR, one or more nucleic acids, or one or more expression vectors.

[0017] This disclosure also provides a method comprising culturing cells according to this disclosure under conditions suitable for cell expression of antigen-binding molecules or CARs.

[0018] This disclosure also provides a composition comprising an antigen-binding molecule according to this disclosure, a CAR, one or more nucleic acids, one or more expression vectors or cells, and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0019] This disclosure also provides methods for using antigen-binding molecules, CARs, one or more nucleic acids, one or more expression vectors, cells, or compositions according to this disclosure for medical treatment or prevention.

[0020] This disclosure also provides antigen-binding molecules, CARs, one or more nucleic acids, one or more expression vectors, cells, or compositions according to this disclosure for the treatment or prevention of the following: pathological inflammation, fibrosis, diseases / conditions characterized as inflammation, diseases / conditions characterized as fibrosis, diseases / conditions characterized as both inflammation and fibrosis, diseases / conditions of pathological significance involving signal transduction via a complex containing gp130, diseases / conditions of pathological significance involving cytokines signaling via a complex containing gp130, autoimmune diseases, metabolic syndrome, neurodegenerative diseases, chronic inflammatory diseases, arthritis, rheumatoid arthritis, juvenile arthritis, systemic juvenile idiopathic arthritis, lupus, systemic lupus erythematosus, pancreatitis, thyroiditis, periodontitis, rhinitis, allergic rhinitis, dermatitis, atopic dermatitis, psoriasis, Hermansky-Pudlak syndrome, Graves' disease. Diseases, obesity, insulin resistance, diabetes, type 1 diabetes, type 2 diabetes, pregnancy-related hyperglycemia, multiple sclerosis, giant cell arteritis, aortitis, cardiovascular disease, atherosclerosis, atrial fibrillation, ventricular fibrillation, cardiac hypertrophy, hypertrophic cardiomyopathy, dilated cardiomyopathy, myocarditis, cardiogenic shock, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, ischemic heart disease, myocardial infarction, Marfan syndrome, systemic sclerosis, keloids, scleroderma, Alzheimer's disease, Parkinson's disease. Diseases including Huntington's disease, amyotrophic lateral sclerosis (ALS), hippocampal atrophy, lung diseases, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, hepatitis, liver fibrosis, cirrhosis, hepatotoxicity, acetaminophen-induced hepatotoxicity, alcoholic liver disease, pancreatitis, fatty degeneration, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, primary biliary cholangitis, primary sclerosing cholangitis, inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, endometriosis, stroke, ischemic stroke, kidney disease, kidney injury, acute kidney injury, nephrotoxicity, glomerulonephritis, chronic kidney disease, renal fibrosis, Alport syndrome, adult-onset Still's disease, and Castleman's disease.Diseases, cytokine release syndrome, sepsis, septic shock, retinal diseases, retinal fibrosis, age-related macular degeneration, wet age-related macular degeneration, retinitis pigmentosa, dry eye, COVID-19, Peutz-Jeghers syndrome, skeletal muscle diseases, muscular dystrophy, muscular atrophy, cachexia, endocrine disorders, polycystic ovary syndrome, cancer, hematologic malignancies, leukemia, plasmacytoma, Hodgkin's lymphoma. Lymphoma, lung cancer, colorectal cancer, bowel cancer, urinary system cancers, bladder cancer, vulvar cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, bone cancer, glioblastoma, breast cancer, stomach cancer, kidney cancer, metastatic renal cell carcinoma, prostate cancer, skin cancer, melanoma, liver cancer, hepatocellular carcinoma, asthenia, age-related increase in fat mass, sarcopenia, age-related hyperlipidemia, age-related hypertriglyceridemia, age-related hypercholesterolemia, age-related hepatic steatosis, age-related non-alcoholic fatty liver disease, age-related non-alcoholic fatty liver disease, age-related non-alcoholic steatohepatitis, age-related cardiovascular disease, age-related hypertension, age-related nephropathy, age-related skin diseases, infectious diseases, viral diseases, viral hepatitis, hepatitis B, HIV infection, influenza infection, malaria, tuberculosis, allergic diseases, transplant rejection and graft-versus-host disease.

[0021] This disclosure also provides in vitro The in vitro complex is optionally isolated and comprises an antigen-binding molecule according to the present disclosure that binds to gp130.

[0022] This disclosure also provides a method for detecting gp130 in a sample, the method comprising contacting a sample containing or suspected of containing gp130 with an antigen-binding molecule according to this disclosure, and detecting the formation of a complex of the antigen-binding molecule and gp130.

[0023] This disclosure also provides a method for selecting or stratifying subjects for treatment with a gp130-targeting agent, the method comprising conjugating a sample from the subject with an antigen-binding molecule according to this disclosure. in vitro Contact, and detection of the formation of complexes between antigen-binding molecules and gp130.

[0024] This disclosure also provides antigen-binding molecules according to this disclosure as... in vitro or in vivo Uses of diagnostic or prognostic agents.

[0025] This disclosure also provides an antigen-binding molecule that binds to gp130 and inhibits IL-6-mediated signaling, IL-11-mediated signaling, OSM-mediated signaling, CNTF-mediated signaling, CT-1-mediated signaling, and LIF-mediated signaling, for the treatment or prevention of the following: pathological inflammation, fibrosis, diseases / conditions characterized as inflammation, diseases / conditions characterized as fibrosis, diseases / conditions characterized as both inflammation and fibrosis, diseases / conditions of pathological significance where signaling occurs via a gp130-containing complex, diseases / conditions of pathological significance where cytokines signal via a gp130-containing complex, autoimmune diseases, metabolic syndrome, neurodegenerative diseases, chronic inflammatory diseases, arthritis, rheumatoid arthritis, juvenile arthritis, systemic juvenile idiopathic arthritis, lupus, systemic lupus erythematosus, pancreatitis, thyroiditis, periodontitis, rhinitis, allergic rhinitis, dermatitis, atopic dermatitis, psoriasis. Hermansky-Pudrag syndrome, Graves' disease, obesity, insulin resistance, diabetes, type 1 diabetes, type 2 diabetes, pregnancy-related hyperglycemia, multiple sclerosis, giant cell arteritis, aortitis, cardiovascular disease, atherosclerosis, atrial fibrillation, ventricular fibrillation, cardiac hypertrophy, hypertrophic cardiomyopathy, dilated cardiomyopathy, myocarditis, cardiogenic shock, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, ischemic heart disease, myocardial infarction, Marfan syndrome Symptoms, systemic sclerosis, keloids, scleroderma, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, hippocampal atrophy, lung diseases, asthma, chronic obstructive pulmonary disease, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, hepatitis, liver fibrosis, cirrhosis, hepatotoxicity, acetaminophen-induced hepatotoxicity, alcoholic liver disease, pancreatitis, fatty degeneration, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cholestasis, primary biliary cholangitis, primary sclerosing cholangitis.Inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, endometriosis, stroke, ischemic stroke, kidney disease, kidney injury, acute kidney injury, nephrotoxicity, glomerulonephritis, chronic kidney disease, renal fibrosis, Allport syndrome, adult-onset Steele disease, Castleman's disease, cytokine release syndrome, sepsis, septic shock, retinopathy, retinal fibrosis, age-related macular degeneration, wet age-related macular degeneration, retinitis pigmentosa, dry eye, COVID-19, Poitz-Yage syndrome, musculoskeletal disorders, muscular dystrophy, amyotrophic lateral sclerosis, cachexia, endocrine disorders, polycystic ovary syndrome, cancer, hematologic malignancies, leukemia, plasmacytoma, Hodgkin's lymphoma, lung cancer, colorectal cancer, bowel cancer, urinary tract cancer, bladder cancer, vulvar cancer, endometrial cancer, ovarian cancer. Cancer, prostate cancer, pancreatic cancer, bone cancer, glioblastoma, breast cancer, gastric cancer, kidney cancer, metastatic renal cell carcinoma, prostate cancer, skin cancer, melanoma, liver cancer, hepatocellular carcinoma, asthenia, age-related increase in fat mass, sarcopenia, age-related hyperlipidemia, age-related hypertriglyceridemia, age-related hypercholesterolemia, age-related hepatic steatosis, age-related non-alcoholic fatty liver disease, age-related non-alcoholic fatty liver, age-related non-alcoholic steatohepatitis, age-related cardiovascular disease, age-related hypertension, age-related nephropathy, age-related skin disease, infectious diseases, viral diseases, viral hepatitis, hepatitis B, HIV infection, influenza infection, malaria, tuberculosis, allergic diseases, transplant rejection and graft-versus-host disease, wherein the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:89.

[0026] This disclosure also provides the use of an antigen-binding molecule that binds to and contacts the gp130 region shown in SEQ ID NO:89 to inhibit IL-6-mediated signaling, IL-11-mediated signaling, OSM-mediated signaling, CNTF-mediated signaling, CT-1-mediated signaling and / or LIF-mediated signaling.

[0027] This disclosure also provides a method for inhibiting IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and / or LIF-mediated signal transduction, the method comprising contacting a cell capable of IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and / or LIF-mediated signal transduction with an antigen-binding molecule, the antigen-binding molecule binding to gp130 and contacting the gp130 region shown in SEQ ID NO:89.

[0028] This disclosure also provides a method for inhibiting IL-6-mediated signaling, IL-11-mediated signaling, OSM-mediated signaling, CNTF-mediated signaling, CT-1-mediated signaling, and LIF-mediated signaling in a subject, the method comprising administering to the subject an antigen-binding molecule that binds to gp130 and contacts the gp130 region shown in SEQ ID NO:89.

[0029] In some implementations, the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12.

[0030] In some implementations, the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence having at least 70% amino acid sequence identity with the VH sequence shown in column A of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VL sequences shown in column B of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C. The VH and VL sequences are selected from the same row of Table C.

[0031] In some implementations, the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:218; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:204; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:20; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:195; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (e) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:214; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (f) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:215; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28.

[0032] In some implementations, the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence that is at least 70% amino acid sequence identical to the VH sequence shown in column A of rows 89, 90, 80-88, 74-79 or 2 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VH sequence shown in column B of rows 89, 90, 80-88, 74-79 or 2 of Table C. The VH and VL sequences are selected from the same row of Table C.

[0033] In some embodiments, the antigen-binding molecule is a multispecific antigen-binding molecule, and wherein the antigen-binding molecule further comprises an antigen-binding domain that binds to antigens other than gp130. Detailed Implementation

[0034] This disclosure provides an antigen-binding molecule that binds to gp130, which has novel biophysical and / or functional properties compared to antigen-binding molecules disclosed in the prior art.

[0035] This disclosure relates to a gp130-binding antigen-binding molecule that inhibits signal transduction mediated by a gp130-containing cytokine receptor complex. The antigen-binding molecule of this disclosure can be used to inhibit signal transduction mediated by IL-6 family cytokines.

[0036] Specifically, this disclosure provides a gp130-binding antigen-binding molecule that binds to the gp130 region, providing protection against... all Effective inhibition of IL-6, IL-11, OSM, CNTF, CT-1, and LIF-mediated signal transduction. In this paper, these antigen-binding molecules are referred to as "PAN" inhibitors of gp130.

[0037] Although the known anti-gp130 antibody B-R3 has previously been described as a PAN inhibitor of gp130-mediated signaling, this disclosure demonstrates that the antibody... in vitro In the assay using 10 ng / ml recombinant human LIF to stimulate A549 cells for 15 min, LIF-mediated signal transduction was not inhibited (see Example 7). Figure 8A ).

[0038] The gp130-binding antigen-binding molecule according to this disclosure also shows cross-reactivity with both human gp130 and mouse gp130. In contrast, BR-3 (see Example 1, Figure 1BThe known anti-gp130 antibodies mAb16692, mAb16680, mAb16673, mAb16646, mAb16636 and mAb16618 (see Tables 9 and 10 of WO 2019 / 126071 A1) did not show binding to mouse gp130.

[0039] gp130 Human gp130 (also known as IL6ST, CD130) is a protein identified by UniProt P40189. The structure and function of gp130 are described in [the original text is missing]. For example Silver and Hunter, J Leukoc Biol. (2010) 88(6):1145-1156 and Rose-John, Cold Spring Harb Perspect Biol. (2018) 10(2):a028415, the entire references of which are hereby incorporated by reference.

[0040] A typical isoform of human gp130 (isoform 1) has the amino acid sequence shown in SEQ ID NO:70. (The last sentence appears to be incomplete and unrelated to the preceding text. It likely refers to a separate topic: human gp130 isoform.) IL6ST Alternative splicing of the gene-encoded mRNA produces three major gp130 isoforms: isoform 1 (SEQ ID NO:70), isoform 2 (also known as gp130-RAPS; SEQ ID NO:71), and isoform 3 (SEQ ID NO:72). Isoform 2 differs from isoform 1 in that positions 325 to 329 of SEQ ID NO:70 are different, and positions 330-918 are missing. Positions 423 to 483 of SEQ ID NO:70 are absent in isoform 3.

[0041] A typical isoform of human gp130 comprises an N-terminal signal peptide (SEQ ID NO:73), followed by an extracellular domain (SEQ ID NO:75), a single-channel transmembrane domain (SEQ ID NO:76), and a C-terminal cytoplasmic domain (SEQ ID NO:77). The mature form of human gp130 isoform 1 is shown in SEQ ID NO:74.

[0042] The extracellular domain comprises an N-terminal Ig-like C2-type domain (SEQ ID NO:78), followed by five fibronectin type III (FNIII) domains (shown in SEQ ID NO:79, 80, 82, 83, and 84, respectively). The cytokine-binding module (CBM) of gp130 is formed by the Ig-like C2-type domain and FNIII domains 1 and 2 (SEQ ID NO:85). FNIII domain 2 contains the WSXWS motif shown in SEQ ID NO:81. The WSXWS motif is conserved in type I cytokine receptor peptides, and the WSXWS motif of gp130 is considered important for cytokine binding.

[0043] gp130 is a constituent polypeptide of all receptors in the IL-6 receptor family, providing signal transduction. Two gp130 polypeptides associate with two IL-6Rα polypeptides to form IL-6 receptors, and similarly, two gp130 polypeptides associate with two IL-11Rα polypeptides to form IL-11 receptors. gp130 associates with OSMRβ to form type II receptors for OSM, or with LIFRβ to form type I receptors for OSM, which also act as receptors for LIF and CT-1. gp130 also associates with LIFRβ and CNTFRα to form receptors for CNTF and CLC. gp130 associates with IL-27Rα to form IL-27 receptors, and with IL-12Rβ2 to form IL-35 receptors.

[0044] Following the formation of a complex with its receptor interacting partner and homologous ligand, gp130 is phosphorylated at tyrosine residues in its cytoplasmic domain (particularly Y767, Y814, Y905, and Y915), thereby triggering downstream signaling via the JAK / STAT and MAPK / ERK signaling pathways. gp130 can also trigger signaling via PI3K / AKT. Receptor binding leads to phosphorylation and activation of JAK1 and JAK2, which in turn phosphorylates STAT1, STAT3, and STAT5. Phosphorylation of gp130 tyrosine residues also leads to the recruitment and activation of SHP2, which in turn activates signaling via the Ras-ERK1 / ERK2 MAPK and PI3K / AKT signaling pathways.

[0045] In this specification, "gp130" means gp130 from any species, and includes gp130 isoforms, fragments, variants, or homologs from any species. In some embodiments, gp130 is derived from mammals ( For examplegp130 refers to gp130 of the subclass Theria, placental mammals, epitherians, preptotheria, archontana, and primates (rhesus monkeys, cynomolgous, non-human primates, or humans). In some embodiments, gp130 is human gp130, rhesus monkey gp130, mouse gp130, rat gp130, or canine gp130. In some embodiments, gp130 is human gp130 or mouse gp130.

[0046] As used in this article, a reference protein ( For example Isotypes, fragments, variants, or homologs of gp130 can be characterized as having at least 70% sequence identity with the amino acid sequence of the reference protein, preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100% amino acid sequence identity.

[0047] A "fragment" typically refers to a portion of a reference protein. A "variant" typically refers to an amino acid sequence that contains one or more amino acid substitutions, insertions, deletions, or other modifications relative to the reference protein's amino acid sequence, but retains a considerable degree of sequence identity with the reference protein's amino acid sequence. For example (At least 60%) of the protein. "Isotype" generally refers to a variant of the reference protein expressed by a species that is the same species as the reference protein. "Homologous" generally refers to a variant of the reference protein produced by a species that is different from the reference protein species. Homologous proteins include orthologous proteins. Homologous proteins of human gp130 include... For example Mouse gp130 (UniProt Q00560) and rat gp130 (UniProt P40190).

[0048] Isotypes, fragments, variants, or homologs of a given reference protein may optionally be characterized as being from a given species ( For example Immature or mature isoforms of related proteins in humans Right now The amino acid sequence in the form of (after processing to remove the signal peptide) has at least 70% (preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity.

[0049] According to this disclosure, isotypes, fragments, variants, or homologs of gp130 may optionally be characterized as being related to those from a given species ( For exampleThe amino acid sequence of immature or mature gp130 isoforms of human has at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) amino acid sequence identity.

[0050] Isotypes, fragments, variants, or homologs may optionally be functional isotypes, fragments, variants, or homologs. For example With reference gp130 ( For example The functional characteristics / activity of human gp130 isoforms 1) are determined by analysis using appropriate functional characteristic / activity assays. For example, isoforms, fragments, variants, or homologs of gp130 may be associated with IL-6Rα, IL-11Rα, OSMRβ, LIFRβ, and / or CNTFRα.

[0051] In some embodiments, gp130 comprises, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 70, 71, or 72.

[0052] In some embodiments, gp130 comprises, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 70 or 74.

[0053] The “fragment” of the reference protein may be of any length (in terms of amino acid number), although it may optionally be at least 25% of the length of the reference protein (i.e., the protein from which the fragment is derived), and the maximum length may be one of 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the length of the reference protein.

[0054] The minimum length of a gp130 fragment can be one of 10, 20, 30, 40, 50, 100, 150, 200, 300, 400, 500, 600, 700, 800, or 900 amino acids, and the maximum length can be one of 20, 30, 40, 50, 100, 150, 200, 300, 400, 500, 600, 700, 800, or 900 amino acids.

[0055] In some embodiments, the fragment of gp130 comprises, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO:74.

[0056] In some embodiments, the fragment of gp130 comprises, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO:75.

[0057] In some embodiments, the fragment of gp130 comprises, or consists of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO:85.

[0058] In this specification, "gp130-mediated signal transduction" refers to signal transduction mediated by gp130 and / or multimeric receptor complexes containing gp130. For example Signal transduction (including gp130 and another member of the IL-6 receptor family) is mediated by other cellular processes that transduce signals and control cellular activity.

[0059] gp130-mediated signal transduction can be mediated by polypeptide complexes containing gp130 ( Right now The association is mediated by a polypeptide complex containing one or more gp130 peptides. The polypeptide complexes according to this disclosure can be characterized by non-covalent, protein:protein interactions between the polypeptides / peptides. In some embodiments, association includes electrostatic interactions (…). For example Ionic bonds, hydrogen bonds) and / or van der Waals forces.

[0060] gp130-mediated signal transduction can be mediated by a heteropolymeric polypeptide complex containing one or more gp130 polypeptides and additionally containing one or more polypeptides from the IL-6 receptor family. For example One or more polypeptides selected from IL-6Rα, IL-11Rα, OSMRβ, LIFRβ, CNTFRα, IL-27Rα, and IL-12Rβ2.

[0061] In some implementations, gp130-mediated signaling can be mediated by polypeptide complexes that form receptors for IL-6 family cytokines. For example, gp130-mediated signaling can be mediated by polypeptide complexes that form receptors for IL-6, IL-11, OSM, LIF, CNTF, CT-1, CLC, IL-27, or IL-35.

[0062] In some implementations, gp130-mediated signal transduction may be mediated by another polypeptide comprising gp130 and the IL-6 receptor family ( For example The signal transduction is mediated by a peptide complex selected from IL-6Rα, IL-11Rα, OSMRβ, LIFRβ, CNTFRα, IL-27Rα, and IL-12Rβ2. In some embodiments, gp130-mediated signal transduction may be mediated by a peptide complex comprising gp130 and IL-6Rα. Right now The signal transduction is mediated by a gp130:IL-6Rα complex. In some embodiments, gp130-mediated signal transduction may be mediated by a peptide complex comprising gp130 and IL-11Rα. Right now gp130:IL-11Rα complex) mediates signal transduction. In some embodiments, gp130-mediated signal transduction may be mediated by a peptide complex comprising gp130 and OSMRβ. Right now gp130:OSMRβ complex) mediates signal transduction. In some embodiments, gp130-mediated signal transduction may be mediated by a peptide complex comprising gp130 and LIFRβ ( Right now The signal transduction is mediated by a gp130:LIFRβ complex. In some embodiments, gp130-mediated signal transduction may be mediated by a polypeptide complex comprising gp130, LIFRβ, and CNTFRα. Right now The signal transduction is mediated by a gp130:LIFRβ:CNTFRα complex. In some embodiments, gp130-mediated signal transduction can be mediated by a peptide complex comprising gp130 and IL-27Rα. Right now gp130:IL-27Rα complex) mediates the signal transduction. In some embodiments, gp130-mediated signal transduction may be mediated by a peptide complex comprising gp130 and IL-12Rβ2 ( Right now Gp130-mediated signaling is mediated by a peptide complex containing more than one gp130 peptide. In some embodiments, gp130-mediated signaling can be mediated by a peptide complex containing two gp130 peptides. In some embodiments, gp130-mediated signaling can be mediated by a peptide complex containing two gp130 peptides and IL-6Rα. Right nowThe signal transduction is mediated by a gp130:gp130:IL-6Rα complex. In some embodiments, gp130-mediated signal transduction can be mediated by a peptide complex comprising two gp130 peptides and IL-11Rα. Right now Mediated by the gp130:gp130:IL-11Rα complex.

[0063] The gp130-mediated signaling via the polypeptide complex described in the preceding paragraph can be triggered by the binding of its homologous cytokines. That is, in some embodiments, gp130-mediated signaling via a polypeptide complex comprising (i) gp130 and (ii) another polypeptide from the IL-6 receptor family can be triggered by the binding of a polypeptide complex formed by protein-protein interactions between cytokines and (i) and (ii). In some embodiments, gp130-mediated signaling occurs via IL-6 and a polypeptide complex comprising gp130 and IL-6Rα. Right now The signaling is triggered by the binding of the gp130:IL-6Rα complex. In some embodiments, gp130-mediated signaling is triggered by the binding of IL-11 to a peptide complex containing gp130 and IL-11Rα. Right now The binding of the gp130:IL-11Rα complex is triggered. In some embodiments, gp130-mediated signaling is triggered by OSM binding to a peptide complex containing gp130 and OSMRβ. Right now The signaling is triggered by the binding of the gp130:OSMRβ complex. In some embodiments, gp130-mediated signaling is triggered by the binding of OSM, LIF, or CT-1 to a peptide complex containing gp130 and LIFRβ. Right now The signaling is triggered by binding to a gp130:LIFRβ complex. In some embodiments, gp130-mediated signaling is triggered by CNTF or CLC with a peptide complex comprising gp130, LIFRβ, and CNTFRα. Right now The signaling is triggered by the binding of the gp130:LIFRβ:CNTFRα complex. In some embodiments, gp130-mediated signaling is triggered by the binding of IL-27 to a peptide complex containing gp130 and IL-27Rα. Right now The signaling is triggered by the binding of the gp130:IL-27Rα complex. In some embodiments, gp130-mediated signaling is triggered by the binding of IL-35 to a peptide complex containing gp130 and IL-12Rβ2. Right now The binding of gp130 (IL-12Rβ2 complex) triggers the process.

[0064] antigen-binding molecules This disclosure provides antigen-binding molecules capable of binding to gp130. Antigen-binding molecules capable of binding to gp130 can also be described as antigen-binding molecules that bind to gp130.

[0065] An "antigen-binding molecule" is a molecule that binds to a given target antigen. Antigen-binding molecules include antibodies (…). Right now Immunoglobulins (Ig) and their antigen-binding fragments. As used herein, "antibody" includes monoclonal antibodies, polyclonal antibodies, monospecific antibodies, and multispecific antibodies. For example Bispecific and trispecific wait Antibodies, and antibody-derived antigen-binding molecules, such as scFv, scFab, biantibodies, triantibodies, scFv-Fc, microantibodies, and single-domain antibodies. For example VhH) wait. Antibody antigen-binding fragments include For example Fv, Fab, F(ab')2, and F(ab') fragments. In some embodiments, the antigen-binding molecule may be an antibody or its antigen-binding fragment.

[0066] The antigen-binding molecules disclosed herein also include antibody-derived molecules. For example Molecules containing antigen-binding regions / domains derived from antibodies. Antibody-derived antigen-binding molecules may contain antigen-binding regions / domains, which include the antigen-binding region of the antibody (…). For example The antigen-binding fragment of the antibody or the antigen-binding region of the antibody may be present. In some embodiments, the antigen-binding region / domain of the antibody-derived antigen-binding molecule may be the Fv of the antibody. For example Provided in the form of scFv or dsFv) or Fab region, or the entire antibody, or containing the Fv or Fab region of the antibody, or the entire antibody. For example, the antigen-binding molecule according to this disclosure includes a (cytotoxic) drug portion ( For example Antibody-drug conjugates (ADCs) as described below. The antigen-binding molecules according to this disclosure also include multispecific antigen-binding molecules, such as immune cell adaptor molecules containing a domain for recruiting (effective) immune cells. For example The antigen-binding molecules disclosed herein also include BiTE, BiKE, and TriKE. These have been reviewed in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17: 418–434 and Ellerman, Methods (2019) 154:102–117 (both articles are hereby incorporated by reference in their entirety). The antigen-binding molecules disclosed herein also include chimeric antigen receptors (CARs), which are recombinant receptors that provide antigen binding and T-cell activation functions (the structure, function, and engineering of CARs are described in detail in the original text). For example Dotti et al. Immunol Rev (2014) 257(1) and Jayaraman et al. These two articles are reviewed in EBioMedicine (2020) 58:102931, and are hereby included in their entirety by citation.

[0067] The antigen-binding molecule disclosed herein comprises one or more portions capable of binding to a target antigen. In some embodiments, the portion capable of binding to the target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) capable of specifically binding to the target antigen. In some embodiments, the portion capable of binding to the target antigen comprises or is composed of an aptamer capable of binding to the target antigen. For example Nucleic acid aptamers (e.g., reviewed in Zhou and Rossi Nat Rev DrugDiscov. 2017 16(3):181-202). In some embodiments, the portion capable of binding to the target antigen comprises or is composed of an antigen-binding peptide / peptide. For example Peptide aptamers, thioreduction proteins, monomers, anticarrier proteins (anticalin), Kunitz domains, high-affinity polymers (avimer), knottin, fynomers, atrimers, DARPin, affibody, nanobodies ( Right now Single-domain antibodies (sdAbs), ubiquitin molecules (affilin), armadillo repeat proteins (ArmRP), OBody or fibronectin – For example In Reverdatto et al. A review was conducted in Curr Top Med Chem. 2015; 15(12): 1082–1101, which is hereby incorporated in its entirety by reference (see also: Curr Top Med Chem. 2015; 15(12): 1082–1101). For example Boersma et al. , J Biol Chem (2011)286:41273-85 and Emanuel et al. , Mabs (2011) 3:38-48).

[0068] As used herein, a "peptide" is a chain of two or more amino acid monomers linked by peptide bonds. Peptides are typically between 2 and 50 amino acids in length. A "polypeptide" is a polymer chain containing two or more peptides. Polypeptides are typically longer than about 50 amino acids.

[0069] The antigen-binding molecules disclosed herein typically include an antigen-binding domain comprising the VH and VL of an antibody capable of specifically binding to a target antigen. The antigen-binding domain formed by the VH and VL may also be referred to herein as the Fv region.

[0070] The antigen-binding molecule may be an antigen-binding polypeptide or an antigen-binding polypeptide complex, or may contain an antigen-binding polypeptide or an antigen-binding polypeptide complex. The antigen-binding molecule may contain more than one polypeptide that together form an antigen-binding domain. The polypeptides may be covalently associated or non-covalently associated. In some embodiments, the polypeptide forms a portion of a larger polypeptide containing said polypeptide ( For example In the case of scFv containing VH and VL, or in the case of scFab containing VH-CH1 and VL-CL).

[0071] Antigen-binding molecules can refer to more than one polypeptide. For example Non-covalent or covalent complexes of 2, 3, 4, 6, or 8 polypeptides. For example An IgG-like antigen-binding molecule comprising two heavy chain polypeptides and two light chain polypeptides.

[0072] The antigen-binding molecules disclosed herein can be designed and prepared using sequences of monoclonal antibodies (mAbs) capable of binding to gp130. Antigen-binding regions of antibodies, such as single-chain variable fragments (scFv), Fab, and F(ab')2 fragments, can also be used / provided. An "antigen-binding region" is any fragment of an antibody that binds to a target to which a given antibody has specificity.

[0073] Antibodies typically contain six complementarity-determining regions (CDRs): three in the heavy chain variable (VH) region (HC-CDR1, HC-CDR2, and HC-CDR3) and three in the light chain variable (VL) region (LC-CDR1, LC-CDR2, and LC-CDR3). Together, the six CDRs define the complementary sites of the antibody, which are the portions of the antibody that bind to the target antigen.

[0074] The VH and VL regions contain frame areas (FRs) on both sides of each CDR, which provide support for the CDR. From the N end to the C end, the VH region contains the following structure: N end - [HC-FR1] - [HC-CDR1] - [HC-FR2] - [HC-CDR2] - [HC-FR3] - [HC-CDR3] - [HC-FR4] - C end; and the VL region contains the following structure: N end - [LC-FR1] - [LC-CDR1] - [LC-FR2] - [LC-CDR2] - [LC-FR3] - [LC-CDR3] - [LC-FR4] - C end.

[0075] There are several different conventions for defining antibody CDRs and FRs, such as those described in Kabat. et al. , Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), Chothia et al. Those mentioned in J. Mol. Biol. 196:901-917 (1987), and VBASE2, as described in Retter et al. Nucl. Acids Res. (2005) 33(suppl 1): D671-D674. The CDR and FR of the VH and VL regions of the antibody clones described in this article are based on the international IMGT (ImMunoGeneTics) information system (LeFrance). et al. Defined in Nucleic Acids Res. (2015) 43 (Databaseissue):D413-22, it uses the IMGT V-DOMAIN numbering rules, as described in Lefranc. et al. , Dev.Comp. Immunol. (2003) 27:55-77. In a preferred embodiment, the CDR and FR of the antigen-binding molecule mentioned herein are defined according to the IMGT information system.

[0076] In some embodiments, the antigen-binding molecule includes a CDR of the antigen-binding molecule that binds to gp130. In some embodiments, the antigen-binding molecule includes a FR of the antigen-binding molecule that binds to gp130. In some embodiments, the antigen-binding molecule includes both a CDR and a FR of the antigen-binding molecule that binds to gp130. That is, in some embodiments, the antigen-binding molecule includes a VH region and a VL region of the antigen-binding molecule that binds to gp130.

[0077] In some implementations, the antigen-binding molecule comprises the antibody described herein. For example The CDR, FR and / or VH and / or VL regions of the antibody described in Table C of this article, or the antibody derived from the antibody described in this article. For exampleThe antibody described in Table C of this document contains the CDR, FR, and / or VH and / or VL regions of those antibodies. In some embodiments, the antigen-binding molecule comprises, or is derived from, the CDR, FR, and / or VH and / or VL regions of the gp130-binding antibody described herein. In some embodiments, the gp130-binding antibody is selected from the antibodies described in Table C of this document.

[0078] In some implementations, the gp130 binding antibody is selected from B035-C03-A3, A3_4.2 VH / A3_3 VL, A3_4.2 VH / A3_4 VL, A3_4.2 VH / A3_5 VL, A3_3.2 VH / A3_3 VL, A3_3.2 VH / A3_4 VL, A3_6.2 VH / A3_3 VL, A3_6.2 VH / A3_4 VL, A3_6.2_I100T / A3_3_G51A, A3_3.2_I100T / A3_4_G51A, CSP-S-5H8, 5H8CVH4_D98G / 5H8CVL3, 5H8CVH4_D98G / 5H8CVL4, 5H8CVH4_D98G / 5H8GVL3, 5H8GVH4_D98G / 5H8CVL4 and 5H8GVH4_D98G / 5H8GVL3.

[0079] In some implementations, the gp130 binding antibody is selected from B035-C03-A3, A3_6.2_I100T / A3_3_G51A, A3_3.2_I100T / A3_4_G51A, CSP-S-5H8, 5H8CVH4_D98G / 5H8CVL3, 5H8CVH4_D98G / 5H8CVL4, 5H8CVH4_D98G / 5H8GVL3, 5H8GVH4_D98G / 5H8CVL4, and 5H8GVH4_D98G / 5H8GVL3.

[0080] In some implementations, the antigen-binding molecule comprises: Includes items 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 as shown in Table A. The HC-CDR1 shown in column A of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. The VH regions of HC-CDR1 (or its variants, wherein 1, 2, or 3 amino acids in HC-CDR1 are replaced by another amino acid), HC-CDR2 (or its variants, wherein 1, 2, or 3 amino acids in HC-CDR2 are replaced by another amino acid), and HC-CDR3 (or its variants, wherein 1, 2, or 3 amino acids in HC-CDR3 are replaced by another amino acid), wherein the HC-CDR1, HC-CDR2, and HC-CDR3 sequences in column A are selected from the same row of Table A.

[0081] For example, in some embodiments, the antigen-binding molecule comprises a VH region containing HC-CDR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:2 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:2, HC-CDR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:3 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:4, and HC-CDR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:4 are substituted with another amino acid). It should be understood that the HC-CDR1, HC-CDR2, and HC-CDR3 sequences in the preceding sentence are selected from column A of the same row (row 1) of Table A.

[0082] In some implementations, the antigen-binding molecule comprises: Includes items 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 4... (as shown in Table B) The HC-FR1 shown in column A of rows 8, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. The VH regions of HC-FR1 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR1 are replaced by another amino acid), HC-FR2 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR2 are replaced by another amino acid), HC-FR3 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR3 are replaced by another amino acid), and HC-FR4 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR4 are replaced by another amino acid), wherein the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences in column A are selected from the same row of Table A.

[0083] For example, in some embodiments, the antigen-binding molecule comprises a VH region containing HC-FR1 (or a variant thereof, wherein 1, 2, or 3 amino acids of SEQ ID NO:5 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:5, HC-FR2 (or a variant thereof, wherein 1, 2, or 3 amino acids of SEQ ID NO:6 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:7, HC-FR3 (or a variant thereof, wherein 1, 2, or 3 amino acids of SEQ ID NO:7 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:8, and HC-FR4 (or a variant thereof, wherein 1, 2, or 3 amino acids of SEQ ID NO:8 are substituted with another amino acid). It should be understood that the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences in the preceding sentence are selected from column A of the same row (row 1) of Table B.

[0084] In some implementations, the antigen-binding molecule comprises: Includes the following VH zones: As shown in Table A, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48... HC-CDR1 shown in column A of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 (or variants thereof, wherein 1, 2, or 3 amino acids in HC-CDR1 are replaced by another amino acid), HC-CDR2 (or variants thereof, wherein 1, 2, or 3 amino acids in HC-CDR2 are replaced by another amino acid), and HC-CDR3 (or variants thereof, wherein 1, 2, or 3 amino acids in HC-CDR3 are replaced by another amino acid), wherein the HC-CDR1, HC-CDR2, and HC-CDR3 sequences in column A are selected from the same row of Table A; and As shown in Table B, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 The HC-FR1 shown in column A of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. (or its variants, wherein 1, 2, or 3 amino acids in HC-FR1 are replaced by another amino acid), HC-FR2 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR2 are replaced by another amino acid), HC-FR3 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR3 are replaced by another amino acid), and HC-FR4 (or its variants, wherein 1, 2, or 3 amino acids in HC-FR4 are replaced by another amino acid), wherein the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences in column A are selected from the same row of Table A.

[0085] In some implementations, the antigen-binding molecule comprises: Includes the following VH zones: As shown in Table A, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48... The following are listed in column A of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90: HC-CDR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-CDR1 are replaced by another amino acid), HC-CDR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-CDR2 are replaced by another amino acid), and HC-CDR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-CDR3 are replaced by another amino acid); and... As shown in Table B, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 The HC-FR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-FR1 are replaced by another amino acid), HC-FR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-FR2 are replaced by another amino acid), HC-FR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-FR3 are replaced by another amino acid), and HC-FR4 (or a variant thereof, wherein 1, 2, or 3 amino acids in HC-FR4 are replaced by another amino acid) shown in column A of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90; The HC-CDR1, HC-CDR2, and HC-CDR3 sequences in column A of Table A and the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences in column B of Table B are selected from rows with the same row number.

[0086] For example, in some embodiments, the antigen-binding molecule comprises a VH region, said VH region comprising: HC-CDR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:2 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:2; HC-FR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:3 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:3; HC-CDR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:4 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:4; HC-FR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:5 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:6; HC-FR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:6 are substituted with another amino acid) having the amino acid sequence of SEQ ID NO:7; and HC-FR3 (or a variant thereof, wherein SEQ ID NO:4 is substituted with another amino acid). HC-FR4 (or a variant thereof, wherein 1, 2, or 3 amino acids in SEQ ID NO:7 are replaced by another amino acid) and having the amino acid sequence of SEQ ID NO:8. It should be understood that the HC-CDR1, HC-CDR2, and HC-CDR3 sequences in the preceding sentence are selected from column A of row 1 in Table A, while the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences are selected from column A of the same row (row 1) in Table B.

[0087] In some embodiments, the antigen-binding molecule includes a VH region, said VH region being selected from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, and 45th regions of Table C. The amino acid sequence of the VH region sequence in column A of row 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 contains at least 70% (preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) amino acid sequence identity.

[0088] In some implementations, the antigen-binding molecule comprises: Includes items 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 as shown in Table A. LC-CDR1 shown in column B of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. The VL regions of LC-CDR1 (or its variants, wherein 1, 2, or 3 amino acids in LC-CDR1 are replaced by another amino acid), LC-CDR2 (or its variants, wherein 1, 2, or 3 amino acids in LC-CDR2 are replaced by another amino acid), and LC-CDR3 (or its variants, wherein 1, 2, or 3 amino acids in LC-CDR3 are replaced by another amino acid), wherein the LC-CDR1, LC-CDR2, and LC-CDR3 sequences in column B are selected from the same row of Table A.

[0089] In some implementations, the antigen-binding molecule comprises: Includes items 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 4... (as shown in Table B) LC-FR1 shown in column B of rows 8, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. The VL regions of LC-FR1 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR1 are replaced by another amino acid), LC-FR2 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR2 are replaced by another amino acid), LC-FR3 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR3 are replaced by another amino acid), and LC-FR4 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR4 are replaced by another amino acid), wherein the LC-FR1, LC-FR2, LC-FR3, and LC-FR4 sequences in column B are selected from the same row of Table A.

[0090] In some implementations, the antigen-binding molecule comprises: Includes the following VL zones: As shown in Table A, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48... LC-CDR1 shown in column B of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 (or variants thereof, wherein 1, 2, or 3 amino acids in LC-CDR1 are replaced by another amino acid), LC-CDR2 (or variants thereof, wherein 1, 2, or 3 amino acids in LC-CDR2 are replaced by another amino acid), and LC-CDR3 (or variants thereof, wherein 1, 2, or 3 amino acids in LC-CDR3 are replaced by another amino acid), wherein the LC-CDR1, LC-CDR2, and LC-CDR3 sequences in column B are selected from the same row of Table A; and As shown in Table B, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 The LC-FR1 shown in column B of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90. (or its variants, wherein 1, 2, or 3 amino acids in LC-FR1 are replaced by another amino acid), LC-FR2 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR2 are replaced by another amino acid), LC-FR3 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR3 are replaced by another amino acid), and LC-FR4 (or its variants, wherein 1, 2, or 3 amino acids in LC-FR4 are replaced by another amino acid), wherein the LC-FR1, LC-FR2, LC-FR3, and LC-FR4 sequences in column B are selected from the same row of Table A.

[0091] In some implementations, the antigen-binding molecule comprises: Includes the following VL zones: As shown in Table A, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48... The LC-CDR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-CDR1 are replaced by another amino acid), LC-CDR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-CDR2 are replaced by another amino acid), and LC-CDR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-CDR3 are replaced by another amino acid) shown in column B of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90; and As shown in Table B, numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, and 48 The LC-FR1 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-FR1 are replaced by another amino acid), LC-FR2 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-FR2 are replaced by another amino acid), LC-FR3 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-FR3 are replaced by another amino acid), and LC-FR4 (or a variant thereof, wherein 1, 2, or 3 amino acids in LC-FR4 are replaced by another amino acid) shown in column B of rows 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90; The LC-CDR1, LC-CDR2, and LC-CDR3 sequences in column B of Table A and the LC-FR1, LC-FR2, LC-FR3, and LC-FR4 sequences in column B of Table B are selected from rows with the same row number.

[0092] In some embodiments, the antigen-binding molecule includes a VL region, said VL region being selected from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, and 45th regions of Table C. The amino acid sequence of the VH region sequence in column B of row 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 contains at least 70% (preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) amino acid sequence identity.

[0093] In some embodiments, the antigen-binding molecule comprises a VH region according to any of the embodiments described herein and a VL region according to any of the embodiments described herein.

[0094] In embodiments according to this disclosure, one or more amino acids are substituted by another amino acid. Substitution includes replacing an amino acid residue with a different "replacement" amino acid residue. The substituted amino acid residue according to this disclosure may be a naturally occurring amino acid residue (…). Right now Encoded by the genetic code, the amino acid residues at the corresponding positions of the equivalent unsubstituted amino acid sequence are different and are selected from: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). In some embodiments, the substituted amino acid may be a non-naturally occurring amino acid residue – Right now Amino acid residues other than those mentioned in the preceding sentence. Examples of non-naturally occurring amino acid residues include ornithine, ornithine, homoserine, aib, and other amino acid residue analogs, such as those described in Ellman, et al.Those of Meth. Enzym. 202 (1991) 301-336.

[0095] In some embodiments, the substitution may be biochemically conserved. In some embodiments, where the amino acid to be substituted is provided in one of rows 1 to 5 of the table below, the substituted amino acid is another, different amino acid provided in the same row: For example, in some embodiments where the substitution is a Met residue, the substituted amino acid may be selected from Ala, Val, Leu, Ile, Trp, Tyr, Phe, and ortholeucine.

[0096] In some embodiments, the substituted amino acid in the substitution may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, the substituted amino acid in the substitution may have the same side chain charge (at pH 7.4) as the amino acid residue it replaces. In other words, in some embodiments, a nonpolar amino acid is replaced by another different nonpolar amino acid. In some embodiments, a polar amino acid is replaced by another different polar amino acid. In some embodiments, an acidic polar amino acid is replaced by another different acidic polar amino acid. In some embodiments, a basic polar amino acid is replaced by another different basic polar amino acid. In some embodiments, an uncharged amino acid is replaced by another different uncharged amino acid. In some embodiments, a positively charged amino acid is replaced by another different positively charged amino acid. In some embodiments, a negatively charged amino acid is replaced by another different negatively charged amino acid.

[0097] In some embodiments, the substitution may be functionally conserved. That is, in some embodiments, the substitution may not affect (or may not significantly affect) one or more functional properties of the antigen-binding molecule containing the substitution compared to an equivalent unsubstituted molecule. For example Target binding).

[0098] In some implementations, substitution can reduce the multireactivity of antigen-binding molecules. As used herein, “multireactivity” refers to low-affinity interactions between antigen-binding molecules and molecules other than the target antigen via general nonspecific chemistry. The multireactivity of antigen-binding molecules can also be referred to as the “stickiness” of antigen-binding molecules. High multireactivity / stickiness of antigen-binding molecules can lead to unacceptably poor PK, potency, bioavailability, and / or immunogenicity.

[0099] In some implementations, one or more amino acids are replaced by another amino acid to reduce / prevent the multireactivity of antigen-binding molecules.

[0100] The multireactivity of therapeutic antibodies For example A review was conducted by Cunningham et al., MAbs, 2021, 13(1):1999-195, which is hereby incorporated in its entirety by reference. The multireactivity of antigen-binding molecules can... For example The assay was performed using a series of structurally different coated antigens. For example Escherichia coli (E. coli) LPS, cardiolipin, insulin, dsDNA, ssDNA, KLH) or via Computer simulation method( For example As reviewed by Harvey et al., Nature Communications, 2022, 13:7554 (the literature is hereby evaluated by reference in its entirety).

[0101] In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:4, which includes replacing I at position 8 of SEQ ID NO:4 with another amino acid.

[0102] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 1, 94, 98, 174, 112, or 117, wherein the VH region includes the I at position 100 (according to Kabat). et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 104 (corresponding to position 104 of SEQ ID NO:1) is replaced with another amino acid.

[0103] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 64, 65, 130, 132, 176, 134, 136, 131, 133, 177, 135, or 137, wherein the VH region includes the I at position 100 (according to Kabat). et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 104 (corresponding to position 104 of SEQ ID NO:64) is replaced with another amino acid.

[0104] In some embodiments according to the preceding three paragraphs, I is substituted with T, S, E, Q, K, H, G, V, Y, A, C, L, F, P, D, N, M, W, or R. In some embodiments, I is substituted with T, S, E, Q, K, or H. In some embodiments, I is substituted with T or K. In some embodiments, I is substituted with T. In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:4, which includes replacing V at position 7 of SEQ ID NO:4 with another amino acid.

[0105] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO:1, 94, 98, 174, 112, or 117, wherein the VH region includes the V at position 99 (according to Kabat). et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 103 (corresponding to position 103 of SEQ ID NO:1) is replaced with another amino acid.

[0106] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 64, 65, 130, 132, 176, 134, 136, 131, 133, 177, 135, or 137, wherein the VH region includes the V at position 99 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 103 (corresponding to position 103 of SEQ ID NO: 64) is replaced with another amino acid.

[0107] In some embodiments according to the preceding three paragraphs, V is replaced by G, S, T, E, Q, K, H, I, C, Y, A, L, F, P, D, N, M, W, or R. In some embodiments, V is replaced by G or S.

[0108] In some implementations, the replacement can prevent / reduce the degradation of antigen-binding molecules. Antigen-binding molecules may degrade during production, processing, storage, handling, and / or application. Degradation can be physical and / or chemical. The degradation process... For example Chemical degradation processes include isomerization, oxidation, cleavage, deamination, hydrolysis, deglycosylation, racemization, disulfide bond cleavage and formation, Maillard reaction, and β-elimination.

[0109] Oxidation is a common chemical modification in monoclonal antibodies and can be caused by reactive oxygen species. Oxidation can occur on cysteine, methionine, tryptophan, tyrosine, lysine, and other amino acid residues. Oxidation of amino acid residues can affect protein structure and / or function. In some embodiments, one or more amino acids are substituted with another amino acid to remove the oxidation site. In some embodiments, one or more easily oxidized amino acids are replaced with another amino acid (…). For example The amino acid is replaced by another amino acid (which is less susceptible to oxidation). In some embodiments, one or more amino acids identified as oxidizable or easily oxidizable are replaced by another amino acid (which is less susceptible to oxidation). For example (Amino acids that are not easily oxidized or are not very easily oxidized) are substituted.

[0110] Can For example pass Computer simulation method( For exampleAs reviewed in Vassa S, Mabs, 2022, 14(1):2023938, the literature mentioned hereby incorporated in its entirety by reference) or by forced oxidation determination ( For example Aspartate (Asp) isomerization is a spontaneous, non-enzymatic post-translational modification that leads to changes in the protein backbone structure, which is common during the production and storage of therapeutic antibodies. Aspartate (Asp) isomerization is often performed in structurally flexible regions such as complementarity-determining regions (CDRs) in antibodies, with higher isomerization rates in Asp–Gly (DG), Asp–Ser (DS), and Asp–Thr (DT) motifs. In some embodiments, one or more isomerizable amino acids are replaced by another amino acid (…). For example The amino acid motif is replaced by an amino acid that is not easily isomerized or isomerized less readily. In some embodiments, one or more easily isomerized amino acids present in the amino acid motif ( For example Asp–Gly (DG), Asp–Ser (DS), or Asp–Thr (DT) are reacted with another amino acid ( For example (Amino acids that reduce the sensitivity of amino acid isomerization in the motif) are substituted.

[0111] The sensitivity of antigen-binding molecules to isomerization can be assessed by in vitro assays. For example As described in Lu et al., MAbs, 2019, 11(1):45-57, the entire reference is hereby incorporated by reference.

[0112] In some embodiments, the VL region according to this disclosure comprises LC-CDR2 according to SEQ ID NO:11, which includes replacing D at position 1 of SEQ ID NO:11 with another amino acid. In some embodiments, the VL region according to this disclosure comprises LC-CDR2 according to SEQ ID NO:192, which includes replacing D at position 1 of SEQ ID NO:192 with another amino acid.

[0113] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106, 109, 186, 187, 188, 189, 190 or 191, wherein the VL region includes the D at position 50 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 50, corresponding to position 50 of SEQ ID NO:9, is replaced by another amino acid.

[0114] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139, 140, 245, 246, 247, 248, 249 or 250), wherein the polypeptide includes D at position 50 (according to Kabat). et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 50, corresponding to position 50 of SEQ ID NO:66, is replaced by another amino acid.

[0115] In some embodiments according to the preceding three paragraphs, D is replaced by A, E, N, C, G, I, L, M, F, P, S, T, W, Y, V, R, K, H, or Q. In some embodiments, D is replaced by A, E, N, C, G, I, L, M, F, P, S, T, W, Y, or V. In some embodiments, D is replaced by A or E. In some embodiments, D is replaced by A.

[0116] In some embodiments, the VL region according to this disclosure comprises LC-CDR2 according to SEQ ID NO:11, which includes replacing the G at position 2 of SEQ ID NO:11 with another amino acid. In some embodiments, the VL region according to this disclosure comprises LC-CDR2 according to SEQ ID NO:149, which includes replacing the G at position 2 of SEQ ID NO:149 with another amino acid. In some embodiments, the VL region according to this disclosure comprises LC-CDR2 according to SEQ ID NO:162, which includes replacing the G at position 2 of SEQ ID NO:162 with another amino acid.

[0117] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106, 109, 143, 144, 145, 146, 147, 148, 156, 157, 158, 159, 160 or 161, wherein the VL region includes the G at position 51 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 51, corresponding to position 51 of SEQ ID NO:9, is replaced by another amino acid.

[0118] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139, 140, 150, 151, 152, 153, 154, 155, 163, 164, 165, 166, 167 or 168), wherein the polypeptide includes G at position 51 (according to Kabat). et al.Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 51, corresponding to position 51 of SEQ ID NO:66, is replaced with another amino acid.

[0119] In some embodiments according to the preceding three paragraphs, G is replaced by A, E, N, C, I, D, L, M, F, P, S, T, W, Y, V, R, K, H, or Q. In some embodiments, G is replaced by A.

[0120] In some embodiments, the VL region according to this disclosure comprises LC-FR3 according to SEQ ID NO:15, which includes replacing L at position 2 of SEQ ID NO:15 with another amino acid.

[0121] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106, 109, 143, 144, 145, 146, 147, 148, 156, 157, 158, 159, 160 or 161, wherein the VL region includes the L at position 54 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 54, corresponding to position 54 of SEQ ID NO:9, is replaced with another amino acid.

[0122] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139, 140, 150, 151, 152, 153, 154, 155, 163, 164, 165, 166, 167 or 168), wherein the polypeptide includes L at position 54 (according to Kabat). et al.Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 54, corresponding to position 54 of SEQ ID NO:66, is replaced with another amino acid.

[0123] In some embodiments according to the preceding three paragraphs, L is replaced by S, A, E, N, C, I, D, G, M, F, P, T, W, Y, V, R, K, H, or Q. In some embodiments, L is replaced by S.

[0124] In some embodiments, the VL region according to this disclosure comprises LC-FR3 according to SEQ ID NO:108, which includes replacing the V at position 6 of SEQ ID NO:108 with another amino acid. In some embodiments, the VL region according to this disclosure comprises LC-FR3 according to SEQ ID NO:111, which includes replacing the V at position 6 of SEQ ID NO:111 with another amino acid.

[0125] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 106, 109, 145, 146, 147, 148, 158, 159, 160 or 161, wherein the VL region includes the V at position 58 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 58, corresponding to position 58 of SEQ ID NO: 106, is replaced by another amino acid.

[0126] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 139, 140, 152, 153, 154, 155, 165, 166, 167 or 168, wherein the polypeptide includes a V at position 58 (according to Kabat). et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 58, corresponding to position 58 of SEQ ID NO:139, is replaced by another amino acid.

[0127] In some embodiments according to the preceding three paragraphs, V is substituted with G, A, E, N, C, I, D, L, M, F, P, S, T, W, Y, R, K, H, or Q. In some embodiments, V is substituted with G. In some embodiments, the VL region according to this disclosure comprises LC-FR3 having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, or 97%) or higher amino acid sequence identity with SEQ ID NO: 124, 129, 15, 104, 108, or 111, wherein the amino acid at position 30 of SEQ ID NO: 15 is D.

[0128] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106, 109, 147, 148, 143, 144, 145, 146, 160, 161, 156, 157, 158 or 159. For example, Contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106 or 109; For exampleIt contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 147, 148, 143, 144, 145 or 146; For example Containing at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 160, 161, 156, 157, 158 or 159, wherein the amino acid corresponding to position 82a (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 82, which corresponds to position 82 of SEQ ID NO:9 is D.

[0129] In some embodiments, the polypeptide according to this disclosure comprises at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139, 140, 154, 155, 150, 151, 152, 153, 167, 168, 163, 164, 165 or 166. For example, Contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139 or 140; For example It contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 154, 155, 150, 151, 152 or 153; For example Containing at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 167, 168, 163, 164, 165 or 166, wherein the amino acid corresponding to position 82a (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 82, which corresponds to position 82 of SEQ ID NO:66, is D.

[0130] In some embodiments, the VL region according to this disclosure comprises LC-FR3 having at least 60% (preferably one of 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%) or higher amino acid sequence identity with SEQ ID NO: 124, 129, 15, 104, 108, or 111, wherein the amino acid at position 30 corresponding to SEQ ID NO: 15 is D, and wherein the amino acid sequence is relative to the reference amino acid sequence ( Right now The amino acid sequences selected from SEQ ID NO: 124, 129, 15, 104, 108, and 111 contain one or more ( For example Amino acid substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14, wherein one or more amino acid substitutions are located at positions corresponding to the following positions in the reference sequence: 22, 23, 24, 25, 26, 27, 28, 29, 31, 32, 33, 34, 35, 36 (i.e., one or more amino acid substitutions are located within the 8 positions immediately adjacent to the N-terminus of position 30 (in the case of the amino acid sequence of LC-FR3), and / or within the 6 positions immediately adjacent to the C-terminus of position 30).

[0131] In some embodiments, the VL region according to this disclosure contains at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106, 109, 147, 148, 143, 144, 145, 146, 160, 161, 156, 157, 158 or 159. For example, Contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 121, 126, 9, 102, 106 or 109; For exampleIt contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 147, 148, 143, 144, 145 or 146; For example Containing at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 160, 161, 156, 157, 158 or 159, wherein the amino acid corresponding to position 82a (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, which corresponds to position 82 of SEQ ID NO:9 is D, and wherein the amino acid sequence contains one or more ( ) relative to the reference amino acid sequence. For example Amino acid substitutions of one or more of the following positions in the reference sequence: 74, 75, 76, 77, 78, 79, 80, 81, 83, 84, 85, 86, 87, 88, 89, 90 (i.e., one or more amino acid substitutions are located within the eight positions immediately adjacent to the N-terminus of position 82 (in the case of the amino acid sequence in the VL region), and / or within the eight positions immediately adjacent to the C-terminus of position 82).

[0132] In some embodiments, the polypeptide according to this disclosure comprises at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139, 140, 154, 155, 150, 151, 152, 153, 167, 168, 163, 164, 165 or 166. For example, Contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 141, 142, 66, 138, 139 or 140; For exampleIt contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 154, 155, 150, 151, 152 or 153; For example Containing at least 70% (preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 167, 168, 163, 164, 165 or 166, wherein the amino acid corresponding to position 82a (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, which corresponds to position 82 of SEQ ID NO:66 is D, and wherein the amino acid sequence contains one or more ( ) relative to the reference amino acid sequence. For example Amino acid substitutions of one or more of the following positions in the reference sequence: 74, 75, 76, 77, 78, 79, 80, 81, 83, 84, 85, 86, 87, 88, 89, 90 (i.e., one or more amino acid substitutions are located within the eight positions immediately adjacent to the N-terminus of position 82 (in the case of the amino acid sequence of the polypeptide), and / or within the eight positions immediately adjacent to the C-terminus of position 82).

[0133] In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:20, which includes replacing the D at position 6 of SEQ ID NO:20 with another amino acid. In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:195, which includes replacing the D at position 6 of SEQ ID NO:195 with another amino acid. In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:204, which includes replacing the D at position 6 of SEQ ID NO:204 with another amino acid.

[0134] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 17, 193, 194, 202 or 203, wherein the polypeptide includes D at position 98 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 101, corresponding to position 101 of SEQ ID NO:17, is replaced by another amino acid.

[0135] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 67, 68, 251, 252, 253 or 254, wherein the VH region includes D at position 98 (according to Kabat) et al. Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 101, corresponding to position 101 of SEQ ID NO:67, is replaced with another amino acid.

[0136] In some embodiments according to the preceding three paragraphs, D is replaced by G, S, E, A, N, C, I, L, M, F, P, T, W, Y, V, R, K, H, or Q. In some embodiments, D is replaced by G, S, E, or A. In some embodiments, D is replaced by G or S. In some embodiments, D is replaced by G.

[0137] In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:20, which includes replacing the G at position 7 of SEQ ID NO:20 with another amino acid. In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:195, which includes replacing the G at position 7 of SEQ ID NO:195 with another amino acid. In some embodiments, the VH region according to this disclosure comprises HC-CDR3 according to SEQ ID NO:204, which includes replacing the G at position 7 of SEQ ID NO:204 with another amino acid.

[0138] In some embodiments, the polypeptide according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) or higher amino acid sequence identity with SEQ ID NO: 17, 193, 194, 202, 203, 211, 212, 213, 216 or 217, wherein the polypeptide includes G at position 99 (according to Kabat) wait people Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 102 (corresponding to position 102 of SEQ ID NO: 17) is replaced with another amino acid.

[0139] In some embodiments, the VH region according to this disclosure contains at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%) or higher amino acid sequence identity with SEQ ID NO: 67, 68, 251, 252, 253, 254, 255, 256, 257, 258 or 259, wherein the VH region includes G at position 99 (according to Kabat) wait people Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, number 102 (corresponding to position 102 of SEQ ID NO: 67) is replaced with another amino acid.

[0140] In some embodiments according to the preceding three paragraphs, G is replaced by A, Y, D, S, E, N, C, I, L, M, F, P, T, W, V, R, K, H, or Q. In some embodiments, G is replaced by A or Y.

[0141] The VH and VL regions of the antigen-binding region of the antibody together constitute the Fv region. In some embodiments, the antigen-binding molecule according to this disclosure comprises or consists of an Fv region that binds to gp130. In some embodiments, the VH and VL regions of the Fv are provided in the form of a single polypeptide linked by a linker sequence. Right now Single-chain Fv (scFv). In some embodiments, the VH and VL regions of the Fv are provided as separate polypeptides, stabilized by introducing one or more cysteine ​​residues to form interchain disulfide bonds. Right now Disulfide bond-stabilized Fv (dsFv).

[0142] The VL region and light chain constant (CL) region of the antigen-binding region of an antibody, together with the VH region and heavy chain constant 1 (CH1) region, constitute the Fab region. In some embodiments, the antigen-binding molecule includes the Fab region, which comprises VH, CH1, VL, and CL (…). For example Cκ or Cλ). In some embodiments, the Fab region contains a polypeptide containing VH and CH1 (Cκ or Cλ). For example VH-CH1 fusion peptide), and peptides containing VL and CL ( For example VL-CL fusion peptide). In some embodiments, the Fab region contains a peptide containing VH and CL. For example VH-CL fusion peptide), and peptides containing VL and CH ( For example VL-CH1 fusion peptide); that is, in some embodiments, the Fab region is a CrossFab region. In some embodiments, the VH, CH1, VL, and CL regions of the Fab or CrossFab are provided as single peptides linked via linker regions. Right now Single-chain Fab (scFab) or single-chain CrossFab (scCrossFab).

[0143] In some embodiments, the antigen-binding molecule described herein comprises or consists of an intact antibody that binds to gp130. As used herein, "intact antibody" means an antibody having a structure substantially similar to that of an immunoglobulin (Ig). Different types of immunoglobulins and their structures are described in [reference needed]. For exampleSchroeder and Cavacini J Allergy Clin Immunol. (2010) 125(202): S41-S52, which are hereby incorporated in their entirety by reference.

[0144] G-type immunoglobulins ( Right now IgG is a glycoprotein of approximately 150 kDa, consisting of two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chain contains the VH domain, followed by the heavy chain constant region containing three constant domains (CH1, CH2, and CH3). Similarly, the light chain contains the VL domain, followed by the CL domain. Immunoglobulins can be classified into IgG based on their heavy chains. For example IgG1, IgG2, IgG3, IgG4), IgA ( For example IgA1, IgA2), IgD, IgE, or IgM. The light chain can be κ (κ) or λ (λ).

[0145] In this article, "CH2 domain" refers to the amino acid sequence corresponding to the CH2 domain of immunoglobulin (Ig). According to the EU numbering system, the CH2 domain is the Ig region formed by positions 231 to 340 of the immunoglobulin constant domain (described in Edelman). et al. , Proc Natl Acad Sci USA (1969) 63(1): 78-85). The “CH3 domain” refers to the amino acid sequence corresponding to the CH3 domain of immunoglobulin (Ig). According to the EU numbering system, the CH3 domain is the region of Ig formed by positions 341 to 447 of the immunoglobulin constant domain. The “CH2-CH3 region” refers to the amino acid sequence corresponding to the CH2 and CH3 domains of immunoglobulin (Ig). According to the EU numbering system, the CH2-CH3 region is the Ig region formed by positions 231 to 447 of the immunoglobulin constant domain.

[0146] In some embodiments, the antigen-binding molecule described herein comprises IgG that binds to gp130. For example IgG1, IgG2, IgG3, IgG4), IgA ( For example IgA1, IgA2), IgD, IgE or IgM, or composed of them.

[0147] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more regions of a constant sequence of the immunoglobulin heavy chain. For example CH1, CH2, CH3 wait In some implementations, the constant sequence of the immunoglobulin heavy chain is or is derived from IgG (…). For example, IgG1, IgG2, IgG3, IgG4), IgA ( For example IgA1, IgA2), IgD, IgE or IgM ( For example Human IgG ( For example , hIgG1, hIgG2, hIgG3, hIgG4), hIgA ( For example The heavy chain constant sequence of immunoglobulin (hIgA1, hIgA2), hIgD, hIgE, or hIgM). In some embodiments, the immunoglobulin heavy chain constant sequence is or is derived from a human IgG1 allotype (hIgA1, hIgA2, hIgD, hIgE, or hIgM). example like The heavy chain constant sequence of G1m1, G1m2, G1m3 or G1m17.

[0148] In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:33 or 38 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:48 or 49 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:41 or 50 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0149] In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:42, 43, 46, or 47 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:44 or 57 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0150] In some embodiments, the antigen-binding molecule comprises a CH1 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:34 or 39 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises a CH1 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:51 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0151] In some embodiments, the antigen-binding molecule includes a hinge region comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:35 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule includes a hinge region comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:52 or 53 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0152] In some embodiments, the antigen-binding molecule comprises a CH2 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:36 or 45 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises a CH2 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:54 or 55 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0153] In some embodiments, the antigen-binding molecule comprises a CH3 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:37 or 40 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In some embodiments, the antigen-binding molecule comprises a CH3 region containing an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:56 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0154] It will be understood that further substitutions can be provided for the CH2 and / or CH3 regions based on modifications to the Fc region of the antigen-binding molecule as described herein.

[0155] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the immunoglobulin light chain constant sequence is the human immunoglobulin κ constant sequence (IGKC; Cκ). In some embodiments, the immunoglobulin light chain constant sequence is the human immunoglobulin λ constant sequence (IGLC; Cλ). For example IGLC1, IGLC2, IGLC3, IGLC6 or IGLC7.

[0156] In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO: 58, 59, 60, 61, 62, or 63 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity). In a preferred embodiment, the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO: 58 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0157] In some implementations, the antigen-binding molecule is a monoclonal antibody or its antigen-binding fragment, or contains a monoclonal antibody or its antigen-binding fragment.

[0158] In some implementations, the antigen-binding molecule is a fully human antibody / antibody fragment or contains a fully human antibody / antibody fragment. A fully human antibody / antibody fragment may be encoded by a human nucleic acid sequence. A fully human antibody / antibody fragment may lack a non-human amino acid sequence. Commonly used techniques for generating fully human antibodies include (i) phage display, in which a human antibody gene is expressed in a phage display library, and (ii) antibody generation in transgenic mice engineered to possess a human antibody gene (described in Park and Smolen, Advances in Protein Chemistry (2001) 56: 369-421). In short, in human antibody gene phage display technology, genes encoding the VH and VL chains are generated from “naive” human lymphocytes via PCR amplification and cloning, and assembled into a library from which they can be expressed as disulfide-linked Fab fragments or as single-stranded Fv (scFv) fragments. The Fab-encoding gene or the scFv-encoding gene is fused to the surface capsid protein of a filamentous phage, and then the Fab or scFv that can bind to the target of interest can be identified by screening a library with an antigen. Molecular evolution or affinity maturation procedures can be used to enhance the affinity of the Fab / scFv fragments. In transgenic mouse technology, mice whose endogenous mouse Ig loci have been replaced by their human homologs through homologous recombination are immunized with an antigen, and monoclonal antibodies are prepared using conventional hybridoma techniques to obtain fully human monoclonal antibodies.

[0159] In some embodiments, the antigen-binding molecule of this disclosure is a mouse antibody / antibody fragment. In some embodiments, the antibody / antibody fragment is obtained from phage display using a human preliminary antibody gene library.

[0160] In some implementations, the antigen-binding molecule is a mouse / human chimeric antigen-binding molecule (…). Right now An antigen-binding molecule comprising a variable domain of a mouse antibody and a constant region of a human antibody. In some embodiments, the antigen-binding molecule is a humanized antigen-binding molecule. Right now Includes humanized non-human animals ( For example The antigen-binding molecule comprises a variable domain derived from the variable domain of a mouse antibody, and includes a mouse antibody variable domain and a human antibody constant region. In some embodiments, the antigen-binding molecule includes a mouse antibody CDR and a human antibody frame region and constant region.

[0161] Mouse / human chimeric antigen-binding molecules can be prepared from mouse antibodies via a chimeric method. For example As described in Human Monoclonal Antibodies: Methods and Protocols, Michael Steinitz (ed.), Methods in Molecular Biology 1060, Springer Protocols, Humana Press (2014), Chapter 8, and in particular Section 3 of Chapter 8.

[0162] Humanized antigen-binding molecules can be prepared from mouse antibodies using humanization methods. For example As described in *Human Monoclonal Antibodies: Methods and Protocols*, Michael Steinitz (ed.), *Methods in Molecular Biology* 1060, Springer Protocols, Humana Press (2014), Chapter 7, and particularly Section 3.1 of Chapter 7, entitled “Antibody Humanization.” Techniques for antibody humanization are also described in… For example Safdari et al. ,Biotechnol Genet Eng Rev (2013) 29:175-86.

[0163] This disclosure relates to multispecific antigen-binding molecules. "Multispecific" means that the antigen-binding molecule exhibits specific binding to more than one target. In some embodiments, the antigen-binding molecule is a bispecific antigen-binding molecule. In some embodiments, the antigen-binding molecule comprises at least two distinct antigen-binding domains (…). Right now At least two antigen-binding domains, For example (Includes different VH and VL).

[0164] In some implementations, the antigen-binding molecule binds to gp130 and another target ( For example It binds to antigens other than gp130, and is therefore at least bispecific. The term "bispecific" means that the antigen-binding molecule is able to bind specifically to at least two different antigenic determinants.

[0165] It will be understood that, according to the antigen-binding molecules disclosed herein ( For example A multispecific antigen-binding molecule may include an antigen-binding molecule capable of binding to a target that is specifically targeted by the antigen-binding molecule. For example, an antigen-binding molecule that binds to antigens other than gp130 may include: (i) an antigen-binding molecule that binds to gp130; and (ii) an antigen-binding molecule that binds to antigens other than gp130.

[0166] It will also be understood that, according to the antigen-binding molecules of this disclosure ( For example Multispecific antigen-binding molecules may include antigen-binding peptides or antigen-binding peptide complexes that are capable of binding to specific targets of the antigen-binding molecule.

[0167] In some implementations, larger antigen-binding molecules ( For example The constituent antigen-binding molecules of a multispecific antigen-binding molecule can be referred to as For example The "antigen-binding domain" or "antigen-binding region" of a larger antigen-binding molecule.

[0168] In some embodiments, the antigen other than gp130 in the multispecific antigen-binding molecule is an immune cell surface molecule. In some embodiments, the antigen is a cancer cell antigen. In some embodiments, the antigen is a receptor molecule. For example Cell surface receptors. In some implementations, antigens are cell signaling molecules. For example Cytokines, chemokines, interferons, interleukins, or lymphokines. In some implementations, the antigen is a growth factor or hormone.

[0169] In some implementations, the antigen-binding molecule is an immune cell adaptor. Immune cell adaptors are... For exampleThese articles have been reviewed in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17: 418–434 and Ellerman, Methods (2019) 154:102-117, and are hereby incorporated in their entirety by citation. Immune cell adaptor molecules contain an antigen-binding region of the target antigen of interest, as well as an antigen-binding region for recruiting / adapting immune cells of interest. Immune cell adaptors recruit / adapt immune cells through antigen-binding regions specific to molecules on the surface of immune cells.

[0170] The most thoroughly studied immune cell adaptors are bispecific T cell adaptors (BiTEs), which contain a target antigen-binding domain and a CD3 polypeptide (usually CD3ε)-binding domain. BiTEs recruit T cells through these binding domains. The binding of BiTEs to their target antigen and to the CD3 polypeptide expressed by T cells leads to T cell activation and ultimately directs T cell effector activity against cells expressing the target antigen. Other types of immune cell adaptors are well-known in the art and include natural killer cell adaptors, such as bispecific killer adaptors (BiKEs), which recruit and activate NK cells.

[0171] In some implementations, the immune cell connected by the immune cell adaptor is a T cell or an NK cell. In some implementations, the immune cell adaptor is a T cell adaptor.

[0172] The multispecific antigen-binding molecules according to this disclosure may be provided in any suitable form, such as those described in Brinkmann and Kontermann, MAbs (2017) 9(2): 182-212, which are hereby incorporated in their entirety by reference. Suitable forms include those described in Brinkmann and Kontermann, MAbs (2017) 9(2): 182-212: antibodyconjugates Figure 2 The forms shown, For example IgG2, F(ab')2, or CovX-Body; IgG or IgG-like molecules, example like IgG, chimeric IgG, and κλ co-HC; CH1 / CL fusion protein, For example scFv2-CH1 / CL, VHH2-CH1 / CL; "variable domain-only" bispecific antigen-binding molecules. For exampleTandem scFv (taFV), trisomy, dimer (Db), dsDb, Db(kih), DART, scDB, dsFv-dsFv, tandAb, trihead, tandem dAb / VHH, tetravalent dAb.VHH; non-Ig fusion proteins. For example scFv2-albumin, scDb-albumin, taFv-albumin, taFv-toxin, microantibodies, DNL-Fab2, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine 2, ImmTAC (TCR-scFv); modified Fc and CH3 fusion proteins, For example scFv-Fc(kih), scFv-Fc(CH3 charge pair), scFv-Fc (EW-RVT), scFv-fc (HA-TF), scFv-Fc Fc fusion, For example Two-dimers, scDb-Fc, taFv-Fc, scFv-Fc-scFv, HCAb-VHH, Fab-scFv-Fc, scFv4-Ig, scFv2-Fcab; CH3 fusions, For example Dual-dimer, scDb-CH3; IgE / IgM CH2 fusion. For example scFv-EHD2-scFv, scFvMHD2-scFv; Fab fusion proteins. For example Fab-scFv (disomole), Fab-scFv2 (triomole), Fab-Fv, Fab-dsFv, Fab-VHH, orthogonal Fab-Fab; non-Ig fusion proteins. For example DNL-Fab3, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine 2; asymmetric IgG or IgG-like molecules For exampleIgG(kih), IgG(kih) co-LC, ZW1 IgG co-LC, Biclonics co-LC, CrossMab, CrossMab(kih), scFab-IgG(kih), Fab-scFab-IgG(kih), orthogonal Fab IgG(kih), DuetMab, CH3 charge pair + CH1 / CL charge pair, hinge / CH3 charge pair, SEED body, Duobody, tetra-CrossMab(kih), LUZ-Y co-LC; LUZ-Y scFab-IgG, FcFc*; IgG with appendage and Fc modification. For example IgG(kih)-Fv, IgG HA-TF-Fv, IgG(kih)scFab, scFab-Fc(kih)-scFv2, scFab-Fc(kih)-scFv, half-DVD-Ig, DVI-Ig (quadruple combination), CrossMab-Fab; modified Fc and CH3 fusion proteins. For example Fab-Fc(kih)-scFv, Fab-scFv-Fc(kih), Fab-scFv-Fc(BEAT), Fab-scFv-Fc-SEEDbody, TriFab; with added IgG-HC fusion compound, For example IgG-HC, scFv, IgG-dAb, IgG-taFV, IgG-CrossFab, IgG-orthogonal Fab, IgG-(CαCβ)Fab, scFv-HC-IgG, tandem Fab-IgG (orthogonal Fab), Fab-IgG (CαCβ Fab), Fab-IgG (CR3), Fab-hinge-IgG (CR3); with added IgG-LC fusions. For example IgG-scFv(LC), scFv(LC)-IgG, dAb-IgG; with added IgG-HC and LC fusion compounds. For example DVD-Ig, TVD-Ig, CODV-Ig, scFv4-IgG, Zybody; Fc fusion compound, For example Fab-scFv-Fc, scFv4-Ig; F(ab')2 fusions For example F(ab')2-scFv2;CH1 / CL fusion protein, For example scFv2-CH1-hinge / CL; modified IgG, For example DAF (dual IgG), DutaMab, Mab 2 ; and non-Ig fusion compounds, For exampleDNL-Fab4-IgG. Those skilled in the art can readily design and generate multispecific antigen-binding molecules.

[0173] This disclosure also provides a chimeric antigen receptor (CAR). A CAR is a recombinant receptor that provides both antigen binding and T cell activation functions. CAR structures and engineering methods are described in, for example, Dotti. et al. A review was conducted in Immunol Rev (2014) 257(1), which is hereby incorporated in its entirety by reference. CARs consist of an antigen-binding region connected to a cell membrane anchoring region and a signal transduction region. Optional hinge regions can provide separation between the antigen-binding region and the cell membrane anchoring region and can act as flexible connectors.

[0174] The antigen-binding domain of the CAR according to this disclosure comprises or is composed of an antigen-binding molecule that binds to gp130 as described herein. Therefore, the CAR according to this disclosure comprises an antigen-binding molecule as described herein.

[0175] It will be understood that the antigen-binding molecules according to this disclosure form or are contained within the antigen-binding domain of a CAR. Therefore, in some embodiments, the antigen-binding molecules of this disclosure are contained within a CAR.

[0176] It will also be understood that the antigen-binding molecule according to this disclosure can be a CAR. Having an antigen-binding molecule comprising this disclosure ( For example A CAR that binds to the Fv of gp130 or consists of an antigen-binding domain composed of the antigen-binding molecule is an antigen-binding molecule. The antigen-binding domain of the CAR disclosed herein may be provided in any suitable form. For example scFv, scFab wait .

[0177] The cell membrane anchoring region is located between the antigen-binding region and the signal transduction region of the CAR, providing a means to anchor the CAR to the cell membrane of a CAR-expressing cell, wherein the antigen-binding region is located in the extracellular space and the signal transduction region is located inside the cell. In some embodiments, the CAR includes a cell membrane anchoring region comprising or consisting of an amino acid sequence comprising, consisting of, or derived from a transmembrane amino acid sequence of one of CD3-ζ, CD4, CD8, or CD28. As used herein, a region “derived from” a reference amino acid sequence comprises having at least 60% ( ) similarity to the reference sequence. For example An amino acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence.

[0178] The signal transduction region of a CAR allows for the activation of T cells. The CAR signal transduction region may contain the amino acid sequence of the CD3-ζ intracellular domain, which provides the immune receptor tyrosine-based activation motif (ITAM) for phosphorylation and activation of CAR-expressing T cells. Signal transduction regions containing sequences of other ITAM-containing proteins (such as FcγRI) have also been used in CARs (Haynes). et al. , 2001 J Immunol 166(1):182-187). The signal transduction region of the CAR may also contain costimulatory sequences derived from the signal transduction regions of costimulatory molecules to promote the activation of CAR-expressing T cells upon binding to target proteins. Suitable costimulatory molecules include CD28, OX40, 4-1BB, ICOS, and CD27. In some cases, the CAR is engineered to provide costimulation for different intracellular signal transduction pathways. For example, signal transduction associated with CD28 costimulation preferentially activates the phosphatidylinositol 3-kinase (PI3K) pathway, while 4-1BB-mediated signal transduction is mediated by TNF receptor-associated factor (TRAF) aptamer proteins. Therefore, the signal transduction region of the CAR sometimes contains costimulatory sequences derived from the signal transduction regions of more than one costimulatory molecule. In some embodiments, the CAR of this disclosure comprises one or more costimulatory sequences comprising or consisting of amino acid sequences comprising, consisting of, or derived from one or more intracellular domains of CD28, OX40, 4-1BB, ICOS, and CD27.

[0179] Optional hinge regions can provide separation between antigen-binding domains and transmembrane domains and can act as flexible linkers. The hinge region may be derived from IgG1 or IgG4. In some embodiments, the CAR of this disclosure includes a hinge region comprising or consisting of an amino acid sequence, said amino acid sequence comprising, consisting of, or derived from the amino acid sequence of the hinge region of IgG1 or IgG4. In some embodiments, the hinge region of the CAR according to this disclosure comprises an amino acid sequence having at least 70% sequence identity (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) with the amino acid sequence of SEQ ID NO:35. In some embodiments, the hinge region of the CAR according to this disclosure comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO:52 or 53 (more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity).

[0180] Cells comprising the CAR according to this disclosure are also provided. The CAR according to this disclosure can be used to generate immune cells expressing the CAR. For example CAR-T or CAR-NK cells. They can be used... in vitro The CAR was engineered into immune cells during the culture process.

[0181] Fc area In some embodiments, the antigen-binding molecule of this disclosure includes an Fc region.

[0182] As used in this article, the “Fc region” refers to a polypeptide complex formed by the interaction between two polypeptides, each polypeptide containing the CH2-CH3 region of the constant sequence of the immunoglobulin (Ig) heavy chain.

[0183] In this article, "CH2 region" refers to the amino acid sequence corresponding to the CH2 region of immunoglobulin (Ig). According to the EU numbering system, the CH2 region is the Ig region formed by positions 231 to 340 of the immunoglobulin constant region (described in Edelman). et al. ,Proc Natl Acad Sci USA (1969) 63(1): 78-85). The “CH3 region” refers to the amino acid sequence corresponding to the CH3 region of immunoglobulin (Ig). According to the EU numbering system, the CH3 region is the Ig region formed by positions 341 to 447 of the immunoglobulin constant region (described in Edelman). et al. , Proc Natl Acad Sci USA (1969) 63(1): 78-85). The “CH2-CH3 region” refers to the amino acid sequence corresponding to the CH2 and CH3 regions of immunoglobulin (Ig). According to the EU numbering system, the CH2-CH3 region is the Ig region formed by positions 231 to 447 of the immunoglobulin constant region (described in Edelman). et al. , Proc NatlAcad Sci USA (1969) 63(1): 78-85).

[0184] In some embodiments, the CH2 region, CH3 region, and / or CH2-CH3 region according to this disclosure correspond to IgG ( For example IgG1, IgG2, IgG3, IgG4), IgA ( For example The CH2 / CH3 / CH2-CH3 regions of IgA1, IgA2, IgD, IgE, or IgM. In some embodiments, the CH2, CH3, and / or CH2-CH3 regions correspond to human IgG ( For example , hIgG1, hIgG2, hIgG3, hIgG4), hIgA ( For example The CH2 / CH3 / CH2-CH3 regions of hIgA1, hIgA2, hIgD, hIgE, or hIgM. In some embodiments, the CH2, CH3, and / or CH2-CH3 regions correspond to human IgG1 allotypes (hIgA1, hIgA2, hIgD, hIgE, or hIgM). For example CH2 region / CH3 region / CH2-CH3 region of G1m1, G1m2, G1m3 or G1m17.

[0185] The Fc region provides the means for interaction with Fc receptors and other molecules in the immune system, thereby exerting functional effects. Fc-mediated effector functions occur in… For example Jefferis et al. A review was conducted in *Immunol Rev 1998*, 163:59-76 (included hereby by reference in its entirety), which discussed the interaction between the Fc region and Fc receptors expressed by immune cells, and the Fc-mediated immune cell activity. For exampleThe complement pathway is mediated by the recruitment and activation of macrophages, dendritic cells, neutrophils, basophils, eosinophils, platelets, mast cells, NK cells, and T cells. This recruitment of complement pathway components occurs through the binding of the Fc region to complement protein C1q, followed by activation of the complement cascade. Fc-mediated functions include Fc receptor binding, antibody-dependent cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), formation of the membrane attack complex (MAC), cell degranulation, production of cytokines and / or chemokines, and antigen processing and presentation.

[0186] Modifications to the Fc region of antibodies that affect Fc-mediated function are known in the art, such as those described in For example Wang wait people The modifications mentioned in Protein Cell (2018) 9(1):63-73 are hereby incorporated in their entirety by reference. Exemplary Fc region modifications known to affect antibody effector function are summarized in Wang... et al. Table 1 of Protein Cell (2018) 9(1):63-73. In some embodiments, the antigen-binding molecule of this disclosure contains an Fc region, which is modified to increase or decrease Fc-mediated function, compared to an antigen-binding molecule containing a corresponding unmodified Fc region.

[0187] In cases where the Fc region / CH2 / CH3 is described as containing modifications "corresponding to" a reference substitution, consider equivalent substitutions in the homologous Fc / CH2 / CH3. For example, the L234A / L235A substitution in human IgG1 (according to the description in Kabat...) et al. (Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991, EU numbering system) corresponds to the L-to-A substitution at positions 117 and 118 in the C region of the mouse Igγ-2A chain (UniProtKB: P01863-1, v1).

[0188] When the Fc region is described as containing a modification, that modification can be present in one or two polypeptide chains that together form the Fc region.

[0189] In some embodiments, the antigen-binding molecule of this disclosure includes a modified Fc region. In some embodiments, the antigen-binding molecule of this disclosure includes a modified Fc region in one or more CH2 and / or CH3 regions.

[0190] In some embodiments, the Fc region contains modifications to enhance Fc-mediated function. In some embodiments, the Fc region contains modifications to enhance ADCC. In some embodiments, the Fc region contains modifications to enhance ADCP. In some embodiments, the Fc region contains modifications to enhance CDC. Compared to antigen-binding molecules containing the corresponding unmodified Fc region, those containing an Fc region (which is modified to enhance Fc-mediated function) are more effective. For example Antigen-binding molecules such as ADCC, ADCP, and CDC induce increased functional levels of related effectors.

[0191] In some embodiments, the Fc region includes modifications to increase binding to the Fc receptor. In some embodiments, the Fc region includes modifications to increase binding to the Fcγ receptor. In some embodiments, the Fc region includes modifications to increase binding to one or more of FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb. In some embodiments, the Fc region includes modifications to increase binding to FcγRIIIa. In some embodiments, the Fc region includes modifications to increase binding to FcγRIIa. In some embodiments, the Fc region includes modifications to increase binding to FcγRIIb. In some embodiments, the Fc region includes modifications to increase binding to FcRn. In some embodiments, the Fc region includes modifications to increase binding to complement proteins. In some embodiments, the Fc region includes modifications to increase binding to C1q. In some embodiments, the Fc region includes modifications to promote hexamerization of the antigen-binding molecule. In some embodiments, the Fc region contains modifications to increase the half-life of the antigen-binding molecule. In some embodiments, the Fc region contains modifications to increase co-conjugation.

[0192] In some embodiments, the Fc region contains modifications corresponding to the substitution combination F243L / R292P / Y300L / V305I / P396L, such as Stavenhagen. et al. Cancer Res. (2007) 67:8882–8890. In some embodiments, the Fc region contains modifications corresponding to the substitutions for combinations of S239D / I332E or S239D / I332E / A330L, as described in Lazar. et al.Proc Natl Acad Sci USA. (2006) 103:4005–4010. In some embodiments, the Fc region includes modifications corresponding to the combinations replacing S298A / E333A / K334A, as described in Shields. et al. , J Biol Chem. (2001) 276:6591–6604. In some embodiments, the Fc region comprises a modification of one of the heavy chain polypeptides corresponding to the combination of substituted L234Y / L235Q / G236W / S239M / H268D / D270E / S298A, and modifications of other heavy chain polypeptides corresponding to the combination of substituted D270E / K326D / A330M / K334E, as described in Mimoto. et al. MAbs. (2013): 5:229–236. In some embodiments, the Fc region includes modifications corresponding to the substitution of combinations of G236A / S239D / I332E, as described in Richards. wait people , Mol Cancer Ther. (2008) 7:2517–2527.

[0193] In some implementations, the Fc region contains modifications corresponding to the substitution combination K326W / E333S, such as Idusogie. et al. J Immunol. (2001) 166(4):2571-5. In some embodiments, the Fc region contains modifications corresponding to the substitution combination S267E / H268F / S324T, such as Moore's. et al. As described in MAbs. (2010) 2(2):181-9. In some embodiments, the Fc region contains modifications corresponding to combinations of substitutions, as described in Natsume. et al. Cancer Res. (2008) 68(10):3863-72. In some embodiments, the Fc region contains modifications corresponding to the substitution combination E345R / E430G / S440Y, such as Diebolder. et al. As described in Science (2014) 343(6176):1260-3.

[0194] In some implementations, the Fc region contains modifications corresponding to the substitution combination M252Y / S254T / T256E, such as Dall'Acqua et al. J Immunol. (2002) 169:5171–5180.

[0195] In some embodiments, the Fc region comprises a CH2-CH3 region, the amino acid sequence of which differs from that of the CH2-CH3 region of a reference Fc region at one or more of the following positions: 252, 254, or 256 (according to the EU numbering system). In some embodiments, the Fc region comprises a CH2-CH3 region containing one or more of the following specified amino acid residues: Y252, T254, or E256 (according to the EU numbering system). In some embodiments, the Fc region comprises a CH2-CH3 region containing Y252, T254, and E256. In some embodiments, the Fc region comprises a CH2-CH3 region, the amino acid sequence of which differs from that of the CH2-CH3 region of a reference Fc region at one or more of the following amino acid substitutions: M252Y, S254T, or T256E (according to the EU numbering system).

[0196] These so-called “YTE” modifications at the CH2-CH3 interface of the Fc region have been shown to increase binding affinity to the MHC class I neonatal Fc receptor (FcRn) at pH 6.0, which is located in the acidic endosomes of endothelial cells and hematopoietic cells, thereby improving the effective recovery rate of mAb administration and prolonging its half-life in plasma.

[0197] In some implementations, the Fc region includes modifications corresponding to the combination replacing M428L / N434S, as described in Zalevsky. et al. As described in Nat Biotechnol. (2010) 28:157–159.

[0198] In some implementations, the Fc region includes modifications corresponding to the combination replacing S267E / L328F, as described in Chu et al. , Mol Immunol. (2008) 45:3926–3933. In some embodiments, the Fc region contains modifications corresponding to the substitution combination N325S / L328F, such as Shang et al. As described in Biol Chem. (2014) 289:15309–15318.

[0199] In some embodiments, the Fc region contains modifications to reduce / prevent Fc-mediated function. In some embodiments, the Fc region contains modifications to reduce / prevent ADCC. In some embodiments, the Fc region contains modifications to reduce / prevent ADCP. In some embodiments, the Fc region contains modifications to reduce / prevent CDC. Compared to antigen-binding molecules containing a corresponding unmodified Fc region, those containing an Fc region (which contains modifications to reduce / prevent Fc-mediated function) are more effective. For exampleAntigen-binding molecules such as ADCC, ADCP, and CDC induce a decrease in the functional level of related effectors.

[0200] In some embodiments, the Fc region contains modifications to reduce / prevent binding to the Fc receptor. In some embodiments, the Fc region contains modifications to reduce / prevent binding to the Fcγ receptor. In some embodiments, the Fc region contains modifications to reduce / prevent binding to one or more of FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb. In some embodiments, the Fc region contains modifications to reduce / prevent binding to FcγRIIIa. In some embodiments, the Fc region contains modifications to reduce / prevent binding to FcγRIIa. In some embodiments, the Fc region contains modifications to reduce / prevent binding to FcγRIIb. In some embodiments, the Fc region contains modifications to reduce / prevent binding to complement proteins. In some embodiments, the Fc region contains modifications to reduce / prevent binding to C1q. In some embodiments, the Fc region contains modifications to reduce / prevent glycosylation of the amino acid residue corresponding to N297.

[0201] In some implementations, the Fc region cannot induce one or more Fc-mediated functions. Right now (They lack the ability to induce relevant Fc-mediated functions). Therefore, antigen-binding molecules containing such Fc regions also lack the ability to induce relevant functions. Such antigen-binding molecules can be described as having no relevant function.

[0202] In some implementations, the Fc region cannot induce ADCC. In some implementations, the Fc region cannot induce ADCP. In some implementations, the Fc region cannot induce CDC. In some implementations, the Fc region cannot induce ADCC and / or cannot induce ADCP and / or cannot induce CDC.

[0203] In some embodiments, the Fc region cannot bind to the Fc receptor. In some embodiments, the Fc region cannot bind to the Fcγ receptor. In some embodiments, the Fc region cannot bind to one or more of FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb. In some embodiments, the Fc region cannot bind to FcγRIIIa. In some embodiments, the Fc region cannot bind to FcγRIIa. In some embodiments, the Fc region cannot bind to FcγRIIb. In some embodiments, the Fc region cannot bind to FcRn. In some embodiments, the Fc region cannot bind to complement proteins. In some embodiments, the Fc region cannot bind to C1q. In some embodiments, the Fc region is not glycosylated at the amino acid residue corresponding to N297.

[0204] In some implementations, the Fc region includes modifications corresponding to N297A, N297Q, or N297G, as described in Leabman. et al. MAbs. (2013) 5:896–903. In some implementations, the Fc region includes modifications corresponding to L235E, as described in Alegre. et al. , J Immunol. (1992) 148:3461–3468. In some embodiments, the Fc region includes modifications corresponding to the substitutions for L234A / L235A or combinations of F234A / L235A, as described in Xu et al. , CellImmunol. (2000) 200:16–26. In some embodiments, the Fc region contains modifications corresponding to P329A or P329G, as described in Schlothauer. et al. , Protein Engineering, Design and Selection (2016), 29(10):457–466. In some embodiments, the Fc region contains modifications corresponding to the substitution combination L234A / L235A / P329G, such as Lo et al. As described in J. Biol. Chem (2017) 292(9):3900-3908. In some embodiments, the Fc region contains modifications corresponding to combinations of substitutions, as described in Rother. et al. Nat Biotechnol. (2007) 25:1256–1264. In some embodiments, the Fc region contains modifications corresponding to the substitution of the S228P / L235E combination, as described in Newman. et al. Clin. Immunol. (2001) 98:164–174. In some embodiments, the Fc region includes modifications corresponding to the substitution of the combination of H268Q / V309L / A330S / P331S, as described in An et al. MAbs. (2009) 1:572–579. In some embodiments, the Fc region includes modifications corresponding to combinations replacing V234A / G237A / P238S / H268A / V309L / A330S / P331S, as described in Vafa. et al. Methods. (2014) 65:114–126. In some embodiments, the Fc region includes modifications corresponding to the substitution combination L234A / L235E / G237A / A330S / P331S, as described in US 2015 / 0044231 A1.

[0205] Known combinations of substitutes for "L234A / L235A" and corresponding substitutes (such as...) For example The F234A / L235A combination in human IgG4 disrupts the binding of Fc to Fcγ receptors and inhibits ADCC and ADCP, while also reducing C1q binding and thus decreasing CDC (Schlothauer's receptor). et al. , Protein Engineering, Design and Selection (2016), 29(10):457–466, which is hereby incorporated in its entirety by reference. Replacing “P329G” and “P329A” reduces C1q binding (and thus reduces CDC). It is known that replacing “N297” with “A”, “G” or “Q” eliminates glycosylation, thereby reducing Fc binding to C1q and Fcγ receptors, and thus reducing CDC and ADCC. et al. J. Biol. Chem (2017) 292(9):3900-3908 (included hereby by reference in its entirety) reported that the combination of replacing L234A / L235A / P329G eliminated complement binding and fixation in both mouse IgG2a and human IgG1, as well as Fcγ receptor-dependent, antibody-dependent, and cell-mediated cytotoxicity.

[0206] US 2015 / 0044231 A1 discloses a combination of substituted L234A / L235E / G237A / A330S / P331S in IgG1 Fc that eliminates phagocytosis, ADCC and CDC induction.

[0207] In some implementations, the Fc region contains modifications corresponding to the substitution of S228P, as described in Silva. et al. , JBiol Chem. (2015) 290(9):5462-5469. Substitution of S228P in IgG4 Fc reduces Fab arm exchange (Fab arm exchange may be undesirable).

[0208] In some embodiments, the Fc region contains modifications corresponding to the substitution combination L234A / L235A. In some embodiments, the Fc region contains modifications corresponding to the substitution of P329G. In some embodiments, the Fc region contains modifications corresponding to the substitution of N297Q.

[0209] In some implementations, the Fc region contains modifications corresponding to the substitution combination L234A / L235A / P329G.

[0210] In some implementations, the Fc region contains modifications corresponding to the substitution combination L234A / L235A / P329G / N297Q.

[0211] In some implementations, the Fc region contains modifications corresponding to the substitution combination L234A / L235E / G237A / A330S / P331S.

[0212] In some implementations, the Fc region contains corresponding to For example The modification of IgG4 by replacing S228P.

[0213] A relatively short half-life of the antigen-binding molecule may be desirable. Since the antigen-binding molecules of this disclosure affect signal transduction mediated by multiple cytokines / through multiple cytokine receptors, a relatively short duration of their functional effect may be desirable. Therefore, in some embodiments, the antigen-binding molecule of this disclosure may include an Fc region modified to shorten the half-life of the antigen-binding molecule.

[0214] Fc modifications that reduce half-life include H345A and the triple substitution I253A / H310A / H345A. These modifications have been reported to reduce the binding of the antigen-binding molecule to the FcRn relative to its unmodified wild-type counterpart, and to decrease the half-life of antigen-binding molecules carrying Fc regions containing such modifications (see [link to FcRn]). For example Kruijsen et al. , JVirol. (2013) 87(13): 7550–7557, the reference of which is hereby incorporated in its entirety by reference.

[0215] In some embodiments, the Fc region contains modifications corresponding to a substitution for H345A. In some embodiments, the Fc region contains modifications corresponding to a substitution for I253A. In some embodiments, the Fc region contains modifications corresponding to a substitution for H310A. In some embodiments, the Fc region contains modifications corresponding to a combination of substitutions for I253A / H310A / H345A.

[0216] In some embodiments, the antigen-binding molecule does not contain an Fc region. In some embodiments, the antigen-binding molecule consists primarily of the Fv portion. For example (scFv or dsFv). In some implementations, the antigen-binding molecule is essentially composed of the Fab moiety ( For example It is Fab, CrossFab, scFab, scCrossFab, or F(ab')2).

[0217] As used herein, an antigen-binding molecule “substantially composed of” a reference polypeptide domain / amino acid sequence (i) consists of a reference domain / amino acid sequence, or (ii) contains a reference domain / amino acid sequence, wherein the reference domain / amino acid sequence constitutes at least 80% of the molecule. For example (One of ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%). It should be understood that an antigen-binding molecule “substantially composed of” a reference polypeptide domain / amino acid sequence may include the reference domain / amino acid sequence, and additional amino acids at one or both of the N-terminus and / or C-terminus of the reference domain / amino acid sequence, provided that the additional amino acids constitute ≤20% of the molecule. For example, an antigen-binding molecule substantially composed of an Fv moiety includes scFv molecules, which contain VH and VL linked via a linker portion.

[0218] In some implementation schemes – particularly where the antigen-binding molecule is multispecific ( For example In an embodiment of a bispecific antigen-binding molecule, the antigen-binding molecule comprises an Fc region containing modifications in one or more of the CH2 and CH3 regions, the modifications promoting association of the Fc region. Recombinant co-expression and subsequent association of the constituent peptides of the antigen-binding molecule result in several possible combinations. To improve the yield of desired peptide combinations in the antigen-binding molecule during recombinant production, it is advantageous to introduce modifications in the Fc region that promote the association of the desired heavy chain peptide combination. These modifications can promote... For example Hydrophobic and / or electrostatic interactions between the CH2 and / or CH3 regions of different polypeptide chains. Appropriate modifications are described in... For example Ha et al. ,Front. Immnol (2016) 7:394, which is hereby incorporated in its entirety by reference.

[0219] In some embodiments, the antigen-binding molecule of this disclosure comprises an Fc region containing paired substitutions in the CH3 region of the Fc region according to one of the following forms, such as Ha. et al. Table 1 of Front. Immnol (2016) 7:394 shows: KiH, KiH s-s , HA-TF, ZW1, 7.8.60, DD-KK, EW-RVT, EW-RVT s-s SEED or A107.

[0220] Specific exemplary peptides and antigen-binding molecules This disclosure also provides the polypeptide composition of the antigen-binding molecule. The polypeptide may be provided in an isolated or substantially purified form.

[0221] The antigen-binding molecule disclosed herein may be a complex of polypeptides or a complex that may contain polypeptides.

[0222] In this specification, when a polypeptide contains more than one domain or region, it will be understood that the multiple domains / regions preferably exist in the same polypeptide chain. That is, a polypeptide containing more than one domain or region is a fusion polypeptide containing domains / regions.

[0223] In some embodiments, the polypeptide according to this disclosure comprises or is composed of VH as described herein. In some embodiments, the polypeptide according to this disclosure comprises VL as described herein or is composed of VH as described herein.

[0224] In some embodiments, the polypeptide additionally includes one or more antibody heavy chain constant regions (CH). In some embodiments, the polypeptide additionally includes one or more antibody light chain constant regions (CL). In some embodiments, the polypeptide includes the CH1, CH2, and / or CH3 regions of an immunoglobulin (Ig).

[0225] In some embodiments, the polypeptide comprises one or more regions of a constant sequence of the immunoglobulin heavy chain. In some embodiments, the polypeptide comprises a CH1 region as described herein. In some embodiments, the polypeptide comprises a CH1-CH2 hinge region as described herein. In some embodiments, the polypeptide comprises a CH2 region as described herein. In some embodiments, the polypeptide comprises a CH3 region as described herein.

[0226] In some embodiments, the polypeptide comprises one or more regions of a constant sequence of an immunoglobulin light chain. In some embodiments, the polypeptide comprises a CL region as described herein.

[0227] In some embodiments, the polypeptide according to this disclosure comprises, from the N-terminus to the C-terminus, a structure according to one of the following: (i) VH (ii) VL (iii) VH-CH1 (iv) VL-CL (v) VL-CH1 (vi) VH-CL (vii) VH-CH1-CH2-CH3 (viii) VL-CL-CH2-CH3 (ix) VL-CH1-CH2-CH3 (x) VH-CL-CH2-CH3 This disclosure also provides antigen-binding molecules composed of the polypeptides of this disclosure. In some embodiments, the antigen-binding molecule of this disclosure comprises one of the following combinations of polypeptides: (A) VH + VL (B) VH-CH1 + VL-CL (C) VL-CH1 + VH-CL (D) VH-CH1-CH2-CH3 + VL-CL (E) VH-CL-CH2-CH3 + VL-CH1 (F) VL-CH1-CH2-CH3 + VH-CL (G) VL-CL-CH2-CH3 + VH-CH1 (H) VH-CH1-CH2-CH3 + VL-CL-CH2-CH3 (I) VH-CL-CH2-CH3 + VL-CH1-CH2-CH3 In some embodiments, the antigen-binding molecule comprises more than one polypeptide in the combinations shown in (A) to (I) above. For example, referring to (D) above, in some embodiments, the antigen-binding molecule comprises two polypeptides containing the structure VH-CH1-CH2-CH3 and two polypeptides containing the structure VL-CL.

[0228] In some embodiments, the antigen-binding molecule of this disclosure comprises one of the following combinations of peptides: (J) VH (anti-gp130) + VL (anti-gp130) (K) VH (anti-gp130)-CH1 + VL (anti-gp130)-CL (L) VL (anti-gp130)-CH1 + VH (anti-gp130)-CL (M) VH (anti-gp130)-CH1-CH2-CH3 + VL (anti-gp130)-CL (N) VH (anti-gp130)-CL-CH2-CH3 + VL (anti-gp130)-CH1 (O) VL (anti-gp130)-CH1-CH2-CH3 + VH (anti-gp130)-CL (P) VL (anti-gp130)-CL-CH2-CH3 + VH (anti-gp130)-CH1 (Q) VH (anti-gp130)-CH1-CH2-CH3 + VL (anti-gp130)-CL-CH2-CH3 Wherein, “VH (anti-gp130)” refers to the VH of an antigen-binding molecule that can bind to gp130 as described herein; and “VL (anti-gp130)” refers to the VL of an antigen-binding molecule that can bind to gp130 as described herein.

[0229] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 183, 184, 112, 117, 179, 1, 180, 94, 181, 98, 182, or 174, or is composed of said amino acid sequence.

[0230] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide that comprises, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 216, 217, 202, 203, 211, 212, 213, 193, 194, or 17.

[0231] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide comprising, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 190, 191, 121, 126, 186, 9, 187, 188, 189, 102, 106, 109, 147, 148, 143, 144, 145, 146, 160, 161, 156, 157, 158, or 159) of the amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 190, 191, 121, 126, 186, 9, 187, 188, 189, 102, 106, or 109, or is composed of said amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 147, 148, 143, 144, 145, or 146, or is composed of said amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 160, 161, 156, 157, 158, or 159, or is composed of said amino acid sequence.

[0232] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 230, 231, 25, 219, 220, or 221, or is composed of said amino acid sequence.

[0233] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide containing, or being composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 243, 244, 135, 137, 134, 136, 239, 65, 240, 131, 241, 133, 242, 177, 130, 132, 176, or 64.

[0234] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 258, 259, 253, 254, 255, 256, 257, 251, 252, 67, or 68, or is composed of said amino acid sequence.

[0235] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide comprising, or is composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 249, 250, 141, 142, 245, 66, 246, 247, 248, 138, 139, 140, 154, 155, 150, 151, 152, 153, 167, 168, 163, 164, 165, or 166) of the amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 249, 250, 141, 142, 245, 66, 246, 247, 248, 138, 139, or 140) of the amino acid sequence, or is composed of said amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 154, 155, 150, 151, 152, or 153) of the amino acid sequence, or is composed of said amino acid sequence. In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide containing, or being composed of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 167, 168, 163, 164, 165, or 166.

[0236] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide having an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with SEQ ID NO: 263, 264, 69, 260, 261, or 262, or is composed of said amino acid sequence.

[0237] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more polypeptides comprising a cloned VH region containing a heavy chain CDR and a VL region containing a light chain CDR, said clone being selected from: B035-C03-A3, CSP-S-5H8, A3_3.2 VH / A3_3 VL, A3_3.2 VH / A3_4 VL, A3_3.2 VH / A3_5 VL, A3_4.2 VH / A3_3 VL, A3_4.2 VH / A3_4 VL, A3_4.2 VH / A3_5 VL, A3_6.2 VH / A3_3 VL, A3_6.2 VH / A3_4 VL, A3_6.2 VH / A3_5 VL, A3-derived VH CON / A3-derived VL CON, A3-Hum VH CON / A3-Hum VL CON, B035-C03-A3_D50X, A3_3.2 VH / A3_3_D50X VL, A3_3.2 VH / A3_4_D50XVL, A3_3.2 VH / A3_5_D50X VL, A3_4.2 VH / A3_3_D50X VL, A3_4.2 VH / A3_4_D50XVL, A3_4.2 VH / A3_5_D50X VL, A3_6.2 VH / A3_3_D50X VL, A3_6.2 VH / A3_4_D50XVL, A3_6.2 VH / A3_5_D50X VL, A3-derivative VH CON / A3-derivative VL_D50X CON, A3-Hum VH CON / A3-Hum VL_D50X CON, B035-C03-A3_D50A, A3_3.2 VH / A3_3_D50A VL, A3_3.2 VH / A3_4_D50A VL, A3_3.2 VH / A3_5_D50A VL, A3_4.2 VH / A3_3_D50A VL, A3_4.2 VH / A3_4_D50A VL, A3_4.2 VH / A3_5_D50A VL, A3_6.2 VH / A3_3_D50A VL, A3_6.2 VH / A3_4_D50A VL, A3_6.2 VH / A3_5_D50A VL, A3 - derived VH CON / A3 - derived VL_D50A CON and A3 - Hum VH CON / A3 - Hum VL_D50A CON, B035 - C03 - A3_I100T VH / B035 - C03 - A3_G51A VL, B035 - C03 - A3_I100T VH / B035 - C03 - A3 VL, B035 - C03 - A3 VH / B035 - C03 - A3_G51A VL, A3_3.2_I100T VH / A3_3_G51A VL, A3_3.2_I100T VH / A3_4_G51A VL, A3_3.2_I100T VH / A3_5_G51A VL, A3_3.2_I100T VH / A3_3 VL, A3_3.2_I100T VH / A3_4 VL, A3_3.2_I100T VH / A3_5 VL, A3_3.2 VH / A3_3_G51A VL, A3_3.2 VH / A3_4_G51A VL, A3_3.2 VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3_G51A VL, A3_4.2_I100T VH / A3_4_G51A VL, A3_4.2_I100T VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3 VL, A3_4.2_I100T VH / A3_4 VL, A3_4.2_I100T VH / A3_5 VL, A3_4.2 VH / A3_3_G51A VL, A3_4.2 VH / A3_4_G51A VL, A3_4.2 VH / A3_5_G51A VL, A3_6.2_I100T VH / A3_3_G51A VL, A3_6.2_I100T VH / A3_4_G51A VL, A2 VH / A3_5_G51A VL, A3-derived_I100T VH CON / A3-derived_G51A VL CON, A3-derived_I100T VH CON / A3-derived VL CON, A3-derived VH CON / A3-derived_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum VL CON, A3-Hum VH CON / A3-Hum_G51A VL CON, 5H8CVH4 VH / 5H8CVL3 VL, 5H8CVH4 VH / 5H8CVL4 VL, 5H8CVH4 VH / 5H8GVL3 VL, 5H8GVH4 VH / 5H8CVL3 VL, 5H8GVH4 VH / 5H8CVL4 VL, 5H8GVH4 VH / 5H8GVL3 VL, 5H8-derivative VH CON / 5H8-derivative VLCON, 5H8-Hum VH CON / 5H8-Hum VL CON, CSP-S-5H8_D98G VH / CSP-S-5H8 VL, 5H8CVH4_D98G VH / 5H8CVL3 VL, 5H8CVH4_D98G VH / 5H8CVL4 VL, 5H8CVH4_D98G VH / 5H8GVL3VL, 5H8GVH4_D98G VH / 5H8CVL3 VL, 5H8GVH4_D98G VH / 5H8CVL4 VL, 5H8GVH4_D98G VH / 5H8GVL3 VL, 5H8-derived_D98G VH CON / 5H8-derived VL CON, or 5H8-Hum_D98G VH CON / 5H8-Hum VL CON, as shown in Table A herein. That is, in some embodiments, the antigen-binding molecule comprises one or more polypeptides, said polypeptides comprising: (i) a VH region comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in column A of Table A, and (ii) a VL region comprising LC-CDR1, LC-CDR2, and LC-CDR3 as shown in column B of Table A, wherein the sequences in columns A and B are selected from the same row of Table A.

[0238] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more polypeptides comprising a cloned VH region containing a heavy chain FR and a VL region containing a light chain FR, said clone being selected from: B035-C03-A3, CSP-S-5H8, A3_3.2 VH / A3_3 VL, A3_3.2 VH / A3_4 VL, A3_3.2 VH / A3_5 VL, A3_4.2 VH / A3_3 VL, A3_4.2 VH / A3_4 VL, A3_4.2 VH / A3_5 VL, A3_6.2 VH / A3_3 VL, A3_6.2 VH / A3_4 VL, A3_6.2 VH / A3_5 VL, A3-derived VH CON / A3-derived VL CON, A3-Hum VH CON / A3-Hum VL CON, B035-C03-A3_D50X, A3_3.2 VH / A3_3_D50X VL, A3_3.2 VH / A3_4_D50XVL, A3_3.2 VH / A3_5_D50X VL, A3_4.2 VH / A3_3_D50X VL, A3_4.2 VH / A3_4_D50XVL, A3_4.2 VH / A3_5_D50X VL, A3_6.2 VH / A3_3_D50X VL, A3_6.2 VH / A3_4_D50XVL, A3_6.2 VH / A3_5_D50X VL, A3-derivative VH CON / A3-derivative VL_D50X CON, A3-Hum VH CON / A3-Hum VL_D50X CON, B035-C03-A3_D50A, A3_3.2 VH / A3_3_D50A VL, A3_3.2 VH / A3_4_D50A VL, A3_3.2 VH / A3_5_D50A VL, A3_4.2 VH / A3_3_D50A VL, A3_4.2 VH / A3_4_D50A VL, A3_4.2 VH / A3_5_D50A VL, A3_6.2 VH / A3_3_D50A VL, A3_6.2 VH / A3_4_D50A VL, A3_6.2 VH / A3_5_D50A VL, A3 - derived VH CON / A3 - derived VL_D50A CON and A3 - Hum VH CON / A3 - Hum VL_D50A CON, B035 - C03 - A3_I100T VH / B035 - C03 - A3_G51A VL, B035 - C03 - A3_I100T VH / B035 - C03 - A3 VL, B035 - C03 - A3 VH / B035 - C03 - A3_G51A VL, A3_3.2_I100T VH / A3_3_G51A VL, A3_3.2_I100T VH / A3_4_G51A VL, A3_3.2_I100T VH / A3_5_G51A VL, A3_3.2_I100T VH / A3_3 VL, A3_3.2_I100T VH / A3_4 VL, A3_3.2_I100T VH / A3_5 VL, A3_3.2 VH / A3_3_G51A VL, A3_3.2 VH / A3_4_G51A VL, A3_3.2 VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3_G51A VL, A3_4.2_I100T VH / A3_4_G51A VL, A3_4.2_I100T VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3 VL, A3_4.2_I100T VH / A3_4 VL, A3_4.2_I100T VH / A3_5 VL, A3_4.2 VH / A3_3_G51A VL, A3_4.2 VH / A3_4_G51A VL, A3_4.2 VH / A3_5_G51A VL, A3_6.2_I100T VH / A3_3_G51A VL, A3_6.2_I100T VH / A3_4_G51A VL, A3_6.2_I100T VH / A3_5_G51A VL, A3_6.2_I100T VH / A3_3 VL, A3_6.2_I100T VH / A3_4 VL, A3_2 VH / A3_5_G51A VL, A3-derived_I100T VH CON / A3-derived_G51A VL CON, A3-derived_I100T VH CON / A3-derived VL CON, A3-derived VH CON / A3-derived_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum VL CON, A3-Hum VH CON / A3-Hum_G51A VL CON, 5H8CVH4 VH / 5H8CVL3 VL, 5H8CVH4 VH / 5H8CVL4 VL, 5H8CVH4 VH / 5H8GVL3 VL, 5H8GVH4 VH / 5H8CVL3 VL, 5H8GVH4 VH / 5H8CVL4 VL, 5H8GVH4 VH / 5H8GVL3 VL, 5H8-derivative VH CON / 5H8-derivative VLCON, 5H8-Hum VH CON / 5H8-Hum VL CON, CSP-S-5H8_D98G VH / CSP-S-5H8 VL, 5H8CVH4_D98G VH / 5H8CVL3 VL, 5H8CVH4_D98G VH / 5H8CVL4 VL, 5H8CVH4_D98G VH / 5H8GVL3VL, 5H8GVH4_D98G VH / 5H8CVL3 VL, 5H8GVH4_D98G VH / 5H8CVL4 VL, 5H8GVH4_D98G VH / 5H8GVL3 VL, 5H8-derived_D98G VH CON / 5H8-derived VL CON, or 5H8-Hum_D98G VH CON / 5H8-Hum VL CON, as shown in Table B herein. That is, in some embodiments, the antigen-binding molecule comprises one or more polypeptides comprising: (i) a VH region containing HC-FR1, HC-FR2, HC-FR3, and HC-FR4 as shown in column A of Table B, and (ii) a VL region containing LC-FR1, LC-FR2, LC-FR3, and LC-FR4 as shown in column B of Table B, wherein the sequences in columns A and B are selected from the same row of Table B.

[0239] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more polypeptides, said polypeptide or polypeptides comprising: (i) an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) amino acid sequence identity with the amino acid sequence shown in column A of Table C, and (ii) an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) amino acid sequence identity with the amino acid sequence shown in column B of Table C, wherein the sequences in columns A and B are selected from the same row of Table C.

[0240] In some embodiments, the antigen-binding molecule of this disclosure comprises one or more polypeptides comprising cloned VH and VL regions, said clones being selected from: B035-C03-A3, CSP-S-5H8, A3_3.2 VH / A3_3 VL, A3_3.2 VH / A3_4 VL, A3_3.2 VH / A3_5 VL, A3_4.2 VH / A3_3 VL, A3_4.2 VH / A3_4 VL, A3_4.2 VH / A3_5 VL, A3_6.2 VH / A3_3 VL, A3_6.2 VH / A3_4 VL, A3_6.2 VH / A3_5 VL, A3-derived VH CON / A3-derived VL CON, A3-Hum VH CON / A3-Hum VL CON, B035-C03-A3_D50X, A3_3.2 VH / A3_3_D50X VL, A3_3.2 VH / A3_4_D50X VL, A3_3.2 VH / A3_5_D50X VL, A3_4.2 VH / A3_3_D50X VL, A3_4.2 VH / A3_4_D50X VL, A3_4.2 VH / A3_5_D50X VL, A3_6.2 VH / A3_3_D50X VL, A3_6.2 VH / A3_4_D50X VL, A3_6.2 VH / A3_5_D50X VL, A3-derivative VH CON / A3-derivative VL_D50X CON, A3-Hum VH CON / A3-Hum VL_D50XCON, B035-C03-A3_D50A, A3_3.2 VH / A3_3_D50A VL, A3_3.2 VH / A3_4_D50A VL, A3_3.2VH / A3_5_D50A VL, A3_4.2 VH / A3_3_D50A VL, A3_4.2 VH / A3_4_D50A VL, A3_4.2 VH / A3_5_D50A VL, A3_6.2 VH / A3_3_D50A VL, A3_6.2 VH / A3_4_D50A VL, A3_6.2 VH / A3_5_D50A VL. B035-C03-A3_I100T VH / A3_4_G51A VL, A3_3.2_I100T VH / A3_5_G51A VL, A3_3.2_I100T VH / A3_3 VL, A3_3.2_I100T VH / A3_4 VL, A3_3.2_I100T VH / A3_5 VL, A3_3.2 VH / A3_3_G51A VL, A3_3.2 VH / A3_4_G51A VL, A3_3.2 VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3_G51A VL, A3_4.2_I100T VH / A3_4_G51A VL, A3_4.2_I100T VH / A3_5_G51A VL、A3_4.2_I100T VH / A3_3 VL、A3_4.2_I100T VH / A3_4 VL、A3_4.2_I100T VH / A3_5 VL、A3_4.2 VH / A3_3_G51A VL、A3_4.2 VH / A3_4_G51A VL、A3_4.2 VH / A3_5_G51A VL、A3_6.2_I100T VH / A3_3_G51A VL、A3_6.2_I100T VH / A3_4_G51A VL、A3_6.2_I100T VH / A3_5_G51A VL、A3_6.2_I100T VH / A3_3 VL、A3_6.2_I100T VH / A3_4 VL、A3_6.2_I100T VH / A3_5 VL、A3_6.2 VH / A3_3_G51A VL、A3_6.2 VH / A3_4_G51A VL、A3_6.2 VH / A3_5_G51A VL, A3-Derivative_I100T VH CON / A3-Derivative_G51A VL CON, A3-Derivative_I100T VH CON / A3-Derivative VL CON, A3-Derivative VH CON / A3-Derivative_G51A VL CON, A3-Hum_I100TVH CON / A3-Hum_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum VL CON, A3-Hum VH CON / A3-Hum_G51A VL CON, 5H8CVH4 VH / 5H8CVL3 VL, 5H8CVH4 VH / 5H8CVL4 VL, 5H8CVH4VH / 5H8GVL3 VL, 5H8GVH4 VH / 5H8CVL3 VL, 5H8GVH4 VH / 5H8CVL4 VL, 5H8GVH4 VH / 5H8GVL3 VL, 5H8-derivative VH CON / 5H8-derivative VL CON, 5H8-Hum VH CON / 5H8-Hum VL CON, CSP-S-5H8_D98G VH / CSP-S-5H8 VL, 5H8CVH4_D98G VH / 5H8CVL3 VL, 5H8CVH4_D98G VH / 5H8CVL4 VL, 5H8CVH4_D98G VH / 5H8GVL3 VL, 5H8GVH4_D98G VH / 5H8CVL3 VL, 5H8GVH4_D98G VH / 5H8CVL4 VL, 5H8GVH4_D98G VH / 5H8GVL3 VL, 5H8-derived_D98G VHCON / 5H8-derived VL CON, or 5H8-Hum_D98G VH CON / 5H8-Hum VL CON, as shown in Table C herein. That is, in some embodiments, the antigen-binding molecule comprises one or more polypeptides comprising: (i) the amino acid sequence shown in column A of Table C, and (ii) the amino acid sequence shown in column B of Table C, wherein the sequences in columns A and B are selected from the same row of Table C.

[0241] In some embodiments, the antigen-binding molecule of this disclosure comprises: (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the amino acid sequence shown in column A of Table D; and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% (preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity with the amino acid sequence shown in column B of Table D, wherein the sequences in columns A and B are selected from the same row of Table D.

[0242] In some embodiments, the antigen-binding molecule of this disclosure comprises a polypeptide of an antigen-binding molecule according to Table D herein. That is, in some embodiments, the antigen-binding molecule comprises: (i) a polypeptide comprising or consisting of the amino acid sequence shown in column A of Table D, and (ii) a polypeptide comprising or consisting of the amino acid sequence shown in column B of Table D, wherein the sequences in columns A and B are selected from the same row of Table D.

[0243] Functional properties of antigen-binding molecules The antigen-binding molecules described herein can be characterized by reference to certain functional properties. In some embodiments, the antigen-binding molecules described herein may have one or more of the following properties: With gp130 ( For example Binding to human gp130, rhesus monkey gp130, mouse gp130 and / or rat gp130); It binds to cells expressing gp130; Inhibits signal transduction mediated by receptors containing gp130; Inhibits signal transduction mediated by gp130:IL-6Rα, gp130:IL-11Rα, gp130:OSMRβ, gp130:LIFRβ, gp130:LIFRβ:CNTFRα, gp130:IL-27Rα and / or gp130:IL-12Rβ2; Inhibits signal transduction mediated by cytokines that bind to receptors containing gp130; Inhibits IL-6, IL-11, OSM, LIF, CNTF, CT-1, CLC, IL-27 and / or IL-35-mediated signal transduction; Reduce inflammation and / or fibrosis; To alleviate the pathology of diseases / disorders characterized by inflammation and / or fibrosis; and / or Increases the killing effect on cells expressing gp130.

[0244] It will be understood that a given antigen-binding molecule may exhibit more than one of the properties described in the preceding paragraphs. Appropriate assays can be used to evaluate the properties of a given antigen-binding molecule listed in the preceding paragraphs. For example, an assay could be... For example, body outside The assay may be cell-based or cell-free. In some embodiments, the assay may be... For example, in the body Measurement ,Right now The assay is performed in non-human animals. In some implementations, the assay can be... For example, isolated Measurement, Right now It is performed using cells / tissues / organs obtained from the subjects.

[0245] When assays are cell-based, they may include treating cells with a given antigen-binding molecule to determine whether the antigen-binding molecule exhibits one or more of the aforementioned properties. Assays may employ substances labeled with detectable entities to facilitate their detection. Assays may include the use of a given antigen-binding molecule in a range of amounts / concentrations ( For example The properties were evaluated after treating cells with the dilution series separately. It will be understood that cells preferably express target antigens that bind to antigen-binding molecules. Right now gp130).

[0246] Analysis of such assay results may include determining the concentration at which 50% of the maximum relevant activity level is achieved. The concentration of a given agent at which 50% of the maximum relevant activity level is achieved may be referred to as the "half-maximum effective concentration" (EC50) of the agent relative to the relevant activity, or "EC50". 50 For example, given an antigen-binding molecule that binds to human gp130, EC... 50 This can be the concentration of antigen-binding molecules that achieves 50% of the maximum binding level with human gp130.

[0247] Based on its characteristics, EC 50 Also known as "half-maximum inhibition concentration" or "IC50". 50 "" refers to the concentration of the agent at which 50% of the maximum inhibition level of a given property is observed.

[0248] The antigen-binding molecule described herein binds to gp130. In some embodiments, the antigen-binding molecule binds to human gp130. In some embodiments, the antigen-binding molecule binds to mouse gp130. In some embodiments, the antigen-binding molecule binds to rat gp130. In some embodiments, the antigen-binding molecule binds to rhesus monkey gp130. In some embodiments, the antigen-binding molecule binds to canine gp130.

[0249] In some implementations, the antigen-binding molecule targets human gp130 and one or more homologs of human gp130. For example, Selected from rhesus monkey gp130, mouse gp130, and rat gp130, cross-reactive binding was observed. It should be understood that cross-reactive binding refers to the ability to bind independently to each antigen, rather than the ability of an antigen-binding molecule's antigen-binding domain to simultaneously bind to two or more antigens.

[0250] In some implementations, the antigen-binding molecule exhibits cross-reactivity with human gp130 and rhesus monkey gp130. Right now It can bind to human gp130 and also to rhesus monkey gp130. In some embodiments, the antigen-binding molecule is cross-reactive to human gp130 and mouse gp130. In some embodiments, the antigen-binding molecule is cross-reactive to human gp130 and rat gp130. In some embodiments, the antigen-binding molecule is cross-reactive to human gp130 and rhesus monkey gp130 and mouse gp130 and rat gp130.

[0251] The antigen-binding molecules and antigen-binding domains described herein preferably exhibit specific binding to gp130. As used herein, "specific binding" means selective binding to an antigen, distinguishable from non-specific binding to non-target antigens. Antigen-binding molecules / domains that specifically bind to target molecules preferably bind to the target with a higher affinity and / or a longer duration than binding to other non-target molecules.

[0252] The ability of a given peptide to specifically bind to a given molecule can be determined by analysis according to methods known in the art, such as by ELISA, surface plasmon resonance (SPR); see For example Hearty et al. Methods Mol Biol (2012) 907:411-442), Biolayer Interference (BLI; see For example Lad et al. (2015) J Biomol Screen20(4): 498-507), flow cytometry, or radiolabeled antigen binding assay (RIA) enzyme-linked immunosorbent assay. Such analyses can measure and quantify binding to a given molecule. In some embodiments, binding can be a reaction detected in a given assay.

[0253] In some implementations, the antigen-binding molecule binds to non-target molecules to a degree less than approximately 10% of the antibody-target molecule's binding degree, such as... For exampleMeasured by ELISA, SPR, BLI, or RIA. Alternatively, binding specificity can be reflected by binding affinity, where the dissociation constant (K0) of the antigen-binding molecule is the value of the binding. D At least compared to the K of antigen-binding molecules for non-target molecules D Larger by 0.1 orders of magnitude Right now 0.1 x 10 n , where n is an integer representing the order of magnitude. This may optionally be at least one of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, or 2.0.

[0254] The binding affinity of the antigen-binding molecules described in this article to a given target antigen can be determined using SPR. For example As described in the embodiments of this disclosure.

[0255] In some embodiments, the antigen-binding molecules described herein bind to gp130 with an affinity in the micromolar range. Right now K D = 9.9 x 10 -4 Up to 1 x 10 -6 M. In some embodiments, the antigen-binding molecule described herein binds to gp130 with sub-micromolar affinity. Right now K D < 1 x 10 -6 M. In some embodiments, the antigen-binding molecule described herein binds to gp130 with an affinity in the nanomolar range. Right now K D = 9.9 x 10 -7 Up to 1 x 10 -9 M. In some embodiments, the antigen-binding molecule described herein binds to gp130 with sub-nanomolar affinity. Right now K D < 1 x 10 -9 M. In some embodiments, the antigen-binding molecule described herein binds to gp130 with an affinity in the picomolar range. Right now K D =9.9 x 10 -10 Up to 1 x 10 -12 M. In some embodiments, the antigen-binding molecule described herein binds to gp130 with subpicomolar affinity. Right now K D < 1 x 10 -12 M.

[0256] In some embodiments, the antigen-binding molecule described herein binds to K-type 1 of human gp130.D For 10 μM or less, preferably ≤5 μM, ≤2 μM, ≤1 μM, ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM or ≤1 pM. For example For example, it can be determined by analyzing surface plasmon resonance. For example As described in Example 2 of this document). In some embodiments, the antigen-binding molecule described herein binds to K-type 1 of human gp130. D For 100 nM or less, preferably ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM or ≤1 pM ( For example For example, it can be determined by analyzing surface plasmon resonance. For example As described in Example 2 of this document). In some embodiments, the antigen-binding molecule described herein binds to K-type 1 of human gp130. D It is 3 nM or less, preferably one of ≤2.5 nM, ≤2 nM, ≤1.5 nM, ≤1 nM, ≤750 pM, ≤500 pM or ≤400 pM.

[0257] In some embodiments, the antigen-binding molecule described herein binds to K-type 1 of human gp130. D≤200 nM, preferably ≤190 nM, ≤180 nM, ≤170 nM, ≤160 nM, ≤150 nM, ≤140 nM, ≤130 nM, ≤120 nM, ≤110 nM, ≤100 nM, ≤95 nM, ≤90 nM, ≤85 nM, ≤80 nM, ≤75 nM, ≤60 One of nM, ≤55nM, ≤50 nM, ≤45 nM, ≤40 nM, ≤35 nM, ≤30 nM, ≤25 nM, ≤20 nM, ≤15 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM or ≤1 nM ( For example For example, it can be determined by analyzing surface plasmon resonance. For example As described in Example 2 of this document). In some embodiments, the antigen-binding molecule described herein binds to K-type 1 of human gp130. D The value is ≤166 nM. In some embodiments, the antigen-binding molecule described herein binds to K+ of human gp130 isoform 1. D It is ≤2.91 nM.

[0258] In some embodiments, the antigen-binding molecule described herein binds to EC130 isoform 1 of human gp130. 50 The value is 10 μM or smaller, preferably ≤5 μM, ≤2 μM, ≤1 μM, ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM or ≤1 pM.

[0259] The antigen-binding molecule of this disclosure can bind to a specific region of interest of gp130. According to this disclosure, the antigen-binding molecule can bind to a linear epitope of gp130, said linear epitope consisting of a continuous amino acid sequence (…). Right now It is composed of an amino acid primary sequence. In some embodiments, the antigen-binding molecule may bind to a conformational epitope of gp130, which is composed of a discontinuous amino acid sequence.

[0260] Those skilled in the art can use various methods well-known in the art to determine the region of a given target molecule to which an antigen-binding molecule binds, including X-ray cocrystallation analysis of antibody-antigen complexes, peptide scanning, mutagenesis mapping, hydrogen-deuterium exchange analysis by mass spectrometry, phage display, competitive ELISA, and proteolysis-based "protective" methods. Such methods are described, for example, in Gershoni... et al. , BioDrugs, 2007, 21(3):145-156, which is hereby incorporated in its entirety by reference.

[0261] In some embodiments, the antigen-binding molecule of this disclosure binds to the extracellular domain of gp130. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:75. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:75.

[0262] In some embodiments, the antigen-binding molecule of this disclosure binds to the cytokine-binding module of gp130. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:85. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:85.

[0263] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:89. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:89. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:89. In some embodiments, the epitope of the antigen-binding molecule comprises or is composed of the amino acid sequence shown in SEQ ID NO:89. In some embodiments, the antigen-binding molecule binds to a polypeptide that comprises or is composed of the amino acid sequence shown in SEQ ID NO:89.

[0264] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:86. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:86. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:86. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:86. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:87. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:87. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:87. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:87. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:88. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:88. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids in the region shown in SEQ ID NO:88. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:88.

[0265] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:86, 87, and 88. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:86, 87, and 88. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:86, 87, and 88. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:86, 87, and 88.

[0266] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:93. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:93. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:93. In some embodiments, the epitope of the antigen-binding molecule comprises or is composed of the amino acid sequence shown in SEQ ID NO:93. In some embodiments, the antigen-binding molecule binds to a polypeptide that comprises or is composed of the amino acid sequence shown in SEQ ID NO:93.

[0267] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:90. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:90. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:90. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:90. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:91. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:91. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:91. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:91. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:92. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:92. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids in the region shown in SEQ ID NO:92. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:92.

[0268] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:90, 91, and 92. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:90, 91, and 92. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:90, 91, and 92. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:90, 91, and 92.

[0269] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:169. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:169. In some embodiments, the antigen-binding molecule binds to gp130 via contacting one or more amino acids of the region shown in SEQ ID NO:169. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:169. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:170. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:170. In some embodiments, the antigen-binding molecule binds to gp130 via contacting one or more amino acids of the region shown in SEQ ID NO:170. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:170.

[0270] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:171. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:171. In some embodiments, the antigen-binding molecule binds to gp130 via contacting one or more amino acids of the region shown in SEQ ID NO:171. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:171. In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:172. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:172. In some embodiments, the antigen-binding molecule binds to gp130 via contacting one or more amino acids of the region shown in SEQ ID NO:172. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:172.

[0271] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:169 and 170. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:169 and 170. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:169 and 170. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:169 and 170.

[0272] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:171 and 172. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:171 and 172. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:171 and 172. In some embodiments, the epitope of the antigen-binding molecule comprises the amino acid sequence shown in SEQ ID NO:171 and 172.

[0273] In some embodiments, the antigen-binding molecule binds to the gp130 region shown in SEQ ID NO:173. In some embodiments, the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:173. In some embodiments, the antigen-binding molecule binds to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:173. In some embodiments, the epitope of the antigen-binding molecule comprises or is composed of the amino acid sequence shown in SEQ ID NO:173. In some embodiments, the antigen-binding molecule binds to a polypeptide that comprises or is composed of the amino acid sequence shown in SEQ ID NO:173.

[0274] In some embodiments, the antigen-binding molecule does not bind to the gp130 region shown in SEQ ID NO:178. In some embodiments, the antigen-binding molecule does not contact the gp130 region shown in SEQ ID NO:178. In some embodiments, the antigen-binding molecule does not bind to gp130 via contact with one or more amino acids of the region shown in SEQ ID NO:178. In some embodiments, the epitope of the antigen-binding molecule does not consist of the amino acid sequence shown in SEQ ID NO:178. In some embodiments, the antigen-binding molecule does not bind to a polypeptide consisting of the amino acid sequence shown in SEQ ID NO:178.

[0275] The ability of an antigen-binding molecule to bind a given peptide / peptide can be analyzed using methods well known to those skilled in the art, including ELISA, Western blotting, etc. For example Analysis was performed using Western blotting, immunoprecipitation, SPR, and BLI.

[0276] In some implementations, the antigen-binding molecule is able to bind to the same region of gp130, or overlapping regions of gp130, with antibodies containing both the VH and VL regions (see [link to implementation details]). For exampleThe antibody that binds to the gp130 region as shown in Table C) is selected from the following: B035-C03-A3, CSP-S-5H8, A3_3.2 VH / A3_3 VL, A3_3.2 VH / A3_4 VL, A3_3.2 VH / A3_5 VL, A3_4.2 VH / A3_3 VL, A3_4.2 VH / A3_4 VL, A3_4.2 VH / A3_5 VL, A3_6.2 VH / A3_3 VL, A3_6.2 VH / A3_4 VL, A3_6.2 VH / A3_5 VL, A3-derived VH CON / A3-derived VLCON, A3-Hum VH CON / A3-Hum VL CON, B035-C03-A3_D50X, A3_3.2 VH / A3_3_D50X VL, A3_3.2 VH / A3_4_D50X VL, A3_3.2 VH / A3_5_D50X VL, A3_4.2 VH / A3_3_D50X VL, A3_4.2 VH / A3_4_D50X VL, A3_4.2 VH / A3_5_D50X VL, A3_6.2 VH / A3_3_D50X VL, A3_6.2 VH / A3_4_D50X VL, A3_6.2 VH / A3_5_D50X VL, A3-derivative VH CON / A3-derivative VL_D50X CON, A3-Hum VH CON / A3-Hum VL_D50X CON, B035-C03-A3_D50A, A3_3.2 VH / A3_3_D50A VL, A3_3.2 VH / A3_4_D50A VL, A3_3.2 VH / A3_5_D50A VL, A3_4.2 VH / A3_3_D50A VL, A3_4.2 VH / A3_4_D50A VL, A3_4.2 VH / A3_5_D50A VL, A3_6.2 VH / A3_3_D50A VL, A3_6.2 VH / A3_4_D50A VL, A3_6.2 VH / A3_5_D50A VL. B035-C03-A3_I100T VH / A3_4_G51A VL, A3_3.2_I100T VH / A3_5_G51A VL, A3_3.2_I100T VH / A3_3 VL, A3_3.2_I100TVH / A3_4 VL, A3_3.2_I100T VH / A3_5 VL, A3_3.2 VH / A3_3_G51A VL, A3_3.2 VH / A3_4_G51A VL, A3_3.2 VH / A3_5_G51A VL, A3_4.2_I100T VH / A3_3_G51A VL, A3_4.2_I100T VH / A3_4_G51A VL, A3_4.2_I100T VH / A3_5_G51A VL、A3_4.2_I100T VH / A3_3VL、A3_4.2_I100T VH / A3_4 VL、A3_4.2_I100T VH / A3_5 VL、A3_4.2 VH / A3_3_G51AVL、A3_4.2 VH / A3_4_G51A VL、A3_4.2 VH / A3_5_G51A VL、A3_6.2_I100T VH / A3_3_G51A VL、A3_6.2_I100T VH / A3_4_G51A VL、A3_6.2_I100T VH / A3_5_G51A VL、A3_6.2_I100T VH / A3_3 VL、A3_6.2_I100T VH / A3_4 VL、A3_6.2_I100T VH / A3_5 VL、A3_6.2VH / A3_3_G51A VL、A3_6.2 VH / A3_4_G51A VL、A3_6.2 VH / A3_5_G51A VL, A3-derived_I100T VH CON / A3-derived_G51A VL CON, A3-derived_I100T VH CON / A3-derived VL CON, A3-derived VH CON / A3-derived_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum_G51A VL CON, A3-Hum_I100T VH CON / A3-Hum VL CON, A3-Hum VH CON / A3-Hum_G51A VL CON, 5H8CVH4VH / 5H8CVL3 VL, 5H8CVH4 VH / 5H8CVL4 VL, 5H8CVH4 VH / 5H8GVL3 VL, 5H8GVH4 VH / 5H8CVL3 VL, 5H8GVH4 VH / 5H8CVL4 VL, 5H8GVH4 VH / 5H8GVL3 VL, 5H8-derivative VH CON / 5H8-derivative VL CON, 5H8-Hum VH CON / 5H8-Hum VL CON, CSP-S-5H8_D98G VH / CSP-S-5H8VL, 5H8CVH4_D98G VH / 5H8CVL3 VL, 5H8CVH4_D98G VH / 5H8CVL4 VL, 5H8CVH4_D98G VH / 5H8GVL3 VL, 5H8GVH4_D98G VH / 5H8CVL3 VL, 5H8GVH4_D98G VH / 5H8CVL4 VL, 5H8GVH4_D98G VH / 5H8GVL3 VL, 5H8-derived_D98G VH CON / 5H8-derivedVL CON, or 5H8-Hum_D98G VH CON / 5H8-Hum VL CON.

[0277] Whether a test antigen-binding molecule binds to the same or overlapping region of a given target as a reference antigen-binding molecule can be assessed, for example, by analyzing (i) the interaction between the test antigen-binding molecule and the target in the absence of a reference antigen-binding molecule, and (ii) the interaction between the test antigen-binding molecule and the target in the presence of a reference antigen-binding molecule, or after incubation of the target with a reference antigen-binding molecule. A reduced level of interaction between the test antigen-binding molecule and the target, as determined by analysis according to (ii), may support the inference that both the test and reference antigen-binding molecules bind to the same or overlapping region of the target. Suitable assays for such analyses include... For example Competitive ELISA assay and epitope binning assay.

[0278] In some implementations, when gp130 is on the cell surface ( Right now When expressed in or at the cell membrane, the antigen-binding molecule disclosed herein binds to the antigen-binding molecule (…). Right now Extracellular antigen-binding molecules (AGMs) bind to gp130 in regions that can be accessed. In some embodiments, the antigen-binding molecule binds to gp130 expressed on the cell surface of cells expressing gp130. In some embodiments, the antigen-binding molecule binds to gp130 expressed on the cell surface of cells expressing gp130. For example (fibroblasts) bind. In some implementations, the antigen-binding molecule does not bind ( Right now Cells lacking gp130 surface expression (basically not bound).

[0279] Antigen-binding molecules and given cell types ( For example The ability of cells expressing gp130 or not to bind antigen-binding molecules can be assessed by contacting the cells with antigen-binding molecules and detecting the antigen-binding molecules bound to the cells. For example The analysis is performed after a washing step to remove unbound antigen-binding molecules. The ability of antigen-binding molecules to bind to specific cell types can be analyzed using methods such as flow cytometry and immunofluorescence microscopy.

[0280] In some embodiments, the antigen-binding molecule of this disclosure inhibits signal transduction mediated by a receptor containing gp130. In some embodiments, the antigen-binding molecule inhibits gp130-mediated signal transduction (…). For example (As described above, gp130-mediated signaling). Signaling mediated by gp130 and / or by receptors containing gp130 can be achieved using cells expressing gp130 / related receptors. For example The analysis is performed using measurements designed to detect and / or quantify gp130-mediated signal transduction. Suitable measurements include... For example An assay used to detect the phosphorylation / activation / expression of factors that are phosphorylated / activated / expressed due to signal transduction via gp130 / a receptor containing gp130.

[0281] Such assays may include contacting cells expressing a given cytokine receptor containing gp130 with an antigen-binding molecule according to the present disclosure. For example In the presence of ligands for cytokine receptors. For example, an assay to study the ability of antigen-binding molecules to inhibit IL-6-mediated signaling and / or the ability of antigen-binding molecules to inhibit signaling mediated by gp130:IL-6Rα may include contacting cells expressing the gp130:IL-6Rα complex with an antigen-binding molecule according to the present disclosure. For exampleIn the presence of IL-6. Further, for example, an assay to study the ability of an antigen-binding molecule to inhibit IL-11-mediated signaling and / or the ability of an antigen-binding molecule to inhibit signaling mediated by gp130:IL-11Rα may include contacting cells expressing the gp130:IL-11Rα complex with an antigen-binding molecule according to the present disclosure. For example In the presence of IL-11.

[0282] For example, this can be achieved by evaluating signal transduction pathways triggered by signal transduction via gp130 / cytokine receptors containing gp130. For example The phosphorylation of one or more signal transduction molecules in the JAK / STAT, MAPK / ERK, or PI3K / AKT pathways can be used to study gp130-mediated signal transduction. For example, the level of gp130-mediated signal transduction can be investigated by detecting and / or quantifying JAK1, JAK2, STAT1, STAT3, STAT5, and / or ERK (JAK / STAT, MAPK / ERK, or PI3K / AKT pathways). For example The phosphorylation levels of STAT3 and / or ERK are analyzed. For example, in experimental embodiments of this disclosure, gp130-mediated signaling (particularly gp130-mediated signaling in response to IL-6, IL-11, OSM, LIF, CT-1, or CNTF stimulation) is analyzed by assessing the phosphorylation of STAT3 or ERK1 / 2 using Western blotting.

[0283] The level of gp130-mediated signaling can also be assessed by analyzing one or more correlations in gp130-mediated signaling. For example, gp130-mediated signaling can be studied by detecting and / or quantifying the expression or activity of factors that are upregulated or downregulated due to gp130-mediated signaling. In some embodiments, gp130-mediated signaling can be studied by detecting and / or quantifying the expression of factors that are upregulated due to gp130-mediated signaling. For example Pro-inflammatory / profibrotic / profibrotic factors. For example, in experimental embodiments of this disclosure, gp130-mediated signaling was analyzed by assessing the expression of αSMA and MMP2.

[0284] Reporter gene-based methods can also be used to analyze gp130-mediated signal transduction levels. For example, gp130-mediated signal transduction can be studied using reporter cell lines that stably express luciferase reporter genes whose expression is driven by gp130-mediated signal transduction. For instance, in experimental embodiments of this disclosure, gp130-mediated signal transduction was studied using the HEK293 reporter cell line (STAT3 reporter gene (Luc)-HEK293 cell line (puromycin), BPS Bioscience) containing a luciferase gene controlled by the STAT3 response element.

[0285] In some embodiments, the antigen-binding molecule inhibits gp130-mediated signaling and / or signaling by a gp130-containing receptor to a less extent than in the absence of the antigen-binding molecule (or in the presence of a suitable control antigen-binding molecule). For example Known to not affect the observed signal transduction level by 1-fold (of antigen-binding molecules that do not affect signal transduction mediated by gp130 and / or signal transduction by receptors containing gp130). For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times or ≤0.01 times.

[0286] In some embodiments, the antigen-binding molecule inhibits IL-6-mediated signaling. In some embodiments, the antigen-binding molecule inhibits IL-11-mediated signaling. In some embodiments, the antigen-binding molecule inhibits OSM-mediated signaling. In some embodiments, the antigen-binding molecule inhibits LIF-mediated signaling. In some embodiments, the antigen-binding molecule inhibits CNTF-mediated signaling. In some embodiments, the antigen-binding molecule inhibits CT-1-mediated signaling. In some embodiments, the antigen-binding molecule inhibits CLC-mediated signaling. In some embodiments, the antigen-binding molecule inhibits IL-27-mediated signaling. In some embodiments, the antigen-binding molecule inhibits IL-35-mediated signaling.

[0287] In some implementations, the antigen-binding molecule inhibits IL-6-mediated signaling, and inhibits IL-11-mediated signaling, and inhibits OSM-mediated signaling, and inhibits LIF-mediated signaling, and inhibits CNTF-mediated signaling, and inhibits CT-1-mediated signaling.

[0288] In some implementations, the antigen-binding molecule inhibits IL-6-mediated signaling, and inhibits IL-11-mediated signaling, and inhibits OSM-mediated signaling, and inhibits LIF-mediated signaling, and inhibits CNTF-mediated signaling, and inhibits CT-1-mediated signaling, and inhibits CLC-mediated signaling, and inhibits IL-27-mediated signaling, and inhibits IL-35-mediated signaling.

[0289] In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-6Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-11Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:OSMRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:LIFRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:LIFRβ:CNTFRα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-27Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-12Rβ2.

[0290] In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-6Rα, and inhibits signal transduction mediated by gp130:IL-11Rα, and inhibits signal transduction mediated by gp130:OSMRβ, and inhibits signal transduction mediated by gp130:LIFRβ, and inhibits signal transduction mediated by gp130:LIFRβ:CNTFRα.

[0291] In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by gp130:IL-6Rα, and inhibits signal transduction mediated by gp130:IL-11Rα, and inhibits signal transduction mediated by gp130:OSMRβ, and inhibits signal transduction mediated by gp130:LIFRβ, and inhibits signal transduction mediated by gp130:LIFRβ:CNTFRα, and inhibits signal transduction mediated by gp130:IL-27Rα, and inhibits signal transduction mediated by gp130:IL-12Rβ2.

[0292] In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of IL-6 to cells expressing gp130:IL-6Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of LIF to cells expressing gp130:LIFRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of CT-1 to cells expressing gp130:LIFRβ. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα. In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2.

[0293] In some embodiments, the antigen-binding molecules inhibit signal transduction mediated by the binding of IL-6 to cells expressing gp130:IL-6Rα, and inhibit signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα, and inhibit signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ, and inhibit signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ, and inhibit signal transduction mediated by the binding of LIF to cells expressing gp130:LIFRβ, and inhibit signal transduction mediated by the binding of CT-1 to cells expressing gp130:LIFRβ, and inhibit signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα.

[0294] In some embodiments, the antigen-binding molecule inhibits signal transduction mediated by the binding of IL-6 to cells expressing gp130:IL-6Rα, and inhibits signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα, and inhibits signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ, and inhibits signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ, and inhibits signal transduction mediated by the binding of LIF to cells expressing gp130:LIFRβ. It inhibits signal transduction mediated by the binding of CT-1 to cells expressing gp130:LIFRβ, and also inhibits signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα, CLC to cells expressing gp130:LIFRβ:CNTFRα, IL-27 to cells expressing gp130:IL-27Rα, and IL-35 to cells expressing gp130:IL-12Rβ2.

[0295] In some implementations, the antigen-binding molecule inhibits IL-6-mediated signaling / gp130:IL-6Rα-mediated signaling / IL-6-binding-mediated signaling by cells expressing gp130:IL-6Rα to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules that do not affect IL-6-mediated signal transduction / signal transduction mediated by gp130:IL-6Rα / signal transduction mediated by the binding of IL-6 to cells expressing gp130:IL-6Rα. For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For exampleKnown antigen-binding molecules that do not affect IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction), but observed IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction) accounts for more than 25% of the observed IL-6-mediated signal transduction. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits IL-6-mediated signaling / gp130:IL-6Rα-mediated signaling / signaling mediated by the binding of IL-6 to cells expressing gp130:IL-6Rα, wherein IC50... 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0296] In some implementations, the antigen-binding molecule inhibits IL-11-mediated signaling / gp130:IL-11Rα-mediated signaling / IL-11 binding-mediated signaling to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules that do not affect IL-11-mediated signal transduction / signal transduction mediated by gp130:IL-11Rα / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα. For example≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect IL-11-mediated signal transduction / signal transduction mediated by gp130:IL-11Rα / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα. Observed IL-11-mediated signal transduction / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits IL-11-mediated signaling / gp130:IL-11Rα-mediated signaling / signaling mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα, wherein IC 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0297] In some implementations, the antigen-binding molecule inhibits OSM-mediated signaling / gp130:OSMRβ-mediated signaling / gp130:LIFRβ-mediated signaling / OSM-mediated signaling by binding to cells expressing gp130:OSMRβ / OSM-mediated signaling by binding to cells expressing gp130:LIFRβ to a level less than that in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The level of signal transduction observed is 1 times that of antigen-binding molecules that do not affect OSM-mediated signal transduction / signal transduction mediated by gp130:OSMRβ / signal transduction mediated by gp130:LIFRβ / signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ. For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ (more than 25% of the observed OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ). For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / OSM binding to cells expressing gp130:OSMRβ-mediated signal transduction / OSM binding to cells expressing gp130:LIFRβ-mediated signal transduction, wherein IC 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0298] In some implementations, the antigen-binding molecule inhibits LIF-mediated signaling / gp130:LIFRβ-mediated signaling / signaling mediated by the binding of LIF to cells expressing gp130:LIFRβ to a level less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules that do not affect LIF-mediated signal transduction / signal transduction mediated by gp130:LIFRβ / signal transduction mediated by the binding of LIF to cells expressing gp130:LIFRβ. For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ (more than 25% of LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits LIF-mediated signaling / gp130:LIFRβ-mediated signaling / signaling mediated by the binding of LIF to cells expressing gp130:LIFRβ, wherein IC 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0299] In some implementations, the antigen-binding molecule inhibits CT-1-mediated signaling / gp130:LIFRβ-mediated signaling / CT-1 binding to cells expressing gp130:LIFRβ-mediated signaling to a level less than that in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules (known to not affect CT-1-mediated signal transduction / signal transduction mediated by gp130:LIFRβ / signal transduction mediated by the binding of CT-1 to cells expressing gp130:LIFRβ). For example≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction (more than 25% of the observed CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / signal transduction mediated by the binding of CT-1 to cells expressing gp130:LIFRβ, wherein IC 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0300] In some implementations, the antigen-binding molecule inhibits CNTF-mediated signaling / signaling mediated by gp130:LIFRβ:CNTFRα / signaling mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). example like Known to not affect CNTF-mediated signal transduction / signal transduction mediated by gp130:LIFRβ:CNTFRα / antigen-binding molecules that mediated signal transduction by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα (1-fold increase in observed signal transduction levels). For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect CNTF-mediated signal transduction / signal transduction mediated by gp130:LIFRβ:CNTFRα / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα. However, CNTF-mediated signal transduction / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα is observed to exceed 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits CNTF-mediated signaling / gp130:LIFRβ:CNTFRα-mediated signaling / signaling mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα, wherein IC 50Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0301] In some implementations, the antigen-binding molecule inhibits CLC-mediated signaling / signaling mediated by gp130:LIFRβ:CNTFRα / signaling mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules that do not affect CLC-mediated signal transduction / signal transduction mediated by gp130:LIFRβ:CNTFRα / signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα. For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect CLC-mediated signal transduction (signal transduction mediated by gp130:LIFRβ:CNTFRα, or signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα) have been observed to account for more than 25% of CLC-mediated signal transduction (signal transduction mediated by gp130:LIFRβ:CNTFRα, or signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα). For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits CLC-mediated signal transduction / gp130:LIFRβ:CNTFRα-mediated signal transduction / signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα, wherein IC 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0302] In some implementations, the antigen-binding molecule inhibits IL-27-mediated signaling / gp130:IL-27Rα-mediated signaling / IL-27 binding to cells expressing gp130:IL-27Rα-mediated signaling to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules that do not affect IL-27-mediated signal transduction / signal transduction mediated by gp130:IL-27Rα / signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα. For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For exampleKnown antigen-binding molecules that do not affect IL-27-mediated signal transduction (or signal transduction mediated by gp130:IL-27Rα, or signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα), but observed IL-27-mediated signal transduction (or signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα) exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits IL-27-mediated signaling / gp130:IL-27Rα-mediated signaling / signaling mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα, wherein IC50... 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0303] In some implementations, the antigen-binding molecule inhibits IL-35-mediated signaling / gp130:IL-12Rβ2-mediated signaling / signaling mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2 to less than in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). For example The observed signal transduction level was 1 times that of antigen-binding molecules known to not affect IL-35-mediated signal transduction / signal transduction mediated by gp130:IL-12Rβ2 / signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2. example like≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times, or ≤0.01 times. In some embodiments, antigen-binding molecule inhibition occurs in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules). For example Known antigen-binding molecules that do not affect IL-35-mediated signal transduction / gp130:IL-12Rβ2-mediated signal transduction / signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2. Observed IL-35-mediated signal transduction / gp130:IL-12Rβ2-mediated signal transduction / signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2 exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%. In some embodiments, the antigen-binding molecule inhibits IL-35-mediated signaling / gp130:IL-12Rβ2-mediated signaling / signaling mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2, wherein IC50... 50 Less than 1 μM, preferably ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤900 pM, ≤800 pM, ≤700 pM, ≤600 pM, ≤500 pM, ≤400 pM, ≤300 pM, ≤200 pM, ≤100 pM, ≤50 pM, ≤40 pM, ≤30 pM, ≤20 pM, ≤10 pM, or ≤1 pM. For example Appropriate antigen-binding molecules' ability to inhibit such signal transduction in vitro Determined during the measurement.

[0304] In some implementations, the antigen-binding molecule: (i) inhibits the binding of antigens in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), For exampleKnown antigen-binding molecules that do not affect IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction), but observed IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction) accounts for more than 25% of the observed IL-6-mediated signal transduction. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (ii) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect IL-11-mediated signal transduction / signal transduction mediated by gp130:IL-11Rα / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα. Observed IL-11-mediated signal transduction / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (iii) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ (more than 25% of the observed OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ). For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (iv) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ (more than 25% of LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (v) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction (more than 25% of the observed CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%; and (vi) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect CNTF-mediated signal transduction / signal transduction mediated by gp130:LIFRβ:CNTFRα / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα. However, CNTF-mediated signal transduction / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα is observed to exceed 25%. For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%.

[0305] In some implementations, the antigen-binding molecule: (i) inhibits the binding of antigens in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), For example Known antigen-binding molecules that do not affect IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction), but observed IL-6-mediated signal transduction (including gp130:IL-6Rα-mediated signal transduction and IL-6 binding to cells expressing gp130:IL-6Rα-mediated signal transduction) accounts for more than 25% of the observed IL-6-mediated signal transduction. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (ii) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect IL-11-mediated signal transduction / signal transduction mediated by gp130:IL-11Rα / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα. Observed IL-11-mediated signal transduction / signal transduction mediated by the binding of IL-11 to cells expressing gp130:IL-11Rα exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (iii) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For exampleKnown antigen-binding molecules that do not affect OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ (more than 25% of the observed OSM-mediated signal transduction / gp130:OSMRβ-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / Signal transduction mediated by the binding of OSM to cells expressing gp130:OSMRβ / Signal transduction mediated by the binding of OSM to cells expressing gp130:LIFRβ). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (iv) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ (more than 25% of LIF-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / LIF-mediated signal transduction by binding to cells expressing gp130:LIFRβ). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (v) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction (more than 25% of the observed CT-1-mediated signal transduction / gp130:LIFRβ-mediated signal transduction / CT-1 binding to cells expressing gp130:LIFRβ-mediated signal transduction). For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (vi) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect CNTF-mediated signal transduction / signal transduction mediated by gp130:LIFRβ:CNTFRα / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα. However, CNTF-mediated signal transduction / signal transduction mediated by the binding of CNTF to cells expressing gp130:LIFRβ:CNTFRα is observed to exceed 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (vii) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect CLC-mediated signal transduction (signal transduction mediated by gp130:LIFRβ:CNTFRα, or signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα) have been observed to account for more than 25% of CLC-mediated signal transduction (signal transduction mediated by gp130:LIFRβ:CNTFRα, or signal transduction mediated by the binding of CLC to cells expressing gp130:LIFRβ:CNTFRα). For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98% or ≥99%; (viii) Inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect IL-27-mediated signal transduction (or signal transduction mediated by gp130:IL-27Rα, or signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα), but observed IL-27-mediated signal transduction (or signal transduction mediated by the binding of IL-27 to cells expressing gp130:IL-27Rα) exceeds 25%. For example≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%; and (ix) inhibition in the absence of antigen-binding molecules (or in the presence of appropriate control antigen-binding molecules, For example Known antigen-binding molecules that do not affect IL-35-mediated signal transduction / gp130:IL-12Rβ2-mediated signal transduction / signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2. Observed IL-35-mediated signal transduction / gp130:IL-12Rβ2-mediated signal transduction / signal transduction mediated by the binding of IL-35 to cells expressing gp130:IL-12Rβ2 exceeds 25%. For example ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%.

[0306] In some embodiments, the antigen-binding molecule according to this disclosure reduces inflammation. In some embodiments, the antigen-binding molecule according to this disclosure reduces fibrosis. In some embodiments, the antigen-binding molecule according to this disclosure reduces both inflammation and fibrosis.

[0307] In appropriate in vitro or in vivo In inflammation / fibrosis models, assess the ability and extent to which a given antigen-binding molecule reduces inflammation and / or fibrosis. For example, in vitro Methods may include enabling cells (in the presence of antigen-binding molecules) For example Cells expressing gp130 (and pro-inflammatory, pro-fibrotic, or pro-fibrotic stimuli) For example IL-6, IL-11, TGFβ1 wait The cells were contacted and subsequently evaluated to determine the level of one or more related conditions of inflammation / fibrosis. For example, in the experimental embodiments of this disclosure, in... in vitro The ability of gp130-specific antigen-binding molecules to alleviate fibrosis was evaluated in the assay, in which cells were stimulated with TGFβ1 or IL-11 and the fibrotic response was subsequently analyzed by assessing MMP2 expression.

[0308] The ability and extent to which a given antigen-binding molecule reduces inflammation and / or fibrosis. in vivoAnalysis can be performed using non-human animal models of diseases characterized as inflammation and / or fibrosis. Such models can also be used to assess the ability of antigen-binding molecules to alleviate the pathology of diseases / disorders characterized as inflammation and / or fibrosis. Such analyses may include administering antigen-binding molecules to subjects with diseases characterized as inflammation and / or fibrosis and subsequently assessing one or more associations of inflammation / fibrosis in the subjects. For example, in experimental embodiments of this disclosure, the reduction of symptoms by gp130-specific antigen-binding molecules in a folic acid-induced mouse model of acute kidney injury was evaluated. in vivo The ability to induce inflammation and fibrosis, through assessment For example Serum IL-6 levels, renal collagen content, and renal expression of pro-inflammatory genes ( For example, Ccl2, Ccl5, Il6, Tnfα and Il1β ) and profibrotic genes ( For example, Col1a1, Col1a2, Col3a1, Fn, Acta2 and Il11 The renal expression of ) is carried out.

[0309] In some embodiments, administration of the antigen-binding molecule according to this disclosure can inhibit the development / progression of a disease / disorder characterized as inflammation and / or fibrosis. In some embodiments, administration of the antigen-binding molecule can reduce the severity of symptoms of a disease / disorder characterized as inflammation and / or fibrosis. For example As determined in the appropriate model.

[0310] In some embodiments, in a given assay, the antigen-binding molecule of this disclosure is able to detect one or more related levels of inflammation and / or fibrosis. For example In inflammation / fibrosis in vitro In the model, or in diseases / disorders characterized as inflammation / fibrosis in vivo In the model, the level decreased to less than one-fold of the level observed in the absence of treatment with antigen-binding molecules (or after treatment with an appropriate control antigen-binding molecule known not to affect the relevant correlation). For example ≤0.99 times, ≤0.95 times, ≤0.9 times, ≤0.85 times, ≤0.8 times, ≤0.75 times, ≤0.7 times, ≤0.65 times, ≤0.6 times, ≤0.55 times, ≤0.5 times, ≤0.45 times, ≤0.4 times, ≤0.35 times, ≤0.3 times, ≤0.25 times, ≤0.2 times, ≤0.15 times, ≤0.1 times, ≤0.05 times or ≤0.01 times.

[0311] The antigen-binding molecule according to this disclosure may include one or more portions for enhancing cell killing of gp130-expressing cells or reducing the number / proportion of gp130-expressing cells. For example, the antigen-binding molecule according to this disclosure may... example like Includes the Fc region and / or the drug portion.

[0312] In some embodiments, the antigen-binding molecule according to this disclosure can enhance ( Right now Upregulation / enhancement of cell killing in cells containing / expressing gp130. In some implementations, antigen-binding molecules do not enhance ( Right now (It will not significantly enhance) the cell killing effect of cells lacking gp130 surface expression.

[0313] For example, it can be used in Zaritskaya et al. Any method used to study cell killing as reviewed in Expert Rev Vaccines (2011), 9(6):601-616 is hereby incorporated in its entirety by reference. Cytotoxicity / cell killing assays in vitro Examples of assays include release assays, such as... 51 Cr release assay, lactate dehydrogenase (LDH) release assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) release assay, and calcein-acetoxymethyl (calcein-AM) release assay. These assays measure cell killing based on detecting factors released from lysed cells. Cell killing of a given effector immune cell type against a given test cell type can be achieved through... For example The test cells were co-cultured with effector immune cells, and the survival / death ratio was measured after an appropriate time period. For example The number / proportion of lysed test cells is analyzed. Other suitable assays include those described in Cerignoli. et al. , PLoS One. (2018) 13(3): e0193498 (included hereby by reference in its entirety) xCELLigence real-time cell lysis in vitro Efficacy determination.

[0314] In some embodiments, the antigen-binding molecule according to this disclosure can reduce the number / proportion of cells expressing gp130. In some embodiments, the antigen-binding molecule according to this disclosure can reduce the number / proportion of cells expressing gp130. In some embodiments, the antigen-binding molecule according to this disclosure can deplete / enhance the depletion of such cells.

[0315] The Fc region provides the means for interaction with Fc receptors and other molecules in the immune system, thereby exerting functional effects. IgG Fc-mediated effector functions are... For example Jefferis et al. A review was conducted in *Immunol Rev 1998*, 163:59-76 (included hereby by reference in its entirety), which discussed the interaction between the Fc region and Fc receptors expressed by immune cells, and the Fc-mediated immune cell activity. For exampleThe complement pathway is mediated by the recruitment and activation of macrophages, dendritic cells, neutrophils, basophils, eosinophils, platelets, mast cells, NK cells, and T cells. This recruitment of complement pathway components occurs through the binding of the Fc region to complement protein C1q, followed by activation of the complement cascade. Fc-mediated functions include Fc receptor binding, antibody-dependent cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), formation of the membrane attack complex (MAC), cell degranulation, production of cytokines and / or chemokines, and antigen processing and presentation.

[0316] In some embodiments, the antigen-binding molecule according to this disclosure comprises the ability to enhance / guide targeting of cells expressing gp130 ( For example One or more of ADCC, ADCP, CDC in cells expressing gp130 on the cell surface, and / or enhance the formation of MAC or cell degranulation Fc regions on cells expressing gp130.

[0317] In some embodiments, the antigen-binding molecule according to this disclosure can enhance / guide ADCC against cells expressing gp130.

[0318] The ability and extent to which a given antigen-binding molecule can induce ADCC in a given target cell type can be determined. For example According to the description in Yamashita et al. Scientific Reports (2016) 6:19772 (included hereby by reference in its entirety), or by description in For example Jedema et al. Blood (2004) 103: 2677–82 (included hereby by reference in its entirety) 51 Cr release assay is used for analysis. The ability and extent to which a given antigen-binding molecule can induce ADCP can be determined. For example According to Kamen et al. The method described in J Immunol (2017) 198 (1 Supplement) 157.17 (incorporated in its entirety by reference) was analyzed. The ability and extent to which a given antigen-binding molecule can induce CDC can be analyzed. For example Determination using C1q binding For example As described in Schlothauer et al. , Protein Engineering, Design and Selection (2016), 29(10):457–466 (included hereby by reference in its entirety) for analysis.

[0319] In some embodiments, the antigen-binding molecule according to this disclosure comprises a pharmaceutical moiety. The antigen-binding molecule may be conjugated to a pharmaceutical moiety. Antibody-drug conjugates are... For example Parslow et al. A review was conducted in Biomedicines. 2016 Sep; 4(3):14 (hereinafter incorporated in its entirety by reference). In some embodiments, the drug component is a cytotoxic agent or contains a cytotoxic agent, such that the antigen-binding molecule is effective against cells expressing gp130 ( For example Cells expressing gp130 on their cell surface exhibited cytotoxicity.

[0320] Connectors and additional sequences The antigen-binding molecules and peptides disclosed herein may further include amino acids or amino acid sequences.

[0321] The antigen-binding molecules and peptides disclosed herein may include one or more adapter sequences between amino acid sequences. For example, the adapter sequence may be provided between a VH sequence and a VL sequence, thereby providing a VH-VL bond. For example In the scFv molecule).

[0322] Connector sequences are known to those skilled in the art and are described, for example, in Chen et al. , Adv Drug DelivRev (2013) 65(10): 1357-1369, which is hereby incorporated in its entirety by reference. In some embodiments, the linker sequence may be a flexible linker sequence. A flexible linker sequence allows relative movement of amino acid sequences linked by the linker sequence. Flexible linkers are known to those skilled in the art, and in Chen et al. Several were identified in Adv Drug Deliv Rev (2013) 65(10):1357-1369. Flexible linker sequences typically contain a high proportion of glycine and / or serine residues.

[0323] In some embodiments, the adapter sequence comprises at least one glycine residue and / or at least one serine residue. In some embodiments, the adapter sequence comprises or consists of glycine and serine residues. In some embodiments, the adapter sequence has the structure (GxS)n or (GxS)nGm; where G = glycine, S = serine, x = 3 or 4, n = 2, 3, 4, 5 or 6, and m = 0, 1, 2 or 3. In some embodiments, the adapter sequence comprises one or more of the sequence motif G4S. For example 1, 2, 3, 4, 5, or 6 copies For example(In tandem). In some embodiments, the adapter sequence comprises (G4S)4 or (G4S)6 or consists of therewith. In some embodiments, the length of the adapter sequence is 1-2, 1-3, 1-4, 1-5, 1-10, 1-15, 1-20, 1-25 or 1-30 amino acids.

[0324] The antigen-binding molecules and peptides disclosed herein may include amino acid sequences to facilitate the expression, folding, transport, processing, purification, or detection of the antigen-binding molecules / peptides. For example, the antigen-binding molecules and peptides disclosed herein may additionally include amino acid sequences forming a detectable moiety. For example As described below.

[0325] The antigen-binding molecules and peptides disclosed herein may additionally include a signal peptide (also known as a leader sequence or signal sequence). A signal peptide typically consists of a sequence of 5-30 hydrophobic amino acids forming a single α-helix. Secretory proteins and proteins expressed on the cell surface often contain signal peptides. Signal peptides are known for many proteins and are documented in databases such as GenBank, UniProt, and Ensembl, and / or For example Amino acid sequence analysis tools such as SignalP (Petersen) can be used. et al. , 2011 Nature Methods 8: 785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24: 2172-2176) to identify / predict.

[0326] Signal peptides can be present at the N-terminus of antigen-binding molecules / peptides, or in newly synthesized antigen-binding molecules / peptides. Signal peptides facilitate efficient transport of antigen-binding molecules / peptides. They are typically removed through cleavage and are therefore not included in mature antigen-binding molecules / peptides.

[0327] Signal peptides are known for many proteins and are documented in databases such as GenBank, UniProt, SwissProt, TrEMBL, Protein Information Resources, Protein Databases, Ensembl, and InterPro, and / or For example Amino acid sequence analysis tools such as SignalP (Petersen) can be used. et al. , 2011 Nature Methods 8:785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24: 2172-2176) to identify / predict.

[0328] Markers and conjugates In some embodiments, the antigen-binding molecule or polypeptide according to this disclosure includes a detectable portion.

[0329] In some implementations, the detectable portion is fluorescently labeled, phosphorescently labeled, luminescently labeled, or immunodetectable labeled. For example Antigen-binding molecules or peptides can be covalently or non-covalently labeled (e.g., epitope labeling, radioactive labeling, chemical labeling, nucleic acid labeling, or enzyme labeling).

[0330] Fluorescent labeling includes For example Fluorescein, rhodamine, allophycocyanin, eosin and NDB, green fluorescent protein (GFP), chelates of rare earth elements (such as europium (Eu), terbium (Tb), and samarium (Sm)), tetramethylrhodamine, Texas red, 4-methylumbelliferone, 7-amino-4-methylcoumarin, Cy3 and Cy5. Radiolabeling includes radioactive isotopes such as hydrogen. 3 ,sulfur 35 ,carbon 14 ,phosphorus 32 ,iodine 123 ,iodine 125 ,iodine 126 ,iodine 131 ,iodine 133 ,bromine 77 ,technetium 99m ,indium 111 ,indium 113m ,gallium 67 ,gallium 68 ,ruthenium 95 ,ruthenium 97 ,ruthenium 103 ,ruthenium 105 ,mercury 207 ,mercury 203 ,rhenium 99m ,rhenium 101 ,rhenium 105 ,scandium 47 ,tellurium 121m ,tellurium 122m ,tellurium 125m ,thulium 165 ,thulium 167 ,thulium 168 ,copper 67 ,fluorine 18 ,yttrium 90 ,palladium 100 ,bismuth 217 and antimony 211 Luminescent tags include radioluminescence and chemiluminescence. For exampleImmunologically detectable markers include acridinium esters, luminol, isoluminol, and bioluminescent labels. Immunologically detectable markers include haptens, peptides / peptides, antibodies, receptors, and ligands such as biotin, avidin, streptavidin, or digoxigenin. Nucleic acid markers include aptamers.

[0331] In some implementations, the antigen-binding molecule / peptide contains an epitope tag. For example His ( For example 6XHis), FLAG, c-Myc, StrepTag, hemagglutinin, E, calmodulin-binding protein (CBP), glutathione S-transferase (GST), maltose-binding protein (MBP), thioredoxin, S-peptide, T7 peptide, SH2 domain, avidin, streptavidin, and hapten ( example like Biotin, digoxin, dinitrophenol), optionally at the N or C terminus of the antigen-binding molecule / peptide.

[0332] In some implementations, the antigen-binding molecule / peptide contains a portion with detectable activity. For example The enzyme component. The enzyme component includes... For example Luciferase, glucose oxidase, galactosidase ( For example β-galactosidase), glucuronidase, phosphatase ( For example alkaline phosphatase), peroxidase ( For example Horseradish peroxidase and cholinesterase.

[0333] In some embodiments, the antigen-binding molecule or polypeptide of this disclosure comprises a chemical moiety. In some embodiments, the antigen-binding molecule / polypeptide of this disclosure is conjugated to a chemical moiety.

[0334] The chemical component can be the part used to provide therapeutic effects. Right now Drug component. The drug component can be a small molecule ( example like Low molecular weight (< 1000 Daltons, typically between about 300 and 700 Daltons) organic compounds. The pharmaceutical section describes... For example Parslow et al. , Biomedicines. Sep 2016; 4(3):14 (included hereby by reference in its entirety). In some embodiments, the pharmaceutical portion may be or contain a cytotoxic agent. In some embodiments, the pharmaceutical portion may be or contain a chemotherapeutic agent. The pharmaceutical portion includes For exampleKazimidic acid, DM1, DM4, methylaurestatin E (MMAE), methylaurestatin F (MMAF), SN-38, doxorubicin, duocarmycin, D6.5, and PBD.

[0335] Nucleic acid and vector This disclosure provides one or more nucleic acids that encode an antigen-binding molecule or polypeptide according to this disclosure. In some embodiments, the nucleic acid comprises or is composed of DNA and / or RNA.

[0336] The antigen-binding molecules or peptides according to this disclosure can be produced intracellularly by translating the RNA encoding the peptide. The antigen-binding molecules or peptides according to this disclosure can also be produced intracellularly by transcribing and subsequently translating the transcribed RNA from the nucleic acid encoding the peptide.

[0337] In some implementations, nucleic acids may be vectors or multiple vectors, or may be contained in or contained within vectors or multiple vectors. As used herein, a "vector" is a nucleic acid molecule used as a medium for transferring exogenous nucleic acids into cells.

[0338] Therefore, this disclosure also provides one or more vectors comprising nucleic acids or nucleic acids according to this disclosure. The vector can facilitate the delivery of nucleic acids encoding polypeptides according to this disclosure to cells. The vector may be an expression vector containing elements required for expressing the polypeptide according to this disclosure. The vector may contain elements that facilitate the integration of the nucleic acid into the genomic DNA of the cell into which the vector is introduced.

[0339] According to this disclosure, nucleic acids and vectors can be provided in purified or isolated forms. Right now Derived from other nucleic acids or naturally occurring biological materials.

[0340] A vector can be a carrier used to express nucleic acids in cells. Right now Expression vectors. Such vectors may include a promoter sequence operatively linked to a nucleotide sequence encoding an antigen-binding molecule or polypeptide according to this disclosure. The vector may also include a stop codon. Right now The vector contains the nucleotide sequence from 3' to the nucleotide sequence encoding the polypeptide and an expression enhancer. Any suitable vector, promoter, enhancer, and stop codon known in the art can be used to express peptides or polypeptides from the vectors disclosed herein.

[0341] The term "operably linked" may include situations where: a nucleic acid encoding a polypeptide according to this disclosure and a regulatory nucleic acid sequence ( For exampleThe promoter and / or enhancer are covalently linked in such a way that the expression of the nucleic acid encoding the polypeptide is under the influence or control of the regulatory nucleic acid sequence (thus forming an expression cassette). Therefore, if the regulatory sequence can influence the transcription of the nucleic acid sequence, the regulatory sequence can be operatively linked to the selected nucleic acid sequence. The resulting transcript can then be translated into the desired polypeptide.

[0342] The vectors considered in conjunction with this disclosure include DNA vectors, RNA vectors, and plasmids. For example conjugated plasmids ( For example F plasmid, non-conjugated plasmid, R plasmid, col plasmid, episome, viral vector ( For example Retroviral vectors, For example γ-retroviral vector ( For example A vector derived from murine leukemia virus (MLV). For example SFG vectors, lentiviral vectors, adenovirus vectors, adeno-associated virus vectors, vaccinia virus vectors, and herpesvirus vectors, transposon-based vectors, and artificial chromosomes ( For example (Yeast artificial chromosome) For example As described in Maus et al. The following two documents are hereby incorporated by reference in their entirety: Annu Rev Immunol (2014) 32:189-225 and Morgan and Boyerinas, Biomedicines (2016) 4:9. In some embodiments, the vector according to this disclosure is a lentiviral vector.

[0343] In some implementations, the vector can be a eukaryotic vector. Right now A vector in a eukaryotic cell contains the elements necessary for protein expression from a vector. In some embodiments, the vector may be a mammalian vector. For example It contains a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0344] According to this disclosure, the constituent polypeptides of the antigen-binding molecule can be encoded by different nucleic acids from multiple nucleic acids, or by different vectors from multiple vectors.

[0345] Generate antigen-binding molecules and peptides The antigen-binding molecules and peptides disclosed herein can be prepared according to methods known to those skilled in the art for producing peptides.

[0346] Antigen-binding molecules and peptides can be chemically synthesized. For example Prepared by liquid-phase or solid-phase synthesis. For example, peptides / polypeptides can be prepared using, for example, Chandrudu. et al.The method of synthesis described in Molecules (2013), 18: 4373-4388, is hereby incorporated in its entirety by reference.

[0347] Alternatively, antigen-binding molecules and peptides can be generated through recombinant expression. Molecular biology techniques suitable for recombinant peptide generation are well-known in the art, such as those listed in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th edition), Cold Spring Harbor Press, 2012, and Nat Methods. (2008); 5(2): 135-146, both of which are hereby incorporated in their entirety by reference. Methods for recombinant generation of antigen-binding molecules are also described in Frenzel. et al. The two references, Front Immunol. (2013); 4: 217 and Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100: 3451–3461, are hereby incorporated in their entirety by reference.

[0348] In some cases, the antigen-binding molecule of this disclosure consists of more than one polypeptide chain. In such cases, the generation of the antigen-binding molecule may include the transcription and translation of more than one polypeptide, and the subsequent association of polypeptide chains to form the antigen-binding molecule.

[0349] For the recombinant production according to this disclosure, any cell suitable for expressing the polypeptide can be used. The cell can be prokaryotic or eukaryotic. In some embodiments, the cell is a prokaryotic cell, such as an archaea or bacterial cell. In some embodiments, the bacteria can be Gram-negative bacteria, such as bacteria of the Enterobacteriaceae family, for example... E. coli (Escherichia coli) In some implementations, the cells are eukaryotic cells, such as yeast cells, plant cells, insect cells, or mammalian cells. For example The cells mentioned above.

[0350] In some cases, the cells are not prokaryotic because some prokaryotic cells do not allow the same folding or post-translational modifications as eukaryotic cells. Additionally, eukaryotes may have very high expression levels, and using appropriate tags can make protein purification from eukaryotes much easier. Specific plasmids can also be used to enhance protein secretion into the culture medium.

[0351] In some implementations, peptides can be prepared via cell-free protein synthesis (CFPS). For example According to descriptions in Zemella et al. The system of Chembiochem (2015) 16(17): 2420-2431 is hereby incorporated in its entirety by reference.

[0352] Production may involve the culture or fermentation of eukaryotic cells modified to express the peptide of interest. Culture or fermentation can be carried out in a bioreactor with a suitable supply of nutrients, air / oxygen, and / or growth factors. Secreted proteins can be collected by separating the cells from the culture medium / fermentation broth, extracting the protein components, and isolating individual proteins to isolate the secreted peptides. Culture, fermentation, and isolation techniques are well known to those skilled in the art and are described, for example, in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th edition; incorporated above by reference).

[0353] A bioreactor comprises one or more containers capable of culturing cells. Cultures in a bioreactor can be continuous, with reactants continuously flowing into the reactor and cultured cells continuously flowing out. Alternatively, cultures can be batch-cultured. Environmental conditions, such as pH, oxygen, inflow and outflow rates, and agitation within the container, are monitored and controlled within the bioreactor to provide optimal conditions for the cells to be cultured.

[0354] After culturing cells expressing a peptide, the peptide of interest can be isolated. Any suitable method known in the art for isolating proteins from cells can be used. To isolate the peptide, it may be necessary to isolate the cells from the nutrient medium. If the peptide is secreted from cells, the cells can be isolated from the medium containing the secreted peptide of interest by centrifugation. If the peptide of interest aggregates intracellularly, protein isolation may include centrifugation to isolate the cells from the cell culture medium, treatment of the cell pellet with lysis buffer, and cell disruption. For example Through acoustic waves, rapid freeze-thaw cycles, or permeation pyrolysis.

[0355] The polypeptide of interest can then be isolated from the supernatant or culture medium, which may contain other protein and non-protein components. A common method for separating protein components from the supernatant or culture medium is by precipitation. Proteins with different solubilities precipitate under different concentrations of precipitant (such as ammonium sulfate). For example, water-soluble proteins are extracted under low concentrations of precipitant. Therefore, proteins with different solubilities can be distinguished by adding different concentrations of precipitant. Dialysis can then be used to remove ammonium sulfate from the separated proteins.

[0356] Other methods for distinguishing different proteins are known in the art, such as ion-exchange chromatography and size chromatography. These can be used as alternatives to precipitation, or can be performed after precipitation.

[0357] Once the polypeptide of interest is isolated from the culture, it may be necessary or essential to concentrate the polypeptide. Many methods for protein concentration are known in the art, such as ultrafiltration or freeze-drying.

[0358] Cells containing / expressing antigen-binding molecules and peptides This disclosure also provides a cell that contains or expresses an antigen-binding molecule or polypeptide according to this disclosure. A cell that contains or expresses a nucleic acid, multiple nucleic acids, a vector, or multiple vectors according to this disclosure is also provided.

[0359] It will be understood that in this article, the singular form ( Right now When referring to a cell as “one / a type / described”, the plural of such cells / groups are also considered.

[0360] This cell can be a eukaryotic cell. For example Mammalian cells. Mammals can be primates (rhesus monkeys, cynomolgus monkeys, non-human primates, or humans) or non-human mammals (…). For example Rabbits, guinea pigs, rats, mice, or other rodents (including any rodent), cats, dogs, pigs, sheep, goats, and cattle (including cows). For example Cows, or any animal of the genus *Bos*, horses (including any animal of the family Equidae), donkeys, and non-human primates.

[0361] In some implementations, the cells are or are derived from cell types commonly used to express peptides for human treatment. Exemplary cells For example The description is in Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100:3451–3461 (incorporated in its entirety by reference), including For example CHO, HEK 293, PER.C6, NSO, and BHK cells. In a preferred embodiment, the cells are or are derived from CHO cells.

[0362] This disclosure also provides a method for generating cells comprising a nucleic acid or vector according to this disclosure, the method comprising introducing a nucleic acid, multiple nucleic acids, a vector, or multiple vectors according to this disclosure into the cells. In some embodiments, introducing the isolated nucleic acid or vector according to this disclosure into the cells includes transformation, transfection, electroporation, or transduction. For example (retroviral transduction).

[0363] This disclosure also provides a method for producing cells expressing / containing an antigen-binding molecule or polypeptide according to this disclosure, the method comprising introducing a nucleic acid, multiple nucleic acids, a vector, or multiple vectors according to this disclosure into the cells. In some embodiments, the method further comprises culturing the cells under conditions suitable for cell expression of the nucleic acid or vector. In some embodiments, the method involves... in vitro conduct.

[0364] This disclosure also provides cells that can be obtained or obtained by the methods according to this disclosure.

[0365] Composition This disclosure also provides compositions comprising the antigen-binding molecules, peptides, nucleic acids, expression vectors, and cells described herein.

[0366] The antigen-binding molecules, peptides, nucleic acids, expression vectors, and cells described herein can be formulated into clinically used pharmaceutical compositions or drugs and may contain pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. Therefore, this disclosure also provides a pharmaceutical composition / drug comprising the antigen-binding molecules, peptides, nucleic acids / multiple nucleic acids, expression vectors / multiple expression vectors, or cells described herein.

[0367] The compositions disclosed herein may comprise one or more pharmaceutically acceptable carriers. For example Liposomes, micelles, microspheres, nanoparticles), diluents / excipients ( For example Starch, cellulose, cellulose derivatives, polyols, glucose, maltodextrin, magnesium stearate), adjuvants, fillers, buffers, preservatives ( For example Vitamin A, Vitamin E, Vitamin C, Retinyl Palmitate, Selenium, Cysteine, Methionine, Citric Acid, Sodium Citrate, Methylparaben, Propylparaben, Antioxidants For example Vitamin A, Vitamin E, Vitamin C, Retinyl Palmitate, Selenium), Lubricants ( For example Magnesium stearate, talc, silica, stearic acid, vegetable stearin, adhesives ( For example Sucrose, lactose, starch, cellulose, gelatin, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), xylitol, sorbitol, mannitol, stabilizers, solubilizers, surfactants ( For example wetting agents, masking agents, or coloring agents For example Titanium dioxide).

[0368] As used herein, the term "pharmaceutically acceptable" refers to compounds, ingredients, materials, compositions, and dosage forms. waitThese are appropriate for the subjects in question, within the bounds of reasonable medical judgment. For example Tissue contact with the compositions (in human subjects) without excessive toxicity, irritation, allergic reactions, or other problems or complications is proportionate to a reasonable benefit / risk ratio. Each carrier, diluent, excipient, adjuvant, filler, buffer, preservative, antioxidant, lubricant, binder, stabilizer, solubilizer, surfactant, masking agent, colorant, flavoring agent, or sweetener in the compositions according to this disclosure must also be “acceptable” in the sense of compatibility with the other components of the formulation. Suitable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, antioxidants, lubricants, binders, stabilizers, solubilizers, surfactants, masking agents, colorants, flavoring agents, or sweeteners can be found in standard pharmaceutical texts, such as Remington, *The Science and Practice of Pharmacy* (edited by A. Adejare), 23rd edition (2020), Academic Press.

[0369] The composition can be formulated for use via topical, parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, intrathecal, oral, or transdermal routes of administration. In some embodiments, the pharmaceutical composition / drug can be formulated for administration by injection or infusion, or by ingestion.

[0370] Suitable formulations may include related products in sterile or isotonic culture media. Drugs and drug compositions may be formulated into fluid forms, including gel forms. Fluid formulations may be formulated for administration by injection or infusion. For example It is applied to a selected area of ​​the human or animal body via a catheter.

[0371] In some embodiments, the composition is formulated for injection or infusion. For example It can be found in blood vessels, the tissue / organ of interest, or tumors.

[0372] This disclosure also provides methods for producing pharmaceutically useful compositions and drugs. Such methods may include one or more steps selected from: producing the antigen-binding molecule, polypeptide, nucleic acid (or multiple nucleic acids), expression vector (or multiple expression vectors) or cell described herein; isolating the antigen-binding molecule, polypeptide, nucleic acid (or multiple nucleic acids), expression vector (or multiple expression vectors) or cell described herein; and / or mixing the antigen-binding molecule, polypeptide, nucleic acid (or multiple nucleic acids), expression vector (or multiple expression vectors) or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent.

[0373] For example, another aspect of this disclosure relates to a formulation or production for treating diseases / diseases ( For example Methods for formulating a pharmaceutical composition or drug for a disease / condition described herein, comprising mixing an antigen-binding molecule, polypeptide, nucleic acid (or multiple nucleic acids), expression vector (or multiple expression vectors) or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[0374] Treatment and preventive applications The antigen-binding molecules, peptides, nucleic acids, expression vectors, cells, and compositions described in this article are found to be useful for therapeutic and preventative approaches.

[0375] This disclosure provides methods for using the antigen-binding molecules, peptides, nucleic acids (or multiple nucleic acids), expression vectors (or multiple expression vectors), cells, or compositions described herein for medical treatment or prevention. It also provides methods for using the antigen-binding molecules, peptides, nucleic acids (or multiple nucleic acids), expression vectors (or multiple expression vectors), cells, or compositions described herein for treating or preventing the diseases or conditions described herein. Furthermore, it provides the use of the antigen-binding molecules, peptides, nucleic acids (or multiple nucleic acids), expression vectors (or multiple expression vectors), cells, or compositions described herein in the manufacture of medicaments for treating or preventing the diseases or conditions described herein. Finally, it provides a method for treating or preventing the diseases or conditions described herein, comprising administering to a subject a therapeutically or preventively effective amount of the antigen-binding molecules, peptides, nucleic acids (or multiple nucleic acids), expression vectors (or multiple expression vectors), cells, or compositions described herein.

[0376] This method can effectively reduce the development or progression of a disease / condition, alleviate its symptoms, or reduce its pathology. It can also effectively prevent the progression of a disease / condition. For example To prevent the worsening of a disease / condition or to slow its progression. In some implementations, this method can lead to an improvement in the disease / condition. For example The reduction of symptoms of the disease / condition or a decrease in some other relevant indicators of the severity / activity of the disease / condition. In some implementation methods, the approach can prevent the disease / condition from progressing to later stages. For example (Chronic phase or metastasis).

[0377] It should be understood that the products of this disclosure can be used to treat / prevent any disease / condition that can be mitigated by reducing the levels of gp130, gp130-containing complexes ( For exampleTherapeutic or preventative benefits may be obtained by selecting from gp130:IL-6Rα complex, gp130:IL-11Rα complex, gp130:OSMRβ complex, gp130:LIFRβ complex, gp130:LIFRβ:CNTFRα complex, gp130:IL-27Rα complex, and gp130:IL-12Rβ2 complex, or by reducing the level / activity of signal transduction mediated by gp130-containing complexes ( For example The therapeutic or preventive benefits can be obtained by assessing the level / activity of signal transduction mediated by cytokines selected from IL-6, IL-11, OSM, LIF, CNTF, CT-1, CLC, IL-27, and IL-35.

[0378] It should also be understood that the products of this disclosure can be used to treat / prevent any disease / condition that can be treated by reducing the content / expression of gp130 or gp130-containing complexes ( For example The therapeutic or preventive benefit may be obtained by increasing the number or activity of cells selected from gp130:IL-6Rα complex, gp130:IL-11Rα complex, gp130:OSMRβ complex, gp130:LIFRβ complex, gp130:LIFRβ:CNTFRα complex, gp130:IL-27Rα complex, and gp130:IL-12Rβ2 complex.

[0379] For example, the disease / disorder may be pathologically associated with gp130, gp130-containing complexes, and / or cytokines that signal through gp130-containing complexes. For example Increased levels / activities of gp130, gp130-containing complexes, and / or cytokines signaling via gp130-containing complexes are positively correlated with the occurrence, development, or progression of a disease / disorder, and / or the severity of one or more symptoms of the disease / disorder. In some embodiments, increased levels / activities of gp130, gp130-containing complexes, and / or cytokines signaling via gp130-containing complexes are risk factors for the occurrence, development, or progression of a disease / disorder.

[0380] In some embodiments, the disease / condition to be treated / prevented according to this disclosure is a disease / condition characterized by increased expression levels or activity of cytokines that are gp130, gp130-containing complexes, and / or signaled by gp130-containing complexes. For exampleThe expression level / activity is compared to the level in the absence of the disease / condition. In some embodiments, the disease / condition to be treated / prevented is characterized by an increased number / proportion / activity of cells expressing gp130, gp130-containing complexes, and / or signaling cytokines via gp130-containing complexes. For example Compared to the level / quantity / proportion / activity in the absence of disease / condition ( For example In healthy subjects, or in equivalent non-disease tissues.

[0381] If the disease / condition is cancer, the expression levels or activity of gp130, gp130-containing complexes, and / or cytokines that signal via gp130-containing complexes may be higher than in equivalent non-cancer / non-tumor tissue. Cancer / its cells may contain one or more mutations ( For example Relative to equivalent non-cancer / non-tumor tissue, the one or more mutations cause upregulation of the expression or activity of gp130, gp130-containing complexes and / or cytokines that signal through gp130-containing complexes.

[0382] Treatment according to the method of this disclosure can achieve a reduction in the activity of gp130, gp130-containing complexes and / or cytokines signaling through gp130-containing complexes in the subject (compared to equivalent untreated subjects or subjects treated with appropriate controls).

[0383] In some aspects and embodiments, the articles of this disclosure provide for the treatment / prevention of diseases / conditions selected from: diseases / conditions pathologically associated with gp130-mediated signaling; and diseases / conditions mediated by gp130-containing complexes ( For example Diseases / conditions pathologically associated with signal transduction via gp130:IL-6Rα complex, gp130:IL-11Rα complex, gp130:OSMRβ complex, gp130:LIFRβ complex, gp130:LIFRβ:CNTFRα complex, gp130:IL-27Rα complex, or gp130:IL-12Rβ2 complex; and diseases / conditions associated with signal transduction via complexes containing gp130 ( For example Cytokines that signal (IL-6, IL-11, OSM, LIF, CNTF, CT-1, CLC, IL-27, or IL-35) that mediate signal transduction in pathologically related diseases / conditions; pathological inflammation; fibrosis; diseases / conditions characterized as inflammation; or diseases / conditions characterized as fibrosis.

[0384] In some aspects and implementations, the products of this disclosure are provided for the treatment / prevention of inflammation, particularly pathological inflammation.

[0385] Inflammation and its role in health and disease For example Chen et al. A review was conducted in Oncotarget (2018) 9(6):7204–7218, which is hereby incorporated in its entirety by reference. Inflammation is the body’s response to cell / tissue damage and is characterized by the response of local immune, vascular, and inflammatory cells to infection or injury, resulting in edema, erythema (redness), fever, pain, and loss of function (rigidity and immobility). The injury may be caused by For example physics( For example Mechanical or chemical damage, trauma, infection, cancer, or an overactive / abnormal immune response ( For example Inflammation is caused by autoimmune diseases. It is part of the innate immune response and plays an important physiological role in wound healing and infection control, and helps restore tissue homeostasis.

[0386] However, many diseases are associated with an overactive inflammatory response ( Right now This refers to inflammation associated with excessive and / or abnormally activated inflammation, and / or chronic (long-term) inflammation. In this article, excessive and / or chronic inflammation may be referred to as "pathological inflammation." Pathological inflammation can refer to inflammation related to the pathology of a disease (…). Right now (Actively promotes inflammation in the pathology of disease).

[0387] According to this disclosure, the inflammation to be treated / prevented can be in any tissue / organ of the body. In some embodiments, the inflammation is in the lungs ( For example bronchioles, alveoli, and airways For example Inflammation can occur in multiple tissues / organs simultaneously, including the nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, heart, kidneys, liver, skeletal muscles, blood vessels, eyes, skin, pancreas, intestines, small intestine, large intestine, colon, joints, brain, or bone marrow.

[0388] In some implementations, the inflammation can be in organs or tissues of the respiratory system. For example lung( For example (bronchioles, alveoli) or airways For example (Nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi). In some implementations, the inflammation can be in organs or tissues of the cardiovascular system. For example The heart or blood vessels. In some implementations, the inflammation can be in organs or tissues of the gastrointestinal system. For example Inflammation may affect the liver, intestines, small intestine, large intestine, colon, or pancreas. In some embodiments, the inflammation may be of the eyes. In some embodiments, the inflammation may be of the skin. In some embodiments, the inflammation may be of organs or tissues of the nervous system. For exampleThe brain. In some embodiments, the inflammation may be in the bone marrow. In some embodiments, the inflammation may be in the joints. In some embodiments, the inflammation may be in organs or tissues of the urinary system. For example Kidneys. In some implementations, inflammation can be in organs or tissues of the musculoskeletal system. For example Muscle tissue. In some implementations, the inflammation may be inflammation of an organ or tissue of one or more organ systems.

[0389] Inflammation can promote angiogenesis through multiple different pathways. Right now New blood vessels grow and develop from the existing vascular system. For example Described in Granger and Senchenkova, 'Chapter 6: Angiogenesis', in 'Inflammation and the Microcirculation', Morgan & Claypool Life Sciences; 2010. For example, inflammation can lead to hypoxia in inflamed tissue, thereby upregulating the expression of the potent angiogenic factor vascular endothelial growth factor (VEGF), which induces new blood vessel growth. Inflammatory cells such as macrophages, lymphocytes, mast cells, and fibroblasts produce angiogenic factors such as VEGF and FGF. Increased blood flow in inflamed tissue can stimulate angiogenesis through shear stress on existing vascular endothelium, and exudated plasma proteins (such as fibrinogen products) may also stimulate new blood vessel formation. In some embodiments, the disease / condition to be treated / prevented according to this disclosure is a disease / condition characterized by angiogenesis. For example Diseases / disorders characterized by inflammation-induced angiogenesis.

[0390] Known angiogenesis disorder, For example In systemic sclerosis, a chronic autoimmune connective tissue disease characterized by inflammation, vascular damage, and fibrosis, the angiogenic response is impaired, failing to ensure effective vascular recovery. For example Cantatore et al.A review was conducted in Biomed Res Int. (2017) 2017:5345673, which is incorporated herein by reference in its entirety. Vascular injury in systemic sclerosis induces hypoxia and tissue ischemia due to tissue avascularization (which is often a major trigger for angiogenesis), and leads to vascular occlusion and thrombosis in larger vessels due to endothelial proliferation, fibrin deposition, and smooth muscle cell hypertrophy. However, when angiogenesis is dysregulated, compensatory angiogenesis is not induced, and vascular injury may induce avascular areas or morphological changes in vessel walls, such as fibrosis. Angiogenic cytokines responsible for angiogenesis and stabilization, such as VEGF, TGFβ, and PDGF, are also associated with fibrosis, further leading to vascular instability and loss of peripheral vascularization.

[0391] Inflammatory responses play a crucial role in triggering fibrosis in many different organ systems. Inflammation can lead to excessive deposition of ECM components in affected tissues. Low-grade but persistent inflammation is also considered a cause of progression in cardiovascular disease and hypertensive fibrosis. In many fibrotic conditions, persistent inflammatory triggering is essential for the upregulation of the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which stimulate the deposition of connective tissue elements, thereby gradually remodeling and destroying normal tissue structure.

[0392] In some aspects and implementations, the articles of this disclosure are provided for the treatment / prevention of fibrosis.

[0393] Fibrosis is a form of pathological tissue remodeling characterized by the excessive deposition of extracellular matrix (ECM) components, including collagen, resulting in an excess of connective tissue. "Excess connective tissue" refers to a given location ( For example The amount of connective tissue in a given tissue / organ (or a portion of a given tissue / organ) is greater than the amount of connective tissue present at the location under normal, non-pathological conditions. Similarly, "excessive deposition of ECM components" refers to the deposition level of one or more ECM components being higher than the deposition level under normal, non-pathological conditions.

[0394] The cellular and molecular mechanisms of fibrosis are described in Wynn, J. Pathol. (2008) 214(2): 199-210, and Wynn and Ramalingam, Nature Medicine (2012) 18:1028-1040, which are hereby incorporated in their entirety by reference.

[0395] Tissue damage can be caused by a variety of stimuli, including infection, autoimmune responses, toxins, radiation, and mechanical injury. Repair typically involves replacing damaged cells with the same type of cells and normal parenchyma with connective tissue. When the repair process is not properly controlled, pathological changes occur, leading to excessive deposition of ECM components, resulting in the replacement of normal parenchyma with connective tissue. In diseases such as idiopathic pulmonary fibrosis, cirrhosis, cardiovascular fibrosis, systemic sclerosis, and nephritis, extensive tissue remodeling and fibrosis can ultimately lead to organ failure and death.

[0396] The primary cellular effector in fibrosis is myofibroblasts. In response to tissue injury, damaged cells and leukocytes produce profibrotic cytokines (such as TGFβ, IL-13, and PDGF), which activate fibroblasts (and other myofibroblast precursor cells) to become αSMA-expressing myofibroblasts and recruit them to the site of injury. Myofibroblasts produce abundant extracellular matrix components (such as collagen and periosteum proteins) for wound contraction and closure, and also produce pro-inflammatory cytokines such as IL-6, as well as tissue remodeling factors such as MMP2 and TIMP1. Persistent / chronic infection and / or inflammation can lead to excessive myofibroblast production, and consequently, excessive extracellular matrix production, resulting in fibrosis. In many diseases and disorders characterized as fibrosis, persistent inflammatory triggers are crucial for the upregulation of the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which stimulate the deposition of connective tissue elements, thereby gradually remodeling and disrupting normal tissue structure.

[0397] Fibrosis can be triggered by pathological conditions. For example This leads to the production of pro-fibrotic factors ( For example TGFβ1 disorders, infections, or disease states. Fibrosis can be caused by physical injury / irritation, chemical injury / irritation, or environmental injury / irritation. Physical injury / irritation may occur during surgery. For example Iatrogenic causes. Chemical damage / irritation can include drug-induced fibrosis. For example Long-term use of drugs (such as bleomycin, cyclophosphamide, amiodarone, procainamide, penicillamine, gold, and nitrofurantoin) (Daba) et al. , Saudi Med J. (2004) 25(6): 700-706). Environmental damage / irritation may include exposure to asbestos fibers or silica.

[0398] Fibrosis can occur in any tissue / organ in the body. In some implementations, fibrosis is in the lungs ( For example bronchioles, alveoli, and respiratory tract For exampleFibrosis can occur in the nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, heart, kidneys, liver, skeletal muscles, blood vessels, eyes, skin, pancreas, intestines, small intestine, large intestine, colon, joints, brain, or bone marrow. Fibrosis can also occur simultaneously in multiple tissues / organs.

[0399] In some implementations, fibrosis can refer to fibrosis of organs or tissues of the respiratory system. For example lung( For example (bronchioles, alveoli) or respiratory tract ( For example Fibrosis of the nasal cavity, oral cavity, pharynx, larynx, trachea, and bronchi. In some implementations, fibrosis may be fibrosis of organs or tissues of the cardiovascular system. For example Fibrosis of the heart or blood vessels. In some implementations, fibrosis may be fibrosis of organs or tissues of the gastrointestinal system. For example Fibrosis of the liver, intestines, small intestine, large intestine, colon, or pancreas. In some embodiments, fibrosis may be fibrosis of the eyes. In some embodiments, fibrosis may be fibrosis of the skin. In some embodiments, fibrosis may be fibrosis of organs or tissues of the nervous system. For example Fibrosis of the brain. In some embodiments, fibrosis may be fibrosis of the bone marrow. In some embodiments, fibrosis may be fibrosis of the joints. In some embodiments, fibrosis may be fibrosis of organs or tissues of the urinary system. For example Kidney fibrosis. In some implementations, fibrosis can refer to fibrosis of organs or tissues of the musculoskeletal system. For example Fibrosis of muscle tissue. In some implementations, fibrosis may be fibrosis of organs or tissues of one or more organ systems.

[0400] In some embodiments, the disease / condition to be treated according to this disclosure is characterized as inflammation. In some embodiments, the disease / condition is characterized as fibrosis. In some embodiments, the disease / condition is characterized as both inflammation and fibrosis.

[0401] As used herein, a disease / condition characterized as inflammation is one in which fibrosis is a symptom of said disease / condition. Diseases / conditions characterized as inflammation include, but are not limited to: Diseases / conditions affecting the respiratory system, such as sinusitis, rhinitis, pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis, pneumonia, pleurisy, and mediastinitis; Diseases / disorders affecting digestive accessory organs, such as hepatitis, ascending cholangitis, cholecystitis, pancreatitis, and peritonitis; Diseases / conditions affecting the cardiovascular system, such as carditis, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, capillary vasculitis, and cardiogenic shock; Diseases / conditions affecting the urinary system, such as nephritis, glomerulonephritis, pyelonephritis, ureteritis, cystitis, and urethritis; Diseases / disorders affecting the nervous system, such as encephalitis, myelitis, meningitis, arachnoiditis, and neuritis; Diseases / disorders affecting the musculoskeletal system, such as arthritis, dermatomyositis, soft tissue, myositis, synovitis / tenosynovitis, bursitis, origin and insertion inflammation, fasciitis, bursitis, epicondylitis, tendinitis, panniculitis, osteochondritis, osteitis / osteomyelitis, spondylitis, periostitis, and chondritis; Diseases / conditions affecting the mouth and throat, such as stomatitis, gingivitis, gingivostomatitis, periodontitis, glossitis, tonsillitis, sialadenitis, parotitis, cheilitis, pulpitis, and jaw inflammation; Diseases / disorders affecting the gastrointestinal system, such as esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, duodenitis, ileitis, cecal inflammation, appendicitis, proctitis, and Peutz-Jeghers syndrome; Diseases / conditions affecting the skin, such as dermatitis, folliculitis, cellulitis, and hidradenitis suppurativa; Diseases / conditions affecting the eyes, such as dacryoadenitis, scleritis, episcleritis, keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis, uveitis, and dry eye syndrome; Diseases / conditions affecting the ear, such as otitis externa, otitis media, labyrinthitis, and mastoiditis; Diseases / disorders of the reproductive system, such as oophoritis, salpingitis, endometritis, endometriosis, parametritis, cervicitis, vaginitis, vulvitis, mastitis, orchitis, epididymitis, prostatitis, seminal vesiculitis, balanitis, posthitis, balanoposthitis, chorioamnionitis, umbilical cord inflammation, and omphalitis. Diseases / disorders of the endocrine system, such as pancreatitis, pituitary inflammation, thyroiditis, parathyroiditis, and adrenalitis; Diseases / disorders of the lymphatic system, such as lymphangitis and lymphadenitis; Cancer, including inflammation-induced and inflammation-related cancers, such as lung cancer ( For example Lung adenocarcinoma, squamous cell carcinoma of the lung), prostate cancer, hematologic malignancies ( For example Multiple myeloma), pancreatic cancer, cervical cancer, stomach cancer, esophageal cancer, head and neck cancer, colorectal cancer, colon cancer, liver cancer ( For example Hepatocellular carcinoma and bile duct cancer.

[0402] It should be understood that many of the diseases / conditions listed above are interconnected.

[0403] It should be understood that diseases / conditions characterized as inflammation may also present with other symptoms / pathologies. For example, a disease / condition characterized as inflammation may further characterize as angiogenesis (…). For example As described in this article, inflammation-induced angiogenesis and / or fibrosis (as in inflammation-induced angiogenesis) and / or fibrosis ( For example As described in this article (fibrosis).

[0404] As used herein, a disease / disorder characterized as fibrosis is a disease / disorder in which fibrosis is a symptom of the disease / disorder. Diseases and disorders characterized as fibrosis include, but are not limited to: Diseases / disorders affecting the respiratory system, such as pulmonary fibrosis, fibrothorax, radiation-induced lung injury, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudler syndrome, asbestosis, silicosis, sarcoidosis, tumor stroma in lung diseases, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma; Diseases / conditions affecting the liver, such as chronic liver disease, liver fibrosis, bridging fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic fatty liver (AFL), alcoholic hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomiasis-related liver disease, and hepatocellular carcinoma (HCC); Diseases / conditions affecting the cardiovascular system, such as hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, myocardial fibrosis, interstitial fibrosis, replacement fibrosis, Brugada syndrome, myocarditis, endocardial myocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmic right ventricular cardiomyopathy (ARVC), atherosclerosis, arteriosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, heart failure with preserved ejection fraction (HFpEF), heart failure with reduced ejection fraction (HFrEF), varicose veins, and cerebral infarction; Diseases / conditions affecting the kidneys, such as tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephrotic syndrome, Alport's syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry's disease, diabetic nephropathy, chronic glomerulonephritis, and nephritis associated with systemic lupus erythematosus; Diseases / disorders affecting the pancreas, such as pancreatic fibrosis, cystic fibrosis, and chronic pancreatitis; Diseases / disorders affecting the nervous system, such as gliosis, Alzheimer's disease, and multiple sclerosis; Diseases / disorders affecting the musculoskeletal system, such as muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker's muscular dystrophy (BMD), and fibrotic myopathy; Diseases / disorders affecting the gastrointestinal system, such as inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, and primary sclerosing cholangitis (PSC); Diseases / conditions affecting the skin, such as scleroderma, renal systemic fibrosis, palmar aponeurosis contracture, and keloids. Diseases / conditions affecting the eyes, such as Graves's opthalmopathy, preretinal fibrosis, retinal fibrosis, subretinal fibrosis, and subretinal fibrosis associated with macular degeneration. For example Wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis ( For example Posterior capsule fibrosis after cataract surgery or fibrosis of filtering blebs after trabeculectomy in glaucoma), conjunctival fibrosis, and subconjunctival fibrosis. Diseases / conditions that affect the joints, such as joint fibrosis, arthritis, and adhesive capsulitis; Diseases / disorders affecting multiple tissue / organ systems, including progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD); pre-fibrotic and neoplastic fibrotic diseases, and fibrosis induced by chemical or environmental damage. For example (Cancer chemotherapy, pesticides, radiation / cancer radiotherapy); Cancers such as hepatocellular carcinoma, stomach cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, and vulvar cancer; Mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease. ...

Claims

1. An antigen-binding molecule, said antigen-binding molecule being optionally isolated, said antigen-binding molecule binding to gp130, wherein said antigen-binding molecule inhibits IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and LIF-mediated signal transduction.

2. The antigen-binding molecule according to claim 1, wherein the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:

89.

3. The antigen-binding molecule according to claim 1 or claim 2, wherein the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:

12.

4. The antigen-binding molecule according to any one of claims 1 to 3, wherein the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence having at least 70% amino acid sequence identity with the VH sequence shown in column A of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VL sequences shown in column B of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C. The VH sequence and the VL sequence are selected from the same row of Table C.

5. The antigen-binding molecule according to claim 1 or claim 2, wherein the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:218; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:204; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:20; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:195; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (e) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:214; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (f) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:215; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:

28.

6. The antigen-binding molecule according to any one of claims 1, 2, or 5, wherein the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence that is at least 70% amino acid sequence identical to the VH sequence shown in column A of rows 89, 90, 80-88, 74-79 or 2 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VH sequence shown in column B of rows 89, 90, 80-88, 74-79 or 2 of Table C. The VH sequence and the VL sequence are selected from the same row of Table C.

7. The antigen-binding molecule according to any one of claims 1 to 6, wherein the antigen-binding molecule is a multispecific antigen-binding molecule, and wherein the antigen-binding molecule further comprises an antigen-binding domain that binds to an antigen other than gp130.

8. A chimeric antigen receptor (CAR) comprising the antigen-binding molecule of any one of claims 1 to 7.

9. One or more nucleic acids, wherein the one or more nucleic acids are optionally isolated, and the one or more nucleic acids encode the antigen-binding molecule of any one of claims 1 to 7 or the CAR of claim 8.

10. An expression vector or multiple expression vectors, wherein the expression vector or multiple expression vectors comprises the nucleic acid or multiple nucleic acids as described in claim 9.

11. A cell comprising the antigen-binding molecule of any one of claims 1 to 7, the CAR of claim 8, one or more nucleic acids of claim 9, or one or more expression vectors of claim 10.

12. A method comprising culturing the cells under conditions suitable for cell expression of an antigen-binding molecule or CAR as described in claim 11.

13. A composition comprising the antigen-binding molecule of any one of claims 1 to 7, the CAR of claim 8, one or more nucleic acids of claim 9, one or more expression vectors of claim 10, or the cell of claim 11, and a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

14. The antigen-binding molecule according to any one of claims 1 to 7, the CAR according to claim 8, one or more nucleic acids according to claim 9, one or more expression vectors according to claim 10, the cell according to claim 11, or the composition according to claim 13, for use in medical treatment or prevention.

15. An antigen-binding molecule according to any one of claims 1 to 7, a CAR according to claim 8, a nucleic acid or multiple nucleic acids according to claim 9, an expression vector or multiple expression vectors according to claim 10, a cell according to claim 11, or a composition according to claim 13, for the treatment or prevention of the following: pathological inflammation, fibrosis, diseases / conditions characterized as inflammation, diseases / conditions characterized as fibrosis, diseases / conditions characterized as both inflammation and fibrosis, diseases / conditions of pathological significance where signal transduction is carried out via a complex containing gp130, diseases / conditions of pathological significance where signaling is carried out via a cytokine containing a complex containing gp130, autoimmune diseases, metabolic syndrome, neurodegenerative diseases, chronic inflammatory diseases, arthritis, rheumatoid arthritis, juvenile arthritis, systemic juvenile idiopathic arthritis, lupus, systemic lupus erythematosus, pancreatitis, thyroiditis, periodontitis, rhinitis, allergic rhinitis, dermatitis, dermatitis, and other related conditions. Heterodermatitis, psoriasis, Hermansky-Pudrag syndrome, Graves' disease, obesity, insulin resistance, diabetes, type 1 diabetes, type 2 diabetes, pregnancy-related hyperglycemia, multiple sclerosis, giant cell arteritis, aortitis, cardiovascular disease, atherosclerosis, atrial fibrillation, ventricular fibrillation, cardiac hypertrophy, hypertrophic cardiomyopathy, dilated cardiomyopathy, myocarditis, cardiogenic shock, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, ischemic heart disease, myocardial infarction, equine disease. Van der Mascheranostomy syndrome, systemic sclerosis, keloids, scleroderma, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), hippocampal atrophy, lung diseases, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, hepatitis, liver fibrosis, cirrhosis, hepatotoxicity, acetaminophen-induced hepatotoxicity, alcoholic liver disease, pancreatitis, fatty degeneration, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, primary biliary cholangitis, primary sclerosing cholangitis.Inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, endometriosis, stroke, ischemic stroke, kidney disease, kidney injury, acute kidney injury, nephrotoxicity, glomerulonephritis, chronic kidney disease, renal fibrosis, Allport syndrome, adult-onset Steele disease, Castleman's disease, cytokine release syndrome, sepsis, septic shock, retinopathy, retinal fibrosis, age-related macular degeneration, wet age-related macular degeneration, retinitis pigmentosa, dry eye syndrome, COVID-19, Boyz-Yage syndrome, musculoskeletal disorders, muscular dystrophy, amyotrophic lateral sclerosis, cachexia, endocrine disorders, polycystic ovary syndrome, cancer, hematologic malignancies, leukemia, plasmacytoma, Hodgkin's lymphoma, lung cancer, colorectal cancer, bowel cancer, urinary tract cancer, bladder cancer, vulvar cancer. Endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, bone cancer, glioblastoma, breast cancer, gastric cancer, kidney cancer, metastatic renal cell carcinoma, prostate cancer, skin cancer, melanoma, liver cancer, hepatocellular carcinoma, asthenia, age-related increase in fat mass, sarcopenia, age-related hyperlipidemia, age-related hypertriglyceridemia, age-related hypercholesterolemia, age-related hepatic steatosis, age-related non-alcoholic fatty liver disease, age-related non-alcoholic fatty liver disease, age-related non-alcoholic steatohepatitis, age-related cardiovascular disease, age-related hypertension, age-related nephropathy, age-related skin diseases, infectious diseases, viral diseases, viral hepatitis, hepatitis B, HIV infection, influenza infection, malaria, tuberculosis, allergic diseases, transplant rejection and graft-versus-host disease.

16. A kind in vitro The in vitro complex is optionally isolated and comprises an antigen-binding molecule of any one of claims 1 to 7 that binds to gp130.

17. A method for detecting gp130 in a sample, the method comprising contacting a sample containing or suspected of containing gp130 with an antigen-binding molecule according to any one of claims 1 to 7, and detecting the formation of a complex of the antigen-binding molecule and gp130.

18. A method for selecting or stratifying subjects for treatment with a gp130-targeting agent, the method comprising conjugating a sample from said subject with an antigen-binding molecule according to any one of claims 1 to 7. in vitro Contact, and detection of the formation of the complex between the antigen-binding molecule and gp130.

19. The antigen-binding molecule according to any one of claims 1 to 7 as... in vitro or in vivo Uses of diagnostic or prognostic agents.

20. An antigen-binding molecule that binds to gp130 and inhibits IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and LIF-mediated signal transduction, for the treatment or prevention of the following: pathological inflammation, fibrosis, diseases / conditions characterized as inflammation, diseases / conditions characterized as fibrosis, diseases / conditions characterized as both inflammation and fibrosis, diseases / conditions of pathological significance involving signal transduction via a gp130-containing complex, diseases / conditions of pathological significance involving cytokines signaling via a gp130-containing complex, autoimmune diseases, metabolic syndrome, neurodegenerative diseases, chronic inflammatory diseases, arthritis, rheumatoid arthritis, juvenile arthritis, systemic juvenile idiopathic arthritis, lupus, systemic lupus erythematosus, pancreatitis, thyroiditis, periodontitis, rhinitis, allergic rhinitis, dermatitis, atopic dermatitis, psoriasis, and eczema. Manskiy-Pudrag syndrome, Graves' disease, obesity, insulin resistance, diabetes, type 1 diabetes, type 2 diabetes, pregnancy-related hyperglycemia, multiple sclerosis, giant cell arteritis, aortitis, cardiovascular disease, atherosclerosis, atrial fibrillation, ventricular fibrillation, cardiac hypertrophy, hypertrophic cardiomyopathy, dilated cardiomyopathy, myocarditis, cardiogenic shock, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, ischemic heart disease, myocardial infarction, Marfan syndrome Systemic sclerosis, keloids, scleroderma, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), hippocampal atrophy, lung diseases, asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, hepatitis, liver fibrosis, cirrhosis, hepatotoxicity, acetaminophen-induced hepatotoxicity, alcoholic liver disease, pancreatitis, fatty degeneration, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, primary biliary cholangitis, primary sclerosing cholangitis.Inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, endometriosis, stroke, ischemic stroke, kidney disease, kidney injury, acute kidney injury, nephrotoxicity, glomerulonephritis, chronic kidney disease, renal fibrosis, Allport syndrome, adult-onset Steele disease, Castleman's disease, cytokine release syndrome, sepsis, septic shock, retinopathy, retinal fibrosis, age-related macular degeneration, wet age-related macular degeneration, retinitis pigmentosa, dry eye, COVID-19, Poitz-Yage syndrome, musculoskeletal disorders, muscular dystrophy, amyotrophic lateral sclerosis, cachexia, endocrine disorders, polycystic ovary syndrome, cancer, hematologic malignancies, leukemia, plasmacytoma, Hodgkin's lymphoma, lung cancer, colorectal cancer, bowel cancer, urinary tract cancer, bladder cancer, vulvar cancer, endometrial cancer, ovarian cancer. Cancer, prostate cancer, pancreatic cancer, bone cancer, glioblastoma, breast cancer, gastric cancer, kidney cancer, metastatic renal cell carcinoma, prostate cancer, skin cancer, melanoma, liver cancer, hepatocellular carcinoma, asthenia, age-related increase in fat mass, sarcopenia, age-related hyperlipidemia, age-related hypertriglyceridemia, age-related hypercholesterolemia, age-related hepatic steatosis, age-related non-alcoholic fatty liver disease, age-related non-alcoholic fatty liver, age-related non-alcoholic steatohepatitis, age-related cardiovascular disease, age-related hypertension, age-related nephropathy, age-related skin disease, infectious diseases, viral diseases, viral hepatitis, hepatitis B, HIV infection, influenza infection, malaria, tuberculosis, allergic diseases, transplant rejection and graft-versus-host disease, wherein the antigen-binding molecule contacts the gp130 region shown in SEQ ID NO:

89.

21. Use of an antigen-binding molecule that binds to and contacts the gp130 region shown in SEQ ID NO:89 to inhibit IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction and / or LIF-mediated signal transduction.

22. A method for inhibiting IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and / or LIF-mediated signal transduction, the method comprising contacting a cell capable of IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and / or LIF-mediated signal transduction with an antigen-binding molecule, said antigen-binding molecule binding to gp130 and contacting the gp130 region shown in SEQ ID NO:

89.

23. A method for inhibiting IL-6-mediated signal transduction, IL-11-mediated signal transduction, OSM-mediated signal transduction, CNTF-mediated signal transduction, CT-1-mediated signal transduction, and LIF-mediated signal transduction in a subject, the method comprising administering to the subject an antigen-binding molecule that binds to gp130 and contacts the gp130 region shown in SEQ ID NO:

89.

24. The antigen-binding molecule of claim 20, the use of the antigen of claim 21, or the method of claim 22 or 23, wherein the antigen-binding molecule comprises: (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:185; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:2 HC-CDR2 with the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 with the amino acid sequence of SEQ ID NO:10 LC-CDR2 with the amino acid sequence SEQ ID NO:192 LC-CDR3 having the amino acid sequence of SEQ ID NO:

12.

25. The antigen-binding molecule used according to claim 20 or claim 24, the use according to claim 21 or claim 24, or the method according to any one of claims 22 to 24, wherein the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence having at least 70% amino acid sequence identity with the VH sequence shown in column A of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VL sequences shown in column B of rows 68, 71, 69, 72, 70, 73, 12, 13, 38-67, 3-11 or 1 of Table C. The VH sequence and the VL sequence are selected from the same row of Table C.

26. The antigen-binding molecule of claim 20, the use of the antigen-binding molecule of claim 21, or the method of claim 22 or 23, wherein the antigen-binding molecule comprises: (a) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:218; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (b) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:204; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (c) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:20; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (d) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:195; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (e) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:214; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:28; or (f) (i) Heavy chain variable (VH) regions incorporated with the following CDRs: HC-CDR1 with the amino acid sequence of SEQ ID NO:18 HC-CDR2 with the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NO:215; and (ii) Light chain variable (VL) regions incorporated with the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:26 LC-CDR2 having the amino acid sequence of SEQ ID NO:27 LC-CDR3 having the amino acid sequence of SEQ ID NO:

28.

27. The antigen-binding molecule used according to claim 20 or claim 26, the use according to claim 21 or claim 26, or the method according to any one of claims 22, 23, or 26, wherein the antigen-binding molecule comprises: (i) A VH region having an amino acid sequence that is at least 70% amino acid sequence identical to the VH sequence shown in column A of rows 89, 90, 80-88, 74-79 or 2 of Table C, and (ii) A VL region having an amino acid sequence that is at least 70% identical to the VH sequence shown in column B of rows 89, 90, 80-88, 74-79 or 2 of Table C. The VH sequence and the VL sequence are selected from the same row of Table C.

28. The antigen-binding molecule used according to any one of claims 20 or 24 to 27, the use according to any one of claims 21 or 24 to 27, or the method according to any one of claims 22 to 27, wherein the antigen-binding molecule is a multispecific antigen-binding molecule, and wherein the antigen-binding molecule further comprises an antigen-binding domain that binds to an antigen other than gp130.

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