Anti-RTK antibodies and uses thereof

Abstract

The present invention relates to RTK agonists, in particular FGFR2b agonists, FGFR1b agonists, and DDR1 agonists, and methods of using the agonists.

Description

Technical Field

[0001] This invention relates to RTK agonists, particularly FGFR2b agonists, FGFR1b agonists and DDR1 agonists, and methods of using the agonists. Background Technology

[0002] Receptor tyrosine kinases (RTKs) play crucial roles in various cellular processes. RTKs are known to be activated through ligand-induced dimerization. Dimerization leads to kinase activation and autophosphorylation of multiple tyrosine residues in the cytoplasmic domain of the receptor, as well as activation of downstream signaling pathways. One family of RTKs includes the fibroblast growth factor receptor (FGFR) family, expressed by various mammalian cells. The FGFR family consists of FGFR1, FGFR2, FGFR3, and FGFR4, and includes high-affinity receptors with many different FGF ligands (NPL1). This receptor is a transmembrane tyrosine kinase, and FGFR signaling drives downstream pathways essential for cell proliferation, differentiation, survival, and migration (NPL2). Binding of FGF ligands to receptors induces dimerization of the FGF:FGFR complex and activation of downstream FGF signaling (NPL3).

[0003] Members of the FGF family are potent stimulants for tissue repair and have been shown to contribute to many tissue regeneration processes, such as maintaining cell stemness, inducing cell dedifferentiation, stimulating cell proliferation, inhibiting cell senescence, suppressing cell death, regulating inflammation, stimulating angiogenesis, and enhancing protease expression. FGF ligands perform these diverse functions by binding to and activating the FGFR family of RTKs.

[0004] FGFR2 has two splicing isoforms: FGFR2b, which is normally located in epithelial cells, and FGFR2c (NPL 1), which is normally located in mesenchymal cells. FGFR2b is a high-affinity receptor for FGF ligands such as FGF7 and FGF10, while FGFR2c is a high-affinity receptor for FGF ligands such as FGF2 (NPL 3). Although various therapeutics targeting FGFR2 have been developed, such as anticancer drugs targeting FGFR2-overexpressing cells, these drugs are inhibitors of FGFR2, and there remains a significant unmet medical need for the development of FGFR2 agonists.

[0005] FGFR1 also has two splice isoforms: FGFR1b is expressed in the brain, tongue, prepuce glands, vas deferens, and skin; FGFR1c is expressed in various tissues (NPL 8). FGFR1b is a receptor for FGF ligands such as FGF1 and FGF2, and is also a weak affinity receptor for FGF7 and FGF10 (NPL 9). FGFR1c shows binding to various types of FGF ligands, including FGF1, FGF2, FGF4, FGF8, FGF9, FGF19, FGF21, and FGF23 (NPL 10, NPL11). In particular, FGF19, FGF21, and FGF23 act on β-Klotho complexed with FGFR1c. FGFR1 has also been evaluated as a therapeutic target for both anticancer and chronic disease (such as obesity and diabetes) inhibitors and agonists. In particular, there are several reports on the agonistic effects of FGFR1, including homodimerization-based agonists targeting both FGFR1b and FGFR1c, and heterodimerization-based agonists targeting FGFR1c and β-Klotho (NPL 12, NPL 13, NPL 14). However, there are no reports on strong FGFR1b-specific agonists that exhibit better agonistic activity than the natural ligands.

[0006] Another member of the RTK family includes the discoidin domain receptor (DDR) family. The DDR family includes DDR1 and DDR2. DDR1 is primarily expressed in epithelial cells, smooth muscle cells, and in some cases in fibroblasts, while DDR2 is expressed in mesenchymal-derived cells (NPL 15). Both DDR1 and DDR2 are activated when collagen binds to the extracellular portion of the receptor, initiating downstream signaling pathways, including receptor phosphorylation (NPL 16). Collagen is an important ECM component that interacts with various cellular receptors in tissues; therefore, DDR participates in a variety of important functions, including regulating cell proliferation, survival, differentiation, cell aggregation, adhesion, migration, and invasion (NPL 16). While the therapeutic applications of DDR have primarily focused on antifibrotic and anticancer therapies due to DDR overexpression in various cancer cells, some reports indicate that DDR activation promotes cartilage regeneration and wound healing (NPL 17, NPL 18), suggesting that DDR agonism also has the potential to regenerate various tissues. However, like FGFR1 and FGFR2, no therapeutic agonist antibody specific to DDR has yet been successfully identified.

[0007] [List of Citations]

[0008] [Patent Literature]

[0009] [PTL 1] WO2012021841A2

[0010] [PTL 2] WO2021247718A1

[0011] [PTL 3] WO2022143728A1

[0012] [PTL 4] WO2022087243A1

[0013] [PTL 5] WO2021157679A1

[0014] [Non-patent literature]

[0015] [NPL 1] Danilo Ranieri, Benedetta Rosato, Monica Nanni, AlessandraMagenta, Francesca Belleudi, Maria Rosaria Torrisi, Expression of the FGFR2mesenchymal splicing variant in epithelial cells drives epithelial-mesenchymal transition. Oncotarget. 2016 Feb 2;7(5):5440-60.

[0016] [NPL 2] Susanne Braun, Ulrich auf dem Keller, Heike Steiling, SabineWerner, Fibroblast growth factors in epithelial repair and cytoprotection. Philos Trans R Soc Lond B Biol Sci. 2004 May 29;359(1445):753-7.

[0017] [NPL 3] Allen Zinkle, Moosa Mohammadi, Structural Biology of the FGF7Subfamily. Front Genet. 2019 Feb 12;10:102.

[0018] [NPL 4] Stefan Dengl, Klaus Mayer, Felix Bormann, Harald Duerr, Eike Hoffmann, Bianca Nussbaum, Michael Tischler, Martina Wagner, Andreas Kuglstatter, Lea Leibrock, Can Buldun, Guy Georges, Ulrich Brinkmann, Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices. Nat Commun. October 2, 2020;11(1):4974.

[0019] [NPL 5] Jun Tashiro, Gustavo A Rubio, Andrew H Limper, Kurt Williams, Sharon J Elliot, Ioanna Ninou, Vassilis Aidinis, Argyrios Tzouvelekis, Marilyn K Glassberg, Exploring Animal Models That Resemble Idiopathic Pulmonary Fibrosis. Front Med (Lausanne). July 28, 2017:4:118.

[0020] [NPL 6] Tianju Liu, Francina Gonzalez De Los Santos, Sem H Phan, The Bleomycin Model of Pulmonary Fibrosis. Methods Mol Biol. 2017:1627:27-42.

[0021] [NPL 7] Dmitri Toren, Hagai Yanai, Reem Abu Taha, Gabriela Bunu,Eugen Ursu, Rolf Ziesche, Robi Tacutu, Vadim E Fraifeld, Systems biology analysis of lung fibrosis-related genes in the bleomycin mouse model. SciRep.2021 Nov 29;11(1):19269.

[0022] [NPL 8] Clementine Fon Tacer, Angie L Bookout, Xunshan Ding, HiroshiKurosu, George B John, Lei Wang, Regina Goetz, Moosa Mohammed, Makoto Kuro-o, David J Mangelsdorf, Steven A Kliewer, Research resource : Comprehensiveexpression atlas of the fibroblast growth factor system in adult mouse. MolEndocrinol. 2010 Jan 10;24(10):2050-64.

[0023] [NPL 9] HD Beer, L Vindevoghel, MJ Gait, JM Revest, DR Duan, IMason, C Dickson, S Werner, Fibroblast growth factor (FGF) receptor 1-IIIb is a naturally occurring functional receptor for FGFs that is preferentiallyexpressed in the skin and the brain. J Biol Chem. 2000 Jan 5 / 26;275(21):16091-7.

[0024] [NPL 10] Ornitz DM, Xu J, Colvin JS, McEwen DG, MacArthur CA, FCoulier, Gao G, Goldfarb M, Receptor specificity of the fibroblast growth factor family. J Biol Chem. 1996 Jan 6 Oct 21;271(25):15292-7.

[0025] [NPL 11] Tian Lan, Donald A Morgan, Kamal Rahmouni, Junichiro Sonoda,Xiaorong Fu, Shawn C Burgess, William L Holland, Steven A Kliewer, David JMangelsdorf. Cell Metab. Nov 7, 2017;26(5):709-718.e3.

[0026] [NPL 12] Ai-Luen Wu, Ganesh Kolumam, Scott Stawicki, Yongmei Chen,Jun Li, Jose Zavala-Solorio, Khanhky Phamluong, Bo Feng, Li Li, ScotMarsters, Lance Kates, Nicholas van Bruggen, Maya Leabman, Anne Wong, DavidWest, Howard Stern, Elizabeth Luis, Hok Seon Kim, Daniel Yansura, Andrew SPeterson, Ellen Filvaroff, Yan Wu, Junichiro Sonoda, Amelioration of type 2diabetes by antibody-mediated activation of fibroblast growth factor receptor1. Sci Transl Med. December 14, 2011;3(113):113ra126.

[0027] [NPL 13] Jocelyn Chan, Joyce Chan, Lily Shao, Scott S Stawicki,Victoria C Pham, Rob W Akita, Marc Hafner, Lisa Crocker, Kebing Yu, James TKoerber, Gabriele Schaefer, Laetitia Comps-Agrar, Systematic pharmacologicalanalysis of agonistic and antagonistic fibroblast growth factor receptor 1Mabs reveals a similar unique mode of action. J Biol Chem. January 2023;299(1):102729.

[0028] [NPL 14] Xiaoshan Min, Jennifer Weiszmann, Sheree Johnstone, WeiWang, Xinchao Yu, William Romanow, Stephen Thibault, Yang Li, Zhulun Wang,Agonistic β-Klotho antibody mimics fibroblast growth factor 21 (FGF21)functions. J Biol Chem. Nov 21, 2018;293(38):14678-14688.

[0029] [NPL 15] Gurubasavaraja Swamy Purawarga Matada, Arka Das, PrasadSanjay Dhiwar & Abhishek Ghara, DDR1 and DDR2 : a review on signaling pathwayand small molecule inhibitors as an anticancer agent. Med Chem Res 30, 535–551 (2021).

[0030] [NPL 16] Coelho NM, Wang A, McCulloch CA, Discoidin domain receptor1 interactions with myosin motors contribute to collagen remodeling andtissue fibrosis. Biochem Biophys Acta Mol Cell Res. May 11, 2019;1866(11):1

[0031] [NPL 17] Arokia Vijaya Anand Mariadoss, Chau-Zen Wang, Exploring theCellular and Molecular Mechanism of Discoidin Domain Receptors (DDR1 andDDR2) in Bone Formation, Regeneration, and Its Associated Disease Conditions.Int J Mol Sci. October 4, 2023;24(19):14895.

[0032] [NPL 18] Muriel Cario, DDR1 and DDR2 in skin. Cell Adh Migr. 2018;12(4): 386-393.

[0033] [NPL 19] D Smedley, A Demiroglu, M Abdul-Rauf, C Heath, C Cooper, JShipley, N C Cross, ZNF198-FGFR1 transforms Ba / F3 cells to growth factorindependence and results in high level tyrosine phosphorylation of STATS 1and 5. Neoplasia. October 1999;1(4):349-55.

[0034] [NPL 20] Victoria Juskaite, David S Corcoran, Birgit Leitinger,Collagen induces activation of DDR1 through lateral dimer association andphosphorylation between dimers. Elife. June 7, 2017:6:e25716.

[0035] [NPL 21] David S Corcoran, Victoria Juskaite, Yuewei Xu, FrederikGorlitz, Yuriy Alexandrov, Christopher Dunsby, Paul MW French, BirgitLeitinger, DDR1 autophosphorylation is a result of aggregation into denseclusters. Sci Rep. 2019 Nov 19;9(1):17104. Summary of the Invention

[0036] [Technical Issues]

[0037] Despite the development of various therapeutic agents targeting receptor tyrosine kinases, there remains a significant medical need for activation of FGFR signaling (particularly FGFR2 and FGFR1). The FGF family is known to be a potent stimulant for tissue repair and has been shown to contribute to many tissue regeneration processes, such as maintaining cell stemness, inducing cell dedifferentiation, stimulating cell proliferation, inhibiting cell senescence, suppressing cell death, regulating inflammation, stimulating angiogenesis, and enhancing protease expression. There is also a significant medical need for activation of DDR signaling (particularly DDR1), where the collagen family is known to be a potential regulator of cell proliferation, survival, cell differentiation, ECM homeostasis, cytokine secretion, cell aggregation, and migration.

[0038] [Solution to the problem]

[0039] This invention provides agonistic antigen-binding molecules, such as antibodies or fragments thereof that bind to receptor tyrosine kinases (RTKs), including the fibroblast growth factor receptor (FGFR) family and the discoid domain receptor (DDR) family. The invention provides antigen-binding molecules, such as antibodies or fragments thereof that bind to human fibroblast growth factor receptor 2 protein (FGFR2), human fibroblast growth factor receptor 1 protein (FGFR1), or human discoid domain receptor 1 (DDR1), preferably wherein the antibody or fragment thereof specifically binds to FGFR2IIIb (FGFR2b), FGFR1IIIb (FGFR1b), or DDR1a, wherein these molecules activate FGFR2IIIb signaling, FGFR1IIIb signaling, or DDR1a signaling, and provides methods of using the antigen-binding molecules. The molecules of the present invention can be used for any disease, ailment or condition in which an increase in FGFR2b signaling, FGFR1b signaling or DDR1a signaling is helpful, and can be used to treat fibrosis or inflammation, particularly pulmonary and skin fibrosis.

[0040] The molecules of the present invention provided herein are agonistic antigen-binding molecules, such as antibodies or fragments thereof that bind to receptor tyrosine kinases (RTKs) and promote their dimerization and activation, and wherein the antigen-binding molecule binds at two sites. In one aspect, the molecules of the present invention are antigen-binding molecules, such as antibodies or fragments thereof that bind to fibroblast growth factor receptor 2 protein (FGFR2), preferably wherein the antibody or fragment thereof specifically binds to FGFR2IIIb. Such antigen-binding molecules (such as antibodies) can be used particularly to target cells expressing FGFR2 (preferably FGFR2IIIb). FGFR2b antigen-binding molecules mimic ligands of FGFR2b (such as FGF7 or FGF10) to selectively activate FGFR2b signaling and are found to be useful in methods for activating downstream FGF signaling. In one aspect, anti-FGFR2b antigen-binding molecules (such as antibodies) and their antigen-binding fragments specifically bind to the IgII, IgIII, or IgIIIb domains of FGFR2 and selectively activate FGFR2IIIb signaling. In one aspect, anti-FGFR2b antigen-binding molecules (such as antibodies) and their antigen-binding fragments comprise a first antigen-binding domain that binds to IgIIIb and a second antigen-binding domain that binds to IgI, IgII, IgIII, or IgIIIb, and selectively activate FGFR2IIIb signaling. In another aspect, anti-FGFR2b antigen-binding molecules (such as antibodies) and their antigen-binding fragments bind to two sites of the IgIIIb and IgI, IgIIIb and IgII, IgIIIb and IgIII, or IgIIIb and IgIIIb domains of FGFR2, and selectively activate FGFR2IIIb signaling. Agonistant antigen-binding molecules (such as antibodies) targeting FGFR2b, including agonist two-site antibodies against FGFR2b, provide a new class of therapeutic agents for treating tissue degenerative diseases.

[0041] In one aspect, the molecules of the present invention are antigen-binding molecules, such as antibodies or fragments thereof that bind to fibroblast growth factor receptor 1 protein (FGFR1), preferably wherein the antibody or fragment thereof specifically binds to FGFR1IIIb. Such antigen-binding molecules (such as antibodies) are particularly useful for targeting cells expressing FGFR1 (preferably FGFR1IIIb). FGFR1b antigen-binding molecules mimic ligands of FGFR1b (such as FGF1 or FGF2) to selectively activate FGFR1b signaling and are found to be useful in methods for activating downstream FGF signaling. In one aspect, anti-FGFR1b antigen-binding molecules (such as antibodies) and their antigen-binding fragments specifically bind to the IgII, IgIII, or IgIIIb domains of FGFR1 and selectively activate FGFR1IIIb signaling. In one aspect, anti-FGFR1b antigen-binding molecules (such as antibodies) and their antigen-binding fragments comprise a first antigen-binding domain that binds to IgIIIb and a second antigen-binding domain that binds to IgI, IgII, IgIII, or IgIIIb, and selectively activate FGFR1IIIb signaling. In another aspect, anti-FGFR1b antigen-binding molecules (such as antibodies) and their antigen-binding fragments bind to two sites of FGFR1: IgIIIb and IgI, IgIIIb and IgII, IgIIIb and IgIII, or IgIIIb and IgIIIb, and selectively activate FGFR1IIIb signaling. Agonistant antigen-binding molecules (such as antibodies) targeting FGFR1b, including agonist two-site antibodies against FGFR1b, provide a new class of therapeutic agents for treating degenerative diseases.

[0042] In one aspect, the molecules of the present invention are antigen-binding molecules, such as antibodies or fragments thereof that bind to the discoid domain receptor 1 protein (DDR1), preferably wherein the antibody or fragment thereof specifically binds to DDR1a. Such antigen-binding molecules (such as antibodies) are particularly useful for targeting cells expressing DDR1 (preferably DDR1a). DDR1a antigen-binding molecules mimic ligands of DDR1a (such as type I, IV, V, VI, or VIII collagen) to selectively activate DDR1a signaling and are found to be useful in methods for activating downstream FGF signaling. In one aspect, anti-DDR1a antigen-binding molecules (such as antibodies) and their antigen-binding fragments specifically bind to the discoid (DS) domain, DS-like domain, or extracellular juxtamembranous region (EJXM) of DDR1a and selectively activate DDR1a signaling. In one aspect, anti-DDR1a antigen-binding molecules (such as antibodies) and their antigen-binding fragments comprise a first antigen-binding domain that binds to the DS domain and a second antigen-binding domain that binds to the DS domain, DS-like domain, or EJXM region, and selectively activate DDR1a signaling. In another aspect, anti-DDR1a antigen-binding molecules (such as antibodies) and their antigen-binding fragments bind to the DS domain and EJXM region, the DS domain and DS-like domain, or the DS domain and DS domain of DDR1 at two sites, and selectively activate DDR1a signaling. Agonistant antigen-binding molecules (such as antibodies) targeting DDR1a, including agonist two-site antibodies against DDR1a, provide a new class of therapeutic agents for treating tissue degenerative diseases.

[0043] Anti-FGFR2b antigen-binding molecules, anti-FGFR1b antigen-binding molecules, or anti-DDR1 antigen-binding molecules (such as antibodies) and their antigen-binding fragments may be used alone in unmodified form or may be included as part of antibody-drug conjugates or fusion proteins. Furthermore, the molecules of the present invention include, but are not limited to, small organic molecules and peptides.

[0044] The present invention also provides methods for using anti-FGFR2b antigen-binding molecules, anti-FGFR1b antigen-binding molecules, or anti-DDR1 antigen-binding molecules (such as antibodies) and their antigen-binding fragments for the treatment or relief of symptoms of a disease or ailment, pharmaceutical compositions comprising anti-FGFR2b antibodies, anti-FGFR1b antibodies, or anti-DDR1 antibodies and their antigen-binding fragments, and uses thereof.

[0045] [Exemplary Example]

[0046] The following aspects and embodiments are part of this invention—whereby those skilled in the art will understand that the embodiments included below are independently intended as embodiments of each general embodiment and aspect set forth below. In aspects and embodiments, the invention relates to any one of the following [A1] and [A1a] through [A53]:

[0047] [A1] An agonistic antigen-binding molecule that binds to human fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of the extracellular domain (ECD) of a first FGFR2 protein and a second antigen-binding domain that binds to a second epitope of the extracellular domain (ECD) of a first FGFR2 protein or a second FGFR2 protein, wherein the first epitope and the second epitope are distinct from each other.

[0048] [A1a] An agonistic antigen-binding molecule that binds to human fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises:

[0049] The first component of the first epitope for binding to the extracellular domain (ECD) of the first FGFR2 protein; and

[0050] A second component for binding to the extracellular domain (ECD) of the first or second FGFR2 protein, a second epitope.

[0051] The first and second epitopes are different from each other.

[0052] [A2] The molecule according to [A1] or [A1a], wherein a first antigen-binding domain or a first component binds a first FGFR2 protein, and a second antigen-binding domain or a second component binds a second FGFR2 protein.

[0053] [A3] The molecule described in [A1], [A1a] or [A2] is a two-site antigen-binding molecule.

[0054] [A4] The molecule according to any one of [A1] and [A1a] to [A3], wherein the human FGFR2 protein is the human FGFR2b protein.

[0055] [A5] The molecule according to any one of [A1] and [A1a] to [A4], wherein each of the extracellular domains is independently selected from the immunoglobulin (Ig)-like domain of the FGFR2 protein.

[0056] [A6] The molecule according to any one of [A1] and [A1a] to [A5], wherein the first antigen-binding domain or the first component binds to the first domain of the FGFR2 protein and the second antigen-binding domain or the second component binds to the second domain of the FGFR2 protein, wherein the first domain and the second domain are identical.

[0057] [A7] The molecule according to any one of [A1] and [A1a] to [A5], wherein a first antigen-binding domain or a first component binds to a first domain of the FGFR2 protein and a second antigen-binding domain or a second component binds to a second domain of the FGFR2 protein, wherein the second domain is different from the first domain.

[0058] [A8] The molecule according to any one of [A1] and [A1a] to [A7], wherein the first antigen-binding domain and the second antigen-binding domain or the first component and the second component independently bind to the Ig-like domain of the FGFR2 protein, the Ig-like domain being selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

[0059] [A9] The molecule according to any one of [A5] to [A8], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR2 in a combination selected from the group consisting of:

[0060] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR2;

[0061] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIII domains of FGFR2;

[0062] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgII domains of FGFR2; and

[0063] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgI domains of FGFR2.

[0064] [A10] The molecule according to any one of [A5] to [A9], wherein (i) the first antigen-binding domain or the first component binds to the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the IgIIIb of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR2.

[0065] [A11] The molecule according to any one of [A1] to [A10], wherein the second antigen-binding domain is selected from the group consisting of: FFB0464, FFB0341, FFB0396, FFB0026, FFB0094, FFB0035, FFB0045, FFB0057 and FFB0080.

[0066] [A12] The molecule according to [A11], wherein the second antigen-binding domain is selected from the group consisting of: FFB0464, FFB0341, FFB0396.

[0067] [A13] The molecule according to [A11], wherein the first antigen-binding domain is selected from the group consisting of: FFB0057, FFB0065, FFB0080, FFB0088, FFB0185, FFB0219, FFB0261, FFB0294, FFB0299, FFB0335, FFB0365, FFB0368, FFB0374 and FFB0453.

[0068] [A14] The molecule according to any one of [A3] to [A13], wherein the binding of the first antigen-binding domain and the second antigen-binding domain or the first component and the second component to the FGFR2b protein results in the following agonist activity:

[0069] (i) wherein the molecule activates signal transduction in the target cell via binding to the FGFR2b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% or more in the expression of an intracellular protein downstream of FGFR2b compared to the expression of said intracellular protein prior to binding of the molecule to the FGFR2b protein expressed on the cell surface of the target cell, particularly wherein the intracellular protein downstream of FGFR2b is selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1 / pERK2, and pMAPK3 / pERK1; or

[0070] (ii) wherein the molecule induces the proliferation of target cells by binding to the FGFR2b protein expressed on the cell surface of the target cells, thereby exhibiting agonist activity, wherein the induction of target cell proliferation is measured by an increase in ATP expression in the target cells in the presence of the molecule by at least 1.5-fold, or 2-fold, or 2.5-fold or greater compared to ATP expression in the target cells in the absence of the molecule, wherein the ATP expression is proportional to the concentration of live target cells.

[0071] In particular, the target cells are lung epithelial cells, optionally selected from type II alveolar (AT2) cells; or

[0072] (iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization indicates agonist activity; or

[0073] (iv) In an in vitro cell assay, phosphorylation of the FGFR2b protein is detected in the presence of the molecule, wherein phosphorylation of the FGFR2b protein indicates agonist activity.

[0074] [A15] The molecule according to any one of [A1] to [A14], wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, wherein the first antigen-binding domain and the second antigen-binding domain are linked by at least one disulfide bond.

[0075] [A16] The molecule according to any one of [A1] to [A15], wherein the first antigen-binding domain and the second antigen-binding domain are characterized by having at least one disulfide bond formed between amino acid residues not in the hinge region.

[0076] [A17] The molecule according to [A16], wherein the at least one disulfide bond is an engineered disulfide bond not present in wild-type IgG, optionally wherein the at least one disulfide bond is formed between the CH1 region, CL region, VH or VHH region, or VL region of the first antigen-binding domain and the CH1 region, CL region, VH or VHH region, or VL region of the second antigen-binding domain, particularly wherein the at least one disulfide bond is formed between regions selected from the group consisting of:

[0077] (i) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CH1 region of the second antigen-binding domain;

[0078] (ii) Amino acid residues in the CL region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0079] (iii) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0080] (iv) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VH or VHH region of the second antigen-binding domain.

[0081] (v) Amino acid residues in the VL region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain; and

[0082] (vi) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain.

[0083] [A18] The molecule according to [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CH1 region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first and second antigen-binding domains originates is located at a position selected from the group consisting of: [the following positions are specified in the original text, but are not translated here: "in the CH1 region according to EU..."] The numbers 119, 122, 123, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 148, 150, 155, 156, 157, 159, 160, 161, 162, 163, 164, 165, 167, 174, 176, 177, 178, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 201, 203, 205, 206, 207, 208, 211, 212, 213, 214, 218, and 219.

[0084] [A19] The molecule according to [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the CL region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 108, 109, 112, 121, 123, 126, 128, 151, 152, 153, 156, 184, 186, 188, 189, 190, 195, 196, 200, 201, 202, 203, 208, 210, 211, 212 and 213 in the CL region according to Kabat numbers.

[0085] [A20] According to the molecule of [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of the following positions: 188, 189, 190, 191, 192, 193, 194, 195, 196 and 197 according to EU numbers, and the amino acid residue in the CL region is selected from the group consisting of the following positions: 121, 122, 123, 124, 125, 126, 127 and 128 according to Kabat numbers.

[0086] [A21] According to the molecule of [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the VH region of the first antigen-binding domain and an amino acid residue in the VH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 6, 8, 16, 20, 25, 26, 28, 74 and 82b of the VH region according to Kabat numbers.

[0087] [A22] According to the molecule of [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the VHH region of the first antigen-binding domain and an amino acid residue in the VHH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 4, 6, 7, 8, 9, 10, 11, 12, 14, 15, 17, 20, 24, 27, 29, 38, 39, 40, 41, 43, 44, 45, 46, 47, 48, 49, 67, 69, 71, 78, 80, 82, 82c, 85, 88, 91, 93, 94 and 107 according to Kabat numbers in the VHH region.

[0088] [A23] According to the molecule described in [A17], wherein the at least one disulfide bond is formed between an amino acid residue in the VL region of the first antigen-binding domain and an amino acid residue in the VL region of the second antigen-binding domain, and

[0089] When VL belongs to the κ subclass, the amino acid residues from which the bond between the first and second antigen-binding domains originates are located at positions selected from the group consisting of: VL regions (κ subclass) at Kabat numbers 21, 27, 58, 77, 100, 105, and 107; or

[0090] When VL belongs to the λ subclass, the amino acid residues from which the bond between the first antigen-binding domain and the second antigen-binding domain originates are located at positions selected from the following groups: VL region (λ subclass) according to Kabat numbers 6, 19, 33 and 34.

[0091] [A24] The molecule according to [A23], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain.

[0092] [A25] The molecule according to [A24], wherein the at least one disulfide bond is formed between an amino acid residue at position 191 (according to Kabat number) in the CH1 region of the first antigen-binding domain and an amino acid residue at position 126 (according to Kabat number) in the CL region of the second antigen-binding domain.

[0093] [A26] The molecule according to [A25], wherein the at least one disulfide bond is formed between S191C (according to Kabat number) in the CH1 region of the first antigen-binding domain and K126C (according to Kabat number) in the CL region of the second antigen-binding domain.

[0094] [A27] The molecule according to any one of [A1] and [A1a] to [A26], wherein the molecule comprises an antibody constant region, particularly (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from the human IgG1, IgG2, IgG3 or IgG4 constant regions.

[0095] [A28] The molecule according to any one of [A1] to [A27], wherein the molecule comprises or is an antibody or an antibody fragment thereof.

[0096] [A29] The molecule according to [A28], wherein the antibody fragment is Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody.

[0097] [A30] The molecule described in [A28] or [A29], wherein the molecule is a monoclonal antibody.

[0098] [A31] The molecule according to any one of [A29] to [A30], wherein the molecule is a human antibody, a humanized antibody or a chimeric antibody.

[0099] [A32] The molecule according to any one of [A1] to [A31], wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a bivalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each monovalent Fab or monovalent VHH, or each monovalent scFab.

[0100] [A33] The molecule according to any one of [A1] and [A1a] to [A32], wherein the molecule comprises more than two antigen-binding domains or more than two binding members, or wherein the molecule is a trivalent, tetravalent or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain that binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, or wherein the molecule comprises a third member for binding to the same epitope on the same domain as the first member or the second member.

[0101] [A34] The molecule according to [A33], wherein the molecule comprises a fourth antigen-binding domain that binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, but not to the same epitope on the third antigen-binding domain, or wherein the molecule comprises a fourth member for binding to the same epitope on the same domain as the first member or the second member, but not to the same epitope on the third member, optionally.

[0102] The molecule contains a third antigen-binding domain and a fourth antigen-binding domain, wherein the third antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds to the same epitope on the same domain as the second antigen-binding domain, or...

[0103] The molecule includes a third component that binds to the same epitope in the same structural domain as the first component, and a fourth component that binds to the same epitope in the same structural domain as the second component.

[0104] [A35] A pharmaceutical composition comprising a molecule according to any one of [A1] and [A1a] to [A34], and a pharmaceutically acceptable carrier.

[0105] [A36] An immune conjugate protein comprising any one of [A1] and [A1a] to [A34].

[0106] [A37] The molecule according to any one of [A1] and [A1a] to [A34], or the pharmaceutical composition according to [A35], or the immunoconjugate according to [A36], for use in the treatment or prevention of a disease, ailment, or condition associated with lung epithelial cell damage in an individual in need.

[0107] [A38] According to [A37] the molecular or pharmaceutical composition or immunoconjugate used for the purpose of the application, wherein the individual's condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0108] [A39] The molecule according to any one of [A1] and [A1a] to [A34], or the pharmaceutical composition according to [A35], or the immunoconjugate according to [A36], for use in the treatment of any disease, ailment, or condition that is readily improved or prevented by one of the following selected from the group consisting of:

[0109] (i) Increase or enhancement of FGFR2b signal conduction;

[0110] (ii) Increased proliferation of lung epithelial cells; and

[0111] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0112] [A40] The molecule according to any one of [A1] and [A1a] to [A34], or the pharmaceutical composition according to [A35], or the immunoconjugate according to [A36], for use in the treatment or prevention of fibrosis or inflammation in an individual in need.

[0113] [A41] The molecular or pharmaceutical composition or immunoconjugate for use according to [A40], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0114] [A42] According to [A41], the molecular or pharmaceutical composition or immunoconjugate used for the purpose thereof, wherein

[0115] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0116] (ii) Fibrosis or inflammation of the liver or bile ducts, selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager syndrome;

[0117] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0118] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0119] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0120] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0121] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0122] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0123] [A43] Use in the manufacture of a medicament of any one of [A1] and [A1a] to [A34], or of a pharmaceutical composition according to [A35], or of an immunoconjugate according to [A36], for the treatment or prevention of a disease, ailment, or condition associated with lung epithelial cell damage in an individual in need.

[0124] [A44] A method of treating an individual in need of a disease, ailment, or condition associated with lung epithelial cell damage, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1] and [A1a] to [A34], or a pharmaceutical composition according to [A35], or an immunoconjugate according to [A36].

[0125] [A45] According to the use described in [A43] or the method described in [A44], wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0126] [A46] According to the use described in [A43] or the method described in [A44], wherein the individual's condition associated with lung epithelial cell damage is a condition that is easily improved or prevented by selecting one of the following groups:

[0127] (i) Increase or enhancement of FGFR2b signal conduction;

[0128] (ii) Increased proliferation of lung epithelial cells; and

[0129] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0130] [A47] Use in the manufacture of a medicament of any one of [A1] and [A1a] to [A34], or of a pharmaceutical composition according to [A35], or of an immunoconjugate according to [A36], for the treatment or prevention of fibrosis or inflammation in an individual in need.

[0131] [A48] A method of treating fibrosis or inflammation in an individual in need, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1] and [A1a] to [A34] or a pharmaceutical composition according to [A35] or an immunoconjugate according to [A36].

[0132] [A49] The use described in [A47] or the method described in [A48], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0133] [A50] According to the use described in [A47] or the method described in [A48], wherein

[0134] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0135] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0136] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0137] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0138] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0139] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0140] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0141] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0142] [A51] An isolated nucleic acid encoding a molecule according to any one of [A1] and [A1a] to [A34]; or multiple isolated nucleic acids encoding a molecule according to any one of [A1] and [A1a] to [A34].

[0143] [A52] A host cell comprising nucleic acid or multiple nucleic acids as described in [A51].

[0144] [A53] A method for producing molecules, or a process for producing molecules, comprising the following steps:

[0145] (a) Culturing host cells as described in [A52], and

[0146] (b) Recovering molecules generated in step (a).

[0147] In various aspects and embodiments, the present invention relates to any one of the following [A1-1] and [A1a-1] to [A52-1]:

[0148] [A1-1] An agonistic antigen-binding molecule that binds to human fibroblast growth factor receptor 1 (FGFR1) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of an extracellular domain (ECD) of a first FGFR1 protein and a second antigen-binding domain that binds to a second epitope of an extracellular domain (ECD) of a first FGFR1 protein or a second FGFR1 protein, wherein the first epitope and the second epitope are distinct from each other.

[0149] [A1a-1] An agonistic antigen-binding molecule that binds to human fibroblast growth factor receptor 1 (FGFR1) protein, wherein the molecule comprises:

[0150] The first component of the first epitope for binding to the extracellular domain (ECD) of the first FGFR1 protein; and

[0151] A second component of a second epitope for binding to the extracellular domain (ECD) of the first or second FGFR1 protein.

[0152] The first and second epitopes are different from each other.

[0153] [A2-1] The molecule according to [A1-1] or [A1a-1], wherein a first antigen-binding domain or a first component binds a first FGFR1 protein, and a second antigen-binding domain or a second component binds a second FGFR1 protein.

[0154] [A3-1] The molecule described in [A1-1], [A1a-1] or [A2-1] is a two-site antigen-binding molecule.

[0155] [A4-1] The molecule according to any one of [A1-1] and [A1a-1] to [A3-1], wherein the human FGFR1 protein is the human FGFR1b protein.

[0156] [A5-1] The molecule according to any one of [A1-1] and [A1a-1] to [A4-1], wherein each of the extracellular domains is independently selected from the immunoglobulin (Ig)-like domain of the FGFR1 protein.

[0157] [A6-1] The molecule according to any one of [A1-1] and [A1a-1] to [A5-1], wherein a first antigen-binding domain or a first component binds to a first domain of the FGFR1 protein and a second antigen-binding domain or a second component binds to a second domain of the FGFR1 protein, wherein the first domain and the second domain are identical.

[0158] [A7-1] The molecule according to any one of [A1-1] and [A1a-1] to [A5-1], wherein a first antigen-binding domain or a first component binds to a first domain of the FGFR1 protein and a second antigen-binding domain or a second component binds to a second domain of the FGFR1 protein, wherein the second domain is different from the first domain.

[0159] [A8-1] The molecule according to any one of [A1-1] and [A1a-1] to [A7-1], wherein the first antigen-binding domain and the second antigen-binding domain or the first component and the second component independently bind to the Ig-like domain of the FGFR1 protein, the Ig-like domain being selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

[0160] [A9-1] The molecule according to any one of [A5-1] to [A8-1], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR1 in a combination selected from the group consisting of:

[0161] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR1.

[0162] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIII domains of FGFR1;

[0163] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgII domains of FGFR1; and

[0164] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgI domains of FGFR1.

[0165] [A10-1] The molecule according to any one of [A5-1] to [A9-1], wherein (i) the first antigen-binding domain or the first component binds to the IgIIIb domain of FGFR1, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the IgIIIb of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR1.

[0166] [A11-1] The molecule according to any one of [A1-1] to [A10-1], wherein the second antigen-binding domain is selected from the group consisting of: FFA0020, FFA0026, FFA0032, FFA0043, FFA0047, FFA0055, FFA0064 and FFA0088.

[0167] [A12-1] According to the molecule described in [A11-1], the first antigen-binding domain is selected from the group consisting of: FFA0029 and FFA0048.

[0168] [A13-1] The molecule according to any one of [A3-1] to [A12-1], wherein the binding of the first antigen-binding domain and the second antigen-binding domain or the first component and the second component to the FGFR1b protein results in the following agonist activity:

[0169] (i) wherein the molecule activates signal transduction in the target cell via binding to the FGFR1b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% or more in the expression of an intracellular protein downstream of FGFR1b compared to the expression of said intracellular protein prior to binding of the molecule to the FGFR1b protein expressed on the cell surface of the target cell, particularly wherein the intracellular protein downstream of FGFR1b is selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1 / pERK2, and pMAPK3 / pERK1; or

[0170] (ii) wherein the molecule induces the proliferation of target cells by binding to the FGFR1b protein expressed on the cell surface of the target cells, thereby exhibiting agonist activity, wherein the induction of target cell proliferation is measured by an increase in ATP expression in the target cells in the presence of the molecule by at least 1.5-fold, 2-fold, 2.5-fold, or greater compared to ATP expression in the target cells in the absence of the molecule, wherein the ATP expression is proportional to the concentration of live target cells.

[0171] In particular, the target cells are lung epithelial cells, optionally selected from type II alveolar (AT2) cells; or

[0172] (iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization indicates agonist activity; or

[0173] (iv) In an in vitro cell assay, phosphorylation of FGFR1b protein is detected in the presence of the molecule, wherein phosphorylation of FGFR1b protein indicates agonist activity.

[0174] [A14-1] The molecule according to any one of [A1-1] to [A13-1], wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, wherein the first antigen-binding domain and the second antigen-binding domain are linked by at least one disulfide bond.

[0175] [A15-1] The molecule according to any one of [A1-1] to [A14-1], wherein the first antigen-binding domain and the second antigen-binding domain are characterized by having at least one disulfide bond formed between amino acid residues not in the hinge region.

[0176] [A16-1] The molecule according to [A15-1], wherein the at least one disulfide bond is an engineered disulfide bond not present in wild-type IgG, optionally wherein the at least one disulfide bond is formed between the CH1 region, CL region, VH or VHH region, or VL region of the first antigen-binding domain and the CH1 region, CL region, VH or VHH region, or VL region of the second antigen-binding domain, particularly wherein the at least one disulfide bond is formed between regions selected from the group consisting of:

[0177] (vii) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CH1 region of the second antigen-binding domain;

[0178] (viii) Amino acid residues in the CL region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0179] (ix) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0180] (x) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VH or VHH region of the second antigen-binding domain.

[0181] (xi) The amino acid residues in the VL region of the first antigen-binding domain and the amino acid residues in the VL region of the second antigen-binding domain; and

[0182] (xii) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain.

[0183] [A17-1] According to the molecule of [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CH1 region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first and second antigen-binding domains originates is located at a position selected from the group consisting of: [the following positions are specified in the original text, but are not translated here: "in the CH1 region according to EU..."] The numbers 119, 122, 123, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 148, 150, 155, 156, 157, 159, 160, 161, 162, 163, 164, 165, 167, 174, 176, 177, 178, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 201, 203, 205, 206, 207, 208, 211, 212, 213, 214, 218, and 219.

[0184] [A18-1] The molecule according to [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the CL region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 108, 109, 112, 121, 123, 126, 128, 151, 152, 153, 156, 184, 186, 188, 189, 190, 195, 196, 200, 201, 202, 203, 208, 210, 211, 212 and 213 in the CL region according to Kabat numbers.

[0185] [A19-1] According to the molecule described in [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of the following positions: 188, 189, 190, 191, 192, 193, 194, 195, 196 and 197 according to EU numbers, and the amino acid residue in the CL region is selected from the group consisting of the following positions: 121, 122, 123, 124, 125, 126, 127 and 128 according to Kabat numbers.

[0186] [A20-1] According to the molecule of [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the VH region of the first antigen-binding domain and an amino acid residue in the VH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 6, 8, 16, 20, 25, 26, 28, 74 and 82b according to Kabat numbers in the VH region.

[0187] [A21-1] According to the molecule of [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the VHH region of the first antigen-binding domain and an amino acid residue in the VHH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 4, 6, 7, 8, 9, 10, 11, 12, 14, 15, 17, 20, 24, 27, 29, 38, 39, 40, 41, 43, 44, 45, 46, 47, 48, 49, 67, 69, 71, 78, 80, 82, 82c, 85, 88, 91, 93, 94 and 107 according to Kabat numbers in the VHH region.

[0188] [A22-1] According to the molecule described in [A16-1], wherein the at least one disulfide bond is formed between an amino acid residue in the VL region of the first antigen-binding domain and an amino acid residue in the VL region of the second antigen-binding domain, and

[0189] When VL belongs to the κ subclass, the amino acid residues from which the bond between the first and second antigen-binding domains originates are located at positions selected from the group consisting of: VL regions (κ subclass) at Kabat numbers 21, 27, 58, 77, 100, 105, and 107; or

[0190] When VL belongs to the λ subclass, the amino acid residues from which the bond between the first antigen-binding domain and the second antigen-binding domain originates are located at positions selected from the following groups: VL region (λ subclass) according to Kabat numbers 6, 19, 33 and 34.

[0191] [A23-1] The molecule according to [A22-1], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain.

[0192] [A24-1] The molecule according to [A23-1], wherein the at least one disulfide bond is formed between an amino acid residue at position 191 (according to Kabat number) in the CH1 region of the first antigen-binding domain and an amino acid residue at position 126 (according to Kabat number) in the CL region of the second antigen-binding domain.

[0193] [A25-1] The molecule according to [A24-1], wherein the at least one disulfide bond is formed between S191C (according to Kabat number) in the CH1 region of the first antigen-binding domain and K126C (according to Kabat number) in the CL region of the second antigen-binding domain.

[0194] [A26-1] A molecule according to any one of [A1-1] and [A1a-1] to [A25-1], wherein the molecule comprises an antibody constant region, particularly (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from the human IgG1, IgG2, IgG3 or IgG4 constant regions.

[0195] [A27-1] The molecule according to any one of [A1-1] to [A26-1], wherein the molecule comprises or is an antibody or an antibody fragment thereof.

[0196] [A28-1] The molecule according to [A27-1], wherein the antibody fragment is Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody.

[0197] [A29-1] The molecule described in [A27-1] or [A28-1], wherein the molecule is a monoclonal antibody.

[0198] [A30-1] The molecule according to any one of [A28-1] to [A29-1], wherein the molecule is a human antibody, a humanized antibody or a chimeric antibody.

[0199] [A31-1] A molecule according to any one of [A1-1] to [A30-1], wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a divalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each monovalent Fab or monovalent VHH, or each monovalent scFab.

[0200] [A32-1] A molecule according to any one of [A1-1] and [A1a-1] to [A31-1], wherein the molecule comprises more than two antigen-binding domains or more than two binding members, or wherein the molecule is a trivalent, tetravalent or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain that binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, or wherein the molecule comprises a third member for binding to the same epitope on the same domain as the first member or the second member.

[0201] [A33-1] The molecule according to [A32-1], wherein the molecule comprises a fourth antigen-binding domain that binds to the same epitope on the same domain as the first or second antigen-binding domain, but not to the same epitope on the third antigen-binding domain, or wherein the molecule comprises a fourth member for binding to the same epitope on the same domain as the first or second member but not to the same epitope on the third member, optionally.

[0202] The molecule contains a third antigen-binding domain and a fourth antigen-binding domain, wherein the third antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds to the same epitope on the same domain as the second antigen-binding domain, or...

[0203] The molecule includes a third component that binds to the same epitope in the same structural domain as the first component, and a fourth component that binds to the same epitope in the same structural domain as the second component.

[0204] [A34-1] A pharmaceutical composition comprising a molecule according to any one of [A1-1] and [A1a-1] to [A33-1], and a pharmaceutically acceptable carrier.

[0205] [A35-1] An immune conjugate protein comprising a molecule according to any one of [A1-1] and [A1a-1] to [A33-1].

[0206] [A36-1] The molecule according to any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition according to [A34-1] or the immunoconjugate according to [A35-1], for use in the treatment or prevention of a disease, ailment or condition associated with lung epithelial cell damage in an individual in need.

[0207] [A37-1] The molecular or pharmaceutical composition or immunoconjugate for use as described in [A36-1], wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0208] [A38-1] The molecule according to any one of [A1-1] and [A1a-1] to [A33-1], or the pharmaceutical composition according to [A34-1], or the immunoconjugate according to [A35-1], for use in the treatment of any disease, ailment, or condition that is readily improved or prevented by one of the following selected from the group consisting of:

[0209] (iv) Increase or enhancement of FGFR1b signal conduction;

[0210] (v) Increased proliferation of lung epithelial cells; and

[0211] (vi) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0212] [A39-1] The molecule according to any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition according to [A34-1] or the immunoconjugate according to [A35-1], for use in the treatment or prevention of fibrosis or inflammation in an individual in need.

[0213] [A40-1] The molecular or pharmaceutical composition or immunoconjugate for use according to [A39-1], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0214] [A41-1] According to [A40-1], the molecular or pharmaceutical composition or immunoconjugate used for the purpose thereof, wherein...

[0215] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0216] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0217] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0218] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0219] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0220] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0221] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0222] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0223] [A42-1] Use in the manufacture of a medicament of any one of [A1-1] and [A1a-1] to [A33-1], or of a pharmaceutical composition according to [A34-1], or of an immunoconjugate according to [A35-1], for the treatment or prevention of a disease, ailment, or condition associated with lung epithelial cell damage in an individual in need.

[0224] [A43-1] A method of treating an individual in need of a disease, ailment, or condition associated with lung epithelial cell damage, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1-1] and [A1a-1] to [A33-1], or a pharmaceutical composition according to [A34-1], or an immunoconjugate according to [A35-1].

[0225] [A44-1] According to the use described in [A42-1] or the method described in [A43-1], wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0226] [A45-1] According to the use described in [A42-1] or the method described in [A43-1], wherein the individual's condition associated with lung epithelial cell damage is a condition that is easily improved or prevented by selecting one of the following groups:

[0227] (i) Increase or enhancement of FGFR1b signal conduction;

[0228] (ii) Increased proliferation of lung epithelial cells; and

[0229] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0230] [A46-1] Use in the manufacture of a medicament of any one of [A1-1] and [A1a-1] to [A33-1], or of a pharmaceutical composition according to [A34-1], or of an immunoconjugate according to [A35-1], for the treatment or prevention of fibrosis or inflammation in an individual in need.

[0231] [A47-1] A method of treating fibrosis or inflammation in an individual in need, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1-1] and [A1a-1] to [A33-1] or a pharmaceutical composition according to [A34-1] or an immunoconjugate according to [A35-1].

[0232] [A48-1] The use described in [A46-1] or the method described in [A47-1], wherein the fibrosis or inflammation is selected from the fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0233] [A49-1] According to the use described in [A46-1] or the method described in [A47-1], wherein

[0234] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0235] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0236] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0237] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0238] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0239] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0240] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0241] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0242] [A50-1] An isolated nucleic acid encoding a molecule according to any one of [A1-1] and [A1a-1] to [A33-1]; or multiple isolated nucleic acids encoding a molecule according to any one of [A1-1] and [A1a-1] to [A33-1].

[0243] [A51-1] A host cell comprising nucleic acids or multiple nucleic acids as described in [A50-1].

[0244] [A52-1] A method for producing molecules, or a process for producing molecules, comprising the following steps:

[0245] (a) Culturing host cells as described in [A51-1], and

[0246] (b) Recovering molecules generated in step (a).

[0247] In various aspects and embodiments, the present invention relates to any one of the following [A1-2] and [A1a-2] to [A52-2]:

[0248] [A1-2] An agonistic antigen-binding molecule that binds to human discoid domain receptor 1 (DDR1) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of the extracellular domain (ECD) of a first DDR1 protein and a second antigen-binding domain that binds to a second epitope of the extracellular domain (ECD) of a first DDR1 protein or a second DDR1 protein, wherein the first epitope and the second epitope are distinct from each other.

[0249] [A1a-2] An agonistic antigen-binding molecule that binds to human discoid domain receptor 1 (DDR1) protein, wherein the molecule comprises:

[0250] The first component for binding the first epitope of the first DDR1 protein's extracellular domain (ECD); and

[0251] A second component for binding to the extracellular domain (ECD) of a first or second DDR1 protein, a second epitope.

[0252] The first and second epitopes are different from each other.

[0253] [A2-2] The molecule according to [A1-2] or [A1a-2], wherein a first antigen-binding domain or a first component binds a first DDR1 protein, and a second antigen-binding domain or a second component binds a second DDR1 protein.

[0254] [A3-2] The molecules described in [A1-2], [A1a-2] or [A2-2] are two-site antigen-binding molecules.

[0255] [A4-2] The molecule according to any one of [A1-2] and [A1a-2] to [A3-2], wherein the human DDR1 protein is the human DDR1a protein.

[0256] [A5-2] The molecule according to any one of [A1-2] and [A1a-2] to [A4-2], wherein each of the extracellular domains is independently selected from the disc-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of the DDR1 protein.

[0257] [A6-2] The molecule according to any one of [A1-2] and [A1a-2] to [A5-2], wherein the first antigen-binding domain or the first component binds to the first domain of the DDR1 protein and the second antigen-binding domain or the second component binds to the second domain of the DDR1 protein, wherein the first domain and the second domain are identical.

[0258] [A7-2] The molecule according to any one of [A1-2] and [A1a-2] to [A5-2], wherein a first antigen-binding domain or a first component binds to a first domain of the DDR1 protein and a second antigen-binding domain or a second component binds to a second domain of the DDR1 protein, wherein the second domain is different from the first domain.

[0259] [A8-2] The molecule according to any one of [A1-2] and [A1a-2] to [A7-2], wherein the first antigen-binding domain and the second antigen-binding domain or the first component and the second component independently bind to the disc-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of the DDR1 protein.

[0260] [A9-2] The molecule according to any one of [A5-2] to [A8-2], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the discoidal (DS) domain, DS-like domain, or extracellular juxtamembrane (EJXM) region of DDR1 in a manner selected from the group consisting of:

[0261] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS and DS domains of DDR1;

[0262] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS and DS-like domains of DDR1;

[0263] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS domain and the EJXM region of DDR1;

[0264] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS-like domain and the DS-like domain of DDR1.

[0265] (vii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS-like domain and the EJXM region of DDR1; and

[0266] (viii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the EJXM region and the EJXM region of DDR1.

[0267] [A10-2] The molecule according to any one of [A5-2] to [A9-2], wherein (i) the first antigen-binding domain or the first component binds to the DS domain of DDR1, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the DS domain of DDR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the DS-like domain of DDR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the EJXM region of DDR1.

[0268] [A11-2] The molecule according to any one of [A1-2] to [A10-2], wherein the second antigen-binding domain is selected from the group consisting of: DAA0007, DAA0012 and DAA0024.

[0269] [A12-2] The molecule according to [A11-2], wherein the first antigen-binding domain is selected from the group consisting of: DAA0011 and DAA0045.

[0270] [A13-2] The molecule according to any one of [A3-2] to [A12-2], wherein the binding of the first antigen-binding domain and the second antigen-binding domain or the first component and the second component to the DDR1 protein results in the following agonist activity:

[0271] (i) wherein the molecule activates signal transduction in the target cell via binding to the DDR1 protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% or more in the expression of an intracellular protein downstream of DDR1 compared to the expression of the intracellular protein prior to the binding of the molecule to the DDR1 protein expressed on the cell surface of the target cell, particularly wherein the intracellular protein downstream of DDR1 is selected from the group consisting of: SHP-2, FAK, NF-κB, Cdc42, MAPK1, AP1, Ras / Raf / ERK, PI3K / AKT, STAT1 / 3 / 5; or

[0272] (ii) wherein the molecule induces the proliferation of the target cells by binding to the DDR1 protein expressed on the cell surface of the target cells, thereby exhibiting agonist activity, wherein the induction of the proliferation of the target cells is measured by an increase in ATP expression in the target cells in the presence of the molecule by at least 1.5-fold, 2-fold, 2.5-fold, or greater compared to ATP expression in the target cells in the absence of the molecule, wherein the ATP expression is proportional to the concentration of the live target cells.

[0273] In particular, the target cells are lung epithelial cells, optionally selected from type II alveolar (AT2) cells; or

[0274] (iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization indicates agonist activity; or

[0275] (iv) In an in vitro cell assay, phosphorylation of the DDR1 protein is detected in the presence of the molecule, wherein phosphorylation of the DDR1 protein indicates agonist activity.

[0276] [A14-2] The molecule according to any one of [A1-2] to [A13-2], wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, wherein the first antigen-binding domain and the second antigen-binding domain are linked by at least one disulfide bond.

[0277] [A15-2] The molecule according to any one of [A1-2] to [A14-2], wherein the first antigen-binding domain and the second antigen-binding domain are characterized by having at least one disulfide bond formed between amino acid residues not in the hinge region.

[0278] [A16-2] The molecule according to [A15-2], wherein the at least one disulfide bond is an engineered disulfide bond not present in wild-type IgG, optionally wherein the at least one disulfide bond is formed between the CH1 region, CL region, VH or VHH region, or VL region of the first antigen-binding domain and the CH1 region, CL region, VH or VHH region, or VL region of the second antigen-binding domain, particularly wherein the at least one disulfide bond is formed between regions selected from the group consisting of:

[0279] (xiii) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CH1 region of the second antigen-binding domain;

[0280] (xiv) Amino acid residues in the CL region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0281] (xv) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0282] (xvi) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VH or VHH region of the second antigen-binding domain.

[0283] (xvii) The amino acid residues in the VL region of the first antigen-binding domain and the amino acid residues in the VL region of the second antigen-binding domain; and

[0284] (xviii) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain.

[0285] [A17-2] According to the molecule of [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CH1 region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: in the CH1 region according to EU The numbers 119, 122, 123, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 148, 150, 155, 156, 157, 159, 160, 161, 162, 163, 164, 165, 167, 174, 176, 177, 178, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 201, 203, 205, 206, 207, 208, 211, 212, 213, 214, 218, and 219.

[0286] [A18-2] The molecule according to [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the CL region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 108, 109, 112, 121, 123, 126, 128, 151, 152, 153, 156, 184, 186, 188, 189, 190, 195, 196, 200, 201, 202, 203, 208, 210, 211, 212 and 213 according to Kabat numbers in the CL region.

[0287] [A19-2] The molecule according to [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of the following positions: 188, 189, 190, 191, 192, 193, 194, 195, 196 and 197 according to EU numbers, and the amino acid residue in the CL region is selected from the group consisting of the following positions: 121, 122, 123, 124, 125, 126, 127 and 128 according to Kabat numbers.

[0288] [A20-2] According to the molecule of [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the VH region of the first antigen-binding domain and an amino acid residue in the VH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 6, 8, 16, 20, 25, 26, 28, 74 and 82b according to Kabat numbers in the VH region.

[0289] [A21-2] The molecule according to [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the VHH region of the first antigen-binding domain and an amino acid residue in the VHH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 4, 6, 7, 8, 9, 10, 11, 12, 14, 15, 17, 20, 24, 27, 29, 38, 39, 40, 41, 43, 44, 45, 46, 47, 48, 49, 67, 69, 71, 78, 80, 82, 82c, 85, 88, 91, 93, 94 and 107 according to Kabat numbers in the VHH region.

[0290] [A22-2] According to the molecule described in [A16-2], wherein the at least one disulfide bond is formed between an amino acid residue in the VL region of the first antigen-binding domain and an amino acid residue in the VL region of the second antigen-binding domain, and

[0291] When VL belongs to the κ subclass, the amino acid residues from which the bond between the first and second antigen-binding domains originates are located at positions selected from the group consisting of: VL regions (κ subclass) at Kabat numbers 21, 27, 58, 77, 100, 105, and 107; or

[0292] When VL belongs to the λ subclass, the amino acid residues from which the bond between the first antigen-binding domain and the second antigen-binding domain originates are located at positions selected from the following groups: VL region (λ subclass) according to Kabat numbers 6, 19, 33 and 34.

[0293] [A23-2] The molecule according to [A22-2], wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain.

[0294] [A24-2] The molecule according to [A23-2], wherein the at least one disulfide bond is formed between an amino acid residue at position 191 (according to Kabat number) in the CH1 region of the first antigen-binding domain and an amino acid residue at position 126 (according to Kabat number) in the CL region of the second antigen-binding domain.

[0295] [A25-2] The molecule according to [A24-2], wherein the at least one disulfide bond is formed between S191C (according to Kabat number) in the CH1 region of the first antigen-binding domain and K126C (according to Kabat number) in the CL region of the second antigen-binding domain.

[0296] [A26-2] A molecule according to any one of [A1-2] and [A1a-2] to [A25-2], wherein the molecule comprises an antibody constant region, particularly (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from the human IgG1, IgG2, IgG3 or IgG4 constant regions.

[0297] [A27-2] The molecule according to any one of [A1-2] to [A26-2], wherein the molecule comprises or is an antibody or an antibody fragment thereof.

[0298] [A28-2] The molecule according to [A27-2], wherein the antibody fragment is Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody.

[0299] [A29-2] The molecule described in [A27-2] or [A28-2], wherein the molecule is a monoclonal antibody.

[0300] [A30-2] The molecule according to any one of [A28-2] to [A29-2], wherein the molecule is a human antibody, a humanized antibody or a chimeric antibody.

[0301] [A31-2] A molecule according to any one of [A1-2] to [A30-2], wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a divalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each monovalent Fab or monovalent VHH, or each monovalent scFab.

[0302] [A32-2] The molecule according to any one of [A1-2] and [A1a-2] to [A31-2], wherein the molecule comprises more than two antigen-binding domains or more than two binding members, or wherein the molecule is a trivalent, tetravalent or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain that binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, or wherein the molecule comprises a third member for binding to the same epitope on the same domain as the first member or the second member.

[0303] [A33-2] The molecule according to [A32-2], wherein the molecule comprises a fourth antigen-binding domain that binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, but not to the same epitope on the third antigen-binding domain, or wherein the molecule comprises a fourth member for binding to the same epitope on the same domain as the first member or the second member, but not to the same epitope on the third member, optionally.

[0304] The molecule contains a third antigen-binding domain and a fourth antigen-binding domain, wherein the third antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds to the same epitope on the same domain as the second antigen-binding domain, or...

[0305] The molecule includes a third component that binds to the same epitope in the same structural domain as the first component, and a fourth component that binds to the same epitope in the same structural domain as the second component.

[0306] [A34-2] A pharmaceutical composition comprising a molecule according to any one of [A1-2] and [A1a-2] to [A33-2], and a pharmaceutically acceptable carrier.

[0307] [A35-2] An immune conjugate protein comprising a molecule according to any one of [A1-2] and [A1a-2] to [A33-2].

[0308] [A36-2] The molecule according to any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition according to [A34-2] or the immunoconjugate according to [A35-2], for use in the treatment or prevention of a disease, ailment or condition associated with lung epithelial cell damage in an individual in need.

[0309] [A37-2] According to [A36-2] the molecular or pharmaceutical composition or immunoconjugate used for the purpose of the application, wherein the individual's condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0310] [A38-2] The molecule according to any one of [A1-2] and [A1a-2] to [A33-2], or the pharmaceutical composition according to [A34-2], or the immunoconjugate according to [A35-2], for use in the treatment of any disease, ailment, or condition that is readily improved or prevented by one of the following selected from the group consisting of:

[0311] (vii) Increase or enhancement of DDR1 signal conduction;

[0312] (viii) Increased proliferation of lung epithelial cells; and

[0313] (ix) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0314] [A39-2] The molecule according to any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition according to [A34-2] or the immunoconjugate according to [A35-2], for use in the treatment or prevention of fibrosis or inflammation in an individual in need.

[0315] [A40-2] The molecular or pharmaceutical composition or immunoconjugate for use according to [A39-2], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0316] [A41-2] According to [A40-2], the molecular or pharmaceutical composition or immunoconjugate used for the purpose of said application, wherein...

[0317] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0318] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0319] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0320] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0321] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0322] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0323] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0324] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0325] [A42-2] Use in the manufacture of a medicament of any one of [A1-2] and [A1a-2] to [A33-2], or of a pharmaceutical composition according to [A34-2], or of an immunoconjugate according to [A35-2], for the treatment or prevention of a disease, ailment, or condition associated with lung epithelial cell damage in an individual in need.

[0326] [A43-2] A method of treating an individual in need of a disease, ailment, or condition associated with lung epithelial cell damage, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1-2] and [A1a-2] to [A33-2], or a pharmaceutical composition according to [A34-2], or an immunoconjugate according to [A35-2].

[0327] [A44-2] According to the use described in [A42-2] or the method described in [A43-2], wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0328] [A45-2] According to the use described in [A42-2] or the method described in [A43-2], wherein the individual's condition associated with lung epithelial cell damage is a condition that is easily improved or prevented by selecting one of the following groups:

[0329] (i) Increase or enhancement of DDR1 signal conduction;

[0330] (ii) Increased proliferation of lung epithelial cells; and

[0331] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0332] [A46-2] Use in the manufacture of a medicament of any one of [A1-2] and [A1a-2] to [A33-2], or of a pharmaceutical composition according to [A34-2], or of an immunoconjugate according to [A35-2], for the treatment or prevention of fibrosis or inflammation in an individual in need.

[0333] [A47-2] A method of treating fibrosis or inflammation in an individual in need, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of [A1-2] and [A1a-2] to [A33-2] or a pharmaceutical composition according to [A34-2] or an immunoconjugate according to [A35-2].

[0334] [A48-2] The use described in [A46-2] or the method described in [A47-2], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0335] [A49-2] According to the use described in [A46-2] or the method described in [A47-2], wherein

[0336] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0337] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0338] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0339] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0340] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0341] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0342] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0343] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0344] [A50-2] An isolated nucleic acid encoding a molecule according to any one of [A1-2] and [A1a-2] to [A33-2]; or multiple isolated nucleic acids encoding a molecule according to any one of [A1-2] and [A1a-2] to [A33-2].

[0345] [A51-2] A host cell comprising nucleic acids or multiple nucleic acids as described in [A50-2].

[0346] [A52-2] A method for producing molecules, or a process for producing molecules, comprising the following steps:

[0347] (a) Culturing host cells as described in [A51-2], and

[0348] (b) Recovering molecules generated in step (a).

[0349] In further aspects and embodiments, the invention relates to any one of the following [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [51-2], [51a-2], [52-2], [52a-2], [53-2] to [61-2],

[62] to

[104] :

[0350] [1] An agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein the molecule is an agonist of the RTK protein.

[0351] [2] The molecule according to [1], wherein the molecule comprises a first antigen-binding domain that binds a first epitope of a first RTK protein and a second antigen-binding domain that binds a second epitope of the first RTK protein or a second RTK protein, wherein the first epitope resides in the first domain of the RTK protein and the second epitope resides in the second domain of the RTK protein.

[0352] [2a] The molecule according to [1], wherein the molecule comprises a first component for binding a first epitope of a first RTK protein and a second component for binding a second epitope of the first RTK protein or a second RTK protein, wherein the first epitope resides in a first domain of the RTK protein and the second epitope resides in a second domain of the RTK protein.

[0353] [3] The molecule according to [2] or [2a], wherein the first antigen-binding domain or the first component binds to the first RTK protein and the second antigen-binding domain or the second component binds to the second RTK protein, optionally wherein the first RTK protein and the second RTK protein form a dimer.

[0354] [4] The molecule according to any one of [2], [2a] and [3], wherein the binding of the first antigen-binding domain and the second antigen-binding domain or the binding of the first component and the second component induces dimerization of the intracellular tyrosine kinase domain of the RTK protein.

[0355] [5] The molecule according to [4], wherein dimerization is detected in the presence of the molecule in an in vitro cell assay, and wherein dimerization indicates agonist activity.

[0356] [6] The molecule according to any one of [2] to [3], wherein the binding of the first antigen-binding domain and the second antigen-binding domain or the binding of the first component and the second component activates phosphorylation of the RTK protein.

[0357] [7] According to the molecule described in [6], wherein phosphorylation of RTK protein is detected in the presence of the molecule in an in vitro cell assay, wherein phosphorylation of RTK protein indicates agonist activity.

[0358] [8] The molecule according to any one of [2] to [7], wherein the first antigen-binding domain and the second antigen-binding domain or the first component and the second component are each bound to the extracellular domain (ECD) of the RTK protein.

[0359] [9] According to the molecule described in [8], wherein for each of the first antigen-binding domain and the second antigen-binding domain or the first component and the second component, the extracellular domain is independently selected from the immunoglobulin (Ig)-like domain of the ECD of the RTK protein.

[0360]

[10] The molecule according to any one of [2] to [9], wherein the first antigen-binding domain or the first component binds to the first domain and the second antigen-binding domain or the second component binds to the second domain, wherein the first domain and the second domain are identical.

[0361]

[11] The molecule according to any one of [2] to [9], wherein a first antigen-binding domain or a first component binds to the first domain and a second antigen-binding domain or a second component binds to the second domain, wherein the second domain is different from the first domain.

[0362]

[12] The molecule according to

[10] , wherein the first epitope and the second epitope are identical.

[0363]

[13] According to the molecule described in

[10] , the first epitope and the second epitope are different.

[0364]

[14] According to the molecule described in

[11] , the first epitope and the second epitope are different.

[0365]

[15] The molecule according to

[13] or

[14] , wherein the molecule binds at two sites.

[0366]

[16] The molecule according to any one of [1] to

[15] , wherein the RTK protein is selected from one of the fibroblast growth factor receptor (FGFR) family, which consists of members of the group consisting of FGFR1, FGFR2, FGFR3 and FGFR4; or one of the discoid domain receptor (DDR) family, which consists of members of the group consisting of DDR1 and DDR2.

[0367]

[17] The molecule according to any one of [1] to

[16] , wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2), preferably the RTK protein is FGFR2b.

[0368]

[18] The molecule according to

[17] , wherein the molecule activates signal transduction in the target cell via binding to the FGFR2b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10% or more in the expression of an intracellular protein downstream of FGFR2b compared to the expression of the intracellular protein prior to the binding of the molecule to the FGFR2b protein expressed on the cell surface of the target cell.

[0369]

[19] According to the molecule described in

[18] , the intracellular proteins downstream of FGFR2b are selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1 / pERK2 and pMAPK3 / pERK1.

[0370]

[20] According to the molecule described in

[19] , the expression of the intracellular proteins was measured using an immunoassay and normalized to the total number of FGFR2b protein and the total number of AKT, PLCG1, FRS2, PAK4, RAS, MAPK1 / ERK2 and MAPK3 / ERK1, respectively.

[0371]

[21] According to the molecule described in

[17] , the molecule induces the proliferation of the target cell by binding to the FGFR2b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity.

[0372]

[22] According to the molecule described in

[21] , the induction of proliferation of the target cell is measured by increasing the ATP expression in the target cell by at least 1.5, 2, 2.5 or more times in the presence of the molecule compared with the ATP expression in the target cell in the absence of the molecule, wherein the ATP expression is proportional to the concentration of the live target cell.

[0373]

[23] According to the molecule described in

[21] , wherein the induction of proliferation of the target cell is measured by increasing BrdU expression in the target cell by at least 1.5-fold, or 2-fold, or 2.5-fold or greater in the presence of the molecule compared with BrdU expression in the target cell in the absence of the molecule, wherein the BrdU expression is proportional to the concentration of the live target cell.

[0374]

[24] The molecule according to any one of

[18] to

[23] , wherein the target cell is a lung epithelial cell.

[0375]

[25] According to the molecule described in

[24] , the lung epithelial cells are selected from alveolar type I (AT1) or alveolar type II (AT2) cells, especially alveolar type II (AT2) cells.

[0376]

[26] The molecule according to any one of

[17] to

[25] , wherein the first antigen-binding domain or the first component binds to a first epitope residing in the extracellular domain (ECD) of FGFR2, or wherein the first antigen-binding domain or the first component binds to a first epitope of the extracellular portion of FGFR2.

[0377]

[27] The molecule according to any one of

[17] to

[26] , wherein the first antigen-binding domain or the first component binds to the Ig-like domain of FGFR2, the Ig-like domain being selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

[0378]

[28] According to the molecule described in

[27] , wherein the first antigen-binding domain or the first component binds the IgIIIb domain of FGFR2 and the second antigen-binding domain or the second component binds the domains of FGFR2 other than the IgIIIb domain.

[0379]

[29] The molecule according to

[27] , wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR2 in a combination of the following selected from the group consisting of:

[0380] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgI domains of FGFR2;

[0381] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgII domains of FGFR2;

[0382] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIII domains of FGFR2;

[0383] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIIIb domains of FGFR2.

[0384] (v) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIIIc domains of FGFR2;

[0385] (vi) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgII domains of FGFR2;

[0386] (vii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgIII domains of FGFR2;

[0387] (viii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgIIIb domains of FGFR2;

[0388] (ix) wherein the first antigen-binding domain or the first component and the second antigen-binding domain or the second component independently bind to the IgII and IgIIIc domains of FGFR2;

[0389] (x) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIII domains of FGFR2;

[0390] (xi) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIIIb domains of FGFR2;

[0391] (xii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIIIc domains of FGFR2;

[0392] (xiii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR2; and

[0393] (xiv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIc domains of FGFR2.

[0394]

[30] The molecule according to any one of

[27] to

[29] , wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR2 in a combination selected from the group consisting of:

[0395] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR2;

[0396] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIII domains of FGFR2;

[0397] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgII domains of FGFR2; and

[0398] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgI domains of FGFR2.

[0399]

[31] The molecule according to any one of

[27] to

[30] , wherein (i) the first antigen-binding domain or the first component binds to the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the IgIIIb of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR2.

[0400]

[32] The molecule according to any one of

[27] to

[30] , wherein (i) the first antigen-binding domain or the first component binds the IgIII domain of FGFR2.

[0401]

[33] The molecule according to any one of

[27] to

[32] , wherein the first antigen-binding domain or the first component binds to a continuous or discontinuous epitope.

[0402]

[34] According to the molecule described in

[33] , wherein continuous or discontinuous epitopes reside in one ECD, or reside in two or three ECDs.

[0403]

[35] The molecule according to any one of

[17] to

[34] , wherein the second antigen-binding domain or the second component binds the second epitope, and wherein the second epitope resides in a domain of FGFR2 that is the same as or different from the first epitope, or wherein the second antigen-binding domain or the second component binds the second epitope of the ECD of FGFR2.

[0404]

[36] The molecule according to any one of

[17] to

[35] , wherein the second antigen-binding domain or the second component binds to the Ig-like domain of FGFR2, the Ig-like domain being selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

[0405]

[37] According to the molecule described in

[36] , wherein (i) the second antigen-binding domain or the second component binds to the IgII of FGFR2, or (ii) the second antigen-binding domain or the second component specifically binds to the IgII of FGFR2 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgI and IgIII of FGFR2.

[0406]

[38] According to the molecule described in

[36] , wherein (i) the second antigen-binding domain or the second component binds to the IgI of FGFR2, or (ii) the second antigen-binding domain or the second component specifically binds to the IgI of FGFR2 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgII and IgIII of FGFR2.

[0407]

[39] The molecule according to

[36] wherein (i) the second antigen-binding domain or the second component binds to the IgIIIb domain of FGFR2, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIIIb of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR2.

[0408]

[40] According to the molecule described in

[36] , wherein (i) the second antigen-binding domain or the first component binds to the IgIIIc domain of FGFR2, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIIIc of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIb of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR2.

[0409]

[41] According to the molecule described in

[36] , wherein (i) the second antigen-binding domain or the second component binds to the IgIII of FGFR2, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIII of FGFR2 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgI and IgII of FGFR2.

[0410]

[42] The molecule according to any one of

[17] to

[41] , wherein the second antigen-binding domain or the second component binds to a continuous or discontinuous epitope.

[0411]

[43] According to the molecule described in

[42] , wherein continuous or discontinuous epitopes reside in one ECD, or reside in two or three ECDs.

[0412]

[44] The molecule according to any one of

[17] to

[43] , wherein the first epitope of the first antigen-binding domain or the first component is different from the second epitope of the second antigen-binding domain or the second component, or wherein the first epitope of the first antigen-binding domain or the first component and the second epitope of the second antigen-binding domain or the second component are non-overlapping.

[0413]

[45] The molecule according to any one of

[27] to

[44] , wherein the first antigen-binding domain or the first component binds to IgIII of FGFR2, and the second antigen-binding domain or the second component binds to IgIII, or IgII, or IgI of FGFR2.

[0414]

[46] The molecule according to any one of

[27] to

[45] , wherein the first antigen-binding domain or the first component binds to IgIIIb of FGFR2, and the second antigen-binding domain or the second component binds to IgIII, or IgIIIb, or IgII, or IgI of FGFR2.

[0415]

[47] The molecule according to

[45] or

[46] , wherein the molecule binds at two sites to the IgIIIb and IgIII of FGFR2.

[0416]

[48] ​​The molecule according to

[46] , wherein the molecule binds to both IgIIIb of FGFR2 and IgIIIb of FGFR2 at two sites.

[0417]

[49] The molecule according to

[46] , wherein the molecule binds at two sites to the IgIIIb and IgII of FGFR2.

[0418]

[50] The molecule according to

[46] , wherein the molecule binds at two sites to the IgIIIb and IgI of FGFR2.

[0419]

[51] The molecule according to any one of [1] to

[50] , wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2), and the molecule comprises a first antigen-binding domain that binds a first epitope and a second antigen-binding domain that binds a second epitope, wherein the first epitope resides in a first domain of a first FGFR2 protein, and the second epitope resides in a second domain of the first FGFR2 protein or a second FGFR2 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0420] [51a] A molecule according to any one of [1] to

[50] , wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first FGFR2 protein, and the second epitope resides in a second domain of the first FGFR2 protein or a second FGFR2 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0421]

[52] The molecule according to any one of [1] to

[50] , wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2), and the molecule comprises a first antigen-binding domain for binding a first epitope and a second antigen-binding domain for binding a second epitope, wherein the first epitope resides in a first domain of a first FGFR2 protein, and the second epitope resides in a second domain of the first FGFR2 protein or a second FGFR2 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0422] [52a] A molecule according to any one of [1] to

[50] , wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first FGFR2 protein, and the second epitope resides in a second domain of the first FGFR2 protein or a second FGFR2 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0423]

[53] The molecule according to any one of

[51] , [51a],

[52] and [52a], wherein the first antigen-binding domain or the first component binds to the first FGFR2 protein and the second antigen-binding domain or the second component binds to the second FGFR2 protein, optionally wherein the first RTK protein and the second RTK protein form a dimer.

[0424]

[54] The molecule according to any one of

[45] to

[53] , wherein the second antigen-binding domain is selected from the group consisting of: FFB0464, FFB0341, FFB0396, FFB0026, FFB0094, FFB0035, FFB0045, FFB0057 and FFB0080.

[0425]

[55] The molecule according to any one of

[45] to

[53] , wherein the first antigen-binding domain is selected from the group consisting of: FFB0057, FFB0065, FFB0080, FFB0088, FFB0185, FFB0219, FFB0261, FFB0294, FFB0299, FFB0335, FFB0365, FFB0368, FFB0374 and FFB0453.

[0426]

[56] According to the molecule described in

[54] or

[55] , wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (a1) to (a14) for binding to FGFR2, or wherein the first antigen-binding domain comprises any one of (a1) to (a14), wherein (a1) to (a14) correspond to (a1) to (a14) referenced in

[0164] .

[0427]

[57] The molecule according to any one of

[54] to

[56] , wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (b1) to (b14) for binding to FGFR2, or wherein the first antigen-binding domain comprises any one of (b1) to (b14), wherein (b1) to (b14) correspond to (b1) to (b14) referenced in

[0165] .

[0428]

[58] The molecule according to any one of

[54] to

[57] , wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (c1) to (c9) for binding to FGFR2, or wherein the second antigen-binding domain comprises any one of (c1) to (c9), wherein (c1) to (c9) correspond to (c1) to (c9) referenced in

[0166] .

[0429]

[59] The molecule according to any one of

[54] to

[58] , wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (d1) to (d9) for binding to FGFR2, or wherein the second antigen-binding domain comprises any one of (d1) to (d9), wherein (d1) to (d9) correspond to (d1) to (d9) referenced in

[0167] .

[0430]

[60] The molecule according to any one of

[54] to

[59] , wherein the first antigen-binding domain and the second antigen-binding domain bind to the same epitope, and wherein the first antigen-binding domain and the second antigen-binding domain compete with an antigen-binding domain comprising any one of (e1) to (e21) for binding to FGFR2, or wherein the first antigen-binding domain and the second antigen-binding domain comprise any one of (e1) to (e21), wherein (e1) to (e21) correspond to (e1) to (e21) referenced in

[0168] .

[0431]

[61] The molecule according to any one of

[54] to

[60] , wherein the first antigen-binding domain and the second antigen-binding domain bind different epitopes, and each of the first antigen-binding domain and the second antigen-binding domain competes with an antigen-binding domain comprising any one of (f1) to (f116) for binding to FGFR2, or wherein each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (f1) to (f116), wherein (f1) to (f116) correspond to (f1) to (f116) referenced in

[0169] .

[0432] [17-1] The molecule according to any one of [1], [2], [2a] and [3] to

[16] , wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1), preferably the RTK protein is FGFR1b.

[0433] [18-1] The molecule according to [17-1], wherein the molecule activates signal transduction in the target cell via binding to the FGFR1b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10% or more in the expression of an intracellular protein downstream of FGFR1b compared to the expression of the intracellular protein prior to the binding of the molecule to the FGFR1b protein expressed on the cell surface of the target cell.

[0434] [19-1] According to the molecule described in [18-1], the intracellular proteins downstream of FGFR1b are selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1 / pERK2 and pMAPK3 / pERK1.

[0435] [20-1] According to the molecule described in [19-1], wherein an immunoassay is used to measure the expression of the intracellular proteins and normalize them to the total number of FGFR1b protein and the total number of AKT, PLCG1, FRS2, PAK4, RAS, MAPK1 / ERK2 and MAPK3 / ERK1, respectively.

[0436] [21-1] The molecule according to [17-1], wherein the molecule induces the proliferation of the target cell by binding to the FGFR1b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity.

[0437] [22-1] According to the molecule described in [21-1], wherein the induction of proliferation of the target cell is measured by an increase of at least 1.5-fold, or 2-fold, or 2.5-fold or greater in the presence of the molecule compared with ATP expression in the target cell in the absence of the molecule, wherein the ATP expression is proportional to the concentration of the live target cell.

[0438] [23-1] According to the molecule described in [21-1], wherein the induction of proliferation of the target cell is measured by an increase of at least 1.5-fold, or 2-fold, or 2.5-fold or greater in the expression of BrdU in the target cell in the absence of the molecule, wherein the expression of BrdU is proportional to the concentration of the live target cell.

[0439] [24-1] The molecule according to any one of [18-1] to [23-1], wherein the target cell is a lung epithelial cell.

[0440] [25-1] According to the molecule described in [24-1], the lung epithelial cells are selected from alveolar type I (AT1) or alveolar type II (AT2) cells, especially alveolar type II (AT2) cells.

[0441] [26-1] The molecule according to any one of [17-1] to [25-1], wherein the first antigen-binding domain or the first component binds to a first epitope residing in the extracellular domain (ECD) of FGFR1, or wherein the first antigen-binding domain or the first component binds to a first epitope of the extracellular portion of FGFR1.

[0442] [27-1] The molecule according to any one of [17-1] to [26-1], wherein the first antigen-binding domain or the first component binds to the Ig-like domain of FGFR1, the Ig-like domain being selected from the group consisting of: IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

[0443] [28-1] The molecule according to [27-1], wherein the first antigen-binding domain or the first component binds the IgIIIb domain of FGFR1 and the second antigen-binding domain or the second component binds the domains of FGFR1 other than the IgIIIb domain.

[0444] [29-1] The molecule according to [27-1], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR1 in a combination of the following, selected from the group consisting of:

[0445] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgI domains of FGFR1;

[0446] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgII domains of FGFR1;

[0447] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIII domains of FGFR1;

[0448] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIIIb domains of FGFR1.

[0449] (v) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgI and IgIIIc domains of FGFR1;

[0450] (vi) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgII domains of FGFR1;

[0451] (vii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgIII domains of FGFR1;

[0452] (viii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgIIIb domains of FGFR1;

[0453] (ix) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgII and IgIIIc domains of FGFR1.

[0454] (x) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIII domains of FGFR1;

[0455] (xi) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIIIb domains of FGFR1.

[0456] (xii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIII and IgIIIc domains of FGFR1.

[0457] (xiii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR1; and

[0458] (xiv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIc domains of FGFR1.

[0459] [30-1] The molecule according to any one of [27-1] to [29-1], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, bind independently to the Ig-like domain of FGFR1 in a combination selected from the group consisting of:

[0460] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIIIb domains of FGFR1.

[0461] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgIII domains of FGFR1;

[0462] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgII domains of FGFR1; and

[0463] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the IgIIIb and IgI domains of FGFR1.

[0464] [31-1] The molecule according to any one of [27-1] to [30-1], wherein (i) the first antigen-binding domain or the first component binds to the IgIIIb domain of FGFR1, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the IgIIIb of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII of FGFR1.

[0465] [32-1] The molecule according to any one of [27-1] to [30-1], wherein (i) the first antigen-binding domain or the first component binds to the IgIII domain of FGFR1.

[0466] [33-1] The molecule according to any one of [27-1] to [32-1], wherein the first antigen-binding domain or the first component binds to a continuous or discontinuous epitope.

[0467] [34-1] According to the molecule described in [33-1], wherein continuous or discontinuous epitopes reside in one ECD, or reside in two or three ECDs.

[0468] [35-1] The molecule according to any one of [17-1] to [34-1], wherein the second antigen-binding domain or the second component binds the second epitope, and wherein the second epitope resides in a domain of FGFR1 that is the same as or different from the first epitope, or wherein the second antigen-binding domain or the second component binds the second epitope of the ECD of FGFR1.

[0469] [36-1] The molecule according to any one of [17-1] to [35-1], wherein the second antigen-binding domain or the second component binds to the Ig-like domain of FGFR1, the Ig-like domain being selected from the group consisting of: IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

[0470] [37-1] The molecule according to [36-1] wherein (i) the second antigen-binding domain or the second component binds to the IgII of FGFR1, or (ii) the second antigen-binding domain or the second component specifically binds to the IgII of FGFR1 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgI and IgIII of FGFR1.

[0471] [38-1] The molecule according to [36-1], wherein (i) the second antigen-binding domain or the second component binds to the IgI of FGFR1, or (ii) the second antigen-binding domain or the second component specifically binds to the IgI of FGFR1 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgII and IgIII of FGFR1.

[0472] [39-1] The molecule according to [36-1] wherein (i) the second antigen-binding domain or the second component binds to the IgIIIb domain of FGFR1, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIIIb domain of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc domain of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII domains of FGFR1.

[0473] [40-1] The molecule according to [36-1], wherein (i) the second antigen-binding domain or the first component binds to the IgIIIc domain of FGFR1, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIIIc domain of FGFR1, and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIb domain of FGFR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII domains of FGFR1.

[0474] [41-1] The molecule according to [36-1] wherein (i) the second antigen-binding domain or the second component binds to the IgIII of FGFR1, or (ii) the second antigen-binding domain or the second component specifically binds to the IgIII of FGFR1 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the IgI and IgII of FGFR1.

[0475] [42-1] The molecule according to any one of [17-1] to [41-1], wherein the second antigen-binding domain or the second component binds to a continuous or discontinuous epitope.

[0476] [43-1] The molecule according to [42-1], wherein continuous or discontinuous epitopes reside in one ECD, or in two or three ECDs.

[0477] [44-1] The molecule according to any one of [17-1] to [43-1], wherein the first epitope of the first antigen-binding domain or the first component is different from the second epitope of the second antigen-binding domain or the second component, or wherein the first epitope of the first antigen-binding domain or the first component and the second epitope of the second antigen-binding domain or the second component are non-overlapping.

[0478] [45-1] The molecule according to any one of [27-1] to [44-1], wherein the first antigen-binding domain or the first component binds to IgIII of FGFR1, and the second antigen-binding domain or the second component binds to IgIII, or IgII, or IgI of FGFR1.

[0479] [46-1] The molecule according to any one of [27-1] to [45-1], wherein the first antigen-binding domain or the first component binds to IgIIIb of FGFR1, and the second antigen-binding domain or the second component binds to IgIII, or IgIIIb, or IgII, or IgI of FGFR1.

[0480] [47-1] The molecule according to [45-1] or [46-1], wherein the molecule binds at two sites to the IgIIIb and IgIII of FGFR1.

[0481] [48-1] The molecule according to [46-1], wherein the molecule binds to both IgIIIb of FGFR1 and IgIIIb of FGFR1 at two sites.

[0482] [49-1] The molecule according to [46-1], wherein the molecule binds at two sites to the IgIIIb and IgII of FGFR1.

[0483] [50-1] The molecule according to [46-1], wherein the molecule binds at two sites to IgIIIb and IgI of FGFR1.

[0484] [51-1] A molecule according to any one of [1] to

[16] and [17-1] to [50-1], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1), and the molecule comprises a first antigen-binding domain that binds a first epitope and a second antigen-binding domain that binds a second epitope, wherein the first epitope resides in a first domain of a first FGFR1 protein, and the second epitope resides in a second domain of the first FGFR1 protein or a second FGFR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0485] [51a-1] A molecule according to any one of [1], [2], [2a] and [3] to

[16] and [17-1] to [50-1], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first FGFR1 protein, and the second epitope resides in a second domain of the first FGFR1 protein or a second FGFR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0486] [52-1] A molecule according to any one of [1] to

[16] and [17-1] to [50-1], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1), and the molecule comprises a first antigen-binding domain that binds a first epitope and a second antigen-binding domain that binds a second epitope, wherein the first epitope resides in a first domain of a first FGFR1 protein, and the second epitope resides in a second domain of the first FGFR1 protein or the second FGFR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0487] [52a-1] A molecule according to any one of [1], [2], [2a] and [3] to

[16] and [17-1] to [50-1], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first FGFR1 protein, and the second epitope resides in a second domain of the first FGFR1 protein or a second FGFR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0488] [53-1] The molecule according to any one of [51-1], [51a-1], [52-1] and [52a-1], wherein the first antigen-binding domain or the first component binds to the first FGFR1 protein and the second antigen-binding domain or the second component binds to the second FGFR1 protein, optionally wherein the first RTK protein and the second RTK protein form a dimer.

[0489] [54-1] The molecule according to any one of [45-1] to [53-1], wherein the second antigen-binding domain is selected from the group consisting of: FFA0020, FFA0026, FFA0032, FFA0043, FFA0047, FFA0055, FFA0064 and FFA0088.

[0490] [55-1] The molecule according to any one of [45-1] to [53-1], wherein the first antigen-binding domain is selected from the group consisting of: FFA0029 and FFA0048.

[0491] [56-1] The molecule according to [54-1] or [55-1], wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (h1) to (h2) for binding to FGFR1, or wherein the first antigen-binding domain comprises any one of (h1) to (h2), wherein (h1) to (h2) correspond to (h1) to (h2) referenced in

[0219] .

[0492] [57-1] The molecule according to any one of [54-1] to [56-1], wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (i1) to (i2) for binding to FGFR1, or wherein the first antigen-binding domain comprises any one of (i1) to (i2), wherein (i2) to (i2) correspond to (i1) to (i2) referenced in

[0220] .

[0493] [58-1] The molecule according to any one of [54-1] to [57-1], wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (j1) to (j8) for binding to FGFR1, or wherein the second antigen-binding domain comprises any one of (j1) to (j8), wherein (j1) to (j8) correspond to (j1) to (j8) referenced in

[0221] .

[0494] [59-1] The molecule according to any one of [54-1] to [58-1], wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (k1) to (k8) for binding to FGFR1, or wherein the second antigen-binding domain comprises any one of (k1) to (k8), wherein (k1) to (k8) correspond to (k1) to (k8) referenced in

[0222] .

[0495] [60-1] The molecule according to any one of [54-1] to [59-1], wherein the first antigen-binding domain and the second antigen-binding domain bind to the same epitope, and wherein the first antigen-binding domain and the second antigen-binding domain compete with an antigen-binding domain comprising any one of (l1) to (l10) for binding to FGFR1, or wherein the first antigen-binding domain and the second antigen-binding domain comprise any one of (l1) to (l10), wherein (l1) to (l10) correspond to (l1) to (l10) referenced in

[0223] .

[0496] [61-1] The molecule according to any one of [54-1] to [60-1], wherein the first antigen-binding domain and the second antigen-binding domain bind different epitopes, and each of the first antigen-binding domain and the second antigen-binding domain competes with an antigen-binding domain comprising any one of (m1) to (m16) for binding to FGFR1, or wherein each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (m1) to (m16), wherein (m1) to (m16) correspond to (m1) to (m16) referenced in

[0224] .

[0497] [17-2] The molecule according to any one of [1], [2], [2a] and [3] to

[16] , wherein the RTK protein is a discoid domain receptor 1 (DDR1).

[0498] [18-2] The molecule according to [17-2], wherein the molecule activates signal transduction in the target cell via binding to the DDR1 protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10% or more in the expression of an intracellular protein downstream of DDR1 compared to the expression of the intracellular protein prior to the binding of the molecule to the DDR1 protein expressed on the cell surface of the target cell.

[0499] [19-2] According to the molecule described in [18-2], the intracellular proteins downstream of DDR1 are selected from the group consisting of: SHP-2, FAK, NF-κB, Cdc42, MAPK1, AP1, Ras / Raf / ERK, PI3K / AKT, STAT1 / 3 / 5.

[0500] [20-2] According to the molecule described in [19-2], wherein the expression of the intracellular proteins is measured using an immunoassay and normalized to the total number of DDR1 protein and the total number of SHP-2, FAK, NF-κB, Cdc42, MAPK1, AP1, Ras / Raf / ERK, PI3K / AKT, STAT1 / 3 / 5, respectively.

[0501] [21-2] The molecule according to [17-2], wherein the molecule induces the proliferation of the target cell by binding to the DDR1 protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity.

[0502] [22-2] According to the molecule described in [21-2], wherein the induction of proliferation of the target cell is measured by an increase of at least 1.5-fold, or 2-fold, or 2.5-fold or greater in the presence of the molecule compared with ATP expression in the target cell in the absence of the molecule, wherein the ATP expression is proportional to the concentration of the live target cell.

[0503] [23-2] According to the molecule described in [21-2], wherein the induction of proliferation of the target cell is measured by an increase of at least 1.5-fold, or 2-fold, or 2.5-fold or greater in the expression of BrdU in the target cell in the absence of the molecule, wherein the expression of BrdU is proportional to the concentration of the live target cell.

[0504] [24-2] The molecule according to any one of [18-2] to [23-2], wherein the target cell is a lung epithelial cell.

[0505] [25-2] According to the molecule described in [24-2], the lung epithelial cells are selected from alveolar type I (AT1) or alveolar type II (AT2) cells, especially alveolar type II (AT2) cells.

[0506] [26-2] The molecule according to any one of [17-2] to [25-2], wherein the first antigen-binding domain or the first component binds to a first epitope residing in the extracellular domain (ECD) of DDR1, or wherein the first antigen-binding domain or the first component binds to a first epitope of the extracellular portion of DDR1.

[0507] [27-2] The molecule according to any one of [17-2] to [26-2], wherein the first antigen-binding domain or the first component binds to the disc-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of DDR1.

[0508] [28-2] The molecule according to [27-2], wherein the first antigen-binding domain or the first component binds the DS domain of DDR1 and the second antigen-binding domain or the second component binds a domain or region of DDR1 other than the DS domain.

[0509] [29-2] The molecule according to [27-2], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the disc-shaped (DS) domain, DS-like domain, or extracellular juxtamembrane (EJXM) region of DDR1 in a manner selected from the group consisting of the following combinations:

[0510] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS and DS domains of DDR1;

[0511] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS and DS-like domains of DDR1;

[0512] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS domain and the EJXM region of DDR1;

[0513] (iv) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS-like domain and the DS-like domain of DDR1.

[0514] (vii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS-like domain and the EJXM region of DDR1; and

[0515] (viii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the EJXM region and the EJXM region of DDR1.

[0516] [30-2] The molecule according to any one of [27-2] to [29-2], wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the discoidal (DS) domain, DS-like domain, or extracellular juxtamembrane (EJXM) region of DDR1 in a manner selected from the group consisting of:

[0517] (i) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS and DS domains of DDR1;

[0518] (ii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind the DS and DS-like domains of DDR1; and

[0519] (iii) wherein the first antigen-binding domain and the second antigen-binding domain, or the first component and the second component, independently bind to the DS and EJXM regions of DDR1.

[0520] [31-2] The molecule according to any one of [27-2] to [30-2], wherein (i) the first antigen-binding domain or the first component binds to the DS domain of DDR1, or wherein (ii) the first antigen-binding domain or the first component specifically binds to the DS domain of DDR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the DS-like domain of DDR1 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the EJXM region of DDR1.

[0521] [32-2] The molecule according to any one of [27-2] to [30-2], wherein (i) the first antigen-binding domain or the first component binds the DS domain of DDR1.

[0522] [33-2] The molecule according to any one of [27-2] to [32-2], wherein the first antigen-binding domain or the first component binds to a continuous or discontinuous epitope.

[0523] [34-2] According to the molecule described in [33-2], wherein continuous or discontinuous epitopes reside in one ECD, or reside in two or three ECDs.

[0524] [35-2] The molecule according to any one of [17-2] to [34-2], wherein the second antigen-binding domain or the second component binds the second epitope, and wherein the second epitope resides in a domain of DDR1 that is the same as or different from the first epitope, or wherein the second antigen-binding domain or the second component binds the second epitope of the ECD of DDR1.

[0525] [36-2] The molecule according to any one of [17-2] to [35-2], wherein the second antigen-binding domain or the second component binds to the disc-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of DDR1.

[0526] [37-2] The molecule according to [36-2] wherein (i) the second antigen-binding domain or the second component binds to the DS domain of DDR1, or (ii) the second antigen-binding domain or the second component specifically binds to the DS domain of DDR1 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the DS-like domain and EJXM region of DDR1.

[0527] [38-2] The molecule according to [36-2] wherein (i) the second antigen-binding domain or the second component binds to the DS-like domain and / or EJXM region of DDR1, or (ii) the second antigen-binding domain or the second component specifically binds to the DS-like domain and / or EJXM region of DDR1 and exhibits less than 10%, or less than 5%, or 0% cross-reactivity with the DS domain of DDR1.

[0528] [39-2] The molecule according to any one of [17-2] to [38-2], wherein the second antigen-binding domain or the second component binds to a continuous or discontinuous epitope.

[0529] [40-2] The molecule according to [39-2], wherein continuous or discontinuous epitopes reside in one ECD, or in two or three ECDs.

[0530] [41-2] The molecule according to any one of [17-2] to [40-2], wherein the first epitope of the first antigen-binding domain or the first component is different from the second epitope of the second antigen-binding domain or the second component, or wherein the first epitope of the first antigen-binding domain or the first component and the second epitope of the second antigen-binding domain or the second component are non-overlapping.

[0531] [42-2] The molecule according to any one of [27-2] to [41-2], wherein the first antigen-binding domain or the first component binds the DS domain of DDR1, and the second antigen-binding domain or the second component binds the DS domain or DS-like domain of DDR1 and / or the EJXM region.

[0532] [43-2] The molecule according to any one of [27-2] to [45-2], wherein the first antigen-binding domain or the first component binds the DS-like domain and / or EJXM region of DDR1, and the second antigen-binding domain or the second component binds the DS-like domain and / or EJXM region of DDR1.

[0533] [44-2] The molecule according to [42-2], wherein the molecule binds to the DS domain of DDR1 at two sites.

[0534] [45-2] The molecule according to [42-2], wherein the molecule binds at two sites to the DS domain of DDR1 and the DS-like domain and / or EJXM region of DDR1.

[0535] [46-2] The molecule according to [43-2], wherein the molecule binds to the DS-like structural domain and / or EJXM region of DDR1 at two sites.

[0536] [47-2] A molecule according to any one of [1] to

[16] and [17-2] to [46-2], wherein the RTK protein is a discoidal domain receptor 1 (DDR1), and the molecule comprises a first antigen-binding domain that binds a first epitope and a second antigen-binding domain that binds a second epitope, wherein the first epitope resides in a first domain of a first DDR1 protein, and the second epitope resides in a second domain of the first DDR1 protein or a second DDR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0537] [47a-2] A molecule according to any one of [1], [2], [2a] and [3] to

[16] and [17-2] to [46-2], wherein the RTK protein is a discoidal domain receptor 1 (DDR1), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first DDR1 protein, and the second epitope resides in a second domain of the first DDR1 protein or a second DDR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are the same.

[0538] [48-2] A molecule according to any one of [1] to

[16] and [17-2] to [46-2], wherein the RTK protein is a discoidal domain receptor 1 (DDR1), and the molecule comprises a first antigen-binding domain that binds a first epitope and a second antigen-binding domain that binds a second epitope, wherein the first epitope resides in a first domain of a first DDR1 protein, and the second epitope resides in a second domain of the first DDR1 protein or a second DDR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0539] [48a-2] A molecule according to any one of [1], [2], [2a] and [3] to

[16] and [17-2] to [46-2], wherein the RTK protein is a discoidal domain receptor 1 (DDR1), and the molecule comprises a first component for binding a first epitope and a second component for binding a second epitope, wherein the first epitope resides in a first domain of a first DDR1 protein, and the second epitope resides in a second domain of the first DDR1 protein or a second DDR1 protein, wherein the first domain and the second domain are the same or different, and wherein the first epitope and the second epitope are different.

[0540] [49-2] The molecule according to any one of [47-2], [47a-2], [48-2] and [48a-2], wherein the first antigen-binding domain or the first component binds to the first DDR1 protein and the second antigen-binding domain or the second component binds to the second DDR1 protein, optionally wherein the first RTK protein and the second RTK protein form a dimer.

[0541] [50-2] The molecule according to any one of [42-2] to [49-2], wherein the second antigen-binding domain is selected from the group consisting of: DAA0007, DAA0012 and DAA0024.

[0542] [51-2] The molecule according to any one of [42-2] to [49-2], wherein the first antigen-binding domain is selected from the group consisting of DAA0011 and DAA0045.

[0543] [52-2] The molecule according to [50-2] or [51-2], wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (o1) to (o2) for binding to DDR1, or wherein the first antigen-binding domain comprises any one of (o1) to (o2), wherein (o1) to (o2) corresponds to (o1) to (o2) referenced in

[0267] .

[0544] [53-2] The molecule according to any one of [50-2] to [52-2], wherein the first antigen-binding domain competes with an antigen-binding domain comprising any one of (p1) to (p2) for binding to DDR1, or wherein the first antigen-binding domain comprises any one of (p1) to (p2), wherein (p1) to (p2) correspond to (p1) to (p2) referenced in

[0268] .

[0545] [54-2] The molecule according to any one of [50-2] to [53-2], wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (q1) to (q3) for binding to DDR1, or wherein the second antigen-binding domain comprises any one of (q1) to (q3), wherein (q1) to (q3) correspond to (q1) to (q3) referenced in

[0269] .

[0546] [55-2] The molecule according to any one of [50-2] to [54-2], wherein the second antigen-binding domain competes with an antigen-binding domain comprising any one of (r1) to (r3) for binding to DDR1, or wherein the second antigen-binding domain comprises any one of (r1) to (r3), wherein (r1) to (r3) correspond to (r1) to (r3) referenced in

[0270] .

[0547] [56-2] The molecule according to any one of [50-2] to [55-2], wherein the first antigen-binding domain and the second antigen-binding domain bind to the same epitope, and wherein the first antigen-binding domain and the second antigen-binding domain compete with an antigen-binding domain comprising any one of (s1) to (s5) for binding to DDR1, or wherein the first antigen-binding domain and the second antigen-binding domain comprise any one of (s1) to (s5), wherein (s1) to (s5) correspond to (s1) to (s5) referenced in

[0271] .

[0548] [57-2] The molecule according to any one of [50-2] to [56-2], wherein the first antigen-binding domain and the second antigen-binding domain bind different epitopes, and each of the first antigen-binding domain and the second antigen-binding domain competes with an antigen-binding domain comprising any one of (t1) to (t6) for binding to DDR1, or wherein each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (t1) to (t6), wherein (t1) to (t6) correspond to (t1) to (t6) referenced in

[0272] .

[0549]

[62] The molecule according to any one of [1] to

[61] , [17-1] to [61-1] and [17-2] to [57-2], wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, wherein the first antigen-binding domain and the second antigen-binding domain are linked by at least one disulfide bond.

[0550]

[63] The molecule according to any one of [1] to

[61] , [17-1] to [61-1], [17-2] to [57-2] and

[62] , wherein the first antigen-binding domain and the second antigen-binding domain are characterized by having at least one disulfide bond formed between amino acid residues not in the hinge region.

[0551]

[64] According to the molecule described in

[63] , wherein the at least one disulfide bond is an engineered disulfide bond that is not present in wild-type IgG.

[0552]

[65] According to the molecule described in

[64] , wherein the at least one disulfide bond is formed between the CH1 region, CL region, VH or VHH region, or VL region of the first antigen-binding domain and the CH1 region, CL region, VH or VHH region, or VL region of the second antigen-binding domain.

[0553]

[66] The molecule according to

[65] , wherein the at least one disulfide bond is formed between the group consisting of:

[0554] (i) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CH1 region of the second antigen-binding domain;

[0555] (ii) Amino acid residues in the CL region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0556] (iii) Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain;

[0557] (iv) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VH or VHH region of the second antigen-binding domain.

[0558] (v) Amino acid residues in the VL region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain; and

[0559] (vi) Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain.

[0560]

[67] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CH1 region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: in the CH1 region according to EU The numbers 119, 122, 123, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 148, 150, 155, 156, 157, 159, 160, 161, 162, 163, 164, 165, 167, 174, 176, 177, 178, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 201, 203, 205, 206, 207, 208, 211, 212, 213, 214, 218, and 219.

[0561]

[68] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the CL region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 108, 109, 112, 121, 123, 126, 128, 151, 152, 153, 156, 184, 186, 188, 189, 190, 195, 196, 200, 201, 202, 203, 208, 210, 211, 212 and 213 in the CL region according to Kabat numbers.

[0562]

[69] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of the following positions: 188, 189, 190, 191, 192, 193, 194, 195, 196 and 197 according to EU numbers, and the amino acid residue in the CL region is selected from the group consisting of the following positions: 121, 122, 123, 124, 125, 126, 127 and 128 according to Kabat numbers.

[0563]

[70] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the VH region of the first antigen-binding domain and an amino acid residue in the VH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 6, 8, 16, 20, 25, 26, 28, 74 and 82b of the VH region according to Kabat numbers.

[0564]

[71] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the VHH region of the first antigen-binding domain and an amino acid residue in the VHH region of the second antigen-binding domain, and wherein the amino acid residue from which the bond between the first antigen-binding domain and the second antigen-binding domain originates is located at a position selected from the group consisting of: 4, 6, 7, 8, 9, 10, 11, 12, 14, 15, 17, 20, 24, 27, 29, 38, 39, 40, 41, 43, 44, 45, 46, 47, 48, 49, 67, 69, 71, 78, 80, 82, 82c, 85, 88, 91, 93, 94 and 107 according to Kabat numbers in the VHH region.

[0565]

[72] According to the molecule of

[66] , wherein the at least one disulfide bond is formed between an amino acid residue in the VL region of the first antigen-binding domain and an amino acid residue in the VL region of the second antigen-binding domain, and

[0566] (i) Wherein, when VL belongs to the κ subclass, the amino acid residues from which the bond between the first and second antigen-binding domains originates are located at positions selected from the group consisting of: VL regions (κ subclass) at Kabat numbers 21, 27, 58, 77, 100, 105, and 107; or

[0567] (ii) Wherein, when VL belongs to the λ subclass, the amino acid residues from which the bond between the first antigen-binding domain and the second antigen-binding domain originates are located at positions selected from the group consisting of the following positions: VL region (λ subclass) according to Kabat numbers 6, 19, 33 and 34.

[0568]

[73] According to the molecule described in

[72] , wherein the at least one disulfide bond is formed between an amino acid residue in the CH1 region of the first antigen-binding domain and an amino acid residue in the CL region of the second antigen-binding domain.

[0569]

[74] According to the molecule described in

[73] , the at least one disulfide bond is formed between an amino acid residue at position 191 (according to Kabat number) in the CH1 region of the first antigen-binding domain and an amino acid residue at position 126 (according to Kabat number) in the CL region of the second antigen-binding domain.

[0570]

[75] The molecule according to any one of [1] to

[61] , [17-1] to [61-1], [17-2] to [57-2] and

[62] to

[74] , wherein the molecule contains an antibody constant region.

[0571]

[76] The molecule according to

[75] , wherein the antibody constant region is a human antibody constant region, or wherein the antibody constant region is selected from the human IgG1, IgG2, IgG3 or IgG4 constant regions.

[0572]

[77] The molecule according to any one of [1] to

[61] , [17-1] to [61-1], [17-2] to [57-2] and

[62] to

[76] , wherein the molecule comprises or is an antibody or an antibody fragment thereof.

[0573]

[78] The molecule according to

[77] wherein the antibody fragment is Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody.

[0574]

[79] The molecule described in

[77] or

[78] , wherein the molecule is a monoclonal antibody.

[0575]

[80] The molecule according to any one of

[77] to

[79] , wherein the molecule is a human antibody, a humanized antibody or a chimeric antibody.

[0576]

[81] A molecule according to any one of [1] to

[61] , [17-1] to [61-1], [17-2] to [57-2] and

[62] to

[80] , wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a bivalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or each is a monovalent scFab.

[0577]

[82] The molecule according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[81] , wherein the molecule comprises more than two antigen-binding domains or more than two binding elements, or wherein the molecule is a trivalent, tetravalent or multivalent molecule.

[0578]

[83] The molecule according to

[82] , wherein the molecule comprises a third antigen-binding domain, wherein the third antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, or wherein the molecule comprises a third member for binding to the same epitope on the same domain as the first member or the second member.

[0579]

[84] The molecule according to

[83] , wherein the molecule comprises a fourth antigen-binding domain, wherein the fourth antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, but not to the same epitope on the third antigen-binding domain, or wherein the molecule comprises a fourth member for binding to the same epitope on the same domain as the first member or the second member, but not to the same epitope on the third member.

[0580]

[85] The molecule according to

[84] , wherein the molecule comprises a third antigen-binding domain and a fourth antigen-binding domain, wherein the third antigen-binding domain binds to the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds to the same epitope on the same domain as the second antigen-binding domain, or wherein the molecule comprises a third member for binding to the same epitope on the same domain as the first member and a fourth member for binding to the same epitope on the same domain as the second member.

[0581]

[86] A pharmaceutical composition comprising a molecule according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , and a pharmaceutically acceptable carrier.

[0582]

[87] An immunoconjugate protein comprising any one of the molecules according to [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] .

[0583]

[88] The molecules according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or the pharmaceutical composition according to

[86] , or the immunoconjugate according to

[87] , are used for the treatment or prevention of diseases, disorders or conditions associated with lung epithelial cell damage in individuals in need.

[0584]

[89] According to

[88] the molecular or pharmaceutical composition or immunoconjugate for use in the application of the invention, wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0585]

[90] The molecule according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or the pharmaceutical composition according to

[86] , or the immunoconjugate according to

[87] , for use in the treatment of any disease, ailment or condition that is readily improved or prevented by one of the following selected from the group consisting of:

[0586] (i) An increase or enhancement of FGFR2b signal conduction, or FGFR1b signal conduction, or DDR1 signal conduction;

[0587] (ii) Increased proliferation of lung epithelial cells; and

[0588] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0589]

[91] The molecules according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or the pharmaceutical composition according to

[86] , or the immunoconjugate according to

[87] , for use in the treatment or prevention of fibrosis or inflammation in an individual in need.

[0590]

[92] The molecular or pharmaceutical composition or immunoconjugate for use according to

[91] , wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0591]

[93] According to

[92] , the molecular or pharmaceutical composition or immunoconjugate used for the purpose thereof, wherein

[0592] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0593] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0594] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0595] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0596] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0597] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0598] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0599] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0600]

[94] Use in the manufacture of a medicament according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or the pharmaceutical composition according to

[86] or the immunoconjugate according to

[87] , for the treatment or prevention of a disease, ailment or condition associated with lung epithelial cell damage in an individual in need.

[0601]

[95] A method of treating an individual in need of a disease, ailment, or condition associated with lung epithelial cell damage, comprising administering to the individual a therapeutically effective amount of any one of the molecules according to [1], [2], [2a], and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or a pharmaceutical composition according to

[86] , or an immunoconjugate according to

[87] .

[0602]

[96] According to the use described in

[94] or the method described in

[95] , wherein the individual’s condition associated with lung epithelial cell damage is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, condition associated with exposure to environmental toxins, condition associated with exposure to bacteria, condition associated with exposure to viruses, cystic fibrosis, lung resection, condition associated with exposure to radiation, condition associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

[0603]

[97] According to the use described in

[94] or the method described in

[95] , wherein the individual's condition associated with lung epithelial cell damage is a condition that is easily improved or prevented by selecting one of the following groups:

[0604] (i) An increase or enhancement of FGFR2b signal conduction, or FGFR1b signal conduction, or DDR1 signal conduction;

[0605] (ii) Increased proliferation of lung epithelial cells; and

[0606] (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

[0607]

[98] Use in the manufacture of a medicament according to [1], or FGFR1b signaling or DDR1 signaling to any one of the molecules in

[85] , or according to the pharmaceutical composition in

[86] , or according to the immunoconjugate in

[87] , for the treatment or prevention of fibrosis or inflammation in an individual in need.

[0608]

[99] A method of treating fibrosis or inflammation in an individual in need, comprising administering to the individual a therapeutically effective amount of any one of the molecules according to [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , or a pharmaceutical composition according to

[86] , or an immunoconjugate according to

[87] .

[0609]

[100] The use according to

[98] or the method according to

[99] , wherein the fibrosis or inflammation is selected from the fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow or muscle tissue.

[0610]

[101] According to the use described in

[98] or the method described in

[99] , wherein

[0611] (i) Pulmonary fibrosis or inflammation is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma.

[0612] (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome;

[0613] (iii) Fibrosis or inflammation of the pancreas is pancreatitis;

[0614] (iv) Renal fibrosis or inflammation, selected from the group consisting of: chronic kidney disease or acute kidney disease;

[0615] (v) Fibrosis or inflammation of the oral mucosa is oral mucositis;

[0616] (vi) Fibrosis or inflammation of the skin, such as diabetic foot ulcers or bullous epidermolysis;

[0617] (vii) Fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and

[0618] (viii) Fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally including ulcerative colitis (UC) or Crohn's disease.

[0619]

[102] An isolated nucleic acid encoding a molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] ; or multiple isolated nucleic acids encoding a molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] . The molecule of any one of

[61] , [17-1], [51-1], [51a-1], [52-1], [52a-1], [53-1], [61-1], [17-2], [47-2], [47a-2], [48-2], [48a-2], [49-2], [57-2],

[62] ,

[85] .

[0620]

[103] A host cell comprising nucleic acid or multiple nucleic acids as described in

[102] .

[0621]

[104] A method for producing the molecule, or a process for producing the molecule, comprising the following steps:

[0622] (a) Culturing host cells as described in

[103] , and

[0623] (b) Recovering molecules generated in step (a).

[0624]

[105] A kit for treating or preventing diseases, disorders or conditions associated with lung epithelial cell damage in an individual in need, comprising at least the molecule according to any one of [1], [2], [2a] and [3]

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , and instructions for use.

[0625] In yet other aspects and embodiments, the present invention relates to any one of the following [B1] to [B41]:

[0626] [B1] A method for activating FGFR2b signaling, or inducing FGFR2b dimerization, or promoting FGFR2b phosphorylation, or enhancing intracellular FGF signaling in FGFR2b-expressing cells, the method comprising contacting the molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] with FGFR2b-expressing cells in vitro, ex vivo or in vivo.

[0627] [B2] A method for activating FGFR2b signaling in FGFR2b-expressing cells, or inducing FGFR2b dimerization, or promoting FGFR2b phosphorylation, or enhancing intracellular FGF signaling, the method comprising the step of contacting the molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] in vitro or in vitro with FGFR2b-expressing cells in lung organoid cultures or skin keratinocyte cultures.

[0628] [B3] A method for maintaining and proliferating progenitor cells, comprising the step of contacting an FGFR2b-expressing cell in vitro or ex vivo with a molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a-1] to [A34].

[0629] [B4] A method for maintaining and proliferating progenitor cells, comprising the step of contacting FGFR2b expressing cells in vitro or ex vivo with a molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] in a lung organoid culture or a skin keratinocyte culture.

[0630] [B5] Use of any one of the molecules according to [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] in FGFR2b-expressing cells to activate FGFR2b signaling in vitro, in vitro or in vivo, or to induce FGFR2b dimerization, or to promote FGFR2b phosphorylation, or to enhance intracellular FGF signaling.

[0631] [B6] Use of any one of the molecules according to [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] in lung organoid cultures or skin keratinocyte cultures to activate FGFR2b signaling in vitro or in vitro, or to induce FGFR2b dimerization, or to promote FGFR2b phosphorylation, or to enhance intracellular FGF signaling.

[0632] [B7] Use of the molecule according to any one of [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] to maintain and proliferate FGFR2b-expressing cells in vitro or in vitro.

[0633] [B8] Use of any one of the molecules described in [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[85] , [A1] and [A1a] to [A34] for maintaining and proliferating FGFR2b-expressing cells in vitro or in vitro in lung organoid cultures or skin keratinocyte cultures.

[0634] [B1-1] A method for activating FGFR1b signaling in FGFR1b-expressing cells, or inducing FGFR1b dimerization, or promoting FGFR1b phosphorylation, or enhancing intracellular FGF signaling, the method comprising contacting a molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] with FGFR1b-expressing cells in vitro, ex vivo or in vivo.

[0635] [B2-1] A method for activating FGFR1b signaling in FGFR1b-expressing cells, or inducing FGFR1b dimerization, or promoting FGFR1b phosphorylation, or enhancing intracellular FGF signaling, the method comprising contacting the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] with FGFR1b-expressing cells in vitro or ex vivo in lung organoid cultures or skin keratinocyte cultures.

[0636] [B3-1] A method for maintaining and proliferating progenitor cells, comprising the step of contacting the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] with FGFR1b expressing cells in vitro or in vitro.

[0637] [B4-1] A method for maintaining and proliferating progenitor cells, comprising the step of contacting FGFR1b expressing cells in vitro or ex vivo with a molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] in lung organoid cultures or skin keratinocyte cultures.

[0638] [B5-1] Use of any one of the molecules described in [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] in FGFR1b-expressing cells in vitro, in vitro or in vivo to activate FGFR1b signaling, or induce FGFR1b dimerization, or promote FGFR1b phosphorylation, or enhance intracellular FGF signaling.

[0639] [B6-1] Use of any one of the molecules described in [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] in lung organoid cultures or skin keratinocyte cultures in FGFR1b-expressing cells in vitro or in vitro to activate FGFR1b signaling, or induce FGFR1b dimerization, or promote FGFR1b phosphorylation, or enhance intracellular FGF signaling.

[0640] [B7-1] Use of the molecules according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] to maintain and proliferate FGFR1b-expressing cells in vitro or in vitro.

[0641] [B8-1] Use of the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1],

[62] to

[85] , [A1-1] and [A1a-1] to [A33-1] in vitro or in vitro for maintaining and proliferating FGFR1b-expressing cells in lung organoid cultures or skin keratinocyte cultures.

[0642] [B1-2] A method for activating DDR1 signaling, inducing DDR1 dimerization, promoting DDR1 phosphorylation, or enhancing intracellular collagen signaling in DDR1-expressing cells, the method comprising contacting the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] with DDR1-expressing cells in vitro, ex vivo, or in vivo.

[0643] [B2-2] A method for activating DDR1 signaling in DDR1-expressing cells, or inducing DDR1 dimerization, or promoting DDR1 phosphorylation, or enhancing intracellular collagen signaling, the method comprising contacting the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] in lung organoid cultures or skin keratinocyte cultures with DDR1-expressing cells in vitro or ex vivo.

[0644] [B3-2] A method for maintaining and proliferating progenitor cells, comprising the step of contacting a molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] with DDR1-expressing cells in vitro or ex vivo.

[0645] [B4-2] A method for maintaining and proliferating progenitor cells, comprising the step of contacting DDR1-expressing cells in vitro or ex vivo with any of the molecules according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] in lung organoid cultures or skin keratinocyte cultures.

[0646] [B5-2] Use of any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] in DDR1-expressing cells in vitro, in vitro or in vivo to activate DDR1 signaling, or induce DDR1 dimerization, or promote DDR1 phosphorylation, or enhance intracellular collagen signaling.

[0647] [B6-2] Use of the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] in lung organoid cultures or skin keratinocyte cultures in DDR1-expressing cells in vitro or in vitro to activate DDR1 signaling, or induce DDR1 dimerization, or promote DDR1 phosphorylation, or enhance intracellular collagen signaling.

[0648] [B7-2] Use of the molecules according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] to maintain and proliferate DDR1-expressing cells in vitro or in vitro.

[0649] [B8-2] Use of the molecule according to any one of [1], [2], [2a] and [3] to

[16] , [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] , [A1-2] and [A1a-2] to [A33-2] in vitro or in vitro for maintaining and proliferating DDR1-expressing cells in lung organoid cultures or skin keratinocyte cultures.

[0650] [B9] A method for generating an agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein the method comprises the following steps:

[0651] (a) Providing a first nucleic acid encoding a polypeptide comprising a first antigen-binding domain that binds to a first epitope in a first domain of an RTK protein and a second nucleic acid encoding a polypeptide comprising a second antigen-binding domain that binds to a second epitope in a second domain of an RTK protein;

[0652] (b) Introduce the nucleic acid encoding the two polypeptides containing the antigen-binding domain from step (a) into a host cell capable of expressing the nucleic acid;

[0653] (c) Cultivate the host cells obtained in step (b) so that two polypeptides containing antigen-binding domains are expressed;

[0654] (d) Obtain an antigen-binding molecule comprising the polypeptide containing the first antigen-binding domain obtained in step (c) and the polypeptide containing the second antigen-binding domain;

[0655] (e) subjecting the antigen-binding molecule obtained in step (d) to a assay capable of determining the activation of the RTK protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the RTK protein; and

[0656] (f) Selecting an agonist antigen-binding molecule that exhibits activating activity or in which receptor dimerization is detected in the assay, in step (e).

[0657] [B10] The method according to [B9], wherein the method includes step (ai) performed after step (a) and before step (b), wherein step (ai) includes selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same domain, or selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to different domains.

[0658] [B11] The method according to [B9] or [B10], wherein the method includes a step (aii) performed after step (ai) and before step (b), wherein step (aii) includes selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same epitope, or selecting a polypeptide containing a first antigen-binding domain and a second antigen-binding domain that bind to different epitopes.

[0659] [B12] The method according to any one of [B9] to [B11], wherein the molecule binds at two sites, and the method comprises:

[0660] (a) Providing a first nucleic acid encoding a polypeptide comprising a first antigen-binding domain that binds to a first epitope in a first domain of an RTK protein and a second nucleic acid encoding a polypeptide comprising a second antigen-binding domain that binds to a second epitope in a second domain of an RTK protein;

[0661] (ai) Select a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same domain, or select a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to different domains.

[0662] (aii) Select peptides containing a first antigen-binding domain and peptides containing a second antigen-binding domain that bind to different epitopes.

[0663] (b) Introduce the nucleic acid encoding two polypeptides containing antigen-binding domains obtained in step (aiii) into a host cell capable of expressing the nucleic acid;

[0664] (c) Cultivate the host cells in step (b) so that the two polypeptides containing the antigen-binding domain are expressed;

[0665] (d) Obtain an antigen-binding molecule comprising the polypeptide containing the first antigen-binding domain obtained in step (c) and the polypeptide containing the second antigen-binding domain;

[0666] (e) subjecting the antigen-binding molecule obtained in step (d) to a assay capable of determining the activation of the RTK protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the RTK protein; and

[0667] (f) Selecting an agonist antigen-binding molecule that exhibits activating activity or in which receptor dimerization is detected in the assay, in step (e).

[0668] [B13] The method according to any one of [B9] to [B12], wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2).

[0669] [B14] According to the method of [B13], wherein the first and second domains in step (a) are independently selected from the Ig-like domains of FGFR2, particularly from IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

[0670] [B15] According to the method described in [B14], wherein the first and second structural domains in step (a) are selected from the group consisting of:

[0671] (i) The first structural domain is IgIIIb and the second structural domain is IgIIIb.

[0672] (ii) The first structural domain is IgIIIb and the second structural domain is IgIII.

[0673] (iii) The first structural domain is IgIIIb and the second structural domain is IgII, and

[0674] (iv) The first structural domain is IgIIIb and the second structural domain is IgI.

[0675] [B13-1] The method according to any one of [B9] to [B12], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1).

[0676] [B14-1] According to the method of [B13-1], wherein the first and second domains in step (a) are independently selected from the Ig-like domains of FGFR1, particularly from IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

[0677] [B15-1] According to the method described in [B14-1], the first and second structural domains in step (a) are selected from the group consisting of:

[0678] (i) The first structural domain is IgIIIb and the second structural domain is IgIIIb.

[0679] (ii) The first structural domain is IgIIIb and the second structural domain is IgIII.

[0680] (iii) The first structural domain is IgIIIb and the second structural domain is IgII, and

[0681] (iv) The first structural domain is IgIIIb and the second structural domain is IgI.

[0682] [B13-2] The method according to any one of [B9] to [B12], wherein the RTK protein is a discoid domain receptor 1 (DDR1).

[0683] [B14-2] According to the method of [B13-2], wherein the first and second domains in step (a) are independently selected from the disk-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of DDR1.

[0684] [B15-2] According to the method described in [B14-2], wherein the first and second structural domains in step (a) are selected from the group consisting of:

[0685] (i) The first structural domain is a DS domain and the second structural domain is a DS domain.

[0686] (ii) The first structural domain is a DS domain and the second structural domain is a DS sample domain, and

[0687] (iii) The first structural domain is the DS structural domain and the second structural domain is the EJXM region.

[0688] [B16] The method according to any one of [B9] to [B15], [B13-1] to [B15-1] and [B13-2] to [B15-2], wherein the method includes step (aiii) performed after step (aii) and before step (b), wherein step (aiii) includes introducing a mutation into a nucleic acid encoding two polypeptides containing two antigen-binding domains, such that at least one bond connecting the two antigen-binding domains in the polypeptides is formed.

[0689] [B17] The method according to [B16], wherein the method further includes step (g) selecting the antigen-binding molecule obtained in step (f), wherein the two antigen-binding domains are located in spatially proximate positions and / or the mobility of the two antigen-binding domains is reduced.

[0690] [B18] A method for screening agonistic antigen-binding molecules that bind to receptor tyrosine kinase (RTK) proteins, wherein the method includes the following steps:

[0691] (a) Providing a first nucleic acid encoding a polypeptide comprising a first antigen-binding domain that binds to a first epitope in a first domain of an RTK protein and a second nucleic acid encoding a polypeptide comprising a second antigen-binding domain that binds to a second epitope in a second domain of an RTK protein;

[0692] (b) Introduce the nucleic acid encoding the two polypeptides containing the antigen-binding domain from step (a) into a host cell capable of expressing the nucleic acid;

[0693] (c) Cultivate the host cells in step (b) so that the two polypeptides containing the antigen-binding domain are expressed;

[0694] (d) Obtain an antigen-binding molecule comprising the polypeptide containing the first antigen-binding domain obtained in step (c) and the polypeptide containing the second antigen-binding domain;

[0695] (e) subjecting the antigen-binding molecule obtained in step (d) to a assay capable of determining the activation of the RTK protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the RTK protein; and

[0696] (f) Selecting an agonist antigen-binding molecule that exhibits activating activity or in which receptor dimerization is detected in the assay, in step (e).

[0697] [B19] The method according to [B18], wherein the method includes step (ai) performed after step (a) and before step (b), wherein step (ai) includes selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same domain, or selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to different domains.

[0698] [B20] The method according to [B18] or [B19], wherein the method includes a step (aii) performed after step (ai) and before step (b), wherein step (aii) includes selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same epitope, or selecting a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to different epitopes.

[0699] [B21] The method according to any one of [B18] to [B20], wherein the molecule binds at two sites, and the method comprises:

[0700] (a) Providing a first nucleic acid encoding a polypeptide comprising a first antigen-binding domain that binds to a first epitope in a first domain of an RTK protein and a second nucleic acid encoding a polypeptide comprising a second antigen-binding domain that binds to a second epitope in a second domain of an RTK protein;

[0701] (ai) Select a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to the same domain, or select a polypeptide containing a first antigen-binding domain and a polypeptide containing a second antigen-binding domain that bind to different domains.

[0702] (aii) Select peptides containing a first antigen-binding domain and peptides containing a second antigen-binding domain that bind to different epitopes.

[0703] (b) Introduce the nucleic acid encoding two polypeptides containing antigen-binding domains obtained in step (aii) into a host cell capable of expressing the nucleic acid;

[0704] (c) Cultivate the host cells in step (b) so that the two polypeptides containing the antigen-binding domain are expressed;

[0705] (d) Obtain an antigen-binding molecule comprising the polypeptide containing the first antigen-binding domain obtained in step (c) and the polypeptide containing the second antigen-binding domain;

[0706] (e) subjecting the antigen-binding molecule obtained in step (d) to a assay capable of determining the activation of the RTK protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the RTK protein; and

[0707] (f) Selecting an agonist antigen-binding molecule that exhibits activating activity or in which receptor dimerization is detected in the assay, in step (e).

[0708] [B22] The method according to any one of [B18] to [B21], wherein the RTK protein is fibroblast growth factor receptor 2 (FGFR2).

[0709] [B23] According to the method of [B22], wherein the first and second domains in step (a) are independently selected from the Ig-like domains of FGFR2, particularly from IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

[0710] [B24] According to the method described in [B23], wherein the first and second structural domains in step (a) are selected from the group consisting of:

[0711] (i) The first structural domain is IgIIIb and the second structural domain is IgIIIb;

[0712] (ii) The first structural domain is IgIIIb and the second structural domain is IgIII;

[0713] (iii) The first structural domain is IgIIIb and the second structural domain is IgII; and

[0714] (iv) The first structural domain is IgIIIb and the second structural domain is IgI.

[0715] [B22-1] The method according to any one of [B18] to [B21], wherein the RTK protein is fibroblast growth factor receptor 1 (FGFR1).

[0716] [B23-1] According to the method of [B22-1], wherein the first and second domains in step (a) are independently selected from the Ig-like domains of FGFR1, particularly from IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

[0717] [B24-1] According to the method described in [B23-1], the first and second structural domains in step (a) are selected from the group consisting of:

[0718] (i) The first structural domain is IgIIIb and the second structural domain is IgIIIb;

[0719] (ii) The first structural domain is IgIIIb and the second structural domain is IgIII;

[0720] (iii) The first structural domain is IgIIIb and the second structural domain is IgII; and

[0721] (iv) The first structural domain is IgIIIb and the second structural domain is IgI.

[0722] [B22-2] The method according to any one of [B18] to [B21], wherein the RTK protein is a discoid domain receptor 1 (DDR1).

[0723] [B23-2] According to the method of [B22-2], wherein the first and second domains in step (a) are independently selected from the disk-shaped (DS) domain, DS-like domain or extracellular juxtamembrane (EJXM) region of DDR1.

[0724] [B24-2] According to the method described in [B23-2], the first and second structural domains in step (a) are selected from the group consisting of:

[0725] (i) The first structural domain is a DS domain and the second structural domain is a DS domain.

[0726] (ii) The first structural domain is a DS domain and the second structural domain is a DS sample domain, and

[0727] (iii) The first domain is a DS domain and the second domain is an EJXM region. [B25] The method according to any one of [B18] to [B24], [B22-1] to [B24-1] and [B22-2] to [B24-2], wherein the method includes step (aiii) performed after step (aii) and before step (b), wherein step (aiii) includes introducing a mutation into a nucleic acid encoding two polypeptides containing two antigen-binding domains, such that at least one bond is formed connecting the two antigen-binding domains in the polypeptides.

[0728] [B26] The method according to [B25], wherein the method further includes step (g) selecting the antigen-binding molecule obtained in step (f), wherein the two antigen-binding domains are located in spatially proximate positions and / or the mobility of the two antigen-binding domains is reduced.

[0729] [B27] The method according to any one of [B9] to [B26], wherein the molecule comprises an antibody constant region, particularly (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from human IgG1, IgG2, IgG3 or IgG4 constant regions.

[0730] [B28] The method according to any one of [B9] to [B15], [B13-1] to [B15-1] and [B13-2] to [B15-2], [B16] to [B24], [B22-1] to [B24-1] and [B22-2] to [B24-2], [B25] to [B27], wherein the antigen-binding domain comprises an antibody variable region, or comprises an antibody fragment independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv or a single-domain antibody.

[0731] [B29] The method according to any one of [B9] to [B15], [B13-1] to [B15-1] and [B13-2] to [B15-2], [B16] to [B24], [B22-1] to [B24-1] and [B22-2] to [B24-2], [B25] to [B28], wherein the molecule comprises an antibody or an antibody fragment thereof.

[0732] [B30] The method according to [B29], wherein the antibody fragment is Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody.

[0733] [B31] The method according to [B29] or [B30], wherein the molecule is a monoclonal antibody.

[0734] [B32] The method according to any one of [B29] to [B31], wherein the molecule is a human antibody, a humanized antibody or a chimeric antibody.

[0735] [B33] The method according to any one of [B29] to [B32], wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a divalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each monovalent Fab or monovalent VHH, or each monovalent scFab.

[0736] [B34] A diagnostic reagent comprising [1], [2], [2a] and [3] to

[51] , [51a],

[52] , [52a],

[53] to

[61] , [17-1] to [51-1], [51a-1], [52-1], [52a-1], [53-1] to [61-1], [17-2] to [47-2], [47a-2], [48-2], [48a-2], [49-2] to [57-2],

[62] to

[85] and [A1], [A1a] to [A34], [A1-1], [A1a-1] to [A33-1], [A1-2] and [A1a-2] The molecule described in any one of [A33-2].

[0737] [B35] A library comprising or substantially consisting of the following:

[0738] A combination of multiple first-group antigen-binding domains having sequences different from each other and second-group antigen-binding domains having sequences different from each other;

[0739] Each antigen-binding domain of the first group of antigen-binding domains binds to immunoglobulin IIIb (IgIIIb) of the FGFR2 protein, and

[0740] The activation of FGFR2b signaling in the FGFR2b protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0741] [B36] The library described in [B35] comprises:

[0742] A combination of a first group of nucleic acids encoding a first group of antigen-binding domains having sequences different from each other, and a second group of nucleic acids encoding a second group of antigen-binding domains having sequences different from each other;

[0743] Each antigen-binding domain of the first group of antigen-binding domains binds to immunoglobulin IIIb (IgIIIb) of the FGFR2 protein, and

[0744] The activation of FGFR2b signaling in the FGFR2b protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0745] [B37] The library according to [B35] or [B36] is generated by a method comprising the following steps:

[0746] (a) Provides a first antigen-binding domain of a first epitope in an IgIIIb domain that binds to FGFR2 and a second antigen-binding domain of a second epitope in a second domain that binds to FGFR2;

[0747] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0748] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR2 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR2 protein; and

[0749] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0750] [B37-1] The library according to [B35] or [B36] is generated by a method comprising the following steps:

[0751] (a) Provides a first antigen-binding domain for a first epitope in IgIIIb that binds to FGFR2 and a second antigen-binding domain for a second epitope in IgIIIb that binds to FGFR2;

[0752] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0753] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR2 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR2 protein; and

[0754] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0755] [B37-2] The library according to [B35] or [B36] is generated by a method comprising the following steps:

[0756] (a) Provides a first antigen-binding domain for a first epitope in an IgIIIb that binds to FGFR2 and a second antigen-binding domain for a second epitope in an IgIII that binds to FGFR2;

[0757] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0758] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR2 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR2 protein; and

[0759] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0760] [B37-3] The library according to [B35] or [B36] is generated by a method comprising the following steps:

[0761] (a) Provides a first antigen-binding domain for a first epitope in IgIIIb that binds to FGFR2 and a second antigen-binding domain for a second epitope in IgII that binds to FGFR2;

[0762] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0763] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR2 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR2 protein; and

[0764] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0765] [B37-4] The library according to [B35] or [B36] is generated by a method comprising the following steps:

[0766] (a) Provides a first antigen-binding domain for a first epitope in an IgIIIb that binds to FGFR2 and a second antigen-binding domain for a second epitope in an IgI that binds to FGFR2;

[0767] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0768] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR2 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR2 protein; and

[0769] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0770] [B38] The library according to any one of [B35] to [B37-4], wherein the method further comprises when the antigen-binding domain is coupled to the IgIIIb of the FGFR2 protein at a value equal to or less than 10 as measured by surface plasmon resonance. -8 The step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when M binds to KD.

[0771] [B39] The library according to any one of [B35] to [B38], wherein the first epitope of the first antigen-binding domain is the same as the second epitope of the second antigen-binding domain.

[0772] [B40] The library according to any one of [B35] to [B38], wherein the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain.

[0773] [B41] The library according to any one of [B35] to [B40], wherein the activation of FGFR2b signaling by the combination of the first antigen-binding domain and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% higher than the activation of FGFR2b signaling by the first antigen-binding domain alone or the second antigen-binding domain alone.

[0774] [B35-1] A library comprising or substantially consisting of the following:

[0775] A combination of multiple first-group antigen-binding domains having sequences different from each other and second-group antigen-binding domains having sequences different from each other;

[0776] Each antigen-binding domain of the first group of antigen-binding domains binds to immunoglobulin IIIb (IgIIIb) of the FGFR1 protein, and

[0777] The activation of FGFR1b signaling in the FGFR1b protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0778] [B36-1] The library according to [B35-1] contains:

[0779] A combination of a first group of nucleic acids encoding a first group of antigen-binding domains having sequences different from each other, and a second group of nucleic acids encoding a second group of antigen-binding domains having sequences different from each other;

[0780] Each antigen-binding domain of the first group of antigen-binding domains binds to immunoglobulin IIIb (IgIIIb) of the FGFR1 protein, and

[0781] The activation of FGFR1b signaling in the FGFR1b protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0782] [B37-1] The library described in [B35-1] or [B36-1] is generated by a method comprising the following steps:

[0783] (a) Provides a first antigen-binding domain for a first epitope in an IgIIIb domain that binds to FGFR1 and a second antigen-binding domain for a second epitope in a second domain that binds to FGFR1;

[0784] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0785] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR1 protein; and

[0786] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0787] [B37-1-1] The library described in [B35-1] or [B36-1] is generated by a method comprising the following steps:

[0788] (a) Provides a first antigen-binding domain for a first epitope in IgIIIb that binds to FGFR1 and a second antigen-binding domain for a second epitope in IgIIIb that binds to FGFR1.

[0789] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0790] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR1 protein; and

[0791] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0792] [B37-2-1] The library described in [B35-1] or [B36-1] is generated by a method comprising the following steps:

[0793] (a) Provides a first antigen-binding domain for a first epitope in an IgIIIb that binds to FGFR1 and a second antigen-binding domain for a second epitope in an IgIII that binds to FGFR1;

[0794] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0795] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR1 protein; and

[0796] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0797] [B37-3-1] The library described in [B35-1] or [B36-1] is generated by a method comprising the following steps:

[0798] (a) Provides a first antigen-binding domain for a first epitope in IgIIIb that binds to FGFR1 and a second antigen-binding domain for a second epitope in IgII that binds to FGFR1;

[0799] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0800] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR1 protein; and

[0801] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0802] [B37-4-1] The library described in [B35-1] or [B36-1] is generated by a method comprising the following steps:

[0803] (a) Provides a first antigen-binding domain for a first epitope in an IgIIIb that binds to FGFR1 and a second antigen-binding domain for a second epitope in an IgI that binds to FGFR1.

[0804] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0805] (c) subjecting one or more antigen-binding molecules in step (b) to an assay capable of determining activation of the FGFR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the FGFR1 protein; and

[0806] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0807] [B38-1] The library according to any one of [B35-1] to [B37-4-1], wherein the method further comprises when the antigen-binding domain is coupled to the IgIIIb of the FGFR1 protein at a value equal to or less than 10 as measured by surface plasmon resonance. -8 The step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when M binds to KD.

[0808] [B39-1] A library according to any one of [B35-1] to [B38-1], wherein the first epitope of the first antigen-binding domain is identical to the second epitope of the second antigen-binding domain.

[0809] [B40-1] The library according to any one of [B35-1] to [B38-1], wherein the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain.

[0810] [B41-1] The library according to any one of [B35-1] to [B40-1], wherein the activation of FGFR1b signaling by the combination of the first antigen-binding domain and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% higher than the activation of FGFR1b signaling by the first antigen-binding domain alone or the second antigen-binding domain alone.

[0811] [B35-2] A library comprising or substantially consisting of the following:

[0812] A combination of multiple first-group antigen-binding domains having sequences different from each other and second-group antigen-binding domains having sequences different from each other;

[0813] Each antigen-binding domain in the first group binds to the DS domain of the DDR1 protein, and

[0814] The activation of DDR1 signaling in the DDR1 protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0815] [B36-2] The library according to [B35-2] contains:

[0816] A combination of a first group of nucleic acids encoding a first group of antigen-binding domains having sequences different from each other, and a second group of nucleic acids encoding a second group of antigen-binding domains having sequences different from each other;

[0817] Each antigen-binding domain in the first group binds to the DS domain of the DDR1 protein, and

[0818] The activation of DDR1 signaling in the DDR1 protein expressed on the cell surface by the combination of the first antigen-binding domain and the second antigen-binding domain selected from (i) is greater than that of the first antigen-binding domain itself or the second antigen-binding domain itself.

[0819] [B37-2] The library described in [B35-2] or [B36-2] is generated by a method comprising the following steps:

[0820] (a) Provides a first antigen-binding domain for a first epitope in a DS domain that binds to DDR1 and a second antigen-binding domain for a second epitope in a second domain that binds to DDR1;

[0821] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0822] (c) subjecting one or more antigen-binding molecules in step (b) to a assay capable of determining activation of the DDR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the DDR1 protein; and

[0823] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0824] [B37-1-2] The library according to [B35-2] or [B36-2] is generated by a method including the following steps:

[0825] (a) Provides a first antigen-binding domain for a first epitope in a DS domain of DDR1 and a second antigen-binding domain for a second epitope in a DS domain of DDR1;

[0826] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0827] (c) subjecting one or more antigen-binding molecules in step (b) to a assay capable of determining activation of the DDR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the DDR1 protein; and

[0828] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0829] [B37-2-2] The library described in [B35-2] or [B36-2] is generated by a method comprising the following steps:

[0830] (a) Provides a first antigen-binding domain for a first epitope in a DS-like domain that binds to DDR1 and a second antigen-binding domain for a second epitope in a DS-like domain that binds to DDR1.

[0831] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0832] (c) subjecting one or more antigen-binding molecules in step (b) to a assay capable of determining activation of the DDR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the DDR1 protein; and

[0833] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0834] [B37-3-2] The library described in [B35-2] or [B36-2] is generated by a method comprising the following steps:

[0835] (a) Provides a first antigen-binding domain for a first epitope in a DS domain that binds to DDR1 and a second antigen-binding domain for a second epitope in an EJXM region that binds to DDR1;

[0836] (b) Obtain one or more antigen-binding molecules comprising a combination of the first antigen-binding domain and the second antigen-binding domain from step (a);

[0837] (c) subjecting one or more antigen-binding molecules in step (b) to a assay capable of determining activation of the DDR1 protein in cells expressing the RTK protein on the cell surface or detecting receptor dimerization of the DDR1 protein; and

[0838] (d) Select one or more antigen-binding molecules that exhibit activation activity or in which receptor dimerization is detected in the assay, from step (c).

[0839] [B38-2] The library according to any one of [B35-2] to [B37-3-2], wherein the method further comprises when the antigen-binding domain is coupled to the DS domain of the DDR1 protein at a ratio equal to or less than 10 as measured by surface plasmon resonance. -8 The step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when M binds to KD.

[0840] [B39-2] The library according to any one of [B35-2] to [B38-2], wherein the first epitope of the first antigen-binding domain is the same as the second epitope of the second antigen-binding domain.

[0841] [B40-2] The library according to any one of [B35-2] to [B38-2], wherein the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain.

[0842] [B41-2] The library according to any one of [B35-2] to [B40-2], wherein the activation of DDR1 signaling by the combination of the first antigen-binding domain and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% higher than the activation of DDR1 signaling by the first antigen-binding domain alone or the second antigen-binding domain alone. Attached Figure Description

[0843] [Figure 1]

[0844] Figure 1 illustrates the process of generating bispecific antibodies using FORCE technology, as described by Dengl S. et al. (“Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices.” Nature Communications 11.1 (2020): 4974).

[0845] [Figure 2]

[0846] Figure 2 shows the molecular format and nomenclature of two-site antibodies.

[0847] [Figure 3]

[0848] Figure 3 illustrates the process of generating two-site antibodies using LINC-Ig technology.

[0849] [Figure 4]

[0850] Figure 4 shows the molecular format and nomenclature of dual-site LINC-Ig and UN-LINC antibodies.

[0851] [Figure 5]

[0852] Figure 5 shows the FreeStyle of the single-site antibody versus the stable expression of the mouse FGFR2IIIb variant. TM Binding activity of the 293-F cell line. The anti-KLH single-site antibody IC17dK showed the level of the negative control (background).

[0853] [Figure 6]

[0854] Figure 6 shows the FreeStyle of a single-site antibody versus a mouse FGFR2IIIb variant stably expressing IgI-deficient antibodies (mouse FGFR2IIIb δ IgI). TM Binding activity of the 293-F cell line. The anti-KLH single-site antibody IC17dK showed the level of the negative control (background).

[0855] [Figure 7]

[0856] Figure 7 shows the FreeStyle of a single-site antibody versus transient expression of IgIII-deficient mouse FGFR2IIIb variant (mouse FGFR2IIIb δ IgIII). TMBinding activity of the 293-F cell line. The anti-KLH single-site antibody IC17dK showed the level of the negative control (background).

[0857] [Figure 8]

[0858] Figure 8 shows the FreeStyle of the single-site antibody versus the stable expression of the mouse FGFR2IIIc variant. TM Binding activity of the 293-F cell line. The anti-KLH single-site antibody IC17dK showed the level of the negative control (background).

[0859] [Figure 9]

[0860] Figure 9 shows the FreeStyle of the single-site antibody and the stable expression of the human FGFR2IIIb variant. TM Binding activity of the 293-F cell line. The anti-KLH single-site antibody IC17dK showed the level of the negative control (background).

[0861] [Figure 10-1]

[0862] Figure 10-1 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (A) FFB0185 / / FFB0396, FFB0185, FFB0396, (B) FFB0219 / / FFB0341, FFB0219, FFB0341. Data points represent mean ± SEM.

[0863] [Figure 10-2]

[0864] Figure 10-2 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (C) FFB0219 / / FFB0396, FFB0219, FFB0396, (D) FFB0261 / / FFB0464, FFB0261, FFB0464. Data points represent mean ± SEM.

[0865] [Figure 10-3]

[0866] Figure 10-3 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (E) FFB0294 / / FFB0464, FFB0294, FFB0464; (F) FFB0299 / / FFB0464, FFB0299, FFB0464. Data points represent mean ± SEM.

[0867] [Figure 10-4]

[0868] Figure 10-4 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (G) FFB0335 / / FFB0464, FFB0335, FFB0464, (H) FFB0365 / / FFB0464, FFB0365, FFB0464. Data points represent mean ± SEM.

[0869] [Figure 10-5]

[0870] Figure 10-5 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (I) FFB0368 / / FFB0341, FFB0341, FFB0368, (J) FFB0374 / / FFB0464, FFB0374, FFB0464. Data points represent mean ± SEM.

[0871] [Figure 10-6]

[0872] Figure 10-6 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open and inverted triangles: single-site antibodies, crosses: N12433). FGF7 and N12433 were used as controls and are shared in each figure. (K) FFB0453 / / FFB0464, FFB0453, FFB0464. Data points represent mean ± SEM.

[0873] [Figure 11]

[0874] Figure 11 shows the results of the assessment of the agonistic activity of anti-FGFR2 single-site and two-site antibodies in lung organoids. Luminescence of lung growth induced by each antibody was measured using CellTiter-Glo (trademarked) 3D (grey: anti-FGFR2 single-site antibody, black: anti-FGFR2 two-site antibody). Data points represent mean ± SEM.

[0875] [Figure 12-1]

[0876] Figure 12-1 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in keratinocytes. The absorbance of BrdU input through each antibody in keratinocytes was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open triangles and inverted triangles: single-site antibodies). FGF7 was used as a control and shared in each figure. (A) FFB0185 / / FFB0396, FFB0185, FFB0396, (B) FFB0219 / / FFB0341, FFB0219, FFB0341. Data points represent mean ± SEM.

[0877] [Figure 12-2]

[0878] Figure 12-2 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in keratinocytes. The absorbance of BrdU input through each antibody in keratinocytes was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open triangles and inverted triangles: single-site antibodies). FGF7 was used as a control and shared in each figure. (C) FFB0219 / / FFB0396, FFB0219, FFB0396, (D) FFB0261 / / FFB0464, FFB0261, FFB0464. Data points represent mean ± SEM.

[0879] [Figure 12-3]

[0880] Figure 12-3 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in keratinocytes. The absorbance of BrdU input through each antibody in keratinocytes was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open triangles and inverted triangles: single-site antibodies). FGF7 was used as a control and shared in each figure. (E) FFB0294 / / FFB0464, FFB0294, FFB0464; (F) FFB0299 / / FFB0464, FFB0299, FFB0464. Data points represent mean ± SEM.

[0881] [Figure 12-4]

[0882] Figure 12-4 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in keratinocytes. The absorbance of BrdU input through each antibody in keratinocytes was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines). Results for each single-site and two-site antibody are shown separately (circles: FGF7, squares: two-site antibodies, open triangles and inverted triangles: single-site antibodies). FGF7 was used as a control and shared in each figure. (G) FFB0368 / / FFB0341, FFB0368, FFB0341, (H) FFB0374 / / FFB0464, FFB0374, FFB0464. Data points represent mean ± SEM.

[0883] [Figure 13-1]

[0884] Figure 13-1 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in NIH3T3. The absorbance of BrdU input through each antibody in NIH3T3 was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines; samples without lines do not converge). (A) shows each single-site antibody, FGF2, FGF18, and FGF7 (circles: FGF2, squares: FGF18, triangles: FFB0185, rhombuses: FFB0219, hexagons: FFB0261, hollow circles: FFB0294, hollow squares: FFB0299, hollow triangles: FFB0335, hollow rhombuses: FFB0365, hollow hexagons: FFB0368, crosses: FGF7). (B) shows each unit spot antibody, FGF2, FGF18, and FGF7 (circle: FGF2, square: FGF18, triangle: FFB0341, rhombus: FFB0374, hexagon: FFB0396, hollow circle: FFB0453, hollow square: FFB0464, cross: FGF7). FGF2, FGF18, and FGF7 were used as controls and shared in each figure. Data points represent mean ± SEM.

[0885] [Figure 13-2]

[0886] Figure 13-2 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR2 single-site and two-site antibodies in NIH3T3. The absorbance of BrdU input through each antibody in NIH3T3 was measured by cell proliferation ELISA, and a four-parameter logistic regression model was used to fit the drug dose-response curves (solid lines, samples without lines do not converge). (C) shows each two-site antibody, FGF2, FGF18, and FGF7 (circles: FGF2, squares: FGF18, triangles: FFB0185 / / FFB0396, rhombuses: FFB0219 / / FFB0341, hexagons: FFB0219 / / FFB0396, hollow circles: FFB0261 / / FFB0464, hollow squares: FFB0294 / / FFB0464, crosses: FGF7). (D) Shows each dual-site antibody, FGF2, FGF18, and FGF7 (circles: FGF2, squares: FGF18, triangles: FFB0299 / / FFB0464, rhombuses: FFB0335 / / FFB0464, hexagons: FFB0365 / / FFB0464, hollow circles: FFB0368 / / FFB0341, hollow squares: FFB0374 / / FFB0464, hollow triangles: FFB0453 / / FFB0464, crosses: FGF7). FGF2, FGF18, and FGF7 were used as controls and shared in each figure. Data points represent mean ± SEM.

[0887] [Figure 14]

[0888] Figure 14 shows the results of the assessment of the agonistic activity of the anti-FGFR2b dual-site LINC-Ig antibody in lung organoids. In this study, 1 μg / mL of each Un-LINC and HL-LINC-Ig or LL-LINC-Ig antibody was used. (A) Luminescence of lung growth with each Un-LINC and HL-LINC-Ig antibody was measured by CellTiter-Glo (registered trademark) 3D and the ratio to each mean in Un-LINC format is shown (grey: FFB0065 / / FFB0045, black: FFB0065 / / FFB0057). (B) Luminescence of lung growth induced by each Un-LINC and LL-LINC-Ig antibody was measured using CellTiter-Glo (registered trademark) 3D and is shown as a ratio to each mean in Un-LINC format (grey: FFB0065 / / FFB0045, white: FFB0065 / / FFB0035, black: FFB0065 / / FFB0057). Data points represent mean ± SEM.

[0889] [Figure 15]

[0890] Figure 15 shows images of lung organoid growth induced by anti-FGFR2b dual-site LINC-Ig antibody. 50 nM FGF7 and each anti-FGFR2b dual-site LINC-Ig antibody were used in this study. Images were obtained via CellVoyager. TM Images were captured using a CV8000, and the Z-stack images were composited using CellPathfinder software. FGF7, control (culture without exogenous FGF7), LL-LINC_FFB0065 / / FFB0035, and LL-LINC_FFB0065 / / FFB0045 are shown. Each scale bar represents 100 μm.

[0891] [Figure 16-1]

[0892] Figure 16-1 shows the results of in vivo induction of proliferation markers using anti-FGFR2b dual-site LINC-Ig antibody. Each proliferation marker gene was normalized by glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean solvent value. Expression of Mki67 in lung tissue (A) and Cdc20 in lung tissue (B) are shown. (Circles: solvent, squares: FGF7, triangles: LL-LINC_FFB0065 / / FFB0035, rhombuses: LL-LINC_FFB0065 / / FFB0045, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM.

[0893] [Figure 16-2]

[0894] Figure 16-2 shows the results of in vivo induction of proliferation markers using anti-FGFR2b dual-site LINC-Ig antibody. Each proliferation marker gene was normalized by glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean solvent value. The expression of Mki67 in ear skin tissue (C) and Cdc20 in ear skin tissue (D) are shown. (Circles: solvent, squares: FGF7, triangles: LL-LINC_FFB0065 / / FFB0035, rhombuses: LL-LINC_FFB0065 / / FFB0045, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM.

[0895] [Figure 17]

[0896] Figure 17 shows the antibody treatment regimen in a bleomycin-induced mouse pulmonary fibrosis model (BLM model).

[0897] [Figure 18]

[0898] Figure 18 shows the results of alveolar type II (AT2) cell marker induction and fibrosis marker reduction by anti-FGFR2b dual-site LINC-Ig antibody in a mouse model of pulmonary fibrosis (BLM). Each marker gene was normalized to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean Sham value. The expression of Sftpc in lung tissue (A), Fn1 in lung tissue (B), and Col1a1 in lung tissue (C) are shown. (Circles: Sham, Squares: Bleomycin-treated control (BLM), Triangles: Bleomycin-treated LL-LINC_FFB0065 / / FFB0035, Rhombuses: Bleomycin-treated LL-LINC_FFB0065 / / FFB0045, ns: not significant, **P<0.01, ****P<0.0001). Data points represent the mean ± SEM.

[0899] [Figure 19]

[0900] Figure 19 shows the results of in vivo induction of proliferation markers using an anti-FGFR2b dual-site antibody. Each proliferation marker gene was normalized to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean solvent value. Expression of Mki67 in lung tissue (A) and Cdc20 in lung tissue (B) are shown. (Circles: solvent, squares: FGF7, triangles: FFB0219 / / FFB0396, rhombuses: FFB0261 / / FFB0464, hexagons: FFB0335 / / FFB0464, hollow circles: FFB0365 / / FFB0464, hollow squares: FFB0368 / / FFB0341, hollow triangles: FFB374 / / FFB0464, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent the mean ± SEM.

[0901] [Figure 20]

[0902] Figure 20 shows the efficacy of anti-FGFR2b dual-site antibody in inducing alveolar type II (AT2) cell markers and reducing fibrosis markers in a mouse model of pulmonary fibrosis (BLM). Each marker gene was normalized to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean Sham+KLH value (with anti-KLH single-site antibody IC17dK used as a negative control). Expression of Sftpc in lung tissue (A), Fn1 in lung tissue (B), and Col1a1 in lung tissue (C) are shown. (Circles: Sham and KLH, Squares: Bleomycin application (BLM) and KLH, Triangles: Bleomycin application and FFB0219 / / FFB0396, Rhombuses: Bleomycin application and FFB0261 / / FFB0464, Hexagons: Bleomycin application and FFB0335 / / FFB0464, ns: not significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM.

[0903] [Figure 21]

[0904] Figure 21 shows the efficacy of anti-FGFR2b dual-site antibody in inducing alveolar type II (AT2) cell markers and reducing fibrosis markers in a mouse model of pulmonary fibrosis (BLM). Each marker gene was normalized to glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression, and each normalized value was calculated as a ratio to the mean Sham+KLH value (with anti-KLH single-site antibody IC17dK used as a negative control). Expression of Sftpc in lung tissue (A), Fn1 in lung tissue (B), and Col1a1 in lung tissue (C) are shown. (Circles: Sham and KLH, Squares: Bleomycin application (BLM) and KLH, Triangles: Bleomycin application and FFB0365 / / FFB0464, Rhombuses: Bleomycin application and FFB0368 / / FFB0341, Hexagons: Bleomycin application and FFB0374 / / FFB0464, ns: not significant, *P<0.05, **P<0.01, ***P<0.001). Data points represent mean ± SEM.

[0905] [Figure 22]

[0906] Figure 22 shows the results of a histopathological assessment of the role of the anti-FGFR2b dual-site antibody in a mouse model of pulmonary fibrosis (BLM). Lung tissue sections were immunohistochemically stained with an antibody against crude surfactant protein C (ProSPC; ab211326, abcam, Cambridge, UK), captured as digital images using a NanoZoomer S360 (Hamamatsu Photonics KK, Shizuoka, Japan), and the number of ProSPC-positive cells was counted using the multiplex IHC module of HALO AI (Indica labs, New Mexico, USA, v3.4). The number of positive cells per area (mm²) is summarized in (A) (circles: Sham and KLH, squares: bleomycin application (BLM) and KLH, triangles: bleomycin application and FFB0219 / / FFB0396, inverted triangles: bleomycin application and FFB0261 / / FFB0464, rhombuses: bleomycin application and FFB0335 / / FFB0464, **P<0.01, ***P<0.001, ****P<0.0001, compared with the BLM + KLH group by Tukey multiple comparison test) and (B) (circles: Sham and KLH, squares: bleomycin application (BLM)). And KLH, triangle: bleomycin application and FFB0365 / / FFB0464, inverted triangle: bleomycin application and FFB0368 / / FFB0341, rhombus: bleomycin application and FFB0374 / / FFB0464, *P<0.05, **P<0.01, ****P<0.0001, compared with the BLM + KLH group by Tukey multiple comparison test). Data points represent mean ± SEM.

[0907] [Figure 23-1]

[0908] Figure 23-1 shows the results of the concentration-dependent agonistic activity assessment of anti-DDR1 single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-DDR1) stably expressing mouse DDR1. Luminescence of Ba / F3-DDR1 growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (A) (Hollow circles: DAA0007, Hollow triangles: DAA0045, Squares: DAA0045 / / DAA0007, Asterisk: Collagen control), (B) (Hollow circles: DAA0024, Hollow triangles: DAA0011, Squares: DAA0011 / / DAA0024, Asterisk: Collagen control). The collagen control was shared in (A) and (B). Data points represent mean ± SEM.

[0909] [Figure 23-2]

[0910] Figure 23-2 shows the results of the concentration-dependent agonistic activity assessment of anti-DDR1 single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-DDR1) stably expressing mouse DDR1. Luminescence of Ba / F3-DDR1 growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (C) (Hollow circles: DAA0012, hollow triangles: DAA0045, squares: DAA0045 / / DAA0012, asterisks: collagen controls). Data points represent mean ± SEM.

[0911] [Figure 24]

[0912] Figure 24 shows the results of the assessment of the agonistic activity of anti-DDR1 single-site and two-site antibodies in Ba / F3-DDR1. The absorbance of DDR1 phosphorylated by each antibody in Ba / F3-DDR1 was measured by ELISA. (A, B) show the results for each single-site and two-site antibody, respectively (grey: single-site antibody, black: two-site antibody). Data points represent mean ± SEM.

[0913] [Figure 25-1]

[0914] Figure 25-1 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR1b single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-FGFR1b) stably expressing the mouse FGFR1IIIb variant. Luminescence of Ba / F3-FGFR1b cell growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (A) (Hollow circles: FFA0026, Hollow triangles: FFA0029, Squares: FFA0029 / / DAA0026, Asterisk: FGF2 control), (B) (Hollow circles: FFA0064, Hollow triangles: FFA0029, Squares: FFA0029 / / FFA0064, Asterisk: FGF2 control). FGF2 was shared in (A) and (B). Data points represent the mean ± SEM.

[0915] [Figure 25-2]

[0916] Figure 25-2 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR1b single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-FGFR1b) stably expressing the mouse FGFR1IIIb variant. Luminescence of Ba / F3-FGFR1b cell growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (C) (Hollow circle: FFA0088, Hollow triangle: FFA0029, Square: FFA0029 / / FFA0088, Asterisk: FGF2 Control), (D) (Hollow circle: FFA0032, Hollow triangle: FFA0048, Square: FFA0048 / / FFA0032, Asterisk: FGF2 Control). FGF2 was shared in (C) and (D). Data points represent the mean ± SEM.

[0917] [Figure 25-3]

[0918] Figure 25-3 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR1b single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-FGFR1b) stably expressing the mouse FGFR1IIIb variant. Luminescence of Ba / F3-FGFR1b cell growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (E) (Hollow circle: FFA0043, Hollow triangle: FFA0048, Square: FFA0048 / / FFA0043, Asterisk: FGF2 Control), (F) (Hollow circle: FFA0047, Hollow triangle: FFA0048, Square: FFA0048 / / FFA0047, Asterisk: FGF2 Control). FGF2 was shared in (E) and (F). Data points represent the mean ± SEM.

[0919] [Figure 25-4]

[0920] Figure 25-4 shows the results of the concentration-dependent agonistic activity assessment of anti-FGFR1b single-site and two-site antibodies in the Ba / F3 cell line (Ba / F3-FGFR1b) stably expressing the mouse FGFR1IIIb variant. Luminescence of Ba / F3-FGFR1b cell growth with each single-site and two-site antibody was measured using a CellTiter-Glo (trademarked) 3D, and a four-parameter logistic model was used to fit the drug dose-response curves (solid lines). (G) (hollow circle: FAA0055, hollow triangle: FFA0048, square: FFA0048 / / FFA0055, asterisk: FGF2 control) and (H) (hollow circle: FFA0020, hollow triangle: FFA0048, square: FFA0048 / / FFA0020, asterisk: FGF2 control). FGF2 was shared in (G) and (H). Data points represent the mean ± SEM. Detailed Implementation

[0921] The following definitions and detailed descriptions are provided to aid in understanding the invention illustrated herein. In particular, these definitions and detailed descriptions serve to explain the foregoing aspects and embodiments.

[0922] definition

[0923] Recipient human framework

[0924] For the purposes of this document, a “recipient human frame” is a frame that comprises an amino acid sequence derived from a light chain variable domain (VL) frame or a heavy chain variable domain (VH) frame of a human immunoglobulin frame or a human common frame as defined below. A recipient human frame “derived from” a human immunoglobulin frame or a human common frame may contain the same amino acid sequence as that human immunoglobulin frame or human common frame, or it may contain amino acid sequence variations. In some embodiments, the number of amino acid variations is 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer. In some embodiments, the VL recipient human frame is sequence-identical to the VL human immunoglobulin frame sequence or the human common frame sequence.

[0925] Affinity

[0926] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise stated, as used herein, "binding affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0927] Mature affinity

[0928] "Affinity-mature" antibodies are those that have one or more alterations in one or more hypervariable regions (HVRs) that result in improved affinity of the antibody for the antigen compared to parental antibodies that do not have such alterations.

[0929] FGFR2

[0930] Unless otherwise specified, the term "FGFR2" as used herein refers to any naturally occurring FGFR2 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term covers "full-length" unprocessed FGFR2, as well as any form of FGFR2 produced through cellular processing. The term also covers naturally occurring FGFR2 variants, such as splice variants or allelic variants.

[0931] FGFR2b / FGFR2IIIb

[0932] As used herein and as interchangeable herein, the terms “FGFR2b” and “FGFR2IIIb” refer to the epithelial FGFR2b isoform and to have the amino acid sequence as listed in SEQ ID NO: 5 and / or have the amino acid sequence as listed in NCBI accession number NP_075259.4, and are encoded by the FGFR2 gene residing on chromosome 10. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for FGFR2 splicing and subsequent expression of the epithelial FGFR2b isoform.

[0933] An exemplary amino acid sequence of the FGFR2IIIb isotype is presented as SEQ ID NO: 5:

[0934] SEQ ID NO: 5 (FGFR2IIIb subtype)

[0935] 1 mvswgrficl vvvtmatlsl arpsfslved ttlepeeppt kyqisqpevy vaapgeslev

[0936] 61 rcllkdaavi swtkdgvhlg pnnrtvlige ylqikgatpr dsglyactas rtvdsetwyf

[0937] 121 mvnvtdaiss gddeddtdga edfvsensnn krapywtnte kmekrlhavp aantvkfrcp

[0938] 181 aggnpmptmr wlkngkefkq ehriggykvr nqhwslimes vvpsdkgnyt cvveneygsi

[0939] 241 nhtyhldvve rsphrpilqa glpanastvv ggdvefvckv ysdaqphiqw ikhvekngsk

[0940] 301 ygpdglpylk vlkhsginss naevlalfnv teadageyic kvsnyigqan qsawltvlpk

[0941] 361 qqapgrekei taspdyleia iycigvflia cmvvtvilcr mknttkkpdf ssqpavhklt

[0942] 421 kriplrrqvt vsaessssmn sntplvritt rlsstadtpm lagvseyelp edpkwefprd

[0943] 481 kltlgkplge gcfgqvvmae avgidkdkpk eavtvavkml kddatekdls dlvsememmk

[0944] 541 migkhkniin llgactqdgp lyviveyask gnlreylrar rppgmeysyd inrvpeeqmt

[0945] 601 fkdlvsctyq largmeylas qkcihrdlaa rnvlvtennv mkiadfglar dinnidyykk

[0946] 661 ttngrlpvkw mapealfdrv ythqsdvwsf gvlmweiftl ggspypgipv eelfkllkeg

[0947] 721 hrmdkpanct nelymmmrdc whavpsqrpt fkqlvedldr iltlttneey ldlsqpleqy

[0948] 781 spsypdtrss cssgddsvfs pdpmpyepcl pqyphingsv kt

[0949] FGFR2c / FGFR2IIIc

[0950] As used herein, the terms “FGFR2c” and “FGFR2IIIc” refer to the epithelial FGFR2c subtype and to the amino acid sequence as listed in SEQ ID NO: 6 and / or have the amino acid sequence as listed in NCBI accession number NP_000132.3.

[0951] An exemplary amino acid sequence of the FGFR2IIIc isotype is shown in SEQ ID NO: 6:

[0952] SEQ ID NO: 6 (FGFR2IIIc subtype)

[0953] 1 mvswgrficl vvvtmatlsl arpsfslved ttlepeeppt kyqisqpevy vaapgeslev

[0954] 61 rcllkdaavi swtkdgvhlg pnnrtvlige ylqikgatpr dsglyactas rtvdsetwyf

[0955] 121 mvnvtdaiss gddeddtdga edfvsensnn krapywtnte kmekrlhavp aantvkfrcp

[0956] 181 aggnpmptmr wlkngkefkq ehriggykvr nqhwslimes vvpsdkgnyt cvveneygsi

[0957] 241 nhtyhldvve rsphrpilqa glpanastvv ggdvefvckv ysdaqphiqw ikhvekngsk

[0958] 301 ygpdglpylk vlkaagvntt dkeievlyir nvtfedagey tclagnsigi sfhsawltvl

[0959] 361 papgrekeit aspdyleiai ycigvfliac mvvtvilcrm knttkkpdfs sqpavhkltk

[0960] 421 riplrrqvtv saessssmns ntplvrittr lsstadtpml agvseyelpe dpkwefprdk

[0961] 481 ltlgkplgeg cfgqvvmaea vgidkdkpke avtvavkmlk ddatekdlsd lvsememmkm

[0962] 541 igkhkniinl lgactqdgpl yviveyaskg nlreylrarr ppgmeysydi nrvpeeqmtf

[0963] 601 kdlvsctyql argmeylasq kcihrdlaar nvlvtennvm kiadfglard innidyykkt

[0964] 661 tngrlpvkwm apealfdrvy thqsdvwsfg vlmweiftlg gspypgipve elfkllkegh

[0965] 721 rmdkpanctn elymmmrdcw havpsqrptf kqlvedldri ltlttneeyl dlsqpleqys

[0966] 781 psypdtrssc ssgddsvfsp dpmpyepclp qyphingsvk t

[0967] FGFR1

[0968] Unless otherwise specified, the term "FGFR1" as used herein refers to any naturally occurring FGFR1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term covers "full-length" unprocessed FGFR1, as well as any form of FGFR1 produced through cellular processing. The term also covers naturally occurring FGFR1 variants, such as splice variants or allelic variants.

[0969] FGFR1b / FGFR1IIIb

[0970] As used herein and as interchangeable herein, the terms “FGFR1b” and “FGFR1IIIb” refer to the epithelial FGFR1b isoform and to the amino acid sequence listed in NCBI accession number NP_001167536.1, encoded by the FGFR1 gene residing on chromosome 8. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for FGFR1 splicing and subsequent expression of the epithelial FGFR1b isoform.

[0971] FGFR1c / FGFR1IIIc

[0972] As used herein, the terms “FGFR1c” and “FGFR1IIIc” refer to the epithelial FGFR1c subtype and to the amino acid sequence listed in NCBI accession number NP_001167534.1.

[0973] DDR1

[0974] Unless otherwise specified, the term "DDR1" as used herein refers to any naturally occurring DDR1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term covers "full-length" unprocessed DDR1, as well as any form of DDR1 produced through cellular processing. The term also covers naturally occurring DDR1 variants, such as splice variants or allele variants.

[0975] DDR1a

[0976] As used herein, the term "DDR1a" refers to the amino acid sequence listed in NCBI accession number NP_001945.3, encoded by the DDR1 gene residing on chromosome 6.

[0977] DDR1b

[0978] As used herein, the term "DDR1b" refers to the amino acid sequence listed in NCBI accession number NP_054699.2, encoded by the DDR1 gene residing on chromosome 6.

[0979] As used herein, the term “DDR1c” refers to the amino acid sequence listed in NCBI accession number NP_054700.2 and encoded by the DDR1 gene residing on chromosome 6.

[0980] As used herein, the term “DDR1d” refers to the amino acid sequence listed in NCBI accession number NP_001189450.1 and encoded by the DDR1 gene residing on chromosome 6.

[0981] As used herein, the term “DDR1e” refers to the amino acid sequence listed in NCBI accession number NP_001189451.1 and encoded by the DDR1 gene residing on chromosome 6.

[0982] Agonist / Agonistic Antigen-Binding Molecule

[0983] As used herein, “agonist antigen-binding molecule,” “agonistic antigen-binding molecule,” or “agonist antibody” refers to a molecule or antibody that significantly enhances the biological activity of the antigen it binds to. In the embodiments, “agonist” or “agonistic” in the context of a cell receptor is understood to “activate” the signaling of the cell receptor. As used herein, the terms “anti-FGFR2 agonist antigen-binding molecule” or “anti-FGFR2 agonist antibody” refer to an antigen-binding molecule or antibody capable of binding its antigen with sufficient affinity, such that the antigen-binding molecule or antibody can be used as a therapeutic agent for activating FGFR2. As used herein, the terms “anti-FGFR1 agonist antigen-binding molecule” or “anti-FGFR1 agonist antibody” refer to an antigen-binding molecule or antibody capable of binding its antigen with sufficient affinity, such that the antigen-binding molecule or antibody can be used as a therapeutic agent for activating FGFR1. As used herein, the terms "anti-DDR1 agonist antigen-binding molecule" or "anti-DDR1 agonist antibody" refer to an antigen-binding molecule or antibody capable of binding with sufficient affinity to its antigen, such that the antigen-binding molecule or antibody can be used as a therapeutic agent for activating DDR1. As used herein, the agonistic activity of the molecule or antibody is observed when the binding of the antigen-binding molecule or agonist antibody to its antigen induces dimerization of the intracellular tyrosine kinase domain of the antigen, or when the binding of the molecule or antibody to its antigen activates phosphorylation of the antigen.

[0984] In one embodiment, as measured, for example by radioimmunoassay (RIA), the binding degree of the anti-FGFR2 antigen-binding molecule or antibody to an unrelated non-FGFR2 protein is less than about 10% of the binding degree of the antigen-binding molecule or antibody to FGFR2. In some embodiments, the antibody binding to FGFR2 has < 1 μM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g., 10 μM). -8 M or smaller, such as 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 The dissociation constant (Kd) of M). In some embodiments, an anti-FGFR2 antigen-binding molecule or an anti-FGFR2 antibody binds to an epitope of FGFR2 that is conserved in FGFR2 from different species.

[0985] In one embodiment, as measured, for example by radioimmunoassay (RIA), the binding degree of the anti-FGFR1 antigen-binding molecule or antibody to an unrelated non-FGFR1 protein is less than about 10% of the binding degree of the antigen-binding molecule or antibody to FGFR1. In some embodiments, the antibody binding to FGFR1 has < 1 μM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g., 10 μM). -8 M or smaller, such as 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 The dissociation constant (Kd) of M). In some embodiments, an anti-FGFR1 antigen-binding molecule or an anti-FGFR1 antibody binds to an epitope of FGFR1 that is conserved in FGFR1 from different species.

[0986] In one embodiment, as measured, for example by radioimmunoassay (RIA), the binding degree of the anti-DDR1 antigen-binding molecule or antibody to an unrelated non-DDR1 protein is less than about 10% of the binding degree of the antigen-binding molecule or antibody to DDR1. In some embodiments, the antibody binding to DDR1 has < 1 μM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g., 10 μM). -8 M or smaller, such as 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 The dissociation constant (Kd) of M). In some embodiments, an anti-DDR1 antigen-binding molecule or an anti-DDR1 antibody binds to an epitope of DDR1 that is conserved in DDR1 from different species.

[0987] In one embodiment, Kd is measured by radiolabeled antigen binding assay (RIA). In one embodiment, RIA is performed using the Fab form of the target antibody and its antigen. For example, by using the minimum concentration in the presence of a series of unlabeled antigen titrations. 125I) The labeled antigen was equilibrated with Fab, and then the bound antigen was captured using a plate coated with anti-Fab antibody to measure the solution-binding affinity of Fab to the antigen (see, for example, Chen et al., J. Mol. Biol. 293:865-881(1999)). To establish the assay conditions, MICROTITER (trademark) multiwell plates (ThermoScientific) were coated overnight with 5 μg / ml capture anti-Fab antibody (CappelLabs) in 50 mM sodium carbonate (pH 9.6) and then blocked at room temperature (approximately 23°C) for two to five hours with 2% (w / v) bovine serum albumin in PBS. In non-adsorbent plates (Nunc #269620), 100 pM or 26 pM [ 125 I] The antigen was mixed with a serially diluted solution of the target Fab (e.g., following the evaluation of anti-VEGF antibody (Fab-12) in Presta et al., Cancer Res. 57:4593-4599 (1997)). The target Fab was then incubated overnight; however, incubation may be prolonged (e.g., about 65 hours) to ensure equilibration. The mixture was then transferred to a capture plate and incubated at room temperature (e.g., one hour). The solution was then removed and the plate was washed eight times with 0.1% polysorbate 20 (TWEEN-20 (registered trademark)) in PBS. When the plate had dried, 150 μl / well of scintillation agent (MICROSCINT-20™; Packard) was added, and the plate was counted for several tens of minutes on a TOPCOUNT™ γ counter (Packard). The concentration of each Fab that yielded a maximum binding of less than or equal to 20% was selected for use in the competitive binding assay.

[0988] According to another embodiment, Kd was measured using the BIACORE (registered trademark) surface plasmon resonance assay. For example, the assay using BIACORE-2000 or BIACORE-3000 (BIAcore, Inc., Piscataway, NJ) was performed at 25°C with an immobilized antigen CM5 chip at approximately 10 response units (RU). In one embodiment, the carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) was activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. The antigen was diluted to 5 μg / ml (approximately 0.2 μM) with 10 mM sodium acetate at pH 4.8, and then injected at a flow rate of 5 μl / min to obtain a conjugated protein of approximately 10 response units (RU). Following antigen injection, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurements, at 25 °C, two serial dilutions (0.78 nM to 500 nM) of Fab in PBS containing 0.05% polysorbate 20 (TWEEN-20™) surfactant (PBST) were injected at a flow rate of approximately 25 μl / min. Associativity (kon) and dissociation (koff) rates were calculated using a simple one-to-one Langmuir binding model (BIACORE evaluation software version 3.2) by simultaneously fitting association and dissociation sensor maps. The equilibrium dissociation constant (Kd) was calculated as the ratio koff / kon. See, for example, Chen et al., J. Mol. Biol. 293:865-881 (1999). If the association rate obtained by the above surface plasmon resonance determination exceeds 106 M⁻¹ s⁻¹, the association rate can be determined by using fluorescence quenching techniques. That is, in a spectrometer such as an Aviv Instruments spectrophotometer equipped with a flow stop or a ThermoSpectronic 8000 series SLM-AMINCO (registered trademark) spectrophotometer, the increase or decrease in fluorescence emission intensity (excitation wavelength = 295 nm; emission wavelength = 340 nm, bandpass = 16 nm) of 20 nM anti-antigen antibody (Fab form) in PBS pH 7.2 at 25°C in the presence of gradually increasing antigen concentration is measured.

[0989] Specific binding

[0990] As used herein, the term "specific binding" refers to the binding interaction that determines the presence of a target protein (FGFR2b, FGFR1b, or DDR1) in a population of proteins including its different isoforms. Those skilled in the art will recognize that isoform-binding molecules, i.e., anti-FGFR2b antigen-binding molecules, anti-FGFR1b antigen-binding molecules, or anti-DDR1 antigen-binding molecules, may exhibit a degree of cross-reactivity between isoforms, depending on the conditions used, such as the target protein concentration, the salt and buffer conditions used, etc. In some embodiments, the terms "specifically binds" or "specifically binding" refer to the nature of the isoform-binding molecule, i.e., the anti-FGFR2b antigen-binding molecule binds to one or more isoform polypeptides or fragments thereof, or nucleic acids encoding one or more isoform polypeptides or fragments thereof, with high affinity (e.g., at least about 10). 7 M -1 Usually about 10 8 M -1 More typically about 10 9 M -1 Up to 10 10 M -1 The anti-FGFR2b antigen-binding molecule binds preferentially to the FGFR2c subtype, with an affinity for the FGFR2b subtype that is at least two, 50, 100, 1000, or greater than that for the FGFR2c subtype. In some embodiments, the anti-FGFR2b antigen-binding molecule preferentially binds to the FGFR2b subtype but substantially does not bind to the FGFR2c subtype (e.g., exhibiting cross-reactivity of less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0%). In some embodiments, the anti-FGFR1b antigen-binding molecule preferentially binds to the FGFR1b subtype but substantially does not bind to the FGFR1c subtype (e.g., exhibiting cross-reactivity of less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0%). In some embodiments, the anti-DDR1a antigen-binding molecule preferentially binds to the DDR1a subtype but substantially does not bind to the DDR1b subtype (e.g., showing cross-reactivity of less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0%).

[0991] FGFR2 expressed on cell surface

[0992] As used herein, the term "cell surface expressed FGFR2" refers to one or more FGFR2 proteins or their extracellular domains, expressed in vitro or in vivo on the cell surface such that at least a portion of the FGFR2 protein is exposed to the extracellular space of the cell membrane and is accessible to the antigen-binding portion of an antibody or antigen-binding fragment. "Cell surface expressed FGFR2" may include or consist of FGFR2 protein expressed on the surface of cells that normally express FGFR2 protein.

[0993] FGFR1 expressed on cell surface

[0994] As used herein, the term "cell surface expressed FGFR1" refers to one or more FGFR1 proteins or their extracellular domains, expressed in vitro or in vivo on the cell surface such that at least a portion of the FGFR1 protein is exposed to the extracellular space of the cell membrane and is accessible to the antigen-binding portion of an antibody or antigen-binding fragment. "Cell surface expressed FGFR1" may include or consist of FGFR1 protein expressed on the surface of cells that normally express FGFR1 protein.

[0995] DDR1 expressed on cell surface

[0996] As used herein, the term "DDR1 expressed on cell surface" refers to one or more DDR1 proteins or their extracellular domains, expressed in vitro or in vivo on the cell surface such that at least a portion of the DDR1 protein is exposed to the extracellular space of the cell membrane and is accessible to the antigen-binding portion of an antibody or antigen-binding fragment. "DDR1 expressed on cell surface" may include or consist of DDR1 protein expressed on the surface of cells that normally express DDR1 protein.

[0997] Two-site antibodies

[0998] As used herein, the term "two-site antibody" refers, for example, an antibody capable of binding to two distinct epitopes on a single target. Without limitation, bispecific antibodies provide the ability to simultaneously recognize and bind to two distinct antigens or epitopes (antigen domains) as a single molecule. A subset of bispecific antibodies contains antigen-binding sites (antibody determinants) that provide the ability to recognize and bind to two distinct epitopes or antigen sites on the same target antigen. In one embodiment, the antigen-binding domain of a two-site antibody recognizes and binds to unique, non-overlapping epitopes on the same target antigen (such as receptors, such as FGFR2, FGFR1, and DDR1).

[0999] and / or

[1000] The term “and / or” in this document is used to indicate any one or any combination of the objects shown before and after “and / or”. For example, “A, B and / or C” includes single objects “A”, “B” and “C”, as well as combinations of “A and B”, “A and C”, “B and C” and “A and B and C”.

[1001] antigen-binding molecules

[1002] As used herein, the term "antigen-binding molecule" can refer to any molecule having any structure, as long as it binds to an antigen. The structure of an "antigen-binding molecule" can include, but is not limited to, polypeptides or portions thereof, small or medium-sized chemical compounds or portions thereof, or polynucleotides or portions thereof. Polypeptides or portions thereof include antigen-binding domains (also referred to as portions, parts, or fragments of antibodies).

[1003] Antigen-binding domain

[1004] The term "antigen-binding domain" includes, but is not limited to, antibody heavy chain variable (VH) region, antibody light chain variable (VL) region (preferably a combination of antibody heavy chain variable (VH) region and antibody light chain variable (VL) region), single-domain antibody (sdAb), VHH, single-chain Fv (scFv), single-chain antibody, Fv, single-chain Fv2 (scFv2), Fab, Fab', single-chain (scFab) and F(ab')2.

[1005] Antibody

[1006] The term “antibody” is used in the broadest sense and includes a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, as long as they exhibit the desired antigen-binding activity.

[1007] antibody fragments

[1008] An "antibody fragment" is a molecule other than a complete antibody that contains a portion of the complete antibody that binds to the antigen bound by the complete antibody. An "antibody fragment" contains at least one "antigen-binding domain." Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; bispecific antibodies; linear antibodies; single-chain antibody molecules (e.g., scFv); VHH, Fab', single-chain Fab (scFab); and multispecific antibodies (such as bispecific antibodies) formed from antibody fragments.

[1009] Antibodies that bind to the same / different epitopes

[1010] "Antibody that binds to the same epitope as the reference antibody" refers to an antibody that blocks the binding of the reference antibody to its antigen by 50% or more in a competitive assay, and conversely, the reference antibody blocks the binding of that antibody to its antigen by 50% or more in a competitive assay. An exemplary competitive assay is provided herein. Alternatively, "antibody that binds to a different epitope" as the reference antibody refers to an antibody that does not bind to the same epitope. The same applies when "antigen-binding domain" is mentioned instead of "antibody."

[1011] Epitope

[1012] An epitope can be defined as one or more sites that define the interaction between an antibody and one or more antigens. Accordingly, in Nilvebrant J, Rockberg J. An Introduction to Epitope Mapping. Methods Mol Biol. 2018;1785:1-10, “Antibody epitopes can be conceptually classified into linear epitopes and nonlinear epitopes. Linear epitopes, also known as continuous or sequential epitopes, can be modeled by short peptide sequences. Epitopes in which the amino acids in the distal portion of the sequence are clustered together by protein folding are called nonlinear epitopes, conformational epitopes, or discontinuous epitopes. For example, discontinuous epitopes typically contain short segments of continuous residues that can be independently bound by antibodies.” As used herein, the terms “conformational epitope,” “nonlinear epitope,” or “discontinuous epitope” are used interchangeably to refer to an epitope consisting of at least two amino acids that are not continuous amino acids in a single protein chain or a single polypeptide chain.

[1013] Significantly reduced

[1014] As used herein, the phrase “significantly reduced” or “significantly different” means that there is a sufficiently high degree of difference between two values ​​(typically one value is associated with a molecule and the other with a reference / comparison molecule) such that a person skilled in the art would consider the difference between the two values ​​to be statistically significant in the context of the biological property measured by the value (e.g., the Kd value).

[1015] Basically similar

[1016] As used herein, the terms “substantially similar” or “substantially identical” mean that two values ​​are similar to each other to a sufficiently high degree (e.g., one value is associated with the antibody of the present invention, while the other value is associated with a reference / contrast antibody), such that those skilled in the art would consider that the difference between the two values ​​is of little biological and / or statistical significance in the context of the biological characteristic measured by the value (e.g., Kd value).

[1017] Inlay

[1018] The term "chimeric" antibody refers to an antibody in which a portion of the heavy chain and / or light chain originates from a specific source or species, while the remainder of the heavy chain and / or light chain originates from a different source or species.

[1019] Antibody categories

[1020] An antibody's "class" refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of them can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The constant domains of the heavy chain corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[1021] Effective dose

[1022] The "therapeutic effective amount" of a drug (e.g., a pharmaceutical preparation) refers to the amount that effectively achieves the desired therapeutic or preventive outcome at the necessary dosage and time period.

[1023] extracellular domain

[1024] "Extracellular domain" or "ECD" is understood as a domain or portion of a membrane-associated protein that is located outside the cell when expressed in the cell. RTK stands for single-pass transmembrane (TM) protein. In RTK, the extracellular domain is typically located at the N-terminus of the transmembrane region. Another term for "extracellular domain" is "extracellular domain" as understood in this art.

[1025] Fc area

[1026] The term “Fc region” used herein is used to define the C-terminal region of an immunoglobulin heavy chain that comprises at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the C-terminus of the heavy chain. However, the C-terminal lysine (Lys447) or glycine-lysine (residues 446-447) of the Fc region may or may not be present. Unless otherwise specified herein, the amino acid residues in the Fc region or constant region are numbered according to the EU numbering system, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[1027] Frame or FR

[1028] "Frame" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. A variable domain FR typically consists of four FR domains: FR1, FR2, FR3, and FR4. Therefore, the HVR and FR sequences usually appear in the VH (or VL) as follows: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[1029] Full-length antibody / Intact antibody / Complete antibody

[1030] The terms “full-length antibody,” “intact antibody,” and “all antibody” are used interchangeably herein to refer to antibodies having a structure substantially similar to that of natural antibodies or having a heavy chain containing an Fc region as defined herein.

[1031] Host cell / host cell line / host cell culture

[1032] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells in which exogenous nucleic acids have been introduced, including progeny cells. Host cells include “transformations” and “transformed cells,” which include primary transformed cells and progeny derived from those primary transformed cells, regardless of passage number. Progeny cells may not have completely identical nucleic acid contents to the parent cells and may contain mutations. This article includes mutant progeny with the same function or biological activity as those screened or selected from the original transformed cells.

[1033] Human antibodies

[1034] A "human antibody" is an antibody whose amino acid sequence corresponds to that of an antibody produced by a human or human cell, or to a non-human antibody derived from a human antibody library or other human antibody-encoding sequences. This definition of a human antibody specifically excludes humanized antibodies containing non-human antigen-binding residues.

[1035] Human shared framework

[1036] The “human common framework” is a framework that represents the most frequently present amino acid residues in the selection of the human immunoglobulin VL or VH framework sequence. Generally, the selection of the human immunoglobulin VL or VH sequence is derived from a subgroup of variable domain sequences. Typically, the subgroups are those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, NIH Publication 91-3242, Bethesda MD (1991), Volumes 1-3. In one embodiment, for VL, this subgroup is subgroup κ I as described in Kabat et al. (ibid.). In one embodiment, for VH, this subgroup is subgroup III as described in Kabat et al. (ibid.).

[1037] Humanized antibodies

[1038] "Humanized" antibodies refer to chimeric antibodies that contain amino acid residues from a non-human HVR and amino acid residues from a human FR. In some embodiments, a humanized antibody will substantially contain at least one of all, typically two, variable domains, wherein all or substantially all HVRs (e.g., CDRs) correspond to the HVRs of the non-human antibody, and all or substantially all FRs correspond to the FRs of the human antibody. Humanized antibodies may optionally contain at least a portion of the antibody constant region derived from a human antibody. Antibodies in a "humanized form," such as non-human antibodies, refer to antibodies that have undergone humanization.

[1039] High-variable zone / HVR

[1040] As used herein, the term "hypervariant region" or "HVR" refers to each of the following: a region of an antibody variable domain that is hypervariable in sequence ("complementarity-determining region" or "CDR") and / or forms a structurally defined loop ("hypervariant loop") and / or contains antigen contact residues ("antigen contact site"). Typically, an antibody contains six HVRs: three in the VH region (H1, H2, H3) and three in the VL region (L1, L2, L3). Exemplary HVRs in this document include:

[1041] (a) Hyperchromatic rings present at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917(1987));

[1042] (b) CDRs located at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991)).

[1043] (c) Antigen contact sites located at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al., J. Mol. Biol. 262:732-745 (1996)); and

[1044] (d) Combinations of (a), (b) and / or (c), including HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3).

[1045] In one embodiment, the HVR residues of the anti-FGFR2b antigen-binding molecule, antibody, or antigen-binding fragment thereof include those identified in Table 1 or elsewhere in this specification.

[1046] In one embodiment, the HVR residues of the anti-FGFR1b antigen-binding molecule, antibody, or antigen-binding fragment thereof include those identified in Table 9 or elsewhere in this specification.

[1047] In one embodiment, the HVR residues of the anti-DDR1 antigen-binding molecule, antibody, or antigen-binding fragment thereof include those identified in Table 10 or elsewhere in this specification.

[1048] Unless otherwise specified, HVR residues and other residues (e.g., FR residues) in the variable domain are referenced in this paper to Kabat et al., with the same reference numbering above.

[1049] Immunoconjugates

[1050] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules. The heterologous molecule conjugated to the antibody may also be referred to as a secondary or adjuvant agent. Exemplary secondary or adjuvant agents include mitotic growth factor, epidermal growth factor (EGF), transforming growth factor α (TGF-α), Wnt family members (including WNT1, WNT2, WNT2B, WNT3, WNT3A, WNT4, WNT5, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, and WNT16), brain-derived neurotrophic factor (BDNF), noggin, granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF) (including IGF-1 and IGF-2), and hepatocyte growth factor (HGF). Further examples target cytokines such as the interleukin family, including IL-22, IL-10, IL-28, IL-26, IL-29, IL-24, and IL-20. Secondary or adjunctive agents may also be used to treat or help treat inflammation or fibrosis and can be nonsteroidal anti-inflammatory agents, steroids, disease-modifying antirheumatic drugs, immunosuppressants, tumor necrosis factor inhibitors, adalimumab, azathioprine, chloroquine, hydroxychloroquine, cyclosporine, D-penicillamine, etanercept, golimumab, gold salts, infliximab, leflunomide, methotrexate, minocycline, sulfasalazine, analeukin, rituximab, and tocilizumab. Secondary or adjunctive agents can also be multi-kinase inhibitors, such as axitinib, nintedanib, pirfenidone, riociguat, sorafenib, sunitinib, lanvatinib, regorafenib, panatinib, and pazopanib.Secondary or adjunctive agents can also be phosphodiesterase inhibitors, such as meribendan, arinone, ciloxamide, BI 1015550, apremilast, mesembranone, ibudilast, pyrrole, luteolin, roflumilast, cilomilast, diazepam, rolipram, YM796, icariin, sildenafil, tadalafil, vardenafil, avanaphile, edenafil, milonafil, udenafil, and zaprinast. Secondary or adjuvant agents can also be integrin inhibitors, such as ProAgio and GSK-3008348. Secondary or adjuvant agents can also be (cytotoxic) agents, including anti-aging compounds such as MDM2 inhibitors (Nutlin 3a, RG-7112); BCL-2 anti-apoptotic protein family members, inhibitors that inhibit the function of at least the anti-apoptotic protein BCL-xL (ABT-263, ABT-737, WEHI-539, A-1155463); GLS1 inhibitors (CB-839); Akt-specific inhibitors (MK-2206); and radioisotopes.

[1051] Individual / Subject

[1052] "Individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., human and non-human primates, such as monkeys), rabbits, and rodents (e.g., mice and rats). In some embodiments, the individual or subject is a human.

[1053] Isolated antibodies

[1054] "Isolated" antibodies are antibodies that have been separated from components in their natural environment. In some embodiments, antibodies are purified to a purity greater than 95% or 99% by means of, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[1055] isolated nucleic acids

[1056] "Isolated" nucleic acids refer to nucleic acid molecules that have been isolated from components of their natural environment. Isolated nucleic acids include nucleic acid molecules that are contained in cells that normally contain nucleic acid molecules, but which are located outside chromosomes or at chromosomal locations different from their natural chromosomal locations.

[1057] Isolated nucleic acids encoding anti-FGFR2 antibody, anti-FGFR1 antibody, or anti-DDR1 antibody.

[1058] "Isolated nucleic acid encoding anti-FGFR2 antibody", "Isolated nucleic acid encoding anti-FGFR1 antibody" or "Isolated nucleic acid encoding anti-DDR1 antibody" refers to one or more nucleic acid molecules encoding the heavy and light chains (or fragments thereof) of an antibody, including such nucleic acid molecules in a single vector or in separate vectors, and such nucleic acid molecules present at one or more locations in a host cell.

[1059] Monoclonal antibodies

[1060] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous group of antibodies, meaning that, apart from possible variant antibodies (e.g., those containing naturally occurring mutations or produced during the production of the monoclonal antibody formulation, such variants are typically present in small quantities), the individual antibodies comprising this group are identical and / or bind to the same epitope. In contrast to polyclonal antibody formulations, which typically comprise different antibodies targeting different determinants (epitaxes), each monoclonal antibody in a monoclonal antibody formulation targets a single determinant on the antigen. Therefore, the modifier "monoclonal" indicates that the antibody is characterized by being obtained from a substantially homogeneous group of antibodies and should not be construed as requiring the antibody to be produced by any particular method. For example, monoclonal antibodies intended for use according to the invention can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci, such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

[1061] naked antibodies

[1062] "Naked antibody" refers to an antibody that is not conjugated to a heterologous part (e.g., a cytotoxic part) or radiolabeled. Naked antibodies can be found in pharmaceutical preparations.

[1063] Natural antibodies

[1064] "Natural antibodies" refer to naturally occurring immunoglobulin molecules with different structures. For example, natural IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 Daltons, composed of two identical light chains and two identical heavy chains bonded by disulfides. Each heavy chain has a variable region (VH), also called a variable heavy chain domain or heavy chain variable domain, from the N-terminus to the C-terminus, followed by three constant domains (CH1, CH2, and CH3). Similarly, each light chain has a variable region (VL), also called a variable light chain domain or light chain variable domain, from the N-terminus to the C-terminus, followed by a constant light chain (CL) domain. The light chains of antibodies can be classified into one of two types based on the amino acid sequence of their constant domains, called kappa (κ) and lamuda (λ).

[1065] Packaging insert

[1066] The term "packaging insert" is used to refer to the instruction leaflet typically included in the commercial packaging of a therapeutic product, which contains information concerning the indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings related to the use of such therapeutic products.

[1067] Amino acid sequence identity percentage (%)

[1068] The percentage of amino acid sequence identical to a reference polypeptide sequence is defined as the percentage of amino acid residues in the candidate sequence that are identical to those in the reference polypeptide sequence after aligning the candidate sequence with the reference polypeptide sequence and introducing vacancies (if necessary) to achieve the maximum percentage of sequence identity, without taking into account any conserved substitutions as part of the sequence identity. Alignment used to determine the percentage of amino acid sequence identity can be performed in various ways within the scope of the art, such as using publicly available computer software, such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR) software, or GENTYX (registered trademark) (Genetyx Co., Ltd.). Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms required to achieve maximum alignment across the full length of the sequences being compared.

[1069] The ALIGN-2 sequence comparison computer program was written by Genentech, Inc., and the source code has been submitted with the user documentation to the US Copyright Office, Washington DC, 20559, where it is registered under US Copyright Registry No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc. (South San Francisco, California), or can be compiled from the source code. The ALIGN-2 program should be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and remain unchanged. When performing amino acid sequence comparisons using ALIGN-2, the percentage of amino acid sequence identity between a given amino acid sequence A and / or with respect to a given amino acid sequence B (which can be alternatively expressed as a given amino acid sequence A having or containing a percentage of amino acid sequence identity with or with respect to a given amino acid sequence B) is calculated as follows:

[1070] 100 multiplied by the fraction X / Y

[1071] Where X represents the number of amino acid residues that are scored as identical matches by the sequence alignment program ALIGN-2 in the alignment of A and B, and Y represents the total number of amino acid residues in B. It should be understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the amino acid sequence identity % between A and B will not be equal to the amino acid sequence identity % between B and A. Unless otherwise specified, all amino acid sequence identity % values ​​used herein were obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[1072] pharmaceutical preparations

[1073] The term "pharmaceutical formulation" refers to a formulation in which the bioactive ingredient contained therein is in a form in which the activity is effective and which does not contain any additional components that would have unacceptable toxicity to a subject to whom the formulation will be administered.

[1074] Pharmaceutically acceptable carriers

[1075] "Pharmaceutically acceptable carriers" refer to components in a pharmaceutical preparation that are non-toxic to the subject, excluding the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffer solutions, excipients, stabilizers, or preservatives.

[1076] Treatment (to treat)

[1077] As used herein, “treatment” (and its grammatical variations such as “treat” or “treating”) refers to the prevention of disease onset or recurrence, the relief of symptoms, the attenuation of any direct or indirect pathological consequences of disease, the prevention of metastasis, the reduction of the rate of disease progression, the improvement or relief of disease symptoms, and the mitigation or improvement of prognosis. In some embodiments, the antibodies of the present invention are used to delay the development of disease or to slow the progression of disease. “Method of treatment” or its equivalents, when applied, for example, to fibrosis, refers to a procedure or process of action designed to reduce or eliminate the number of fibrotic cells in a patient or to alleviate the symptoms of fibrosis. It does not necessarily mean that fibrotic cells or other disease will actually be eliminated, that the number of cells or disease will actually be reduced, or that fibrosis or other disease will actually be relieved. Generally, treatments for fibrosis, even if the probability of success is low, are still considered to elicit a generally beneficial process of action, taking into account the patient’s medical history and estimated survival expectations.

[1078] Variable region / variable structural domain

[1079] The term "variable region" or "variable domain" refers to a domain of the antibody heavy or light chain involved in antibody-antigen binding. The variable domains (VH and VL, respectively) of the heavy and light chains of natural antibodies typically have similar structures, with each domain containing four conserved frame regions (FRs) and three hypervariable regions (HVRs). (See, for example, Kindt et al., Kuby Immunology, 6th ed., WH Freeman and Co., p. 91 (2007)). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies binding to a specific antigen can be isolated using either the VH or VL domain from the antibody binding that antigen to screen libraries of complementary VL or VH domains. See, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[1080] vector

[1081] As used herein, the term "vector" refers to a nucleic acid molecule capable of carrying another nucleic acid linked to it. This term includes vectors that function as self-replicating nucleic acid structures, as well as vectors incorporated into the genome of a host cell into which they have been introduced. Some vectors can direct the expression of nucleic acids operatively linked to them. Such vectors are referred to herein as "expression vectors." Vectors can be introduced into host cells using methods such as viral methods and electroporation, but the introduction of vectors is not limited to in vitro introduction, and direct in vivo introduction is also possible.

[1082] Detailed Description of Embodiments of the Invention

[1083] Activation of receptor tyrosine kinases (RTKs) plays a crucial role in regulating kinase activity and activating downstream signaling pathways in cells. RTK autophosphorylation is mediated by a sequential and precisely ordered intermolecular reaction following binding to its natural ligand. The aspects and embodiments described herein are based, at least in part, on the finding that RTK activation can be achieved using antigen-binding molecules that mimic the binding of natural ligands to RTKs, including ligand-receptor interactions involving the binding of the ligand to different domains or to unique, non-overlapping epitopes on the same domain of the same receptor protein. Without wishing to be bound by theory, it is assumed that the antigen-binding molecules described herein at least in part mimic the binding of the ligand to its receptor and induce receptor dimerization and autophosphorylation, thereby activating its downstream intracellular signaling.

[1084] In one aspect, an antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein is provided, wherein the molecule is an agonist. Hereinafter referred to herein as an anti-RTK agonist antigen-binding molecule.

[1085] In one embodiment, the anti-RTK ...

Claims

1. An agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein the molecule is an agonist of the RTK protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of a first RTK protein and a second antigen-binding domain that binds to a second epitope of the first RTK protein or a second RTK protein, wherein the first epitope resides in the first domain of the RTK protein and the second epitope resides in the second domain of the RTK protein.

2. The agonist antigen-binding molecule of claim 1, wherein the RTK protein is a human fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of the extracellular domain (ECD) of a first FGFR2 protein and a second antigen-binding domain that binds to a second epitope of the extracellular domain (ECD) of the first FGFR2 protein or a second FGFR2 protein, wherein the first epitope and the second epitope are different from each other.

3. The molecule according to claim 2, wherein the first antigen-binding domain binds to a first FGFR2 protein, and the second antigen-binding domain binds to a second FGFR2 protein.

4. The molecule according to claim 2 or 3, wherein it is a two-site antigen-binding molecule.

5. The molecule according to any one of claims 2 to 4, wherein the human FGFR2 protein is the human FGFR2b protein.

6. The molecule according to any one of claims 2 to 5, wherein each of the extracellular domains is independently selected from the immunoglobulin (Ig)-like domain of the FGFR2 protein.

7. The molecule according to any one of claims 2 to 6, wherein the first antigen-binding domain binds to a first domain of the FGFR2 protein and the second antigen-binding domain binds to a second domain of the FGFR2 protein, wherein the first domain and the second domain are identical.

8. The molecule according to any one of claims 2 to 6, wherein the first antigen-binding domain binds to a first domain of the FGFR2 protein and the second antigen-binding domain binds to a second domain of the FGFR2 protein, wherein the second domain is different from the first domain.

9. The molecule according to any one of claims 2 to 8, wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the Ig-like domain of the FGFR2 protein, wherein the Ig-like domain is selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

10. The molecule according to any one of claims 6 to 9, wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the Ig-like domain of the FGFR2 in a combination selected from the group consisting of: (i) wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the IgIIIb and IgIIIb domains of FGFR2; (ii) wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the IgIIIb and IgIII domains of FGFR2; (iii) wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the IgIIIb and IgII domains of FGFR2; and (iv) wherein the first antigen-binding domain and the second antigen-binding domain independently bind to the IgIIIb and IgI domains of FGFR2.

11. The molecule according to any one of claims 6 to 10, wherein (i) the first antigen-binding domain binds to the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain specifically binds to the IgIIIb domain of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgIIIc domain of FGFR2 and exhibits less than 10% or less than 5% or 0% cross-reactivity with the IgI and IgII domains of FGFR2.

12. The molecule according to any one of claims 4 to 11, wherein the binding of the first antigen-binding domain and the second antigen-binding domain to the FGFR2b protein results in the following agonist activity: (i) wherein the molecule activates signal transduction in the target cell via binding to the FGFR2b protein expressed on the cell surface of the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% or more in the expression of an intracellular protein downstream of FGFR2b compared to the expression of the intracellular protein prior to the binding of the molecule to the FGFR2b protein expressed on the cell surface of the target cell, particularly wherein the intracellular protein downstream of FGFR2b is selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1 / pERK2, and pMAPK3 / pERK1; or (ii) wherein the molecule induces the proliferation of the target cells via binding to the FGFR2b protein expressed on the cell surface of the target cells, thereby exhibiting agonist activity, wherein the induction of target cell proliferation is measured by an increase in ATP expression in the target cells in the presence of the molecule by at least 1.5-fold, or 2-fold, or 2.5-fold or greater compared to ATP expression in the target cells in the absence of the molecule, wherein the ATP expression is proportional to the concentration of live target cells. In particular, the target cells are lung epithelial cells, optionally selected from type II alveolar (AT2) cells; or (iii) wherein receptor dimerization is detected in the presence of said molecule in an in vitro cell assay, wherein receptor dimerization indicates agonist activity; or (iv) wherein phosphorylation of the FGFR2b protein is detected in the presence of the molecule in an in vitro cell assay, wherein phosphorylation of the FGFR2b protein indicates agonist activity.

13. The agonist antigen-binding molecule of claim 1, wherein the RTK protein is a human fibroblast growth factor receptor 1 (FGFR1) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of the extracellular domain (ECD) of a first FGFR1 protein and a second antigen-binding domain that binds to a second epitope of the extracellular domain (ECD) of the first FGFR1 protein or a second FGFR1 protein, wherein the first epitope and the second epitope are distinct from each other.

14. The agonist antigen-binding molecule of claim 1, wherein the RTK protein is a human discoid domain receptor 1 (DDR1) protein, wherein the molecule comprises a first antigen-binding domain that binds to a first epitope of the extracellular domain (ECD) of a first DDR1 protein and a second antigen-binding domain that binds to a second epitope of the extracellular domain (ECD) of the first DDR1 protein or a second DDR1 protein, wherein the first epitope and the second epitope are distinct from each other.

15. The molecule according to any one of claims 1 to 14, wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each Fab, Fab', F(ab)'2, scFab, scFv, Fv, VHH or a single-domain antibody, wherein the first antigen-binding domain and the second antigen-binding domain are linked by at least one disulfide bond.

16. The molecule according to any one of claims 1 to 15, wherein the first antigen-binding domain and the second antigen-binding domain are characterized by having at least one disulfide bond formed between amino acid residues not in the hinge region.

17. The molecule of claim 16, wherein the at least one disulfide bond is an engineered disulfide bond not present in wild-type IgG, optionally wherein the at least one disulfide bond is formed between the CH1 region, CL region, VH or VHH region, or VL region of the first antigen-binding domain and the CH1 region, CL region, VH or VHH region, or VL region of the second antigen-binding domain, particularly wherein the at least one disulfide bond is formed between regions selected from the group consisting of: [i] Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CH1 region of the second antigen-binding domain; [ii] Amino acid residues in the CL region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain; [iii] Amino acid residues in the CH1 region of the first antigen-binding domain and amino acid residues in the CL region of the second antigen-binding domain; [iv] Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VH or VHH region of the second antigen-binding domain; [v] Amino acid residues in the VL region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain; and [vi] Amino acid residues in the VH or VHH region of the first antigen-binding domain and amino acid residues in the VL region of the second antigen-binding domain.

18. The molecule according to any one of claims 1 to 17, wherein the molecule comprises an antibody constant region, particularly (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from human IgG1, IgG2, IgG3 or IgG4 constant regions.

19. The molecule according to any one of claims 1 to 18, wherein the molecule is characterized by two antigen-binding domains, or wherein the molecule is a bivalent molecule, or wherein the first antigen-binding domain and the second antigen-binding domain are each monovalent Fab or monovalent VHH, or each is monovalent scFab.

20. The molecule according to any one of claims 1 to 19, wherein the molecule comprises more than two antigen-binding domains, or wherein the molecule is a trivalent, tetravalent, or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain, the third antigen-binding domain binding to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain.

21. The molecule of claim 20, wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binding to the same epitope on the same domain as the first antigen-binding domain or the second antigen-binding domain, but not binding to the same epitope on the third antigen-binding domain, optionally wherein the molecule comprises a third antigen-binding domain and a fourth antigen-binding domain, wherein the third antigen-binding domain binding to the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binding to the same epitope on the same domain as the second antigen-binding domain.

22. A pharmaceutical composition comprising: a molecule according to any one of claims 1 to 21, and a pharmaceutically acceptable carrier.

23. An immune conjugate protein comprising the molecule according to any one of claims 1 to 21.

24. The molecule according to any one of claims 1 to 21, or the pharmaceutical composition according to claim 22, or the immunoconjugate according to claim 23, for use in the treatment or prevention of a disease, ailment, or condition associated with lung epithelial cell damage in an individual in need.

25. The molecular or pharmaceutical composition or immunoconjugate for use according to claim 24, wherein the condition associated with the lung epithelial cell damage in an individual is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, conditions associated with exposure to environmental toxins, conditions associated with exposure to bacteria, conditions associated with exposure to viruses, cystic fibrosis, lung resection, conditions associated with exposure to radiation, conditions associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

26. The molecule according to any one of claims 1 to 21, the pharmaceutical composition according to claim 22, or the immunoconjugate according to claim 23, for use in the treatment of any disease, ailment, or condition that is readily improved or prevented by one of the following selected from the group consisting of: (i) Increase or enhancement of FGFR2b signal conduction; (ii) Increased proliferation of lung epithelial cells; and (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

27. The molecule according to any one of claims 1 to 21, or the pharmaceutical composition according to claim 22, or the immunoconjugate according to claim 23, for use in the treatment or prevention of fibrosis or inflammation in an individual in need.

28. The molecular or pharmaceutical composition or immunoconjugate for use according to claim 27, wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lung, heart, blood vessels, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.

29. The molecular or pharmaceutical composition or immunoconjugate for use according to claim 28, wherein... (i) The fibrosis or inflammation of the lungs is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma; (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome; (iii) The aforementioned fibrosis or inflammation of the pancreas is pancreatitis; (iv) The aforementioned fibrosis or inflammation of the kidneys is selected from the group consisting of: chronic kidney disease or acute kidney disease; (v) The aforementioned fibrosis or inflammation of the oral mucosa is oral mucositis; (vi) The fibrosis or inflammation of the skin is diabetic foot ulcer or bullous epidermolysis; (vii) The aforementioned fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and (viii) The fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn's disease.

30. Use of the molecule according to any one of claims 1 to 21, or the pharmaceutical composition according to claim 22, or the immunoconjugate according to claim 23, in the manufacture of a medicament for the treatment or prevention of a disease or ailment or condition associated with lung epithelial cell damage in an individual in need.

31. A method of treating an individual in need of a disease or ailment related to lung epithelial cell damage, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of claims 1 to 21, or a pharmaceutical composition according to claim 22, or an immunoconjugate according to claim 23.

32. The use according to claim 30 or the method according to claim 31, wherein the condition associated with the lung epithelial cell damage in an individual is selected from the group consisting of: pulmonary fibrosis, pneumonia, age-related pulmonary fibrosis, asthma, conditions associated with exposure to environmental toxins, conditions associated with exposure to bacteria, conditions associated with exposure to viruses, cystic fibrosis, lung resection, conditions associated with exposure to radiation, conditions associated with exposure to tobacco smoke or chemicals, and bleomycin-mediated epithelial damage.

33. The use according to claim 30 or the method according to claim 31, wherein the condition associated with lung epithelial cell damage in an individual is a condition that is easily improved or prevented by selecting one of the following: (i) Increase or enhancement of FGFR2b signal conduction; (ii) Increased proliferation of lung epithelial cells; and (iii) Increased proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

34. Use of the molecule according to any one of claims 1 to 21, or the pharmaceutical composition according to claim 22, or the immunoconjugate according to claim 23, in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need.

35. A method for treating fibrosis or inflammation in an individual in need, the method comprising administering to the individual a therapeutically effective amount of a molecule according to any one of claims 1 to 21, a pharmaceutical composition according to claim 22, or an immunoconjugate according to claim 23.

36. The use according to claim 34 or the method according to claim 35, wherein the fibrosis or inflammation is selected from fibrosis or inflammation of the lungs, heart, blood vessels, liver, bile ducts, small intestine, large intestine, pancreas, kidneys, eyes, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.

37. The use according to claim 34 or the method according to claim 35, wherein... (i) The fibrosis or inflammation of the lungs is selected from the group consisting of: pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, chronic obstructive pulmonary disease (COPD), obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, interstitial lung tumors, and asthma; (ii) The fibrosis or inflammation of the liver or bile duct is selected from the group consisting of: non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Arager's syndrome; (iii) The aforementioned fibrosis or inflammation of the pancreas is pancreatitis; (iv) The aforementioned fibrosis or inflammation of the kidneys is selected from the group consisting of: chronic kidney disease or acute kidney disease; (v) The aforementioned fibrosis or inflammation of the oral mucosa is oral mucositis; (vi) The fibrosis or inflammation of the skin is diabetic foot ulcer or bullous epidermolysis; (vii) The aforementioned fibrosis or inflammation of the thymus is graft-versus-host disease (GvHD); and (viii) The fibrosis or inflammation of the small or large intestine is inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn's disease.

38. An isolated nucleic acid encoding a molecule according to any one of claims 1 to 21; or multiple isolated nucleic acids encoding a molecule according to any one of claims 1 to 21.

39. A host cell comprising the nucleic acid or a plurality of nucleic acids according to claim 38.

40. A method for producing a molecule or a process for producing a molecule, said method or process comprising the steps of: (a) culturing a host cell according to claim 39, and (b) recovering the molecule produced in step (a).

Images