EP4784624A1

EP4784624A1 - Anti-rtk antibodies and uses thereof

Info

Publication number: EP4784624A1
Application number: EP24872515.2A
Authority: EP
Inventors: Noriyuki Takahashi; Takaaki NOGUCHI; Chiew Ying YEO
Original assignee: Chugai Pharmaceutical Co Ltd
Current assignee: Chugai Pharmaceutical Co Ltd
Priority date: 2023-09-29
Filing date: 2024-09-27
Publication date: 2026-08-05
Also published as: WO2025070733A1; CN121986114A

Abstract

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

Description

ANTI-RTK ANTIBODIES AND USES THEREOF

Receptor tyrosine kinases (RTKs) play crucial roles in various cellular processes. RTKs are known to be activated by ligand-induced dimerization. Dimerization leads to kinase activation and autophosphorylation of multiple tyrosine residues in the cytoplasmic domain of the receptor, and activation of downstream signaling of various pathways. One of the members of RTKs includes the family of fibroblast growth factor receptors (FGFRs) expressed by various mammalian cells. The family of FGFRs consists of FGFR1, FGFR2, FGFR3, and FGFR4, and comprises high affinity receptors for many different FGF ligands (NPL 1). The receptor is a transmembrane tyrosine kinase, and FGFR signaling drives downstream pathways that are essential for cell proliferation, differentiation, survival and migration (NPL 2). Binding of the FGF ligand to the receptor induces dimerization of the FGF:FGFR complex, and activation of FGF downstream signaling (NPL 3).

The members of the FGF family are potent stimulators of tissue repairs, and have been shown to be helpful in numerous tissue regenerative processes, such as maintenance of cell stemness, induction of cell de-differentiation, stimulation of cell proliferation, suppression of cell senescence, suppression of cell death, regulation of inflammation, stimulation of angiogenesis, and enhancement of proteases expression etc. The FGF ligands carry out these diverse functions by binding and activating the FGFR family of RTKs.

FGFR2 has two splice isoforms, FGFR2b that is normally localized in the epithelial cells, and FGFR2c which is normally localized in the mesenchymal cells (NPL 1). 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). While various therapeutics have been developed that target FGFR2, such as anti-cancer drugs that target FGFR2 over-expressing cells, these are inhibitors to FGFR2, and there remains a significant unmet medical need for the development of FGFR2 agonists.

FGFR1 also has two splice isoforms: FGFR1b is expressed in the cerebrum, tongue, preputial gland, vas deferens, and skin; and FGFR1c is expressed in various tissue (NPL 8). FGFR1b is a receptor for FGF ligands such as FGF1 and FGF2, and FGFR1b 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, NPL 11). Especially, FGF19, FGF21, and FGF23 act on β-Klotho in complex with FGFR1c. FGFR1 is also evaluated as therapeutic targets by both inhibitor and agonist for anti-cancer and chronic diseases such as obesity and diabetes. Particularly, there are several reports for agonisms of FGFR1, including homodimerization-based agonist for both FGFR1b and FGFR1c, and heterodimerization-based agonist for FGFR1c and β-Klotho (NPL 12, NPL 13, NPL 14), however, there are no reports about strong FGFR1b specific agonist that could show better agonistic activity than natural ligands.

Another one of the members of RTKs includes the family of discoidin domain receptors (DDRs). The family of DDRs consists of DDR1 and DDR2. DDR1 is expressed in mainly in epithelial cells, smooth muscle cells, and in certain condition in fibroblasts, and DDR2 is expressed in cells of mesenchymal origin (NPL 15). Both DDR1 and DDR2 are activated upon collagen binding at the extracellular portion of the receptors, and then initiates downstream signaling pathways including receptor phosphorylation (NPL 16). Collagen is an important ECM component that interacts with various cellular receptors in the tissue, thus DDRs are involved in various important functions including regulation of cell proliferation, survival, differentiation, cellular aggregation, adhesion, migration, and invasions (NPL 16). Although therapeutic applications of DDRs are mainly focused on anti-fibrosis and anti-cancer treatment because various cancer cells overexpress DDRs, some of reports indicated that DDRs activation regenerates cartilage tissue and wound healing (NPL 17, NPL 18), suggesting that DDRs agonism also has potential to regenerate various tissue. As well as FGFR1 and FGFR2, however, therapeutic agonist antibody specific for DDRs has not been successfully identified.

[PTL 1] WO2012021841A2
[PTL 2] WO2021247718A1
[PTL 3] WO2022143728A1
[PTL 4] WO2022087243A1
[PTL 5] WO2021157679A1

Non-Patent Literature

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[NPL 2] Susanne Braun, Ulrich auf dem Keller, Heike Steiling, Sabine Werner, Fibroblast growth factors in epithelial repair and cytoprotection. Philos Trans R Soc Lond B Biol Sci. 2004 May 29;359(1445):753-7.
[NPL 3] Allen Zinkle, Moosa Mohammadi, Structural Biology of the FGF7 Subfamily. Front Genet. 2019 Feb 12;10:102.
[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. 2020 Oct 2;11(1):4974.
[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). 2017 Jul 28:4:118.
[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.
[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. Sci Rep. 2021 Sep 29;11(1):19269.
[NPL 8] Klementina Fon Tacer, Angie L Bookout, Xunshan Ding, Hiroshi Kurosu, George B John, Lei Wang, Regina Goetz, Moosa Mohammadi, Makoto Kuro-o, David J Mangelsdorf, Steven A Kliewer, Research resource : Comprehensive expression atlas of the fibroblast growth factor system in adult mouse. Mol Endocrinol. 2010 Oct;24(10):2050-64.
[NPL 9] H D Beer, L Vindevoghel, M J Gait, J M Revest, D R Duan, I Mason, C Dickson, S Werner, Fibroblast growth factor (FGF) receptor 1-IIIb is a naturally occurring functional receptor for FGFs that is preferentially expressed in the skin and the brain. J Biol Chem. 2000 May 26;275(21):16091-7.
[NPL 10] D M Ornitz, J Xu, J S Colvin, D G McEwen, C A MacArthur, F Coulier, G Gao, M Goldfarb, Receptor specificity of the fibroblast growth factor family. J Biol Chem. 1996 Jun 21;271(25):15292-7.
[NPL 11] Tian Lan, Donald A Morgan, Kamal Rahmouni, Junichiro Sonoda, Xiaorong Fu, Shawn C Burgess, William L Holland, Steven A Kliewer, David J Mangelsdorf, FGF19, FGF21, and an FGFR1/β-Klotho-Activating Antibody Act on the Nervous System to Regulate Body Weight and Glycemia. Cell Metab. 2017 Nov 7;26(5):709-718.e3.
[NPL 12] Ai-Luen Wu, Ganesh Kolumam, Scott Stawicki, Yongmei Chen, Jun Li, Jose Zavala-Solorio, Khanhky Phamluong, Bo Feng, Li Li, Scot Marsters, Lance Kates, Nicholas van Bruggen, Maya Leabman, Anne Wong, David West, Howard Stern, Elizabeth Luis, Hok Seon Kim, Daniel Yansura, Andrew S Peterson, Ellen Filvaroff, Yan Wu, Junichiro Sonoda, Amelioration of type 2 diabetes by antibody-mediated activation of fibroblast growth factor receptor 1. Sci Transl Med. 2011 Dec 14;3(113):113ra126.
[NPL 13] Jocelyn Chan, Joyce Chan, Lily Shao, Scott S Stawicki, Victoria C Pham, Rob W Akita, Marc Hafner, Lisa Crocker, Kebing Yu, James T Koerber, Gabriele Schaefer, Laetitia Comps-Agrar, Systematic pharmacological analysis of agonistic and antagonistic fibroblast growth factor receptor 1 Mabs reveals a similar unique mode of action. J Biol Chem. 2023 Jan;299(1):102729.
[NPL 14] Xiaoshan Min, Jennifer Weiszmann, Sheree Johnstone, Wei Wang, Xinchao Yu, William Romanow, Stephen Thibault, Yang Li, Zhulun Wang, Agonistic β-Klotho antibody mimics fibroblast growth factor 21 (FGF21) functions. J Biol Chem. 2018 Sep 21;293(38):14678-14688.
[NPL 15] Gurubasavaraja Swamy Purawarga Matada, Arka Das, Prasad Sanjay Dhiwar & Abhishek Ghara, DDR1 and DDR2 : a review on signaling pathway and small molecule inhibitors as an anticancer agent. Med Chem Res 30, 535-551 (2021).
[NPL 16] N M Coelho, A Wang, C A McCulloch, Discoidin domain receptor 1 interactions with myosin motors contribute to collagen remodeling and tissue fibrosis. Biochim Biophys Acta Mol Cell Res. 2019 Nov;1866(11):118510.
[NPL 17] Arokia Vijaya Anand Mariadoss, Chau-Zen Wang, Exploring the Cellular and Molecular Mechanism of Discoidin Domain Receptors (DDR1 and DDR2) in Bone Formation, Regeneration, and Its Associated Disease Conditions. Int J Mol Sci. 2023 Oct 4;24(19):14895.
[NPL 18] Muriel Cario, DDR1 and DDR2 in skin. Cell Adh Migr. 2018;12(4):386-393.
[NPL 19] D Smedley, A Demiroglu, M Abdul-Rauf, C Heath, C Cooper, J Shipley, N C Cross, ZNF198-FGFR1 transforms Ba/F3 cells to growth factor independence and results in high level tyrosine phosphorylation of STATS 1 and 5. Neoplasia. 1999 Oct;1(4):349-55.
[NPL 20] Victoria Juskaite, David S Corcoran, Birgit Leitinger, Collagen induces activation of DDR1 through lateral dimer association and phosphorylation between dimers. Elife. 2017 Jun 7:6:e25716.
[NPL 21] David S Corcoran, Victoria Juskaite, Yuewei Xu, Frederik Gorlitz, Yuriy Alexandrov, Christopher Dunsby, Paul M W French, Birgit Leitinger, DDR1 autophosphorylation is a result of aggregation into dense clusters. Sci Rep. 2019 Nov 19;9(1):17104.

While various therapeutics have been developed to target receptor tyrosine kinases, there remains a significant medical need for activating FGFR signaling, in particular FGFR2 and FGFR1, where the FGF family are known to be potent stimulators of tissue repairs, and have been shown to be helpful in numerous tissue regenerative processes, such as maintenance of cell stemness, induction of cell de-differentiation, stimulation of cell proliferation, suppression of cell senescence, suppression of cell death, regulation of inflammation, stimulation of angiogenesis, and enhancement of proteases expression etc. There also remains a significant medical need for activating DDR signaling, in particular DDR1, where the collagen family are known to be potent regulator of cell proliferation, survival, cellular differentiation, ECM homeostasis, cytokine secretions, cellular aggregation and migration etc.

The present invention provides agonistic antigen-binding molecules, such as antibodies or fragments thereof that bind to receptor tyrosine kinases (RTKs), including the family of fibroblast growth factor receptors (FGFRs) and the family of discoidin domain receptors (DDRs). The present invention provides antigen-binding molecules, such as antibodies or fragments thereof that bind human fibroblast growth factor receptor 2 protein (FGFR2), human fibroblast growth factor receptor 1 protein (FGFR1), or human discoidin domain receptor 1 (DDR1), preferably wherein the antibodies or fragments 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 same. The molecules of the invention are useful in any disease, disorder or condition, where an increase in FGFR2b signaling, FGFR1b signaling, or DDR1a signaling is helpful and can be used to treat fibrosis or inflammation, in particular lung and skin fibrosis.

The molecules of the invention provided herein are agonistic antigen-binding molecules, such as antibodies or fragments thereof that bind receptor tyrosine kinases (RTKs) and promote their dimerization and activation, and wherein the antigen-binding molecules bind biparatopically. In one aspect, the molecules of the invention are antigen-binding molecules, such as antibodies or fragments thereof that bind fibroblast growth factor receptor 2 protein (FGFR2), preferably wherein the antibodies or fragments thereof specifically bind to FGFR2IIIb. The antigen-binding molecules, such as antibodies, are useful, inter alia, for targeting cells that express FGFR2, preferably FGFR2IIIb. The FGFR2b antigen-binding molecules mimic the ligands of FGFR2b such as FGF7 or FGF10 to selectively activate FGFR2b signaling and find uses in methods of activating FGF downstream signaling. In one aspect, the anti-FGFR2b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind specifically to the IgII, IgIII, or IgIIIb domain of FGFR2, and selectively activate FGFR2IIIb signaling. In one aspect, the anti-FGFR2b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, 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 one aspect, the anti-FGFR2b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind biparatopically to the IgIIIb and IgI, IgIIIb and IgII, IgIIIb and IgIII, or IgIIIb and IgIIIb domains of FGFR2, and selectively activate FGFR2IIIb signaling. Agonistic antigen-binding molecules, such as antibodies, against FGFR2b, including agonistic biparatopic antibodies against FGFR2b provide a new class of therapeutics for the treatment of tissue-degenerative diseases.

In one aspect, the molecules of the invention are antigen-binding molecules, such as antibodies or fragments thereof that bind fibroblast growth factor receptor 1 protein (FGFR1), preferably wherein the antibodies or fragments thereof specifically bind to FGFR1IIIb. The antigen-binding molecules, such as antibodies, are useful, inter alia, for targeting cells that express FGFR1, preferably FGFR1IIIb. The FGFR1b antigen-binding molecules mimic the ligands of FGFR1b such as FGF1 or FGF2 to selectively activate FGFR1b signaling and find uses in methods of activating FGF downstream signaling. In one aspect, the anti-FGFR1b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind specifically to the IgII, IgIII, or IgIIIb domain of FGFR1, and selectively activate FGFR1IIIb signaling. In one aspect, the anti-FGFR1b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, 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 one aspect, the anti-FGFR1b antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind biparatopically to the IgIIIb and IgI, IgIIIb and IgII, IgIIIb and IgIII, or IgIIIb and IgIIIb domains of FGFR1, and selectively activate FGFR1IIIb signaling. Agonistic antigen-binding molecules, such as antibodies, against FGFR1b, including agonistic biparatopic antibodies against FGFR1b provide a new class of therapeutics for the treatment of tissue-degenerative diseases.

In one aspect, the molecules of the invention are antigen-binding molecules, such as antibodies or fragments thereof that bind discoidin domain receptor 1 protein (DDR1), preferably wherein the antibodies or fragments thereof specifically bind to DDR1a. The antigen-binding molecules, such as antibodies, are useful, inter alia, for targeting cells that express DDR1, preferably DDR1a. The DDR1a antigen-binding molecules mimic the ligands of DDR1a such as collagen type I, IV, V, VI, or VIII to selectively activate DDR1a signaling and find uses in methods of activating FGF downstream signaling. In one aspect, the anti-DDR1a antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind specifically to a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1a, and selectively activate DDR1a signaling. In one aspect, the anti-DDR1a antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, comprises a first antigen-binding domain that binds to a DS domain and a second antigen-binding domain that binds to a DS domain, a DS-like domain, or an EJXM region and selectively activate DDR1a signaling. In one aspect, the anti-DDR1a antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, bind biparatopically to the DS domain and the EJXM region, the DS domain and the DS-like domain, or the DS domain and the DS domain of DDR1, and selectively activate DDR1a signaling. Agonistic antigen-binding molecules, such as antibodies, against DDR1a, including agonistic biparatopic antibodies against DDR1a provide a new class of therapeutics for the treatment of tissue-degenerative diseases.

The anti-FGFR2b antigen-binding molecules, the anti-FGFR1b antigen-binding molecules, or the anti-DDR1 antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, may be used alone in unmodified form, or may be included as part of an antibody-drug conjugate, or a fusion protein. In addition, molecules of the invention include, without limitation, small organic molecules and polypeptides.

The invention also provides herein methods for the use of the anti-FGFR2b antigen-binding molecules, the anti-FGFR1b antigen-binding molecules, or the anti-DDR1 antigen-binding molecules, such as antibodies, and antigen-binding fragments thereof, in the treatment or alleviation of symptoms of diseases or disorders, pharmaceutical compositions comprising the anti-FGFR2b antibodies, the anti-FGFR1b antibodies, or the anti-DDR1 antibodies, and antigen-binding fragments thereof and their uses thereof.

Exemplary embodiments

The following aspects and embodiments are part of the invention - wherein it will be appreciated by the skilled reader that the embodiments included in the following are independently intended as the embodiments for each and every general embodiments and aspects set forth below. In aspects and embodiments, the invention relates to any one of [A1] and [A1a] to [A53] below:
[A1] An agonistic antigen-binding molecule that binds to a human Fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first FGFR2 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein, wherein the first epitope and the second epitope are different from each other.
[A1a] An agonistic antigen-binding molecule that binds to a human Fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises:
a first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR2 protein; and
a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein,
wherein the first epitope and the second epitope are different from each other.
[A2] The molecule of [A1] or [A1a], wherein the first antigen-binding domain or the first means binds a first FGFR2 protein and the second antigen-binding domain or the second means binds a second FGFR2 protein.
[A3] The molecule of [A1], [A1a], or [A2], which is a biparatopic antigen-binding molecule.
[A4] The molecule of any one of [A1] and [A1a] to [A3], wherein the human FGFR2 protein is human FGFR2b protein.
[A5] The molecule of any one of [A1] and [A1a] to [A4], wherein each of the extracellular domains is independently selected from an Immunoglobulin (Ig)-like domain of the FGFR2 protein.
[A6] The molecule of any one of [A1] and [A1a] to [A5], wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR2 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR2 protein, wherein the first domain and the second domain is the same.
[A7] The molecule of any one of [A1] and [A1a] to [A5], wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR2 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR2 protein, wherein the second domain is different from the first domain.
[A8] The molecule of any one of [A1] and [A1a] to [A7], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind an Ig-like domain of the FGFR2 protein, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.
[A9] The molecule of any one of [A5] to [A8], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR2;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR2; and
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR2.
[A10] The molecule of any one of [A5] to [A9], wherein (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
[A11] The molecule of 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.
[A12] The molecule of [A11], wherein the second antigen-binding domain is selected from the group consisting of FFB0464, FFB0341, FFB0396.
[A13] The molecule of [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.
[A14] The molecule of any one of [A3] to [A13], wherein binding of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means to FGFR2b protein leads to agonist activity as follows:
(i) wherein binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR2b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR2b protein in the target cell, in particular 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 binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of 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 FGFR2b protein phosphorylation is indicative of agonist activity.
[A15] The molecule of any one of [A1] to [A14], wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.
[A16] The molecule of 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 which are not in a hinge region.
[A17] The molecule of [A16], wherein the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, optionally wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain, in particular wherein said at least one disulfide bond is formed between the group selected from the following:
(i) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
(ii) an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iii) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iv) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
(v) an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
(vi) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.
[A18] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a 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 originate is present at a position selected from the group consisting of positions 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, according to EU numbering, in the CH1 region.
[A19] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL 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 originate is present at a position selected from the group consisting of positions 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 numbering in the CL region.
[A20] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197 according to EU numbering, and the amino acid residue in the CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128 according to Kabat numbering.
[A21] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a VH region of a first antigen-binding domain and an amino acid residue in a VH 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 originate is present at a position selected from the group consisting of positions 6, 8, 16, 20, 25, 26, 28, 74, and 82b according to Kabat numbering in the VH region.
[A22] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a VHH region of a first antigen-binding domain and an amino acid residue in a VHH 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 originate is present at a position selected from the group consisting of positions 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 numbering in the VHH region.
[A23] The molecule of [A17], wherein said at least one disulfide bond is formed between an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of the second antigen-binding domain, and
wherein, when the VL is of a subclass kappa, the amino acid residue from which the bond between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 21, 27, 58, 77, 100, 105, and 107 according to Kabat numbering in the VL region (subclass kappa); or
wherein, when the VL is of a subclass lambda, the amino acid residue from which the bonds between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 6, 19, 33, and 34 according to Kabat numbering in the VL region (subclass lambda).
[A24] The molecule of [A23], wherein the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain.
[A25] The molecule of [A24], wherein the at least one disulfide bond is formed between the amino acid residue at position 191 in a CH1 region of a first antigen-binding domain (according to Kabat numbering) and the amino acid residue at position 126 in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A26] The molecule of [A25], wherein the at least one disulfide bond is formed between S191C in the CH1 region of a first antigen-binding domain (according to Kabat numbering) and K126C in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A27] The molecule of any one of [A1] and [A1a] to [A26], wherein the molecule comprises an antibody constant region, in particular (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.
[A28] The molecule of any one of [A1] to [A27], wherein the molecule comprises or is an antibody or antibody fragment thereof.
[A29] The molecule of [A28], wherein the antibody fragment is a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody.
[A30] The molecule of [A28] or [A29] wherein the molecule is a monoclonal antibody.
[A31] The molecule of any one of [A29] to [A30], wherein the molecule is human, humanized, or chimeric antibody.
[A32] The molecule of 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 second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.
[A33] The molecule of any one of [A1] and [A1a] to [A32], wherein the molecule comprises more than two antigen-binding domains or more than two means for binding, or wherein the molecule is tri-valent, tetra-valent, or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain, or wherein the molecule comprises a third means for binding the same epitope on the same domain as the first or the second means.
[A34] The molecule of [A33], wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain, or wherein the molecule comprises a fourth means for binding the same epitope on the same domain as the first or the second means but not the same epitope as the third means, optionally
wherein the molecule comprises a third and a fourth antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain, or
wherein the molecule comprises a third means for binding the same epitope on the same domain as the first means, and a fourth means binding the same epitope on the same domain as the second means.
[A35] A pharmaceutical composition comprising the molecule of any one of [A1] and [A1a] to [A34], and a pharmaceutically acceptable carrier.
[A36] An immunoconjugate protein comprising the molecule of any one of [A1] and [A1a] to [A34].
[A37] The molecule of any one of [A1] and [A1a] to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36] for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A38] The molecule or pharmaceutical composition or immunoconjugate for the use of [A37], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A39] The molecule of any one of [A1] and [A1a]to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36] for use in the treatment of any disease, disorder, or condition susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A40] The molecule of any one of [A1] and [A1a] to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36] for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A41] The molecule or pharmaceutical composition or immunoconjugate for the use of [A40], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A42] The molecule or pharmaceutical composition or immunoconjugate for the use of [A41], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A43] Use of the molecule of any one of [A1] and [A1a] to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36], in the manufacture of a medicament for the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A44] A method of treating a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1] and [A1a] to [A34], or the pharmaceutical composition of [A35], or the immunoconjugate of [A36].
[A45] The use of [A43], or the method of [A44], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A46] The use of [A43], or the method of [A44], wherein the condition related to the injury of lung epithelial cells in an individual is one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A47] Use of the molecule of any one of [A1] and [A1a] to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36] in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A48] A method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1] and [A1a] to [A34] or the pharmaceutical composition of [A35] or the immunoconjugate of [A36].
[A49] The use of [A47], or the method of [A48], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A50] The use of [A47], or the method of [A48], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A51] An isolated nucleic acid encoding the molecule of any one of [A1] and [A1a] to [A34] or a plurality of isolated nucleic acids encoding the molecule of any one of [A1] and [A1a] to [A34].
[A52] A host cell comprising the nucleic acid or the plurality of nucleic acids of [A51].
[A53] A method of producing the molecule, or a process for producing the molecule comprising the following steps:
(a) culturing the host cell of [A52], and
(b) recovering the molecule that is produced in step (a).

In aspects and embodiments, the invention relates to any one of [A1-1] and [A1a-1] to [A52-1] below:
[A1-1] An agonistic antigen-binding molecule that binds to a human Fibroblast growth factor receptor 1 (FGFR1) protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first FGFR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein, wherein the first epitope and the second epitope are different from each other.
[A1a-1] An agonistic antigen-binding molecule that binds to a human Fibroblast growth factor receptor 1 (FGFR1) protein, wherein the molecule comprises:
a first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR1 protein; and
a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein,
wherein the first epitope and the second epitope are different from each other.
[A2-1] The molecule of [A1-1] or [A1a-1], wherein the first antigen-binding domain or the first means binds a first FGFR1 protein and the second antigen-binding domain or the second means binds a second FGFR1 protein.
[A3-1] The molecule of [A1-1], [A1a-1], or [A2-1], which is a biparatopic antigen-binding molecule.
[A4-1] The molecule of any one of [A1-1] and [A1a-1] to [A3-1], wherein the human FGFR1 protein is human FGFR1b protein.
[A5-1] The molecule of any one of [A1-1] and [A1a-1] to [A4-1], wherein each of the extracellular domains is independently selected from an Immunoglobulin (Ig)-like domain of the FGFR1 protein.
[A6-1] The molecule of any one of [A1-1] and [A1a-1] to [A5-1], wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR1 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR1 protein, wherein the first domain and the second domain is the same.
[A7-1] The molecule of any one of [A1-1] and [A1a-1] to [A5-1], wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR1 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR1 protein, wherein the second domain is different from the first domain.
[A8-1] The molecule of 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 means and the second means, independently of each other, bind an Ig-like domain of the FGFR1 protein, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.
[A9-1] The molecule of any one of [A5-1] to [A8-1], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR1; and
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR1.
[A10-1] The molecule of any one of [A5-1] to [A9-1], wherein (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR1, or wherein (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
[A11-1] The molecule of 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.
[A12-1] The molecule of [A11-1], wherein the first antigen-binding domain is selected from the group consisting of FFA0029, and FFA0048.
[A13-1] The molecule of any one of [A3-1] to [A12-1], wherein binding of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means to FGFR1b protein leads to agonist activity as follows:
(i) wherein binding of the molecule to FGFR1b protein via a cell-surface expressed FGFR1b in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR1b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR1b protein in the target cell, in particular 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
(ii) wherein binding of the molecule to FGFR1b protein via a cell-surface expressed FGFR1b in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of agonist activity; or
(iv) wherein phosphorylation of the FGFR1b protein is detected in the presence of the molecule in an in vitro cell assay, wherein FGFR1b protein phosphorylation is indicative of agonist activity.
[A14-1] The molecule of 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 a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.
[A15-1] The molecule of 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 which are not in a hinge region.
[A16-1] The molecule of [A15-1], wherein the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, optionally wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain, in particular wherein said at least one disulfide bond is formed between the group selected from the following:
(vii) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
(viii) an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(ix) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(x) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
(xi) an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
(xii) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.
[A17-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a 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 originate is present at a position selected from the group consisting of positions 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, according to EU numbering, in the CH1 region.
[A18-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL 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 originate is present at a position selected from the group consisting of positions 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 numbering in the CL region.
[A19-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197 according to EU numbering, and the amino acid residue in the CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128 according to Kabat numbering.
[A20-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a VH region of a first antigen-binding domain and an amino acid residue in a VH 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 originate is present at a position selected from the group consisting of positions 6, 8, 16, 20, 25, 26, 28, 74, and 82b according to Kabat numbering in the VH region.
[A21-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a VHH region of a first antigen-binding domain and an amino acid residue in a VHH 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 originate is present at a position selected from the group consisting of positions 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 numbering in the VHH region.
[A22-1] The molecule of [A16-1], wherein said at least one disulfide bond is formed between an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of the second antigen-binding domain, and
wherein, when the VL is of a subclass kappa, the amino acid residue from which the bond between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 21, 27, 58, 77, 100, 105, and 107 according to Kabat numbering in the VL region (subclass kappa); or
wherein, when the VL is of a subclass lambda, the amino acid residue from which the bonds between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 6, 19, 33, and 34 according to Kabat numbering in the VL region (subclass lambda).
[A23-1] The molecule of [A22-1], wherein the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain.
[A24-1] The molecule of [A23-1], wherein the at least one disulfide bond is formed between the amino acid residue at position 191 in a CH1 region of a first antigen-binding domain (according to Kabat numbering) and the amino acid residue at position 126 in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A25-1] The molecule of [A24-1], wherein the at least one disulfide bond is formed between S191C in the CH1 region of a first antigen-binding domain (according to Kabat numbering) and K126C in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A26-1] The molecule of any one of [A1-1] and [A1a-1] to [A25-1], wherein the molecule comprises an antibody constant region, in particular (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.
[A27-1] The molecule of any one of [A1-1] to [A26-1], wherein the molecule comprises or is an antibody or antibody fragment thereof.
[A28-1] The molecule of [A27-1], wherein the antibody fragment is a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody.
[A29-1] The molecule of [A27-1] or [A28-1] wherein the molecule is a monoclonal antibody.
[A30-1] The molecule of any one of [A28-1] to [A29-1], wherein the molecule is human, humanized, or chimeric antibody.
[A31-1] The molecule of any one of [A1-1] to [A30-1], 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 second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.
[A32-1] The molecule of 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 means for binding, or wherein the molecule is tri-valent, tetra-valent, or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain, or wherein the molecule comprises a third means for binding the same epitope on the same domain as the first or the second means.
[A33-1] The molecule of [A32-1], wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain, or wherein the molecule comprises a fourth means for binding the same epitope on the same domain as the first or the second means but not the same epitope as the third means, optionally
wherein the molecule comprises a third and a fourth antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain, or
wherein the molecule comprises a third means for binding the same epitope on the same domain as the first means, and a fourth means binding the same epitope on the same domain as the second means.
[A34-1] A pharmaceutical composition comprising the molecule of any one of [A1-1] and [A1a-1] to [A33-1], and a pharmaceutically acceptable carrier.
[A35-1] An immunoconjugate protein comprising the molecule of any one of [A1-1] and [A1a-1] to [A33-1].
[A36-1] The molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1] for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A37-1] The molecule or pharmaceutical composition or immunoconjugate for the use of [A36-1], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A38-1] The molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1] for use in the treatment of any disease, disorder, or condition susceptible of being improved or prevented by one of the following selected from the group consisting of:
(iv) an increase or enhancement in FGFR1b signaling;
(v) an increase in the proliferation of lung epithelial cells; and
(vi) an increase in the proliferation alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A39-1] The molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1] for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A40-1] The molecule or pharmaceutical composition or immunoconjugate for the use of [A39-1], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A41-1] The molecule or pharmaceutical composition or immunoconjugate for the use of [A40-1], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A42-1] Use of the molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1], in the manufacture of a medicament for the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A43-1] A method of treating a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1-1] and [A1a-1] to [A33-1], or the pharmaceutical composition of [A34-1], or the immunoconjugate of [A35-1].
[A44-1] The use of [A42-1], or the method of [A43-1], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A45-1] The use of [A42-1], or the method of [A43-1], wherein the condition related to the injury of lung epithelial cells in an individual is one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR1b signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A46-1] Use of the molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1] in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A47-1] A method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1-1] and [A1a-1] to [A33-1] or the pharmaceutical composition of [A34-1] or the immunoconjugate of [A35-1].
[A48-1] The use of [A46-1], or the method of [A47-1], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A49-1] The use of [A46-1], or the method of [A47-1], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A50-1] An isolated nucleic acid encoding the molecule of any one of [A1-1] and [A1a-1] to [A33-1] or a plurality of isolated nucleic acids encoding the molecule of any one of [A1-1] and [A1a-1] to [A33-1].
[A51-1] A host cell comprising the nucleic acid or the plurality of nucleic acids of [A50-1].
[A52-1] A method of producing the molecule, or a process for producing the molecule comprising the following steps:
(a) culturing the host cell of [A51-1], and
(b) recovering the molecule that is produced in step (a).

In aspects and embodiments, the invention relates to any one of [A1-2] and [A1a-2] to [A52-2] below:
[A1-2] An agonistic antigen-binding molecule that binds to a human discoidin domain receptor1 (DDR1) protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first DDR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein, wherein the first epitope and the second epitope are different from each other.
[A1a-2] An agonistic antigen-binding molecule that binds to a human discoidin domain receptor1 (DDR1) protein, wherein the molecule comprises:
a first means for binding a first epitope of an extracellular domain (ECD) of a first DDR1 protein; and
a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein,
wherein the first epitope and the second epitope are different from each other.
[A2-2] The molecule of [A1-2] or [A1a-2], wherein the first antigen-binding domain or the first means binds a first DDR1 protein and the second antigen-binding domain or the second means binds a second DDR1 protein.
[A3-2] The molecule of [A1-2], [A1a-2], or [A2-2], which is a biparatopic antigen-binding molecule.
[A4-2] The molecule of any one of [A1-2] and [A1a-2] to [A3-2], wherein the human DDR1 protein is human DDR1a protein.
[A5-2] The molecule of any one of [A1-2] and [A1a-2] to [A4-2], wherein each of the extracellular domains is independently selected from a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 protein.
[A6-2] The molecule of any one of [A1-2] and [A1a-2] to [A5-2], wherein the first antigen-binding domain or the first means binds to a first domain of the DDR1 protein and the second antigen-binding domain or the second means binds to a second domain of the DDR1 protein, wherein the first domain and the second domain is the same.
[A7-2] The molecule of any one of [A1-2] and [A1a-2] to [A5-2], wherein the first antigen-binding domain or the first means binds to a first domain of the DDR1 protein and the second antigen-binding domain or the second means binds to a second domain of the DDR1 protein, wherein the second domain is different from the first domain.
[A8-2] The molecule of 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 means and the second means, independently of each other, bind a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 protein.
[A9-2] The molecule of any one of [A5-2] to [A8-2], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS domain of DDR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS-like domain of DDR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and EJXM region of DDR1;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and DS-like domain of DDR1;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and EJXM region of DDR1; and
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the EJXM region and EJXM region of DDR1.
[A10-2] The molecule of any one of [A5-2] to [A9-2], wherein (i) the first antigen-binding domain or the first means binds the DS domain of DDR1, or wherein (ii) the first antigen-binding domain or the first means binds specifically to DS domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with DS-Like domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with EJXM region of DDR1.
[A11-2] The molecule of any one of [A1-2] to [A10-2], wherein the second antigen-binding domain is selected from the group consisting of DAA0007, DAA0012, and DAA 0024.
[A12-2] The molecule of [A11-2], wherein the first antigen-binding domain is selected from the group consisting of DAA0011, and DAA0045.
[A13-2] The molecule of any one of [A3-2] to [A12-2], wherein binding of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means to DDR1 protein leads to agonist activity as follows:
(i) wherein binding of the molecule to DDR1 protein via a cell-surface expressed DDR1 in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of DDR1 by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed DDR1 protein in the target cell, in particular wherein the intracellular protein downstream of DDR1 is selected from the group consisting of: SHP-2, FAK, NF-kappa B, Cdc42, MAPK1, AP1, Ras/Raf/ERK, PI3K/AKT, STAT1/3/5; or
(ii) wherein binding of the molecule to DDR1 protein via a cell-surface expressed DDR1 in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of agonist activity; or
(iv) wherein phosphorylation of the DDR1 protein is detected in the presence of the molecule in an in vitro cell assay, wherein DDR1 protein phosphorylation is indicative of agonist activity.
[A14-2] The molecule of 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 a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.
[A15-2] The molecule of 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 which are not in a hinge region.
[A16-2] The molecule of [A15-2], wherein the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, optionally wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain, in particular wherein said at least one disulfide bond is formed between the group selected from the following:
(xiii) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
(xiv) an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(xv) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(xvi) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
(xvii) an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
(xviii) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.
[A17-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a 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 originate is present at a position selected from the group consisting of positions 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, according to EU numbering, in the CH1 region.
[A18-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL 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 originate is present at a position selected from the group consisting of positions 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 numbering in the CL region.
[A19-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197 according to EU numbering, and the amino acid residue in the CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128 according to Kabat numbering.
[A20-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a VH region of a first antigen-binding domain and an amino acid residue in a VH 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 originate is present at a position selected from the group consisting of positions 6, 8, 16, 20, 25, 26, 28, 74, and 82b according to Kabat numbering in the VH region.
[A21-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a VHH region of a first antigen-binding domain and an amino acid residue in a VHH 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 originate is present at a position selected from the group consisting of positions 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 numbering in the VHH region.
[A22-2] The molecule of [A16-2], wherein said at least one disulfide bond is formed between an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of the second antigen-binding domain, and
wherein, when the VL is of a subclass kappa, the amino acid residue from which the bond between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 21, 27, 58, 77, 100, 105, and 107 according to Kabat numbering in the VL region (subclass kappa); or
wherein, when the VL is of a subclass lambda, the amino acid residue from which the bonds between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 6, 19, 33, and 34 according to Kabat numbering in the VL region (subclass lambda).
[A23-2] The molecule of [A22-2], wherein the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain.
[A24-2] The molecule of [A23-2], wherein the at least one disulfide bond is formed between the amino acid residue at position 191 in a CH1 region of a first antigen-binding domain (according to Kabat numbering) and the amino acid residue at position 126 in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A25-2] The molecule of [A24-2], wherein the at least one disulfide bond is formed between S191C in the CH1 region of a first antigen-binding domain (according to Kabat numbering) and K126C in a CL region of a second antigen-binding domain (according to Kabat numbering).
[A26-2] The molecule of any one of [A1-2] and [A1a-2] to [A25-2], wherein the molecule comprises an antibody constant region, in particular (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.
[A27-2] The molecule of any one of [A1-2] to [A26-2], wherein the molecule comprises or is an antibody or antibody fragment thereof.
[A28-2] The molecule of [A27-2], wherein the antibody fragment is a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody.
[A29-2] The molecule of [A27-2] or [A28-2] wherein the molecule is a monoclonal antibody.
[A30-2] The molecule of any one of [A28-2] to [A29-2], wherein the molecule is human, humanized, or chimeric antibody.
[A31-2] The molecule of any one of [A1-2] to [A30-2], 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 second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.
[A32-2] The molecule of 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 means for binding, or wherein the molecule is tri-valent, tetra-valent, or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain, or wherein the molecule comprises a third means for binding the same epitope on the same domain as the first or the second means.
[A33-2] The molecule of [A32-2], wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain, or wherein the molecule comprises a fourth means for binding the same epitope on the same domain as the first or the second means but not the same epitope as the third means, optionally
wherein the molecule comprises a third and a fourth antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain, or
wherein the molecule comprises a third means for binding the same epitope on the same domain as the first means, and a fourth means binding the same epitope on the same domain as the second means.
[A34-2] A pharmaceutical composition comprising the molecule of any one of [A1-2] and [A1a-2] to [A33-2], and a pharmaceutically acceptable carrier.
[A35-2] An immunoconjugate protein comprising the molecule of any one of [A1-2] and [A1a-2] to [A33-2].
[A36-2] The molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2] for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A37-2] The molecule or pharmaceutical composition or immunoconjugate for the use of [A36-2], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A38-2] The molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2] for use in the treatment of any disease, disorder, or condition susceptible of being improved or prevented by one of the following selected from the group consisting of:
(vii) an increase or enhancement in DDR1 signaling;
(viii) an increase in the proliferation of lung epithelial cells; and
(ix) an increase in the proliferation alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A39-2] The molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2] for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A40-2] The molecule or pharmaceutical composition or immunoconjugate for the use of [A39-2], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A41-2] The molecule or pharmaceutical composition or immunoconjugate for the use of [A40-2], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A42-2] Use of the molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2], in the manufacture of a medicament for the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[A43-2] A method of treating a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1-2] and [A1a-2] to [A33-2], or the pharmaceutical composition of [A34-2], or the immunoconjugate of [A35-2].
[A44-2] The use of [A42-2], or the method of [A43-2], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[A45-2] The use of [A42-2], or the method of [A43-2], wherein the condition related to the injury of lung epithelial cells in an individual is one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in DDR1 signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[A46-2] Use of the molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2] in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[A47-2] A method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of [A1-2] and [A1a-2] to [A33-2] or the pharmaceutical composition of [A34-2] or the immunoconjugate of [A35-2].
[A48-2] The use of [A46-2], or the method of [A47-2], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[A49-2] The use of [A46-2], or the method of [A47-2], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[A50-2] An isolated nucleic acid encoding the molecule of any one of [A1-2] and [A1a-2] to [A33-2] or a plurality of isolated nucleic acids encoding the molecule of any one of [A1-2] and [A1a-2] to [A33-2].
[A51-2] A host cell comprising the nucleic acid or the plurality of nucleic acids of [A50-2].
[A52-2] A method of producing the molecule, or a process for producing the molecule comprising the following steps:
(a) culturing the host cell of [A51-2], and
(b) recovering the molecule that is produced in step (a).

In further aspects and embodiments, the invention relates 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 [51-2], [51a-2], [52-2], [52a-2], [53-2] to [61-2], [62] to [104] below:
[1] An agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, and wherein the molecule is an agonist of the RTK protein.
[2] The molecule of [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 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.
[2a] The molecule of [1], wherein the molecule comprises a first means for binding a first epitope of a first RTK protein and a second means for binding a second epitope of the first 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.
[3] The molecule of [2] or [2a], wherein the first antigen-binding domain or the first means binds a first RTK protein and the second antigen-binding domain or the second means binds a second RTK protein, optionally wherein the first and the second RTK proteins form a dimer.
[4] The molecule of any one of [2], [2a] and [3], wherein binding of the first and second antigen-binding domains or binding of the first and second means induces dimerization of the intracellular tyrosine kinase domains of the RTK protein.
[5] The molecule of [4], wherein dimerization is detected in the presence of the molecule in an in vitro cell assay, and wherein dimerization is indicative of agonist activity.
[6] The molecule of any one of [2] to [3], wherein binding of the first and second antigen-binding domains or binding of the first and second means activates phosphorylation of the RTK protein.
[7] The molecule of [6], wherein phosphorylation of the RTK protein is detected in the presence of the molecule in an in vitro cell assay, wherein RTK protein phosphorylation is indicative of agonist activity.
[8] The molecule of any one of [2] to [7], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means each bind to an extracellular domain (ECD) of the RTK protein.
[9] The molecule of [8], wherein the extracellular domain is independently selected from an Immunoglobulin (Ig)-like domain of the ECD of the RTK protein for each of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means.
[10] The molecule of any one of [2] to [9], wherein the first antigen-binding domain or the first means binds to a first domain and the second antigen-binding domain or the second means binds to a second domain, wherein the first domain and the second domain is the same.
[11] The molecule of any one of [2] to [9], wherein the first antigen binding-domain or the first means binds to a first domain and the second antigen-binding domain or the second means binds to a second domain, wherein the second domain is different from the first domain.
[12] The molecule of [10], wherein the first and the second epitope is the same.
[13] The molecule of [10], wherein the first epitope and the second epitope is different
[14] The molecule of [11], wherein the first epitope and the second epitope is different.
[15] The molecule of [13] or [14], wherein the molecule binds biparatopically.
[16] The molecule of any one of [1] to [15], wherein the RTK protein is selected from one of the Fibroblast Growth Factor Receptor (FGFR) family consisting of the members in the group consisting of: FGFR1, FGFR2, FGFR3 and FGFR4 or one of the Discoidin Domain Receptor (DDR) family consisting of the members in the group consisting of: DDR1 and DDR2.
[17] The molecule of any one of [1] to [16], wherein the RTK protein is Fibroblast growth factor receptor 2 (FGFR2), preferably the RTK protein is FGFR2b.
[18] The molecule of [17], wherein binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR2b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR2b protein in the target cell.
[19] The molecule of [18], wherein the intracellular protein downstream of FGFR2b is selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1/pERK2, and pMAPK3/pERK1.
[20] The molecule of [19], wherein the expression of said intracellular protein is measured using an immunoassay and normalized to the total number of FGFR2b protein and total number of AKT, PLCG1, FRS2, PAK4, RAS, MAPK1/ERK2, and MAPK3/ERK1 respectively.
[21] The molecule of [17], wherein binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity.
[22] The molecule of [21], wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells.
[23] The molecule of [21], wherein the induction in proliferation of the target cell is measured by an increase in BrdU expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the BrdU expression is directly proportional to the concentration of viable target cells.
[24] The molecule of any one of [18] to [23], wherein the target cell is a lung epithelial cell.
[25] The molecule of [24], wherein the lung epithelial cell is selected from alveolar type I (AT1) or alveolar type II (AT2) cells, in particular alveolar type II (AT2) cells.
[26] The molecule of any one of [17] to [25], wherein the first antigen-binding domain or the first means binds a first epitope residing in a domain of an extracellular domain (ECD) of FGFR2, or wherein the first antigen-binding domain or the first means binds a first epitope of the extracellular portion of FGFR2.
[27] The molecule of any one of [17] to [26], wherein the first antigen-binding domain or the first means binds an Ig-like domain of FGFR2, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.
[28] The molecule of [27], wherein the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR2 and the second antigen-binding domain or the second means binds a domain of FGFR2 other than the IgIIIb domain.
[29] The molecule of [27], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgI domain of FGFR2;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgII domain of FGFR2;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIII domain of FGFR2;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIb domain of FGFR2;
(v) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIc domain of FGFR2;
(vi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgII domain of FGFR2;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIII domain of FGFR2;
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIb domain of FGFR2;
(ix) wherein the first antigen-binding domain or the first means and the second antigen-binding domain or the second means, independently of each other, bind the IgII and IgIIIc domain of FGFR2;
(x) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIII domain of FGFR2;
(xi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIb domain of FGFR2;
(xii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIc domain of FGFR2;
(xiii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2; and
(xiv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIc domain of FGFR2.
[30] The molecule of any one of [27] to [29], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR2;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR2; and
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR2.
[31] The molecule of any one of [27] to [30], wherein (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
[32] The molecule of any one of [27] to [30], wherein (i) the first antigen-binding domain or the first means binds the IgIII domain of FGFR2.
[33] The molecule of any one of [27] to [32], wherein the first antigen-binding domain or the first means binds to a continuous or discontinuous epitope.
[34] The molecule of [33], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECDs.
[35] The molecule of any one of [17] to [34], wherein the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the FGFR2 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of FGFR2.
[36] The molecule of any one of [17] to [35], wherein the second antigen-binding domain or the second means binds an Ig-like domain of FGFR2, selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR2.
[37] The molecule of [36], wherein the (i) second antigen-binding domain or the second means binds to IgII of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgII of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI, and IgIII of FGFR2.
[38] The molecule of [36], wherein the (i) second antigen-binding domain or the second means binds to IgI of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgI of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgII, and IgIII of FGFR2.
[39] The molecule of [36], wherein the (i) second antigen-binding domain or the second means binds to IgIIIb of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIb of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
[40] The molecule of [36], wherein the (i) second antigen-binding domain or the second means binds to IgIIIc of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIc of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIb of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
[41] The molecule of [36], wherein the (i) second antigen-binding domain or the second means binds to IgIII of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIII of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI, and IgII of FGFR2.
[42] The molecule of any one of [17] to [41], wherein the second antigen-binding domain or the second means binds to a continuous or discontinuous epitope.
[43] The molecule of [42], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECD.
[44] The molecule of any one of [17] to [43], wherein the first epitope of the first antigen-binding domain or the first means is different from the second epitope of the second antigen-binding domain or the second means, or wherein the first epitope of the first antigen-binding domain or the first means and the second epitope of the second antigen-binding domain or the second means are non-overlapping.
[45] The molecule of any one of [27] to [44], wherein the first antigen-binding domain or the first means binds IgIII of FGFR2, and the second antigen-binding domain or the second means binds IgIII, or IgII, or IgI of FGFR2.
[46] The molecule of any one of [27] to [45], wherein the first antigen-binding domain or the first means binds IgIIIb of FGFR2, and the second antigen-binding domain or the second means binds IgIII, or IgIIIb, or IgII, or IgI of FGFR2.
[47] The molecule of [45] or [46], wherein the molecule binds biparatopically to IgIIIb of FGFR2 and IgIII of FGFR2.
[48] The molecule of [46], wherein the molecule binds biparatopically to IgIIIb of FGFR2 and IgIIIb of FGFR2.
[49] The molecule of [46], wherein the molecule binds biparatopically to IgIIIb of FGFR2 and IgII of FGFR2.
[50] The molecule of [46], wherein the molecule binds biparatopically to IgIIIb of FGFR2 and IgI of FGFR2.
[51] The molecule of any one of [1] to [50], wherein the RTK protein is Fibroblast growth factor receptor2 (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 or a second FGFR2 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[51a] The molecule of any one of [1] to [50], wherein the RTK protein is Fibroblast growth factor receptor2 (FGFR2), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second FGFR2 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[52] The molecule of any one of [1] to [50], wherein the RTK protein is Fibroblast growth factor receptor2 (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 or a second FGFR2 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[52a] The molecule of any one of [1] to [50], wherein the RTK protein is Fibroblast growth factor receptor2 (FGFR2), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second FGFR2 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[53] The molecule of any one of [51], [51a], [52], and [52a], wherein the first antigen-binding domain or the first means binds a first FGFR2 protein and the second antigen-binding domain or the second means binds a second FGFR2 protein, optionally wherein the first and the second RTK proteins form a dimer.
[54] The molecule of 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.
[55] The molecule of 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.
[56] The molecule of [54] or [55], wherein the first antigen binding domain competes for binding FGFR2 with an antigen binding domain comprising any one of (a1) to (a14), or wherein the first antigen-binding domain comprises any one of (a1) to (a14), wherein (a1) to (a14) corresponds to (a1) to (a14) recited in [0164].
[57] The molecule of any one of [54] to [56], wherein the first antigen binding domain competes for binding FGFR2 with an antigen binding domain comprising any one of (b1) to (b14), or wherein the first antigen-binding domain comprises any one of (b1) to (b14), wherein (b1) to (b14) corresponds to (b1) to (b14) recited in [0165].
[58] The molecule of any one of [54] to [57], wherein the second antigen binding domain competes for binding FGFR2 with an antigen binding domain comprising any one of (c1) to (c9), or wherein the second antigen-binding domain comprises any one of (c1) to (c9), wherein (c1) to (c9) corresponds to (c1) to (c9) recited in [0166].
[59] The molecule of any one of [54] to [58], wherein the second antigen binding domain competes for binding FGFR2 with an antigen binding domain comprising any one of (d1) to (d9), or wherein the second antigen-binding domain comprises any one of (d1) to (d9), wherein (d1) to (d9) corresponds to (d1) to (d9) recited in [0167].
[60] The molecule of any one of [54] to [59], wherein the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to FGFR2 with an antigen-binding domain comprising any one of (e1) to (e21), or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (e1) to (e21), wherein (e1) to (e21) corresponds to (e1) to (e21) recited in [0168].
[61] The molecule 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 compete for binding to FGFR2 with an antigen-binding domain comprising any one of (f1) to (f116), 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) corresponds to (f1) to (f116) recited in [0169].
[17-1] The molecule of 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.
[18-1] The molecule of [17-1], wherein binding of the molecule to FGFR1b protein via a cell-surface expressed FGFR1b in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR1b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR1b protein in the target cell.
[19-1] The molecule of [18-1], wherein the intracellular protein downstream of FGFR1b is selected from the group consisting of: pAKT, pPLCG1, pFRS2, pPAK4, pRAS, pMAPK1/pERK2, and pMAPK3/pERK1.
[20-1] The molecule of [19-1], wherein the expression of said intracellular protein is measured using an immunoassay and normalized to the total number of FGFR1b protein and total number of AKT, PLCG1, FRS2, PAK4, RAS, MAPK1/ERK2, and MAPK3/ERK1 respectively.
[21-1] The molecule of [17-1], wherein binding of the molecule to FGFR1b protein via a cell-surface expressed FGFR1b in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity.
[22-1] The molecule of [21-1], wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells.
[23-1] The molecule of [21-1], wherein the induction in proliferation of the target cell is measured by an increase in BrdU expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the BrdU expression is directly proportional to the concentration of viable target cells.
[24-1] The molecule of any one of [18-1] to [23-1], wherein the target cell is a lung epithelial cell.
[25-1] The molecule of [24-1], wherein the lung epithelial cell is selected from alveolar type I (AT1) or alveolar type II (AT2) cells, in particular alveolar type II (AT2) cells.
[26-1] The molecule of any one of [17-1] to [25-1], wherein the first antigen-binding domain or the first means binds a first epitope residing in a domain of an extracellular domain (ECD) of FGFR1, or wherein the first antigen-binding domain or the first means binds a first epitope of the extracellular portion of FGFR1.
[27-1] The molecule of any one of [17-1] to [26-1], wherein the first antigen-binding domain or the first means binds an Ig-like domain of FGFR1, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.
[28-1] The molecule of [27-1], wherein the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR1 and the second antigen-binding domain or the second means binds a domain of FGFR1 other than the IgIIIb domain.
[29-1] The molecule of [27-1], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgI domain of FGFR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgII domain of FGFR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIII domain of FGFR1;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIb domain of FGFR1;
(v) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIc domain of FGFR1;
(vi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgII domain of FGFR1;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIII domain of FGFR1;
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIb domain of FGFR1;
(ix) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIc domain of FGFR1;
(x) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIII domain of FGFR1;
(xi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIb domain of FGFR1;
(xii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIc domain of FGFR1;
(xiii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR1; and
(xiv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIc domain of FGFR1.
[30-1] The molecule of any one of [27-1] to [29-1], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR1; and
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR1.
[31-1] The molecule of any one of [27-1] to [30-1], wherein (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR1, or wherein (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
[32-1] The molecule of any one of [27-1] to [30-1], wherein (i) the first antigen-binding domain or the first means binds the IgIII domain of FGFR1.
[33-1] The molecule of any one of [27-1] to [32-1], wherein the first antigen-binding domain or the first means binds to a continuous or discontinuous epitope.
[34-1] The molecule of [33-1], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECDs.
[35-1] The molecule of any one of [17-1] to [34-1], wherein the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the FGFR1 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of FGFR1.
[36-1] The molecule of any one of [17-1] to [35-1], wherein the second antigen-binding domain or the second means binds an Ig-like domain of FGFR1, selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR1.
[37-1] The molecule of [36-1], wherein the (i) second antigen-binding domain or the second means binds to IgII of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgII of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI, and IgIII of FGFR1.
[38-1] The molecule of [36-1], wherein the (i) second antigen-binding domain or the second means binds to IgI of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgI of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgII, and IgIII of FGFR1.
[39-1] The molecule of [36-1], wherein the (i) second antigen-binding domain or the second means binds to IgIIIb of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIb of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
[40-1] The molecule of [36-1], wherein the (i) second antigen-binding domain or the second means binds to IgIIIc of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIc of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIb of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
[41-1] The molecule of [36-1], wherein the (i) second antigen-binding domain or the second means binds to IgIII of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIII of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI, and IgII of FGFR1.
[42-1] The molecule of any one of [17-1] to [41-1], wherein the second antigen-binding domain or the second means binds to a continuous or discontinuous epitope.
[43-1] The molecule of [42-1], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECD.
[44-1] The molecule of any one of [17-1] to [43-1], wherein the first epitope of the first antigen-binding domain or the first means is different from the second epitope of the second antigen-binding domain or the second means, or wherein the first epitope of the first antigen-binding domain or the first means and the second epitope of the second antigen-binding domain or the second means are non-overlapping.
[45-1] The molecule of any one of [27-1] to [44-1], wherein the first antigen-binding domain or the first means binds IgIII of FGFR1, and the second antigen-binding domain or the second means binds IgIII, or IgII, or IgI of FGFR1.
[46-1] The molecule of any one of [27-1] to [45-1], wherein the first antigen-binding domain or the first means binds IgIIIb of FGFR1, and the second antigen-binding domain or the second means binds IgIII, or IgIIIb, or IgII, or IgI of FGFR1.
[47-1] The molecule of [45-1] or [46-1], wherein the molecule binds biparatopically to IgIIIb of FGFR1 and IgIII of FGFR1.
[48-1] The molecule of [46-1], wherein the molecule binds biparatopically to IgIIIb of FGFR1 and IgIIIb of FGFR1.
[49-1] The molecule of [46-1], wherein the molecule binds biparatopically to IgIIIb of FGFR1 and IgII of FGFR1.
[50-1] The molecule of [46-1], wherein the molecule binds biparatopically to IgIIIb of FGFR1 and IgI of FGFR1.
[51-1] The molecule of any one of [1] to [16] and [17-1] to [50-1], wherein the RTK protein is Fibroblast growth factor receptor1 (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 or a second FGFR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[51a-1] The molecule of any one of [1], [2], [2a], and [3] to [16] and [17-1] to [50-1], wherein the RTK protein is Fibroblast growth factor receptor1 (FGFR1), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second FGFR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[52-1] The molecule of any one of [1] to [16] and [17-1] to [50-1], wherein the RTK protein is Fibroblast growth factor receptor1 (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 or a second FGFR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[52a-1] The molecule of any one of [1], [2], [2a], and [3] to [16] and [17-1] to [50-1], wherein the RTK protein is Fibroblast growth factor receptor1 (FGFR1), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second FGFR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[53-1] The molecule of any one of [51-1], [51a-1], [52-1], and [52a-1], wherein the first antigen-binding domain or the first means binds a first FGFR1 protein and the second antigen-binding domain or the second means binds a second FGFR1 protein, optionally wherein the first and the second RTK proteins form a dimer.
[54-1] The molecule of 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.
[55-1] The molecule of any one of [45-1] to [53-1], wherein the first antigen-binding domaini is selected from the group consisting of FFA0029, and FFA0048.
[56-1] The molecule of [54-1] or [55-1], wherein the first antigen binding domain competes for binding FGFR1 with an antigen binding domain comprising any one of (h1) to (h2), or wherein the first antigen-binding domain comprises any one of (h1) to (h2), wherein (h1) to (h2) corresponds to (h1) to (h2) recited in [0219].
[57-1] The molecule of any one of [54-1] to [56-1], wherein the first antigen binding domaini competes for binding FGFR1 with an antigen binding domain comprising any one of (i1) to (i2), or wherein the first antigen-binding domain comprises any one of (i1) to (i2), wherein (i2) to (i2) corresponds to (i1) to (i2) recited in [0220].
[58-1] The molecule of any one of [54-1] to [57-1], wherein the second antigen binding domain competes for binding FGFR1 with an antigen binding domain comprising any one of (j1) to (j8), or wherein the second antigen-binding domain comprises any one of (j1) to (j8), wherein (j1) to (j8) corresponds to (j1) to (j8) recited in [0221].
[59-1] The molecule of any one of [54-1] to [58-1], wherein the second antigen binding domain competes for binding FGFR1 with an antigen binding domain comprising any one of (k1) to (k8), or wherein the second antigen-binding domain comprises any one of (k1) to (k8), wherein (k1) to (k8) corresponds to (k1) to (k8) recited in [0222].
[60-1] The molecule of any one of [54-1] to [59-1], wherein the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to FGFR1 with an antigen-binding domain comprising any one of (l1) to (l10), or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (l1) to (l10), wherein (l1) to (l10) corresponds to (l1) to (l10) recited in [0223].
[61-1] The molecule 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 compete for binding to FGFR1 with an antigen-binding domain comprising any one of (m1) to (m16), 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) corresponds to (m1) to (m16) recited in [0224].
[17-2] The molecule of any one of [1], [2], [2a], and [3] to [16], wherein the RTK protein is discoidin domain receptor1 (DDR1).
[18-2] The molecule of [17-2], wherein binding of the molecule to DDR1 protein via a cell-surface expressed DDR1 in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of DDR1 by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed DDR1 protein in the target cell.
[19-2] The molecule of [18-2], wherein the intracellular protein downstream of DDR1 is selected from the group consisting of: SHP-2, FAK, NF-kappa B, Cdc42, MAPK1, AP1, Ras/Raf/ERK, PI3K/AKT, STAT1/3/5.
[20-2] The molecule of [19-2], wherein the expression of said intracellular protein is measured using an immunoassay and normalized to the total number of DDR1 protein and total number of SHP-2, FAK, NF-kappa B, Cdc42, MAPK1, AP1, Ras/Raf/ERK, PI3K/AKT, STAT1/3/5 respectively.
[21-2] The molecule of [17-2], wherein binding of the molecule to DDR1 protein via a cell-surface expressed DDR1 in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity.
[22-2] The molecule of [21-2], wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells.
[23-2] The molecule of [21-2], wherein the induction in proliferation of the target cell is measured by an increase in BrdU expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the BrdU expression is directly proportional to the concentration of viable target cells.
[24-2] The molecule of any one of [18-2] to [23-2], wherein the target cell is a lung epithelial cell.
[25-2] The molecule of [24-2], wherein the lung epithelial cell is selected from alveolar type I (AT1) or alveolar type II (AT2) cells, in particular alveolar type II (AT2) cells.
[26-2] The molecule of any one of [17-2] to [25-2], wherein the first antigen-binding domain or the first means binds a first epitope residing in a domain of an extracellular domain (ECD) of DDR1, or wherein the first antigen-binding domain or the first means binds a first epitope of the extracellular portion of DDR1.
[27-2] The molecule of any one of [17-2] to [26-2], wherein the first antigen-binding domain or the first means binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1.
[28-2] The molecule of [27-2], wherein the first antigen-binding domain or the first means binds the DS domain of DDR1 and the second antigen-binding domain or the second means binds a domain or region of DDR1 other than the DS domain.
[29-2] The molecule of [27-2], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS domain of DDR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS-like domain of DDR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and EJXM region of DDR1;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and DS-like domain of DDR1;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and EJXM region of DDR1; and
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the EJXM region and EJXM region of DDR1.
[30-2] The molecule of any one of [27-2] to [29-2], wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS domain of DDR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS-like domain of DDR1; and
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and EJXM region of DDR1.
[31-2] The molecule of any one of [27-2] to [30-2], wherein (i) the first antigen-binding domain or the first means binds the DS domain of DDR1, or wherein (ii) the first antigen-binding domain or the first means binds specifically to the DS domain of DDR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS-like domain of DDR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with the EJXM region of DDR1.
[32-2] The molecule of any one of [27-2] to [30-2], wherein (i) the first antigen-binding domain or the first means binds the DS domain of DDR1.
[33-2] The molecule of any one of [27-2] to [32-2], wherein the first antigen-binding domain or the first means binds to a continuous or discontinuous epitope.
[34-2] The molecule of [33-2], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECDs.
[35-2] The molecule of any one of [17-2] to [34-2], wherein the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the DDR1 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of DDR1.
[36-2] The molecule of any one of [17-2] to [35-2], wherein the second antigen-binding domain or the second means binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1.
[37-2] The molecule of [36-2], wherein the (i) second antigen-binding domain or the second means binds to the DS domain of DDR1, or (ii) the second antigen-binding domain or the second means binds specifically to the DS domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS-like domain and the EJXM region of DDR1.
[38-2] The molecule of [36-2], wherein the (i) second antigen-binding domain or the second means binds to the DS-like domain and/or the EJXM region of DDR1, or (ii) the second antigen-binding domain or the second means binds specifically to the DS-like domain and/or the EJXM region of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS domain of DDR1.
[39-2] The molecule of any one of [17-2] to [38-2], wherein the second antigen-binding domain or the second means binds to a continuous or discontinuous epitope.
[40-2] The molecule of [39-2], wherein the continuous or discontinuous epitope resides in one ECD, or in two or three ECD.
[41-2] The molecule of any one of [17-2] to [40-2], wherein the first epitope of the first antigen-binding domain or the first means is different from the second epitope of the second antigen-binding domain or the second means, or wherein the first epitope of the first antigen-binding domain or the first means and the second epitope of the second antigen-binding domain or the second means are non-overlapping.
[42-2] The molecule of any one of [27-2] to [41-2], wherein the first antigen-binding domain or the first means binds the DS domain of DDR1, and the second antigen-binding domain or the second means binds the DS domain or the DS-like domain and/or the EJXM region of DDR1.
[43-2] The molecule of any one of [27-2] to [45-2], wherein the first antigen-binding domain or the first means binds the DS-like domain and/or the EJXM region of DDR1, and the second antigen-binding domain or the second means binds the DS-like domain and/or the EJXM region of DDR1.
[44-2] The molecule of [42-2], wherein the molecule binds biparatopically to the DS domain of DDR1 and the DS domain of DDR1.
[45-2] The molecule of [42-2], wherein the molecule binds biparatopically to the DS domain of DDR1 and the DS-like domain and/or the EJXM region of DDR1.
[46-2] The molecule of [43-2], wherein the molecule binds biparatopically to the DS-like domain and/or the EJXM region of DDR1 and the DS-like domain and/or the EJXM region of DDR1.
[47-2] The molecule of any one of [1] to [16] and [17-2] to [46-2], wherein the RTK protein is Discoidin domain receptor1 (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 or a second DDR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[47a-2] The molecule of any one of [1], [2], [2a], and [3] to [16] and [17-2] to [46-2], wherein the RTK protein is Discoidin domain receptor1 (DDR1), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second DDR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is the same.
[48-2] The molecule of any one of [1] to [16] and [17-2] to [46-2], wherein the RTK protein is Discoidin domain receptor1 (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 or a second DDR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[48a-2] The molecule of any one of [1], [2], [2a], and [3] to [16] and [17-2] to [46-2], wherein the RTK protein is Discoidin domain receptor1 (DDR1), and the molecule comprises a first means for binding a first epitope and a second means 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 or a second DDR1 protein, wherein the first and the second domain is the same or different, and wherein the first epitope and the second epitope is different.
[49-2] The molecule of any one of [47-2], [47a-2], [48-2], and [48a-2], wherein the first antigen-binding domain or the first means binds a first DDR1 protein and the second antigen-binding domain or the second means binds a second DDR1 protein, optionally wherein the first and the second RTK proteins form a dimer.
[50-2] The molecule of 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.
[51-2] The molecule of any one of [42-2] to [49-2], wherein the first antigen-binding domain is selected from the group consisting of DAA0011, and DAA0045.
[52-2] The molecule of [50-2] or [51-2], wherein the first antigen binding domain competes for binding DDR1 with an antigen binding domain comprising any one of (o1) to (o2), or wherein the first antigen-binding domain comprises any one of (o1) to (o2), wherein (o1) to (o2) corresponds to (o1) to (o2) recited in [0267].
[53-2] The molecule of any one of [50-2] to [52-2], wherein the first antigen binding domain competes for binding DDR1 with an antigen binding domain comprising any one of (p1) to (p2), or wherein the first antigen-binding domain comprises any one of (p1) to (p2), wherein (p1) to (p2) corresponds to (p1) to (p2) recited in [0268].
[54-2] The molecule of any one of [50-2] to [53-2], wherein the second antigen binding domain competes for binding DDR1 with an antigen binding domain comprising any one of (q1) to (q3), or wherein the second antigen-binding domain comprises any one of (q1) to (q3), wherein (q1) to (q3) corresponds to (q1) to (q3) recited in [0269].
[55-2] The molecule of any one of [50-2] to [54-2], wherein the second antigen binding domain competes for binding DDR1 with an antigen binding domain comprising any one of (r1) to (r3), or wherein the second antigen-binding domain comprises any one of (r1) to (r3), wherein (r1) to (r3) corresponds to (r1) to (r3) recited in [0270].
[56-2] The molecule of any one of [50-2] to [55-2], wherein the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to DDR1 with an antigen-binding domain comprising any one of (s1) to (s5), or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (s1) to (s5), wherein (s1) to (s5) corresponds to (s1) to (s5) recited in [0271].
[57-2] The molecule 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 compete for binding to DDR1 with an antigen-binding domain comprising any one of (t1) to (t6), 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) corresponds to (t1) to (t6) recited in [0272].
[62] The molecule of 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 an Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.
[63] The molecule of 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 which are not in a hinge region.
[64] The molecule of [63], wherein the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG.
[65] The molecule of [64], wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain.
[66] The molecule of [65], wherein said at least one disulfide bond is formed between the group selected from the following:
(i) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
(ii) an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iii) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iv) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
(v) an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
(vi) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.
[67] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a 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 originate is present at a position selected from the group consisting of positions 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, according to EU numbering, in the CH1 region.
[68] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL 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 originate is present at a position selected from the group consisting of positions 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 numbering in the CL region.
[69] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197 according to EU numbering, and the amino acid residue in the CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128 according to Kabat numbering.
[70] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a VH region of a first antigen-binding domain and an amino acid residue in a VH 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 originate is present at a position selected from the group consisting of positions 6, 8, 16, 20, 25, 26, 28, 74, and 82b according to Kabat numbering in the VH region.
[71] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a VHH region of a first antigen-binding domain and an amino acid residue in a VHH 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 originate is present at a position selected from the group consisting of positions 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 numbering in the VHH region.
[72] The molecule of [66], wherein said at least one disulfide bond is formed between an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of the second antigen-binding domain, and
(i) wherein when the VL is of a subclass kappa, the amino acid residue from which the bond between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 21, 27, 58, 77, 100, 105, and 107 according to Kabat numbering in the VL region (subclass kappa); or
(ii) wherein when the VL is of a subclass lambda, the amino acid residue from which the bonds between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 6, 19, 33, and 34 according to Kabat numbering in the VL region (subclass lambda).
[73] The molecule of [72], wherein the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain.
[74] The molecule of [73], wherein the at least one disulfide bond is formed between the amino acid residue at position 191 in a CH1 region of a first antigen-binding domain (according to Kabat numbering) and the amino acid residue at position 126 in a CL region of a second antigen-binding domain (according to Kabat numbering).
[75] The molecule of any one of [1] to [61], [17-1] to [61-1], [17-2] to [57-2], and [62] to [74], wherein the molecule comprises an antibody constant region.
[76] The molecule of [75] wherein the antibody constant region is a human antibody constant region, or wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.
[77] The molecule of 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 antibody fragment thereof.
[78] The molecule of [77], wherein the antibody fragment is a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody.
[79] The molecule of [77] or [78] wherein the molecule is a monoclonal antibody.
[80] The molecule of any one of [77] to [79], wherein the molecule is human, humanized, or chimeric antibody.
[81] The molecule of 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 second antigen-binding domain are each a monovalent Fab, or a monovalent VHH, or are each a monovalent scFab.
[82] The molecule of 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 means for binding, or wherein the molecule is tri-valent, tetra-valent, or multivalent molecule.
[83] The molecule of [82], wherein the molecule comprises a third antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain, or wherein the molecule comprises a third means for binding the same epitope on the same domain as the first or the second means.
[84] The molecule of [83], wherein the molecule comprises a fourth antigen-binding domain, wherein the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain, or wherein the molecule comprises a fourth means for binding the same epitope on the same domain as the first or the second means but not the same epitope as the third means.
[85] The molecule of [84], wherein the molecule comprises a third and a fourth antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain, or wherein the molecule comprises a third means for binding the same epitope on the same domain as the first means, and a fourth means for binding the same epitope on the same domain as the second means.
[86] A pharmaceutical composition comprising the molecule of 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.
[87] An immunoconjugate protein comprising the molecule of 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].
[88] The molecule of 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 of [86], or the immunoconjugate of [87], for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[89] The molecule or pharmaceutical composition or immunoconjugate for the use of [88], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[90] The molecule of 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 of [86], or the immunoconjugate of [87], for use in the treatment of any disease, disorder, or condition susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling, or FGFR1b signaling, or DDR1 signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[91] The molecule of 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 of [86], or the immunoconjugate of [87], for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[92] The molecule or pharmaceutical composition or immunoconjugate for the use of [91], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[93] The molecule or pharmaceutical composition or immunoconjugate for the use of [92], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[94] Use of the molecule of 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 of [86], or the immunoconjugate of [87], in the manufacture of a medicament for the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
[95] A method of treating a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of 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 of [86], or the immunoconjugate of [87].
[96] The use of [94], or the method of [95], wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
[97] The use of [94], or the method of [95], wherein the condition related to the injury of lung epithelial cells in an individual is one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling, or FGFR1b signaling, or DDR1 signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
[98] Use of the molecule of any one of [1], or FGFR1b signaling, or DDR1 signaling to [85], or the pharmaceutical composition of [86], or the immunoconjugate of [87], in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
[99] A method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of 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 the pharmaceutical composition of [86], or the immunoconjugate of [87].
[100] The use of [98], or the method of [99], wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
[101] The use of [98], or the method of [99], wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
[102] An isolated nucleic acid encoding the molecule of 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 a plurality of isolated nucleic acids encoding the molecule of 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].
[103] A host cell comprising the nucleic acid or the plurality of nucleic acids of [102].
[104] A method of producing the molecule, or a process for producing the molecule comprising the following steps:
(a) culturing the host cell of [103], and
(b) recovering the molecule that is produced in step (a).
[105] A kit for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, which comprises at least the the molecule of 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.

In further aspects and embodiments, the invention relates to any one of [B1] to [B41] below:
[B1] A method to activate FGFR2b signaling, or induce dimerization of FGFR2b, or promote phosphorylation of FGFR2b, or enhance intracellular FGF signaling, in an FGFR2b-expressing cell comprising a step of contacting the molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a] to [A34] with an FGFR2b-expressing cell in vitro, ex vivo, or in vivo.
[B2] A method to activate FGFR2b signaling, or induce dimerization of FGFR2b, or promote phosphorylation of FGFR2b, or enhance intracellular FGF signaling, in an FGFR2b-expressing cell comprising a step of contacting the molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a] to [A34] with an FGFR2b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B3] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a-1] to [A34] with an FGFR2b-expressing cell in vitro, or ex vivo.
[B4] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a] to [A34] with an FGFR2b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B5] Use of a molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a] to [A34] to activate FGFR2b signaling, or induce dimerization of FGFR2b, or promote phosphorylation of FGFR2b, or enhance intracellular FGF signaling, in an FGFR2b-expressing cell in vitro, ex vivo, or in vivo.
[B6] Use of a molecule of any one of [1], [2], [2a], and [3] to [51], [51a], [52], [52a], [53] to [85], [A1], and [A1a] to [A34] to activate FGFR2b signaling, or induce dimerization of FGFR2b, or promote phosphorylation of FGFR2b, or enhance intracellular FGF signaling, in an FGFR2b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B7] Use of a molecule of 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 ex vivo.
[B8] Use of a molecule of 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 ex vivo in lung organoid culture, or skin keratinocyte culture.
[B1-1] A method to activate FGFR1b signaling, or induce dimerization of FGFR1b, or promote phosphorylation of FGFR1b, or enhance intracellular FGF signaling, in an FGFR1b-expressing cell comprising a step of contacting the molecule of 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 an FGFR1b-expressing cell in vitro, ex vivo, or in vivo.
[B2-1] A method to activate FGFR1b signaling, or induce dimerization of FGFR1b, or promote phosphorylation of FGFR1b, or enhance intracellular FGF signaling, in an FGFR1b-expressing cell comprising a step of contacting the molecule of 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 an FGFR1b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B3-1] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule of 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 an FGFR1b-expressing cell in vitro, or ex vivo.
[B4-1] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule of 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 an FGFR1b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B5-1] Use of a molecule of 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 activate FGFR1b signaling, or induce dimerization of FGFR1b, or promote phosphorylation of FGFR1b, or enhance intracellular FGF signaling, in an FGFR1b-expressing cell in vitro, ex vivo, or in vivo.
[B6-1] Use of a molecule of 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 activate FGFR1b signaling, or induce dimerization of FGFR1b, or promote phosphorylation of FGFR1b, or enhance intracellular FGF signaling, in an FGFR1b-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B7-1] Use of a molecule of 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 ex vivo.
[B8-1] Use of a molecule of 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 ex vivo in lung organoid culture, or skin keratinocyte culture.
[B1-2] A method to activate DDR1 signaling, or induce dimerization of DDR1, or promote phosphorylation of DDR1, or enhance intracellular collagen signaling, in a DDR1-expressing cell comprising a step of contacting the molecule 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] with a DDR1-expressing cell in vitro, ex vivo, or in vivo.
[B2-2] A method to activate DDR1 signaling, or induce dimerization of DDR1, or promote phosphorylation of DDR1, or enhance intracellular collagen signaling, in a DDR1-expressing cell comprising a step of contacting the molecule 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] with a DDR1-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B3-2] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule 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] with a DDR1-expressing cell in vitro, or ex vivo.
[B4-2] A method to maintain and proliferate progenitor cells, comprising a step of contacting the molecule 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] with a DDR1-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B5-2] Use of a molecule 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] to activate DDR1 signaling, or induce dimerization of DDR1, or promote phosphorylation of DDR1, or enhance intracellular collagen signaling, in a DDR1-expressing cell in vitro, ex vivo, or in vivo.
[B6-2] Use of a molecule 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] to activate DDR1 signaling, or induce dimerization of DDR1, or promote phosphorylation of DDR1, or enhance intracellular collagen signaling, in an DDR1-expressing cell in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B7-2] Use of a molecule 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] to maintain and proliferate DDR1-expressing cells in vitro or ex vivo.
[B8-2] Use of a molecule 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] to maintain and proliferate DDR1-expressing cells in vitro, or ex vivo in lung organoid culture, or skin keratinocyte culture.
[B9] A method of producing an agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein the method comprises the steps of:
(a) providing a first nucleic acid encoding a polypeptide comprising the first antigen-binding domain that binds a first epitope in a first domain in the RTK protein, and a second nucleic acid encoding a polypeptide comprising the second antigen-binding domain that binds a second epitope in a second domain in the RTK protein;
(b) introducing the nucleic acids encoding the two polypeptides comprising the antigen-binding domains of step (a) into a host cell capable of expressing said nucleic acids;
(c) culturing the host cell obtained in step (b) such that the two polypeptides comprising the antigen-binding domains are expressed;
(d) obtaining the antigen-binding molecule comprising the polypeptide comprising the first antigen-binding domain and the polypeptide comprising the second antigen-binding domain obtained in step (c);
(e) subjecting the antigen-binding molecule obtained in step (d) to an assay capable of determining activation, or detecting receptor dimerization of, the RTK protein in a cell expressing said RTK protein on the cell surface; and
(f) selecting the antigen-binding molecule in step (e) that exhibits activation activity or wherein receptor dimerization was detected in said assay, thereby selecting an agonistic antigen-binding molecule.
[B10] The method of [B9], wherein the method comprises a step (ai) which follows step (a) and is carried out prior to step (b), wherein step (ai) comprises selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to the same domain, or selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domains that bind to different domains.
[B11] The method of [B9] or [B10], wherein the method comprises a step (aii) which follows step (ai) and is carried out prior to step (b), wherein step (aii) comprises selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to the same epitope, or selecting the polypeptides comprising the first antigen-binding domain and second antigen-binding domain that bind to different epitopes.
[B12] The method of any one of [B9] to [B11], wherein the molecule binds biparatopically, and the method comprises:
(a) providing a first nucleic acid encoding a polypeptide comprising the first antigen-binding domain that binds a first epitope in a first domain in the RTK protein, and a second nucleic acid encoding a polypeptide comprising the second antigen-binding domain that binds a second epitope in a second domain in the RTK protein;
(ai) selecting the polypeptide comprising the first antigen-binding domains and polypeptide comprising the second antigen-binding domain that bind to the same domain, or selecting the polypeptides comprising the first antigen-binding domains and polypeptides comprising the second antigen-binding domain that bind to different domains;
(aii) selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to different epitopes;
(b) introducing the nucleic acids encoding the two polypeptides comprising the antigen-binding domains obtained in step (aiii) into a host cell capable of expressing said nucleic acids;
(c) culturing the host cell in step (b) such that the two polypeptides comprising the antigen-binding domains are expressed;
(d) obtaining the antigen-binding molecule comprising the polypeptide comprising first antigen-binding domain and the polypeptide comprising the second antigen-binding domain obtained in step (c);
(e) subjecting the antigen-binding molecule obtained in step (d) to an assay capable of determining activation, or detecting receptor dimerization of, the RTK protein in a cell expressing said RTK protein on the cell surface; and
(f) selecting the antigen-binding molecule in step (e) that exhibits activation activity or wherein receptor dimerization was detected in said assay, thereby selecting an agonistic antigen-binding molecule.
[B13] The method of any one of [B9] to [B12], wherein the RTK protein is Fibroblast growth factor receptor 2 (FGFR2).
[B14] The method of [B13], wherein first and second domain in step (a) is / are independently selected from an Ig-like domain of FGFR2, in particular selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR2.
[B15] The method of [B14], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is IgIIIb, and the second domain is IgIIIb,
(ii) the first domain is IgIIIb, and the second domain is IgIII,
(iii) the first domain is IgIIIb, and the second domain is IgII, and
(iv) the first domain is IgIIIb, and the second domain is IgI,
[B13-1] The method of any one of [B9] to [B12], wherein the RTK protein is Fibroblast growth factor receptor 1 (FGFR1).
[B14-1] The method of [B13-1], wherein first and second domain in step (a) is / are independently selected from an Ig-like domain of FGFR1, in particular selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR1.
[B15-1] The method of [B14-1], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is IgIIIb, and the second domain is IgIIIb,
(ii) the first domain is IgIIIb, and the second domain is IgIII,
(iii) the first domain is IgIIIb, and the second domain is IgII, and
(iv) the first domain is IgIIIb, and the second domain is IgI,
[B13-2] The method of any one of [B9] to [B12], wherein the RTK protein is discoidin domain receptor1 (DDR1).
[B14-2] The method of [B13-2], wherein first and second domain in step (a) is / are independently selected from a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1.
[B15-2] The method of [B14-2], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is a DS domain, and the second domain is a DS domain,
(ii) the first domain is a DS domain, and the second domain is a DS-like domain, and
(iii) the first domain is a DS domain, and the second domain is an EJXM region.
[B16] The method of any one of [B9] to [B15], [B13-1] to [B15-1], and [B13-2] to [B15-2], wherein method comprises a step (aiii) which follows step (aii) and is carried out prior to step (b), wherein step (aiii) comprises introducing a mutation into the nucleic acids encoding the two polypeptides comprising the two antigen-binding domains such that at least one bond linking the two antigen-binding domains in the polypeptides is formed.
[B17] The method of [B16], wherein the method further comprises step (g) selecting the antigen-binding molecule obtained in step (f) wherein the two antigen-binding domains are present at spatially close positions and/or the mobility of the two antigen-binding domains is reduced.
[B18] A method of screening an agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein method comprises a step of:
(a) providing a first nucleic acid encoding a polypeptide comprising the first antigen-binding domain that binds a first epitope in a first domain in the RTK protein, and a second nucleic acid encoding a polypeptide comprising the second antigen-binding domain that binds a second epitope in a second domain in the RTK protein;
(b) introducing the nucleic acids encoding the two polypeptides comprising the antigen-binding domains of step (a) into a host cell capable of expressing said nucleic acids;
(c) culturing the host cell in step (b) such that the two polypeptides comprising the antigen-binding domains are expressed;
(d) obtaining the antigen-binding molecule comprising the polypeptide comprising the first antigen-binding domain and polypeptide comprising the second antigen-binding domain obtained in step (c);
(e) subjecting the antigen-binding molecule obtained in step (d) to an assay capable of determining activation, or detecting receptor dimerization of, the RTK protein in a cell expressing said RTK protein on the cell surface; and
(f) selecting the antigen-binding molecule in step (e) that exhibits activation activity or wherein receptor dimerization was detected in said assay, thereby selecting an agonistic antigen-binding molecule.
[B19] The method of [B18], wherein the method comprises a step (ai) which follows step (a) and is carried out prior to step (b), wherein step (ai) comprises selecting the polypeptides comprising the first antigen-binding domain and the polypeptides comprising the second antigen-binding domains that bind to the same domain, or selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to different domains.
[B20] The method of [B18] or [B19], wherein the method comprises a step (aii) which follows step (ai) and is carried out prior to step (b), wherein step (aii) comprises selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to the same epitope, or selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to different epitopes.
[B21] The method of any one of [B18] to [B20], wherein the molecule binds biparatopically, and the method comprises:
(a) providing a first nucleic acid encoding a polypeptide comprising the first antigen-binding domain that binds a first epitope in a first domain in the RTK protein, and a second nucleic acid encoding a polypeptide comprising the second antigen-binding domain that binds a second epitope in a second domain in the RTK protein;
(ai) comprises selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to the same domain, or selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain that bind to different domains;
(aii) selecting the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domains that bind to different epitopes;
(b) introducing the nucleic acids encoding the two polypeptides comprising the antigen-binding domains obtained in step (aii) into a host cell capable of expressing said nucleic acids;
(c) culturing the host cell in step (b) such that the two polypeptides comprising the antigen-binding domains are expressed;
(d) obtaining the antigen-binding molecule comprising the polypeptides comprising the first antigen-binding domain and polypeptides comprising the second antigen-binding domain obtained in step (c);
(e) subjecting the antigen-binding molecule obtained in step (d) to an assay capable of determining activation, or detecting receptor dimerization of, the RTK protein in a cell expressing said RTK protein on the cell surface; and
(f) selecting the antigen-binding molecule in step (e) that exhibits activation activity or wherein receptor dimerization was detected in said assay, thereby selecting an agonistic antigen-binding molecule.
[B22] The method of any one of [B18] to [B21], wherein the RTK protein is Fibroblast growth factor receptor 2 (FGFR2).
[B23] The method of [B22], wherein first and second domain in step (a) is/are independently selected from an Ig-like domain of FGFR2, in particular selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR2.
[B24] The method of [B23], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is IgIIIb and the second domain is IgIIIb;
(ii) the first domain is IgIIIb and the second domain is IgIII;
(iii) the first domain is IgIIIb and the second domain is IgII; and
(iv) the first domain is IgIIIb and the second domain is IgI.
[B22-1] The method of any one of [B18] to [B21], wherein the RTK protein is Fibroblast growth factor receptor 1 (FGFR1).
[B23-1] The method of [B22-1], wherein first and second domain in step (a) is/are independently selected from an Ig-like domain of FGFR1, in particular selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR1.
[B24-1] The method of [B23-1], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is IgIIIb and the second domain is IgIIIb;
(ii) the first domain is IgIIIb and the second domain is IgIII;
(iii) the first domain is IgIIIb and the second domain is IgII; and
(iv) the first domain is IgIIIb and the second domain is IgI.
[B22-2] The method of any one of [B18] to [B21], wherein the RTK protein is discoidin domain receptor1 (DDR1).
[B23-2] The method of [B22-2], wherein first and second domain in step (a) is/are independently selected from a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1.
[B24-2] The method of [B23-2], wherein the first domain and second domain in step (a) is selected from the group consisting of the following:
(i) the first domain is a DS domain, and the second domain is a DS domain,
(ii) the first domain is a DS domain, and the second domain is a DS-like domain, and
(iii) the first domain is a DS domain, and the second domain is an EJXM region.[B25] The method of any one of [B18] to [B24], [B22-1] to [B24-1], and [B22-2] to [B24-2], wherein method comprises a step (aiii) which follows step (aii) and is carried out prior to step (b), wherein step (aiii) comprises introducing a mutation into the nucleic acids encoding the two polypeptides comprising the two antigen-binding domains such that at least one bond linking the two antigen-binding domains in the polypeptides is formed.
[B26] The method of [B25], wherein the method further comprises step (g) selecting the antigen-binding molecule obtained in step (f) wherein the two antigen-binding domains are present at spatially close positions and/or the mobility of the two antigen-binding domains is reduced.
[B27] The method of any one of [B9] to [B26], wherein the molecule comprises an antibody constant region, in particular (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions
[B28] The method of 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 domains comprise an antibody variable region, or comprise an antibody fragment independently selected from a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, or single-domain antibody.
[B29] The method of 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 antibody fragment thereof.
[B30] The method of [B29], wherein the antibody fragment is a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody.
[B31] The method of [B29] or [B30] wherein the molecule is a monoclonal antibody.
[B32] The method of any one of [B29] to [B31], wherein the molecule is human, humanized, or chimeric antibody.
[B33] The method of any one of [B29] to [B32], 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 second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.
[B34] A diagnostic reagent comprising the molecule of 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] and [A1], [A1a] to [A34], [A1-1], [A1a-1] to [A33-1], [A1-2], and [A1a-2] to [A33-2].
[B35] A library that comprises or essentially consists of:
a combination of a plurality of a first group of antigen-binding domains which have different sequences from one another, and a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to Immunoglobulin IIIb (IgIIIb) of FGFR2 protein, and
wherein activation of FGFR2b signaling in a cell-surface expressed FGFR2b protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B36] The library of [B35] comprising:
a combination of a plurality of a first group of nucleic acids that encode a first group of antigen-binding domains which have different sequences from one another, and a plurality of a second group of nucleic acids that encode a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to Immunoglobulin IIIb (IgIIIb) of FGFR2 protein, and
wherein activation of FGFR2b signaling in a cell-surface expressed FGFR2b protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B37] The library of [B35] or [B36], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR2, and a second antigen-binding domain that binds a second epitope in a second domain of FGFR2;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR2 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-1] The library of [B35] or [B36], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR2, and a second antigen-binding domain that binds a second epitope in IgIIIb of FGFR2;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR2 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-2] The library of [B35] or [B36], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR2, and a second antigen-binding domain that binds a second epitope in IgIII of FGFR2;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR2 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-3] The library of [B35] or [B36], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR2, and a second antigen-binding domain that binds a second epitope in IgII of FGFR2;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR2 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-4] The library of [B35] or [B36], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR2, and a second antigen-binding domain that binds a second epitope in IgI of FGFR2;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR2 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B38] The library of any one of [B35] to [B37-4], wherein the method further comprises a step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when the antigen-binding domain binds to IgIIIb of FGFR2 protein with a KD equal to or less than 10^-8M as measured by surface plasmon resonance.
[B39] The library of 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.
[B40] The library of 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.
[B41] The library of any one of [B35] to [B40], wherein the activation of FGFR2b signaling by the combination of the first and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% greater than the activation of FGFR2b signaling by the first antigen-binding domain on its own, or the second antigen-binding domain on its own.
[B35-1] A library that comprises or essentially consists of:
a combination of a plurality of a first group of antigen-binding domains which have different sequences from one another, and a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to Immunoglobulin IIIb (IgIIIb) of FGFR1 protein, and
wherein activation of FGFR1b signaling in a cell-surface expressed FGFR1b protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B36-1] The library of [B35-1] comprising:
a combination of a plurality of a first group of nucleic acids that encode a first group of antigen-binding domains which have different sequences from one another, and a plurality of a second group of nucleic acids that encode a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to Immunoglobulin IIIb (IgIIIb) of FGFR1 protein, and
wherein activation of FGFR1b signaling in a cell-surface expressed FGFR1b protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B37-1] The library of [B35-1] or [B36-1], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR1, and a second antigen-binding domain that binds a second epitope in a second domain of FGFR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-1-1] The library of [B35-1] or [B36-1], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR1, and a second antigen-binding domain that binds a second epitope in IgIIIb of FGFR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-2-1] The library of [B35-1] or [B36-1], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR1, and a second antigen-binding domain that binds a second epitope in IgIII of FGFR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-3-1] The library of [B35-1] or [B36-1], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR1, and a second antigen-binding domain that binds a second epitope in IgII of FGFR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-4-1] The library of [B35-1] or [B36-1], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in IgIIIb of FGFR1, and a second antigen-binding domain that binds a second epitope in IgI of FGFR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the FGFR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B38-1] The library of any one of [B35-1] to [B37-4-1], wherein the method further comprises a step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when the antigen-binding domain binds to IgIIIb of FGFR1 protein with a KD equal to or less than 10^-8M as measured by surface plasmon resonance.
[B39-1] The library of any one of [B35-1] to [B38-1], wherein the first epitope of the first antigen-binding domain is the same as the second epitope of the second antigen-binding domain.
[B40-1] The library of 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.
[B41-1] The library of any one of [B35-1] to [B40-1], wherein the activation of FGFR1b signaling by the combination of the first and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% greater than the activation of FGFR1b signaling by the first antigen-binding domain on its own, or the second antigen-binding domain on its own.
[B35-2] A library that comprises or essentially consists of:
a combination of a plurality of a first group of antigen-binding domains which have different sequences from one another, and a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to a DS domain of DDR1 protein, and
wherein activation of DDR1 signaling in a cell-surface expressed DDR1 protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B36-2] The library of [B35-2] comprising:
a combination of a plurality of a first group of nucleic acids that encode a first group of antigen-binding domains which have different sequences from one another, and a plurality of a second group of nucleic acids that encode a second group of antigen-binding domains which have different sequences from one another;
wherein each of the antigen-binding domain of the first group of antigen-binding domains binds to a DS domain of DDR1 protein, and
wherein activation of DDR1 signaling in a cell-surface expressed DDR1 protein by a combination of a first and a second antigen-binding domain selected from the plurality of the first and second groups of antigen-binding domains in (i) is greater than a first antigen-binding domain on its own, or a second antigen-binding domain on its own.
[B37-2] The library of [B35-2] or [B36-2], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in a DS domain of DDR1, and a second antigen-binding domain that binds a second epitope in a second domain of DDR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the DDR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-1-2] The library of [B35-2] or [B36-2], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in a DS domain of DDR1, and a second antigen-binding domain that binds a second epitope in a DS domain of DDR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the DDR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-2-2] The library of [B35-2] or [B36-2], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in a DS domain of DDR1, and a second antigen-binding domain that binds a second epitope in a DS-like domain of DDR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the DDR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B37-3-2] The library of [B35-2] or [B36-2], which is produced by a method comprising the steps of:
(a) providing a first antigen-binding domain that binds a first epitope in a DS domain of DDR1, and a second antigen-binding domain that binds a second epitope in an EJXM region of DDR1;
(b) obtaining an antigen-binding molecule or antigen-binding molecule comprising the combination of the first and the second antigen-binding domain in step (a);
(c) subjecting the antigen-binding molecule or antigen-binding molecule in step (b) to an assay capable of determining activation, or detecting receptor dimerization of, the DDR1 protein in a cell expressing said RTK protein on the cell surface; and
(d) selecting the antigen-binding molecule or antigen-binding molecule in step (c) that exhibits activation activity or wherein receptor dimerization was detected in said assay.
[B38-2] The library of any one of [B35-2] to [B37-3-2], wherein the method further comprises a step (ai) of selecting the antigen-binding domain as the first antigen-binding domain when the antigen-binding domain binds to a DS domain of DDR1 protein with a KD equal to or less than 10^-8M as measured by surface plasmon resonance.
[B39-2] The library of 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.
[B40-2] The library of 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.
[B41-2] The library of any one of [B35-2] to [B40-2], wherein the activation of DDR1 signaling by the combination of the first and the second antigen-binding domain is at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% greater than the activation of DDR1 signaling by the first antigen-binding domain on its own, or the second antigen-binding domain on its own.

Figure 1 shows the process of generating biparatopic antibody using FORCE technology as described in 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). Figure 2 shows molecular format and nomenclature of biparatopic antibody. Figure 3 shows the process of generating biparatopic antibody using LINC-Ig technology. Figure 4 shows molecular format and nomenclature of biparatopic LINC-Ig and UN-LINC antibody. Figure 5 shows binding activity of monoparatopic antibodies to mouse FGFR2IIIb variant stably expressing FreeStyle^TM 293-F cell line. Anti-KLH monoparatopic antibody IC17dK shows the level of a negative control (background). Figure 6 shows binding activity of monoparatopic antibodies to IgI deleted mouse FGFR2IIIb variant (mouse FGFR2IIIb delta IgI) stably expressing FreeStyle^TM 293-F cell line. Anti-KLH monoparatopic antibody IC17dK shows the level of a negative control (background). Figure 7 shows binding activity of monoparatopic antibodies to IgIII deleted mouse FGFR2IIIb variant (mouse FGFR2IIIb delta IgIII) transiently expressing FreeStyle^TM 293-F cell line. Anti-KLH monoparatopic antibody IC17dK shows the level of a negative control (background). Figure 8 shows binding activity of monoparatopic antibodies to mouse FGFR2IIIc variant stably expressing FreeStyle^TM 293-F cell line. Anti-KLH monoparatopic antibody IC17dK shows the level of a negative control (background). Figure 9 shows binding activity of monoparatopic antibodies to human FGFR2IIIb variant stably expressing FreeStyle^TM 293-F cell line. Anti-KLH monoparatopic antibody IC17dK shows the level of a negative control (background). Figure 10-1 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (A) FFB0185//FFB0396, FFB0185, FFB0396, (B) FFB0219//FFB0341, FFB0219, FFB0341. Data points represent mean ± SEM. Figure 10-2 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (C) FFB0219//FFB0396, FFB0219, FFB0396, (D) FFB0261//FFB0464, FFB0261, FFB0464. Data points represent mean ± SEM. Figure 10-3 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (E) FFB0294//FFB0464, FFB0294, FFB0464, (F) FFB0299//FFB0464, FFB0299, FFB0464. Data points represent mean ± SEM. Figure 10-4 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (G) FFB0335//FFB0464, FFB0335, FFB0464, (H) FFB0365//FFB0464, FFB0365, FFB0464. Data points represent mean ± SEM. Figure 10-5 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (I) FFB0368//FFB0341, FFB0341, FFB0368, (J) FFB0374//FFB0464, FFB0374, FFB0464. Data points represent mean ± SEM. Figure 10-6 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody, cross: N12433). FGF7 and N12433 were used as control and are shared in each figure. (K) FFB0453//FFB0464, FFB0453, FFB0464. Data points represent mean ± SEM. Figure 11 shows the result of agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid. Luminescence of lung growth by each antibody was measured by CellTiter-Glo (registered trademark) 3D (gray: anti-FGFR2 monoparatopic antibodies, black: anti-FGFR2 biparatopic antibodies). Data points represent mean ± SEM. Figure 12-1 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in keratinocyte. Absorbance of imported BrdU in keratinocyte by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody). FGF7 was used as a control and is shared in each figure. (A) FFB0185//FFB0396, FFB0185, FFB0396, (B) FFB0219//FFB0341, FFB0219, FFB0341. Data points represent mean ± SEM. Figure 12-2 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in keratinocyte. Absorbance of imported BrdU in keratinocyte by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody). FGF7 was used as a control and is shared in each figure. (C) FFB0219//FFB0396, FFB0219, FFB0396, (D) FFB0261//FFB0464, FFB0261, FFB0464. Data points represent mean ± SEM. Figure 12-3 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in keratinocyte. Absorbance of imported BrdU in keratinocyte by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody). FGF7 was used as a control and is shared in each figure. (E) FFB0294//FFB0464, FFB0294, FFB0464, (F) FFB0299//FFB0464, FFB0299, FFB0464. Data points represent mean ± SEM. Figure 12-4 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in keratinocyte. Absorbance of imported BrdU in keratinocyte by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (solid line). Result of each monoparatopic and biparatopic antibody are shown, respectively (circle: FGF7, square: biparatopic antibody, open triangle and inverted triangle: monoparatopic antibody). FGF7 was used as a control and is shared in each figure. (G) FFB0368//FFB0341, FFB0368, FFB0341, (H) FFB0374//FFB0464, FFB0374, FFB0464. Data points represent mean ± SEM. Figure 13-1 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in NIH3T3. Absorbance of imported BrdU in NIH3T3 by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (dotted line, samples with no lines were not converged). (A) Each monoparatopic antibody, FGF2, FGF18, and FGF7 are shown (circle: FGF2, square: FGF18, triangle: FFB0185, diamond: FFB0219, hexagon: FFB0261, open circle: FFB0294, open square: FFB0299, open triangle: FFB0335, open diamond: FFB0365, open hexagon: FFB0368, cross: FGF7). (B) Each monoparatopic antibody, FGF2, FGF18, and FGF7 are shown (circle: FGF2, square: FGF18, triangle: FFB0341, diamond: FFB0374, hexagon: FFB0396, open circle: FFB0453, open square: FFB0464, cross: FGF7). FGF2, FGF18, and FGF7 were used as control and are shared in each figure. Data points represent mean ± SEM. Figure 13-2 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR2 monoparatopic and biparatopic antibodies in NIH3T3. Absorbance of imported BrdU in NIH3T3 by each antibody was measured by Cell Proliferation ELISA, and four-parameter logistic model was used to fit drug dose-response curves (dotted line, samples with no lines were not converged). (C) Each biparatopic antibody, FGF2, FGF18, and FGF7 are shown (circle: FGF2, square: FGF18, triangle: FFB0185//FFB0396, diamond: FFB0219//FFB0341, hexagon: FFB0219//FFB0396, open circle: FFB0261//FFB0464, open square: FFB0294//FFB0464, cross: FGF7). (D) Each biparatopic antibody, FGF2, FGF18, and FGF7 are shown (circle: FGF2, square: FGF18, triangle: FFB0299//FFB0464, diamond: FFB0335//FFB0464, hexagon: FFB0365//FFB0464, open circle: FFB0368//FFB0341, open square: FFB0374//FFB0464, open triangle: FFB0453//FFB0464, cross: FGF7). FGF2, FGF18, and FGF7 were used as control and are shared in each figure. Data points represent mean ± SEM. Figure 14 shows the result of agonistic activity evaluation of anti-FGFR2b biparatopic LINC-Ig antibodies in lung organoid. 1 micro g/mL of each Un-LINC and HL-LINC-Ig or LL-LINC-Ig antibody were used in this study. (A) Luminescence of lung growth by each Un-LINC and HL-LINC-Ig antibody were measured by CellTiter-Glo (registered trademark) 3D, and the ratio to each average value of Un-LINC format is shown (gray: FFB0065//FFB0045, black: FFB0065//FFB0057). (B) Luminescence of lung growth by each Un-LINC and LL-LINC-Ig antibody were measured by CellTiter-Glo (registered trademark) 3D, and the ratio to each average value of Un-LINC format is shown (gray: FFB0065//FFB0045,white: FFB0065//FFB0035, black: FFB0065//FFB0057). Data points represent mean ± SEM. Figure 15 shows the image of lung organoid growth by anti-FGFR2b biparatopic LINC-Ig antibodies. 50nM of FGF7 and each anti-FGFR2b biparatopic LINC-Ig antibody were used in this study. Image was captured by CellVoyager^TMCV8000 and Z stack image was combined by CellPathfinder software. FGF7, Control (culture without exogenous FGF7), LL-LINC_FFB0065//FFB0035, and LL-LINC_FFB0065//FFB0045 are shown. Each scale bar represents 100 micro m. Figure 16-1 shows the result of proliferation marker induction by anti-FGFR2b biparatopic LINC-Ig antibodies in vivo. Each proliferation marker gene was normalized by Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value calculated as ratio to average Vehicle value. Mki67 expression in lung tissue (A), and Cdc20 expression in lung tissue (B) are shown. (circle: Vehicle, square: FGF7, triangle: LL-LINC_FFB0065//FFB0035, diamond: LL-LINC_FFB0065//FFB0045, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM. Figure 16-2 shows the result of proliferation marker induction by anti-FGFR2b biparatopic LINC-Ig antibodies in vivo. Each proliferation marker gene was normalized by Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value calculated as ratio to average Vehicle value. Mki67 expression in ear skin tissue (C), and Cdc20 expression in ear skin tissue (D) are shown. (circle: Vehicle, square: FGF7, triangle: LL-LINC_FFB0065//FFB0035, diamond: LL-LINC_FFB0065//FFB0045, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM. Figure 17 shows the antibody treatment regimen in bleomycin induced mouse lung fibrosis model (BLM model). Figure 18 shows the result of alveolar type II (AT2) cells marker induction and fibrotic markers reduction by anti-FGFR2b biparatopic LINC-Ig antibodies in mouse lung fibrosis model (BLM model). Each marker gene was normalized to Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value calculated as ratio to average Sham value. Sftpc expression in lung tissue (A), Fn1 expression in lung tissue (B), and Col1a1 expression in lung tissue (C) are shown. (circle: Sham, square: bleomycin administration control (BLM), triangle: bleomycin administration LL-LINC_FFB0065//FFB0035, diamond: bleomycin administration with LL-LINC_FFB0065//FFB0045, ns: not significant, **P<0.01, ****P<0.0001). Data points represent mean ± SEM. Figure 19 shows the result of proliferation marker induction by anti-FGFR2b biparatopic antibodies in vivo. Each proliferation marker gene was normalized to Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value was calculated as ratio to average Vehicle value. Mki67 expression in lung tissue (A), and Cdc20 expression in lung tissue (B) are shown. (circle: Vehicle, square: FGF7, triangle: FFB0219//FFB0396, diamond: FFB0261//FFB0464, hexagon: FFB0335//FFB0464, open circle: FFB0365//FFB0464, open square: FFB0368//FFB0341, open triangle: FFB374//FFB0464, ns: not significant, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM. Figure 20 shows the efficacy study result of alveolar type II (AT2) cells marker induction and fibrotic markers reduction by anti-FGFR2b biparatopic antibodies in mouse lung fibrosis model (BLM model). Each marker gene was normalized to Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value was calculated as ratio to average Sham + KLH value (Anti-KLH monoparatopic antibody IC17dK was used as a negative control). Sftpc expression in lung tissue (A), Fn1 expression in lung tissue (B), and Col1a1 expression in lung tissue (C) are shown. (circle: Sham with KLH, square: bleomycin administration (BLM) with KLH, triangle: bleomycin administration with FFB0219//FFB0396, diamond: bleomycin administration with FFB0261//FFB0464, hexagon: bleomycin administration with FFB0335//FFB0464, ns: not significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). Data points represent mean ± SEM. Figure 21 shows the efficacy study result of alveolar type II (AT2) cells marker induction and fibrotic markers reduction by anti-FGFR2b biparatopic antibodies in mouse lung fibrosis model (BLM model). Each marker gene was normalized to Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) expression and each normalized value was calculated as ratio to average Sham + KLH value (Anti-KLH monoparatopic antibody IC17dK was used as a negative control). Sftpc expression in lung tissue (A), Fn1 expression in lung tissue (B), and Col1a1 expression in lung tissue (C) are shown. (circle: Sham with KLH, square: bleomycin administration (BLM) with KLH, triangle: bleomycin administration with FFB0365//FFB0464, diamond: bleomycin administration with FFB0368//FFB0341, hexagon: bleomycin administration with FFB0374//FFB0464, ns: not significant, *P<0.05, **P<0.01, ***P<0.001). Data points represent mean ± SEM. Figure 22 shows the results of histopathological evaluation for effects by anti-FGFR2b biparatopic antibodies in mouse lung fibrosis model (BLM model). Lung tissue sections were immunohistochemically stained with antibody against Prosurfactant Protein C (ProSPC; ab211326, abcam, Cambridge, UK), captured as digital images with NanoZoomer S360 (Hamamatsu Photonics K.K., 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) were summarized in (A) (circle: Sham with KLH, square: bleomycin administration (BLM) with KLH, triangle: bleomycin administration with FFB0219//FFB0396, inverted triangle: bleomycin administration with FFB0261//FFB0464, diamond: bleomycin administration with FFB0335//FFB0464, **P<0.01, ***P< 0.001, ****P< 0.0001 vs. BLM + KLH group compared by Tukey’s multiple comparisons test) and (B) (circle: Sham with KLH, square: bleomycin administration (BLM) with KLH, triangle: bleomycin administration with FFB0365//FFB0464, inverted triangle: bleomycin administration with FFB0368//FFB0341, diamond: bleomycin administration with FFB0374//FFB0464, *P<0.05, **P<0.01, ****P< 0.0001 vs. BLM + KLH group compared by Tukey’s multiple comparisons test). Data points represent mean ± SEM. Figure 23-1 shows the result of concentration-dependent agonistic activity evaluation of anti-DDR1 monoparatopic and biparatopic antibodies in mouse DDR1 stably expressing Ba/F3 cell line (Ba/F3-DDR1). Luminescence of Ba/F3-DDR1 growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (A) (open circle: DAA0007, open triangle: DAA0045, square: DAA0045//DAA0007, star: collagen control), (B) (open circle: DAA0024, open triangle: DAA0011, square: DAA0011//DAA0024, star: collagen control). Collagen control is shared in (A) and (B). Data points represent mean ± SEM. Figure 23-2 shows the result of concentration-dependent agonistic activity evaluation of anti-DDR1 monoparatopic and biparatopic antibodies in mouse DDR1 stably expressing Ba/F3 cell line (Ba/F3-DDR1). Luminescence of Ba/F3-DDR1 growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (C) (open circle: DAA0012, open triangle: DAA0045, square: DAA0045//DAA0012, star: collagen control). Data points represent mean ± SEM. Figure 24 shows the result of agonistic activity evaluation of anti-DDR1 monoparatopic and biparatopic antibodies in Ba/F3-DDR1. Absorbance of phosphorylated DDR1 in Ba/F3-DDR1 by each antibody was measured by ELISA. (A, B) Result of each monoparatopic and biparatopic antibody are shown, respectively (gray: monoparatopic antibody, black: biparatopic antibody). Data points represent mean ± SEM. Figure 25-1 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR1b monoparatopic and biparatopic antibodies in mouse FGFR1IIIb variant stably expressing Ba/F3 cell line (Ba/F3-FGFR1b). Luminescence of Ba/F3-FGFR1b cells growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (A) (open circle: FFA0026, open triangle: FFA0029, square: FFA0029//DAA0026, star: FGF2 control), (B) (open circle: FFA0064, open triangle: FFA0029, square: FFA0029//FFA0064, star: FGF2 control). FGF2 is shared in (A) and (B). Data points represent mean ± SEM. Figure 25-2 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR1b monoparatopic and biparatopic antibodies in mouse FGFR1IIIb variant stably expressing Ba/F3 cell line (Ba/F3-FGFR1b). Luminescence of Ba/F3-FGFR1b cells growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (C) (open circle: FFA0088, open triangle: FFA0029, square: FFA0029//FFA0088, star: FGF2 control), (D) (open circle: FFA0032, open triangle: FFA0048, square: FFA0048//FFA0032, star: FGF2 control). FGF2 is shared in (C) and (D). Data points represent mean ± SEM. Figure 25-3 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR1b monoparatopic and biparatopic antibodies in mouse FGFR1IIIb variant stably expressing Ba/F3 cell line (Ba/F3-FGFR1b). Luminescence of Ba/F3-FGFR1b cells growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (E) (open circle: FFA0043, open triangle: FFA0048, square: FFA0048//FFA0043, star: FGF2 control), (F) (open circle: FFA0047, open triangle: FFA0048, square: FFA0048//FFA0047, star: FGF2 control). FGF2 is shared in (E) and (F). Data points represent mean ± SEM. Figure 25-4 shows the result of concentration-dependent agonistic activity evaluation of anti-FGFR1b monoparatopic and biparatopic antibodies in mouse FGFR1IIIb variant stably expressing Ba/F3 cell line (Ba/F3-FGFR1b). Luminescence of Ba/F3-FGFR1b cells growth by each monoparatopic and biparatopic antibody was measured by CellTiter-Glo (registered trademark) 3D, and four-parameter logistic model was used to fit drug dose-response curves (solid line). (G) (open circle: FAA0055, open triangle: FFA0048, square: FFA0048//FFA0055, star: FGF2 control), and (H) (open circle: FFA0020, open triangle: FFA0048, square: FFA0048//FFA0020, star: FGF2 control). FGF2 is shared in (G) and (H). Data points represent mean ± SEM.

The definitions and detailed description below are provided to facilitate understanding of the present invention illustrated herein. Especially, these definitions and detailed description serve to interpret the aspects and embodiments above.

DEFINITIONS
Acceptor human framework
An "acceptor human framework" for the purposes herein is a framework comprising the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. An acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence thereof, or it may contain amino acid sequence changes. In some embodiments, the number of amino acid changes are 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or human consensus framework sequence.

Affinity
"Affinity" refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, 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 a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described in the following.

Affinity matured
An "affinity matured" antibody refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs), compared to a parent antibody which does not possess such alterations, such alterations resulting in an improvement in the affinity of the antibody for antigen.

FGFR2
The term "FGFR2" and the like, as used herein, refers to any native FGFR2 from any vertebrate source, including mammals such as primates (e.g. humans) and rodents (e.g. mice and rats), unless otherwise indicated. The term encompasses "full-length" unprocessed FGFR2 as well as any form of FGFR2 that results from processing in the cell. The term also encompasses naturally occurring variants of FGFR2, e.g. splice variants or allelic variants.

FGFR2b/FGFR2IIIb
The terms "FGFR2b" and "FGFR2IIIb", as used herein, and as used herein interchangeably, refer to the epithelial FGFR2b isoform, and refers to the amino acid sequence as set forth in SEQ ID NO: 5, and/or having the amino acid sequence as set forth in NCBI accession No. NP_075259.4, and is encoded by the FGFR2 gene residing on chromosome 10. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for the FGFR2 splicing and consequent expression of the epithelial FGFR2b isoform.
An exemplary amino acid sequence for the FGFR2IIIb isoform is presented as SEQ ID NO:5:
SEQ ID NO: 5 (FGFR2IIIb isoform)
1 mvswgrficl vvvtmatlsl arpsfslved ttlepeeppt kyqisqpevy vaapgeslev
61 rcllkdaavi swtkdgvhlg pnnrtvlige ylqikgatpr dsglyactas rtvdsetwyf
121 mvnvtdaiss gddeddtdga edfvsensnn krapywtnte kmekrlhavp aantvkfrcp
181 aggnpmptmr wlkngkefkq ehriggykvr nqhwslimes vvpsdkgnyt cvveneygsi
241 nhtyhldvve rsphrpilqa glpanastvv ggdvefvckv ysdaqphiqw ikhvekngsk
301 ygpdglpylk vlkhsginss naevlalfnv teadageyic kvsnyigqan qsawltvlpk
361 qqapgrekei taspdyleia iycigvflia cmvvtvilcr mknttkkpdf ssqpavhklt
421 kriplrrqvt vsaessssmn sntplvritt rlsstadtpm lagvseyelp edpkwefprd
481 kltlgkplge gcfgqvvmae avgidkdkpk eavtvavkml kddatekdls dlvsememmk
541 migkhkniin llgactqdgp lyviveyask gnlreylrar rppgmeysyd inrvpeeqmt
601 fkdlvsctyq largmeylas qkcihrdlaa rnvlvtennv mkiadfglar dinnidyykk
661 ttngrlpvkw mapealfdrv ythqsdvwsf gvlmweiftl ggspypgipv eelfkllkeg
721 hrmdkpanct nelymmmrdc whavpsqrpt fkqlvedldr iltlttneey ldlsqpleqy
781 spsypdtrss cssgddsvfs pdpmpyepcl pqyphingsv kt

FGFR2c/FGFR2IIIc
The terms "FGFR2c" and "FGFR2IIIc", as used herein, refer to the epithelial FGFR2c isoform, and refers to the amino acid sequence as set forth in SEQ ID NO: 6, and/or having the amino acid sequence as set forth in NCBI accession No. NP_000132.3.
An exemplary amino acid sequence for the FGFR2IIIc isoform is presented as SEQ ID NO:6:
SEQ ID NO: 6 (FGFR2IIIc isoform)
1 mvswgrficl vvvtmatlsl arpsfslved ttlepeeppt kyqisqpevy vaapgeslev
61 rcllkdaavi swtkdgvhlg pnnrtvlige ylqikgatpr dsglyactas rtvdsetwyf
121 mvnvtdaiss gddeddtdga edfvsensnn krapywtnte kmekrlhavp aantvkfrcp
181 aggnpmptmr wlkngkefkq ehriggykvr nqhwslimes vvpsdkgnyt cvveneygsi
241 nhtyhldvve rsphrpilqa glpanastvv ggdvefvckv ysdaqphiqw ikhvekngsk
301 ygpdglpylk vlkaagvntt dkeievlyir nvtfedagey tclagnsigi sfhsawltvl
361 papgrekeit aspdyleiai ycigvfliac mvvtvilcrm knttkkpdfs sqpavhkltk
421 riplrrqvtv saessssmns ntplvrittr lsstadtpml agvseyelpe dpkwefprdk
481 ltlgkplgeg cfgqvvmaea vgidkdkpke avtvavkmlk ddatekdlsd lvsememmkm
541 igkhkniinl lgactqdgpl yviveyaskg nlreylrarr ppgmeysydi nrvpeeqmtf
601 kdlvsctyql argmeylasq kcihrdlaar nvlvtennvm kiadfglard innidyykkt
661 tngrlpvkwm apealfdrvy thqsdvwsfg vlmweiftlg gspypgipve elfkllkegh
721 rmdkpanctn elymmmrdcw havpsqrptf kqlvedldri ltlttneeyl dlsqpleqys
781 psypdtrssc ssgddsvfsp dpmpyepclp qyphingsvk t

FGFR1
The term "FGFR1" and the like, as used herein, refers to any native FGFR1 from any vertebrate source, including mammals such as primates (e.g. humans) and rodents (e.g. mice and rats), unless otherwise indicated. The term encompasses "full-length" unprocessed FGFR1 as well as any form of FGFR1 that results from processing in the cell. The term also encompasses naturally occurring variants of FGFR1, e.g. splice variants or allelic variants.

FGFR1b/FGFR1IIIb
The terms "FGFR1b" and "FGFR1IIIb", as used herein, and as used herein interchangeably, refer to the epithelial FGFR1b isoform, and refers to the amino acid sequence as set forth in NCBI accession No. NP_001167536.1, and is encoded by the FGFR1 gene residing on chromosome 8. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for the FGFR1 splicing and consequent expression of the epithelial FGFR1b isoform.

FGFR1c/FGFR1IIIc
The terms "FGFR1c" and "FGFR1IIIc", as used herein, refer to the epithelial FGFR1c isoform, and refers to the amino acid sequence as set forth in NCBI accession No. NP_001167534.1.

DDR1
The term "DDR1" and the like, as used herein, refers to any nativeDDR1 from any vertebrate source, including mammals such as primates (e.g. humans) and rodents (e.g. mice and rats), unless otherwise indicated. The term encompasses "full-length" unprocessed DDR1 as well as any form of DDR1 that results from processing in the cell. The term also encompasses naturally occurring variants of DDR1, e.g. splice variants or allelic variants.

DDR1a
The term "DDR1a", as used herein, refers to the amino acid sequence as set forth in NCBI accession No. NP_001945.3, and is encoded by the DDR1 gene residing on chromosome 6

DDR1b
The term "DDR1b", as used herein, refers to the amino acid sequence as set forth in NCBI accession No. NP_054699.2, and is encoded by the DDR1 gene residing on chromosome 6

The term "DDR1c", as used herein, refers the amino acid sequence as set forth in NCBI accession No. NP_054700.2, and is encoded by the DDR1 gene residing on chromosome 6.

The term "DDR1d", as used herein, refers the amino acid sequence as set forth in NCBI accession No. NP_001189450.1, and is encoded by the DDR1 gene residing on chromosome 6.

The term "DDR1e", as used herein, refers the amino acid sequence as set forth in NCBI accession No. NP_001189451.1, and is encoded by the DDR1 gene residing on chromosome 6.

Agonist/agonistic antigen-binding molecule
An "agonist antigen-binding molecule", "agonistic antigen-binding molecule" or "agonist antibody" or "agonistic antibody" as used herein, is a molecule or antibody which significantly potentiates a biological activity of the antigen it binds. In embodiments, "agonist" or "agonistic" in the context of a cell receptor is understood as "activating" the signaling of said cell receptor. The term "anti-FGFR2 agonist antigen-binding molecule" or "anti-FGFR2 agonist antibody", as used herein, refers to an antigen-binding molecule or an antibody that is capable of binding its antigen with sufficient affinity such that the antigen-binding molecule or antibody is useful as a therapeutic agent in activating FGFR2. The term "anti-FGFR1 agonist antigen-binding molecule" or "anti-FGFR1 agonist antibody", as used herein, refers to an antigen-binding molecule or an antibody that is capable of binding its antigen with sufficient affinity such that the antigen-binding molecule or antibody is useful as a therapeutic agent in activating FGFR1. The term "anti-DDR1 agonist antigen-binding molecule" or "anti- DDR1 agonist antibody", as used herein, refers to an antigen-binding molecule or an antibody that is capable of binding its antigen with sufficient affinity such that the antigen-binding molecule or antibody is useful as a therapeutic agent in activating DDR1. As used herein, the agonistic activity of the antigen-binding molecule or agonist antibody is observed when the binding of the molecule or antibody to its antigen induces dimerization of the intracellular tyrosine kinase domains of the antigen, or wherein the binding of the molecule or antibody to its antigen activates phosphorylation of the antigen.

In one embodiment, the extent of binding of an anti-FGFR2 antigen-binding molecule or antibody to an unrelated, non-FGFR2 protein is less than about 10% of the binding of the antigen-binding molecule or antibody to FGFR2 as measured, e.g. by a radioimmunoassay (RIA). In certain embodiments, an antibody that binds to FGFR2 has a dissociation constant (Kd) of < 1 micro Μ, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g. 10^-8 M or less, e.g. from 10^-8 M to 10^-13 M, e.g., from 10^-9 M to 10^-13 M). In certain embodiments, an anti-FGFR2 antigen-binding molecule or anti-FGFR2 antibody binds to an epitope of FGFR2 that is conserved among FGFR2 from different species.

In one embodiment, the extent of binding of an anti-FGFR1 antigen-binding molecule or antibody to an unrelated, non-FGFR1 protein is less than about 10% of the binding of the antigen-binding molecule or antibody to FGFR1 as measured, e.g. by a radioimmunoassay (RIA). In certain embodiments, an antibody that binds to FGFR1 has a dissociation constant (Kd) of < 1 micro Μ, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g. 10^-8 M or less, e.g. from 10^-8 M to 10^-13 M, e.g., from 10^-9 M to 10^-13 M). In certain embodiments, an anti-FGFR1 antigen-binding molecule or anti-FGFR1 antibody binds to an epitope of FGFR1 that is conserved among FGFR1 from different species.

In one embodiment, the extent of binding of an anti-DDR1 antigen-binding molecule or antibody to an unrelated, non- DDR1 protein is less than about 10% of the binding of the antigen-binding molecule or antibody to DDR1 as measured, e.g. by a radioimmunoassay (RIA). In certain embodiments, an antibody that binds to DDR1 has a dissociation constant (Kd) of < 1 micro Μ, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g. 10^-8 M or less, e.g. from 10^-8 M to 10^-13 M, e.g., from 10^-9 M to 10^-13 M). In certain embodiments, an anti- DDR1 antigen-binding molecule or anti- DDR1 antibody binds to an epitope of DDR1 that is conserved among DDR1 from different species.

In one embodiment, Kd is measured by a radiolabeled antigen binding assay (RIA). In one embodiment, an RIA is performed with the Fab version of an antibody of interest and its antigen. For example, solution binding affinity of Fabs for antigen is measured by equilibrating Fab with a minimal concentration of (¹²⁵I)-labeled antigen in the presence of a titration series of unlabeled antigen, then capturing bound antigen with an anti-Fab antibody-coated plate (see, e.g., Chen et al., J. Mol. Biol. 293:865-881(1999)). To establish conditions for the assay, MICROTITER (registered trademark) multi-well plates (Thermo Scientific) are coated overnight with 5 micro g/ml of a capturing anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6), and subsequently blocked with 2% (w/v) bovine serum albumin in PBS for two to five hours at room temperature (approximately 23 degrees C). In a non-adsorbent plate (Nunc #269620), 100 pM or 26 pM [¹²⁵I]-antigen are mixed with serial dilutions of a Fab of interest (e.g., consistent with assessment of the anti-VEGF antibody, Fab-12, in Presta et al., Cancer Res. 57:4593-4599 (1997)). The Fab of interest is then incubated overnight; however, the incubation may continue for a longer period (e.g., about 65 hours) to ensure that equilibrium is reached. Thereafter, the mixtures are transferred to the capture plate for incubation at room temperature (e.g., for one hour). The solution is then removed and the plate washed eight times with 0.1% polysorbate 20 (TWEEN-20 (registered trademark)) in PBS. When the plates have dried, 150 micro l/well of scintillant (MICROSCINT-20 TM; Packard) is added, and the plates are counted on a TOPCOUNTTM gamma counter (Packard) for ten minutes. Concentrations of each Fab that give less than or equal to 20% of maximal binding are chosen for use in competitive binding assays.

According to another embodiment, Kd is measured using a BIACORE (registered trademark) surface plasmon resonance assay. For example, an assay using a BIACORE (registered trademark)-2000 or a BIACORE(registered trademark)-3000 (BIAcore, Inc., Piscataway, NJ) is performed at 25 degrees C with immobilized antigen CM5 chips at ~10 response units (RU). In one embodiment, carboxymethylated dextran biosensor chips (CM5, BIACORE, Inc.) are activated with N-ethyl-N’- (3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier’s instructions. Antigen is diluted with 10 mM sodium acetate, pH 4.8, to 5 micro g/ml (~0.2 micro M) before injection at a flow rate of 5 micro l/minute to achieve approximately 10 response units (RU) of coupled protein. Following the injection of antigen, 1 M ethanolamine is injected to block unreacted groups. For kinetics measurements, two-fold serial dilutions of Fab (0.78 nM to 500 nM) are injected in PBS with 0.05% polysorbate 20 (TWEEN-20TM) surfactant (PBST) at 25 degrees C at a flow rate of approximately 25 micro l/min. Association rates (kon) and dissociation rates (koff) are calculated using a simple one-to-one Langmuir binding model (BIACORE (registered trademark) Evaluation Software version 3.2) by simultaneously fitting the association and dissociation sensorgrams. The equilibrium dissociation constant (Kd) is calculated as the ratio koff/kon. See, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). If the on-rate exceeds 106 M-1 s-1 by the surface plasmon resonance assay above, then the on-rate can be determined by using a fluorescent quenching technique that measures the increase or decrease in fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm band-pass) at 25 degrees C of a 20 nM anti-antigen antibody (Fab form) in PBS, pH 7.2, in the presence of increasing concentrations of antigen as measured in a spectrometer, such as a stop-flow equipped spectrophotometer (Aviv Instruments) or a 8000-series SLM-AMINCO (registered trademark) spectrophotometer (ThermoSpectronic) with a stirred cuvette.

Specific binding
As used herein, the term "specifically binds" refers to a binding interaction that is determinative of the presence of a target (FGFR2b, FGFR1b, or DDR1) protein in a population of proteins including proteins in their different isoforms. As the skilled artisan will recognize that isoform-binding molecule, i.e. anti-FGFR2b antigen binding molecule, anti-FGFR1b antigen binding molecule, or anti-DDR1 antigen binding molecule, may show some degree of cross-reactivity between isoforms depending on the conditions used, e.g. target protein concentration, salt and buffer conditions used, among others. In certain embodiments, the term "specifically binds", or "specific binding" refers to a property of the isoform-binding molecule, i.e. anti-FGFR2b antigen binding molecule to bind 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., with an affinity constant of at least about 10⁷ M^-1, typically about 10⁸ M^-1, and more typically, about 10⁹ M^-1 to 10¹⁰ M^-1 or stronger, and preferentially bind to the isoform with an affinity that is at least two-fold, 50-fold, 100-fold, 1000-fold, or more greater than its affinity for binding to the another isoform. In certain embodiments, anti-FGFR2b antigen binding molecule binds preferentially to FGFR2b isoform, but does not substantially bind to (e.g., shows less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0% cross-reactivity with) to FGFR2c isoform. In certain embodiments, anti-FGFR1b antigen binding molecule binds preferentially to FGFR1b isoform, but does not substantially bind to (e.g., shows less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0% cross-reactivity with) to FGFR1c isoform. In certain embodiments, anti-DDR1a antigen binding molecule binds preferentially to DDR1a isoform, but does not substantially bind to (e.g., shows less than 10%, 8%, 5%, 4%, 3%, 2%, 1%, or 0% cross-reactivity with) to DDR1b isoform.

Cell surface-expressed FGFR2
As used herein, the expression "cell surface-expressed FGFR2" or the like, means one or more FGFR2 protein(s), or the extracellular domain thereof, that is/are expressed on the surface of a cell in vitro or in vivo, such that at least a portion of a FGFR2 protein is exposed to the extracellular side of the cell membrane and is accessible to an antigen-binding portion of an antibody or antigen-binding fragment. A "cell surface-expressed FGFR2" can comprise or consist of an FGFR2 protein expressed on the surface of a cell which normally expresses FGFR2 protein.

Cell surface-expressed FGFR1
As used herein, the expression "cell surface-expressed FGFR1" or the like, means one or more FGFR1 protein(s), or the extracellular domain thereof, that is/are expressed on the surface of a cell in vitro or in vivo, such that at least a portion of a FGFR1 protein is exposed to the extracellular side of the cell membrane and is accessible to an antigen-binding portion of an antibody or antigen-binding fragment. A "cell surface-expressed FGFR1" can comprise or consist of an FGFR1 protein expressed on the surface of a cell which normally expresses FGFR1 protein.

Cell surface-expressed DDR1
As used herein, the expression "cell surface-expressed DDR1" or the like, means one or more DDR1 protein(s), or the extracellular domain thereof, that is/are expressed on the surface of a cell in vitro or in vivo, such that at least a portion of a DDR1 protein is exposed to the extracellular side of the cell membrane and is accessible to an antigen-binding portion of an antibody or antigen-binding fragment. A "cell surface-expressed DDR1" can comprise or consist of an DDR1 protein expressed on the surface of a cell which normally expresses DDR1 protein.

Biparatopic antibody
As used herein, the term "biparatopic antibody" refers to, for example, an antibody that is capable of binding to two different epitopes on a single target. Without intending to be limiting, bispecific antibodies provide the ability to recognize and bind to two different antigens or epitopes (antigen domains) simultaneously as a single molecule. Biparatopic antibodies, which constitute a subset of bispecific antibodies, comprise antigen binding sites (paratopes) that provide the ability to recognize and bind to two different epitopes or antigenic sites on the same target antigen. In an embodiment, the antigen binding domains of biparatopic antibodies recognize and bind unique, non-overlapping epitopes on the same target antigen, such as a receptor, such as FGFR2, FGFR1, and DDR1.

And/or
The term "and/or" herein is used to indicate any one of the subjects shown before and after "and/or", or any combination thereof. For example, "A, B, and/or C" includes the individual subjects "A", "B", and "C", and also combinations "A and B", "A and C", "B and C", and "A and B and C".

Antigen-binding molecule
The term "antigen-binding molecule" as used herein may refer to any molecule having any structure as long as it binds to an antigen. The structure of the "antigen-binding molecule" may include but is not limited to, a polypeptide or a portion thereof, or a small or medium chemical compound or a portion thereof, or a polynucleotide or a portion thereof. The polypeptide or a portion thereof includes an antigen-binding domain (also referred as a portion, a part or a fragment of an antibody).

Antigen-binding domain
The term "antigen-binding domain" includes but is not limited to an antibody heavy chain variable (VH) region, an antibody light chain variable (VL) region (preferably a combination of an antibody heavy chain variable (VH) region and an antibody light chain variable (VL) region,), a single-domain antibody (sdAb), a VHH, a single-chain Fv (scFv), a single-chain antibody, a Fv, a single-chain Fv2 (scFv2), a Fab, Fab’, single chain Fab (scFab), and F (ab')2.

Antibody
The term "antibody" herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired antigen-binding activity.

Antibody fragment
An "antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. An "antibody fragment" comprises at least one "antigen-binding domain". Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab’-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g. scFv); VHH, Fab’, single chain Fab (scFab), and multispecific antibodies formed from antibody fragments such as bispecific antibodies.

Antibody that binds to the same/different epitope
An "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks binding of the reference antibody to its antigen in a competition assay by 50% or more, and conversely, the reference antibody blocks binding of the antibody to its antigen in a competition assay by 50% or more. An exemplary competition assay is provided herein. In the alternative, an "antibody that binds to a different epitope" as a reference antibody refers to an antibody which does not bind to the same epitope. The same applies where reference is made to "an antigen binding domain" instead of "an antibody".

Epitope
An "epitope" may be referred to as defining the site or sites of interaction between an antibody and its antigen(s). Accordingly, in Nilvebrant J, Rockberg J. An Introduction to Epitope Mapping. Methods Mol Biol. 2018;1785:1-10, "Antibody epitopes can conceptually be divided into linear and non-linear epitopes. Linear epitopes, which are also known as continuous or sequential epitopes, can be mimicked by short peptide sequences. Epitopes in which amino acids in distant parts of the sequence are brought together by protein folding are called non-linear, conformational, or discontinuous epitopes. For example, discontinuous epitopes often contain short segments of contiguous residues that can be independently bound by an antibody". As used herein, the terms "conformational epitope" or "non-linear epitope" or "discontinuous epitope" are used interchangeably to refer to an epitope which is composed of at least two amino acids which are not consecutive amino acids in a single protein chain or single polypeptide chain.

Substantially reduced
The phrase "substantially reduced" or "substantially different", as used herein, refers to a sufficiently high degree of difference between two numeric values (generally one associated with a molecule and the other associated with a reference/comparator molecule) such that one of skill in the art would consider the difference between the two values to be of statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values).

Substantially similar
The term "substantially similar" or "substantially the same," as used herein, refers to a sufficiently high degree of similarity between two numeric values (for example, one associated with an antibody of the invention and the other associated with a reference/comparator antibody), such that one of skill in the art would consider the difference between the two values to be of little or no biological and/or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values).

Chimeric
The term "chimeric" antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

Class of an antibody
The "class" of an antibody 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 several of these may be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively.

Effective amount
An "effective amount" of an agent, e.g., a pharmaceutical formulation, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

Extracellular domain
An "extracellular domain" or "ECD" is understood as a domain or portion of a membrane-associated protein which is located extracellularly when expressed in a cell. RTKs are single-pass transmembrane (TM) proteins. In RTK, the extracellular domains are typically located N-terminal of the transmembrane region. Another term for an "extracellular domain" is "ectodomain" as understood in the art.

Fc region
The term "Fc region" herein is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226, or from Pro230, to the carboxyl-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, numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

Framework or FR
"Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Accordingly, the HVR and FR sequences generally appear in the following sequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

Full length antibody/intact antibody/whole antibody
The terms "full length antibody," "intact antibody," and "whole antibody" are used herein interchangeably to refer to an antibody having a structure substantially similar to a native antibody structure or having heavy chains that contain an Fc region as defined herein.

Host cell/host cell line/host cell culture
The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and progeny derived therefrom without regard to the number of passages. Progeny may not be completely identical in nucleic acid content to a parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

Human antibody
A "human antibody" is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human or a human cell or derived from a non-human source that utilizes human antibody repertoires or other human antibody-encoding sequences. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues.

Human consensus framework
A "human consensus framework" is a framework which represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In one embodiment, for the VL, the subgroup is subgroup kappa I as in Kabat et al., supra. In one embodiment, for the VH, the subgroup is subgroup III as in Kabat et al., supra.

Humanized antibody
A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization.

Hypervariable region / HVR
The term "hypervariable region" or "HVR" as used herein refers to each of the regions of an antibody variable domain which are hypervariable in sequence ("complementarity determining regions" or "CDRs") and/or form structurally defined loops ("hypervariable loops") and/or contain the antigen-contacting residues ("antigen contacts"). Generally, antibodies comprise six HVRs: three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3). Exemplary HVRs herein include:
(a) hypervariable loops occurring 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));
(b) CDRs occurring 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 Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991));
(c) antigen contacts occurring 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
(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).

In one embodiment, HVR residues of anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof comprise those identified in Table 1 or elsewhere in the specification.

In one embodiment, HVR residues of anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof comprise those identified in Table 9 or elsewhere in the specification.

In one embodiment, HVR residues of anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof comprise those identified in Table 10 or elsewhere in the specification.

Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered herein according to Kabat et al., supra.

Immunoconjugate
An "immunoconjugate" is an antibody conjugated to one or more heterologous molecule(s). The heterologous molecule conjugated to the antibody can also be termed a secondary or adjunctive agent. Exemplary secondary or adjunctive agent are mitogenic growth factor, epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), Wnt family member including WNT1, WNT2, WNT2B, WNT3, WNT3A, WNT4, WNT5, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, 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, IGF-2, and hepatocyte growth factor (HGF). Further examples are directed at cytokines, such as interleukin family including IL-22, IL-10, IL-28, IL-26, IL-29, IL-24, and IL-20. The secondary or adjunctive agent can also be suitable to treat or help in treating inflammation or fibrosis, and can be a non-steroidal anti-inflammatory agent, steroid, a disease-modifying antirheumatic drug, an immunosuppressant, tumor necrosis factor inhibitor, adalimumab, azathioprine, chloroquine, hydroxychloroquine, ciclosporin, D-penicillamine, etanercept, golimumab, gold salts, infliximab, leflunomide, methotrexate, minocycline, sulfasalazine, anakinra, abatacept, rituximab, and tocilizumab. The secondary or adjunctive agent can also be a multi-kinase inhibitor, such as axitinib, nintedanib, pirfenidone, riociguat, sorafenib, sunitinib, lenvatinib, regorafenib, ponatinib, and pazopanib. The secondary or adjunctive agent can also be a phosphodiesterase inhibitor, such as meribendan, arinone Xi Luo acyl Amine, BI 1015550, Apremilast (apremilast), mesembranone, Ibudilast, pyrrole Rummy department spy, luteolin, roflumilast, cilomilast, diazepam, rolipram, YM796, icariin, silaenafil, he Da Lafei, Vardenafil, avanaphil, Yi Dina be non-, meter Luo Nafei, udenafil and Zaprinast. The secondary or adjunctive agent can also be an integrin inhibitor, such as ProAgio, and GSK-3008348. The secondary or adjunctive agent can also be a (cyto)toxic reagent, including a senolytic compound, such as MDM2 inhibitors (Nutlin 3a, RG-7112), BCL-2 anti-apoptotic protein family members, which inhibitors inhibit a function of at least the anti-apoptotic protein, BCL-xL (ABT-263, ABT-737, WEHI-539, A-1155463), GLS1 inhibitor (CB-839), Akt specific inhibitors (MK-2206), and radioisotopes.

Individual/subject
An "individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the individual or subject is a human.

Isolated antibody
An "isolated" antibody is one which has been separated from a component of its natural environment. In some embodiments, an antibody is purified to greater than 95% or 99% purity as determined by, for example, electrophoretic (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatographic (e.g., ion exchange or reverse phase HPLC). For review of methods for assessment of antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007).

Isolated nucleic acid
An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

Isolated nucleic acid encoding an anti-FGFR2 antibody, an anti-FGFR1 antibody, or an anti-DDR1 antibody
"Isolated nucleic acid encoding an anti-FGFR2 antibody", "isolated nucleic acid encoding an anti-FGFR1 antibody", or "isolated nucleic acid encoding an anti-DDR1 antibody" refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including such nucleic acid molecule(s) in a single vector or separate vectors, and such nucleic acid molecule(s) present at one or more locations in a host cell.

Monoclonal antibody
The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies composing the population are identical and/or bind the same epitope, except for possible variant antibodies, e.g., containing naturally occurring mutations or arising during production of a monoclonal antibody preparation, such variants generally being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including but not limited to the hybridoma method, 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 making monoclonal antibodies being described herein.

Naked antibody
A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or radiolabel. The naked antibody may be present in a pharmaceutical formulation.

Native antibodies
"Native antibodies" refer to naturally occurring immunoglobulin molecules with varying structures. For example, native IgG antibodies are heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light chains and two identical heavy chains that are disulfide-bonded. From N- to C-terminus, each heavy chain has a variable region (VH), also called a variable heavy domain or a heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from N- to C-terminus, each light chain has a variable region (VL), also called a variable light domain or a light chain variable domain, followed by a constant light (CL) domain. The light chain of an antibody may be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

Package insert
The term "package insert" is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and/or warnings concerning the use of such therapeutic products.

Percent (%) amino acid sequence identity
"Percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR) software, or GENETYX (registered trademark) (Genetyx Co., Ltd.). Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

The ALIGN-2 sequence comparison computer program was authored by Genentech, Inc., and the source code has been filed with user documentation in the U.S. Copyright Office, Washington D.C., 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or may be compiled from the source code. The ALIGN-2 program should be compiled for use on a UNIX operating system, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary. In situations where ALIGN-2 is employed for amino acid sequence comparisons, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be phrased as a given amino acid sequence A that has or comprises a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows:
100 times the fraction X/Y
where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program’s alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A. Unless specifically stated otherwise, all % amino acid sequence identity values used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

Pharmaceutical formulation
The term "pharmaceutical formulation" refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered.

Pharmaceutically acceptable carrier
A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, which is nontoxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.

Treatment/treating/to treat
As used herein, "treatment" (and grammatical variations thereof such as "treat" or "treating") refers to preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. In some embodiments, antibodies of the invention are used to delay development of a disease or to slow the progression of a disease. A "method of treating" or its equivalent, when applied to, for example, fibrosis, refers to a procedure or course of action that is 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 the fibrotic cells or other disorder will, in fact, be eliminated, that the number of cells or disorder will, in fact, be reduced, or that the symptoms of fibrosis or other disorder will, in fact, be alleviated. Often, a method of treatment of fibrosis will be performed even with a low likelihood of success, but which, given the medical history and estimated survival expectancy of a patient, is nevertheless deemed to induce an overall beneficial course of action.

Variable region/variable domain
The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs). (See, e.g., Kindt et al. Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007).) A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind a particular antigen may be isolated using a VH or VL domain from an antibody that binds the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

Vector
The term "vector," as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a self-replicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors." The vector can be introduced into a host cell by a method using a virus, an electroporation method, or the like, but the introduction of a vector is not limited to in vitro introduction, and it is also possible to directly introduce a vector into a body.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION
The activation of receptor tyrosine kinases (RTKs) plays an important role in the regulation of kinase activity and activation of downstream signaling pathways in the cell. Autophosphorylation of RTKs is mediated by sequential and precisely ordered intermolecular reaction upon binding to their natural ligands. The aspects and embodiments described herein are based, at least in part, on the discovery that activation of RTK can be achieved with the use of an antigen-binding molecule that mimics the binding of the natural ligands to the RTKs, including wherein the ligand-receptor interaction includes binding of the ligand to different domains, or binding to unique, non-overlapping epitopes on the same domain, on the same receptor protein. Without wishing to be bound by theory, it is hypothesized that the antigen-binding molecule described herein, at least in part, mimic the binding of the ligands to their receptors and induce receptor dimerization and autophosphorylation, thereby activating the downstream intracellular signals thereof.

In one aspect, an antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, wherein the molecule is an agonist is provided. Henceforth, referred herein as the anti-RTK agonist antigen-binding molecule.

In one embodiment, the anti-RTK agonist antigen-binding 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 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.

In embodiments, the first antigen-binding domain binds a first RTK protein and a second antigen-binding domain binds a second RTK protein, optionally wherein the first and the second RTK proteins form a dimer.

In such embodiments, activation, and optionally dimerization, of the RTK protein takes place in trans, between different RTK proteins.

In other embodiments, the first antigen-binding domain binds a first RTK protein and a second antigen-binding domain binds the first RTK protein.

In such embodiments, activation, and optionally dimerization, of the RTK protein takes place in cis, within one RTK protein.

In embodiments of the molecules herein, binding of the first and second antigen-binding domains induces dimerization of the intracellular tyrosine kinase domains of the RTK protein.

Dimerization of the intracellular tyrosine kinase domains of RTK proteins typically results in activation of intracellular signaling of the RTK domain. An agonist molecule for an RTK may therefore induce dimerization of the intracellular tyrosine kinase domains. Dimerization may by detected in vitro using appropriate assays.

Accordingly, in embodiments, dimerization is detected in the presence of the molecule in an in vitro cell assay, and wherein dimerization is indicative of agonist activity.

Further, activation of an RTK typically results in phosphorylation of the RTK via the tyrosine kinase domain of the RTK. Accordingly, in embodiments, an agonist molecule or agonist antibody provided herein, when binding to the RTK, activates phosphorylation of the RTK protein. Dimerization may be detected in vitro using appropriate assays. Phosphorylation of the RTK may by detected in vitro using appropriate assays.

Accordingly, in embodiments, binding of the first and second antigen-binding domains activates phosphorylation of the RTK protein.

Further, in embodiments, phosphorylation of the RTK protein is detected in the presence of the molecule in an in vitro cell assay, wherein RTK protein phosphorylation is indicative of agonist activity.

In vitro assays for detecting in vitro RTK phosphorylation are well known in the art.

Moreover, it is preferred that the molecules provided herein bind to an extracellular domain (ECD) of the RTK protein.

Binding of the antigen-binding domain(s) of the agonist molecules provided herein to ECD(s) of an RTK protein may activate intracellular signaling.

Accordingly, in embodiments, the first antigen-binding domain and the second antigen-binding domain each bind to an extracellular domain (ECD) of the RTK protein.

Several RTK proteins, such as the members of Fibroblast Growth Factor Receptor (FGFR) family of RTK comprise a plurality of Ig-like domains as extracellular domains.

Accordingly, in embodiments, the extracellular domain is independently selected from an Immunoglobulin (Ig)-like domain of the ECD of the RTK protein for each of the first antigen-binding domain and the second antigen-binding domain.

It is possible that the first and second antigen-binding domains bind to the same or to different domain. Moreover, in case the first and second antigen-binding domains bind to the same domain, the epitope of the first and second antigen-binding domains may be the same epitope or may be a different epitope.

In certain embodiments, the first antigen-binding domain binds to a first domain and the second antigen-binding domain binds to a second domain, wherein the first domain and the second domain is the same.

In embodiments thereof, the first and the second epitope is the same. In such embodiment, the molecule provided herein binds monoparatopically.

In different embodiments thereof, the first epitope and the second epitope are different. In such embodiment, the molecule provided herein binds biparatopically.

In different embodiments thereof, the first epitope and the second epitopes are different, and wherein the first epitope and second epitope are unique, non-overlapping epitopes. In such embodiment, the molecule provided herein binds biparatopically.

In further embodiments of molecules provided herein, the first antigen binding-domain binds to a first domain and the second antigen-binding domain binds to a second domain, wherein the second domain is different from the first domain.

In such embodiment, the first epitope and the second epitope are different. In such embodiment, the molecule provided herein binds biparatopically.

In one embodiment, the RTK protein is selected from the group consisting of Fibroblast growth factor receptor 2 (FGFR2), Fibroblast growth factor receptor 1 (FGFR1), Fibroblast growth factor receptor 3 (FGFR3), Fibroblast growth factor receptor 4 (FGFR4), Epidermal growth factor receptor (EGFR), Hepatocyte growth factor receptor (HGFR) or MET proto-oncogene receptor tyrosine kinase (MET), Bone morphogenetic protein receptor 1 (BMR1), Bone morphogenetic protein receptor 2 (BMR2), HER2(Erb-B2) receptor, HER3(Erb-B3) receptor, HER4(Erb-B4) receptor, insulin receptor (InsR), insulin-like growth factor I receptor (IGF1R), insulin receptor-related receptor (IRR), Platelet derived growth factor receptor alpha (PDGFRa), Platelet derived growth factor receptor beta (PDGFRb), Colony stimulating factor 1 receptor (CSFR), KIT proto-oncogene receptor tyrosine kinase (Kit), FMS related receptor tyrosine kinase (FLT3), vascular endothelial growth factor receptor (VEGFR), fms related receptor tyrosine kinase 1 (VEGFR-1), kinase insert domain receptor (VEGFR-2), fms related receptor tyrosine kinase 4 (VEGFR-3), Protein tyrosine kinase 7 (PTK7/CCK4), Neurotrophin receptor A (TrkA), Neurotrophin receptor B (TrkB), Neurotrophin receptor C (TrkC), Receptor tyrosine kinase like orphan receptor 1 (ROR1), Receptor tyrosine kinase like orphan receptor 2 (ROR2), Muscle associated receptor tyrosine kinase (MuSK), macrophage stimulating 1 receptor (Ron), TYRO3 protein tyrosine kinase (Tyro3), AXL receptor tyrosine kinase (AXL), and MER proto-oncogene tyrosine kinase (Mer), Tyrosine kinase with immunoglobulin like and EGF like domain 1 (TIE1) TEK receptor tyrosine kinase (TIE2), Ephrin receptor A1 (EphA1), Ephrin receptor A2 (EphA2), Ephrin receptor A3 (EphA3), Ephrin receptor A4 (EphA4), Ephrin receptor A5 (EphA5), Ephrin receptor A6 (EphA6), Ephrin receptor A7 (EphA7), Ephrin receptor A8 (EphA8), Ephrin receptor A9 (EphA9), Ephrin receptor A10 (EphA10), Ephrin receptor B1 (EphB1), Ephrin receptor B2 (EphB2), Ephrin receptor B3 (EphB3), Ephrin receptor B4 (EphB4), Ephrin receptor B6 (EphB6), Ret proto-oncogene (RET), receptor like tyrosine kinase (RYK), Discoidin domain receptor tyrosine kinase 1 (DDR1), Discoidin domain receptor tyrosine kinase 2 (DDR2), c-ros oncogene 1, receptor tyrosine kinase (ROS), apoptosis associated tyrosine kinase (Lmr1), lemur tyrosine kinase 2 (Lmr2), lemur tyrosine kinase 3 (Lmr3), leukocyte receptor tyrosine kinase (LTK), ALK receptor tyrosine kinase (ALK) and serine/threonine/tyrosine kinase 1 (STYK1).

In certain embodiments, the RTK protein is a protein of the Fibroblast Growth Factor Receptor (FGFR) family. Members of this family include FGFR1, FGFR2, FGFR3 and FGFR4.

In one embodiment, the RTK protein is selected from one of the Fibroblast Growth Factor Receptor (FGFR) family consisting of the members in the group consisting of: FGFR1, FGFR2, FGFR3 and FGFR4.

The family of FGFRs consists of FGFR1, FGFR2, FGFR3 and FGFR4. FGFRs contain three extracellular domains that belong to the immunoglobulin superfamily (IgI, IgII, and IgIII), a single transmembrane helix, and an intracellular tyrosine kinase domain. The IgIII domains of FGFR2 is produced by the IIIa exon being spliced to one of two other exons - IIIb or IIIc, to form FGFR2IIIb that is normally localized in the epithelial cells, and FGFR2IIIc which is normally localized in the mesenchymal cells and is tightly regulated in a tissue-specific expression manner (NPL 1).

An extracellular domain that belongs to the immunoglobulin superfamily, such as IgI, IgII, or IgIII in FGFR2 or FGFR1 is also referred herein as "Ig-like domain".
In a preferred embodiment, the RTK protein is Fibroblast growth factor receptor 2 (FGFR2) or Fibroblast growth factor receptor 1 (FGFR1).

anti-FGFR2 agonistic antigen-binding molecules
FGFR2 refers to a tyrosine protein kinase that acts as a cell-surface receptor for fibroblast growth factors. The protein is also known as keratinocyte growth factor receptor and CD332. Structurally, a FGFR2 amino acid sequence is a receptor tyrosine kinase protein having a signal peptide, at least one or more immunoglobulin (Ig)-like domains, an acidic box, a transmembrane domain, and a split tyrosine kinase domain and has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of NCBI accession No. NP_000132.3, NP_001138385.1, NP_001138386.1, NP_001138387.1, NP_001138388.1. Structurally, a FGFR2 nucleic acid sequence has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the nucleic acid sequence of NCBI accession No. NM_000141.4, NM_001144913.1, NM_001144914.1, NM_001144915.1, NM_001144916.1. The FGFR2 signal peptide may be retained or cleaved off.

FGFR2 plays an essential role in the regulation of cell proliferation, differentiation, migration and apoptosis, and in the regulation of embryonic development. Activation of FGFR2 results in phosphorylation of PLCG1, FRS2, and PAK4 and activation of several signaling cascades. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. Phosphorylation of FRS2 triggers recruitment of GRB2, GAB1, PIK3R1 and SOS1, and mediates activation of RAS, MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling pathway, as well as of the AKT1 signaling pathway. FGFR2 signaling is down-regulated by ubiquitination, internalization and degradation. Mutations that lead to constitutive kinase activation or impair normal FGFR2 maturation, internalization and degradation lead to aberrant signaling. Likewise, over-expressed FGFR2 promotes activation of STAT1. In some of these cases, such as in some cancers, FGFR2 aberrations result in the formation of FGFR2 fusion proteins, or FGFR2 oncogenic isoforms, such as FGFR2-BICC, FGFR2-PHGDH, FGFR2-AHCYL, FGFR2-NDC80, FGFR2-CIT, or FGFR2-OFD1 (Nita-et al; Xu et al).

Fibroblast growth factor (FGF) 7 (FGF-7) and its homologue FGF-10 act as a specific agonist for epithelial IIIb FGFR2 (FGFR2b) by binding to IgIII domain of epithelial IIIb in FGFR2 (NPL 2). In addition to the binding to IgIII domain of epithelial IIIb, binding of FGF-7 or FGF-10 to IgII domain in FGFR2 with heparin or heparan sulfate proteoglycans (HSPGs) is thought to be essential for agonistic FGF signaling activation (NPL 3), however, mechanisms of agonistic signal activation are not completely understood yet.

Without wishing to be bound by theory, the present inventors hypothesized that the antigen-binding molecules described herein in part mimic the binding of FGF7 with FGFR2, wherein binding of FGF7 to FGFR2 has been elucidated to involve binding to more than one domain of FGFR2. The aspects and embodiments described herein are based, at least in part, on the discovery that antigen-binding molecules that bind FGFR2 on different domains or bind to FGFR2 on different, or different (i.e. unique and non-overlapping) epitopes on the same domain is capable of activating FGFR2. In a preferred embodiment, an anti-FGFR2b antigen-binding molecule that binds FGFR2 on different domains or binds to FGFR2 on different, or different (i.e. unique and non-overlapping) epitopes on the same domain, wherein the antigen-binding molecule is an agonist, is provided.

In aspects of the present invention, an anti-FGFR2b antigen-binding molecule described herein does not substantially bind to mutated FGFR2, such as FGFR2 fusion proteins, or FGFR2 oncogenic isoforms.

In yet further embodiments, the RTK protein is FGFR2b.

The terms "FGFR2b" and "FGFR2IIIb", as used herein, refer to the amino acid sequence as set forth in SEQ ID NO: 5, and/or having the amino acid sequence as set forth in NCBI accession No. NP_075259.4, and is encoded by the FGFR2 gene residing on chromosome 10. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for the FGFR2 splicing and consequent expression of the epithelial FGFR2b isoform.

An exemplary amino acid sequence of FGFR2b (FGFR2IIIb) is provided as UniProtKB P21802-3, which is reproduced as SEQ ID NO: 5 shown below.
MVSWGRFICLVVVTMATLSLARPSFSLVEDTTLEPEEPPTKYQISQPEVYVAAPGESLEVRCLLKDAAVISWTKDGVHLGPNNRTVLIGEYLQIKGATPRDSGLYACTASRTVDSETWYFMVNVTDAISSGDDEDDTDGAEDFVSENSNNKRAPYWTNTEKMEKRLHAVPAANTVKFRCPAGGNPMPTMRWLKNGKEFKQEHRIGGYKVRNQHWSLIMESVVPSDKGNYTCVVENEYGSINHTYHLDVVERSPHRPILQAGLPANASTVVGGDVEFVCKVYSDAQPHIQWIKHVEKNGSKYGPDGLPYLKVLKHSGINSSNAEVLALFNVTEADAGEYICKVSNYIGQANQSAWLTVLPKQQAPGREKEITASPDYLEIAIYCIGVFLIACMVVTVILCRMKNTTKKPDFSSQPAVHKLTKRIPLRRQVTVSAESSSSMNSNTPLVRITTRLSSTADTPMLAGVSEYELPEDPKWEFPRDKLTLGKPLGEGCFGQVVMAEAVGIDKDKPKEAVTVAVKMLKDDATEKDLSDLVSEMEMMKMIGKHKNIINLLGACTQDGPLYVIVEYASKGNLREYLRARRPPGMEYSYDINRVPEEQMTFKDLVSCTYQLARGMEYLASQKCIHRDLAARNVLVTENNVMKIADFGLARDINNIDYYKKTTNGRLPVKWMAPEALFDRVYTHQSDVWSFGVLMWEIFTLGGSPYPGIPVEELFKLLKEGHRMDKPANCTNELYMMMRDCWHAVPSQRPTFKQLVEDLDRILTLTTNEEYLDLSQPLEQYSPSYPDTRSSCSSGDDSVFSPDPMPYEPCLPQYPHINGSVKT (SEQ ID NO: 5)

The terms "FGFR2c" and "FGFR2IIIc", as used herein, refer to the epithelial FGFR2c isoform, and refer to the amino acid sequence as set forth in SEQ ID NO: 6, and/or having the amino acid sequence as set forth in NCBI accession No. NP_000132.3.

In the present disclosure, an anti-FGFR2b antigen binding molecule comprising a first antigen-binding domain and a second antigen-binding domain, and wherein the molecule is an agonist is provided.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human FGFR2 protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first FGFR2 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein, wherein the first epitope and the second epitope are different from each other.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human FGFR2 protein, wherein the molecule comprises a first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR2 protein and a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein, wherein the first epitope and the second epitope are different from each other.

According to one aspect, the first antigen-binding domain or the first means binds a first FGFR2 protein and the second antigen-binding domain or the second means binds a second FGFR2 protein.

According to one aspect, the agonistic antigen-binding molecule of the present invention is a biparatopic antigen-binding molecule.

According to one aspect, the human FGFR2 protein is human FGFR2b protein.

According to one aspect, each of the extracellular domains is independently selected from an Immunoglobulin (Ig)-like domain of the FGFR2 protein.

According to one aspect of the agonistic antigen-binding molecule of the present invention, wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR2 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR2 protein, the first domain and the second domain is the same.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first epitope in a first domain of the FGFR2 protein and the second antigen-binding domain or the second means binds to a second epitope in a second domain of the FGFR2 protein, and the first domain and the second domain is the same, but the first epitope and the second epitopes are different. In a further aspect, the first epitope and the second epitope are unique, non-overlapping epitopes.

According to another aspect of the agonistic antigen-binding molecule of the present invention, wherein the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR2 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR2 protein, and the second domain is different from the first domain.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind an Ig-like domain of the FGFR2 protein, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgI domain of FGFR2;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgII domain of FGFR2;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIII domain of FGFR2;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIb domain of FGFR2;
(v) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIc domain of FGFR2;
(vi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgII domain of FGFR2;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIII domain of FGFR2;
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIb domain of FGFR2;
(ix) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIc domain of FGFR2;
(x) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIII domain of FGFR2;
(xi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIb domain of FGFR2;
(xii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIc domain of FGFR2;
(xiii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2; and
(xiv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIc domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2;
(ii) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR2;
(iii) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR2; and
(iv) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR2, or (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain or the first means binds to a continuous or discontinuous epitope, and optionally, the continuous or discontinuous epitope resides in one ECD, or in two or three ECDs.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the FGFR2 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds an Ig-like domain of FGFR2, selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR2.
In one aspect, the second antigen binding domain or the second means binds to IgI of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgI of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgII and IgIII of FGFR2.
In one aspect, the second antigen binding domain or the second means binds to IgII of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgII of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgIII of FGFR2.
In one aspect, the second antigen binding domain or the second means binds to IgIII of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIII of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
In one aspect, the second antigen binding domain or the second means binds to IgIIIb of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIb of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
In one aspect, the second antigen binding domain or the second means binds to IgIIIc of FGFR2, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIc of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIb of FGFR2, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds to a continuous or discontinuous epitope, and optionally, the continuous or discontinuous epitope resides in one ECD, or in two or three ECDs.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain, or the first epitope of the first antigen-binding domain and the second epitope of the second antigen-binding domain are non-overlapping. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same or different domain of FGFR2, in particular the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgIII of FGFR2. According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgIIIb of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgI of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain or the first means binds a first epitope in the IgIII domain of FGFR2, and the second antigen-binding domain or the second means binds a second epitope in the IgIII domain of FGFR2, and wherein the first and the second epitope are different. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgIII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgI of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgI and IgII of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain is selected from the group consisting of FFB0464, FFB0341, FFB0396, FFB0026, FFB0094, FFB0035, FFB0045, FFB0057, and FFB0080.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain is the same, and is selected from the group consisting of FFB0026, FFB0035, FFB0045, FFB0057, FFB0065, FFB0080, FFB0088, FFB0094, FFB0185, FFB0219, FFB0261, FFB0294, FFB0299, FFB0335, FFB0341, FFB0365, FFB0368, FFB0374, FFB0396, FFB0453, and FFB0464.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain are different, and are each independently selected from the group consisting of FFB0026, FFB0035, FFB0045, FFB0057, FFB0065, FFB0080, FFB0088, FFB0094, FFB0185, FFB0219, FFB0261, FFB0294, FFB0299, FFB0335, FFB0341, FFB0365, FFB0368, FFB0374, FFB0396, FFB0453, and FFB0464.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain complementarity determining regions (HCDR1, HCDR2, HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, LCDR3) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 1, the amino acid sequence identifiers (SEQ ID NOs) of the variable region of the heavy chain (VH), and the variable region of the light chain (VL) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 1.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain variable regions (VH), light chain variable regions (VL), heavy chains (HC), and light chains (LC) of the biparatopic anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 2.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain variable regions (VH), light chain variable regions (VL), heavy chains (HC), and light chains (LC) of the biparatopic LINC-Ig anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 3.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to FGFR2 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (a1) to (a14) below, or as described in Tables 1 to 4, or comprises any one of (a1) to (a14) below:
(a1) [FFB0185] HCDR1 comprising the amino acid sequence of SEQ ID NO:11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO:13;
(a2) [FFB0219] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 21;
(a3) [FFB0261] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 29;
(a4) [FFB0294] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 37;
(a5) [FFB0299] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 45;
(a6) [FFB0335] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 53;
(a7) [FFB0365] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 61;
(a8) [FFB0368] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 69;
(a9) [FFB0374] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 77;
(a10) [FFB0453] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 85;
(a11) [FFB0065] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 117;
(a12) [FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125;
(a13) [FFB0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 133; and
(a14) [FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to FGFR2 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (b1) to (b14) below, or as described in Tables 1 to 4, or comprises any one of (b1) to (b14) below:
(b1) [FFB0185] LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16;
(b2) [FFB0219] LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 24;
(b3) [FFB0261] LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 32;
(b4) [FFB0294] LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 40;
(b5) [FFB0299] LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 48;
(b6) [FFB0335] LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 56;
(b7) [FFB0365] LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 64;
(b8) [FFB0368] LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 72;
(b9) [FFB0374] LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 80;
(b10) [FFB0453] LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 88;
(b11) [FFB0065] LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 120;
(b12) [FFB0080] LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(b13) [FFB0088] LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 136; and
(b14) [FFB0057] LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to FGFR2 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (c1) to (c9) below, or as described in Tables 1 to 4, or wherein the second antigen-binding domain comprises any one of (c1) to (c9) below:
(c1) [FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93;
(c2) [FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101;
(c3) [FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109;
(c4) [FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141;
(c5) [FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149;
(c6) [FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157;
(c7) [FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165;
(c8) [FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173; and
(c9) [FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to FGFR2 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (d1) to (d9) below, or as described in Tables 1 to 4, or wherein the second antigen-binding domain comprises any one of (d1) to (d9) below:
(d1) [FFB0341] LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(d2) [FFB0396] LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(d3) [FFB0464] LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(d4) [FFB0026] LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(d5) [FFB0035] LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(d6) [FFB0094] LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(d7) [FFB0045] LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(d8) [FFB0057] LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176; and
(d9) [FFB0080] LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to FGFR2 with an antigen-binding domain comprising any one of (e1) to (e21) below, or as described in Tables 1 to 4, or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (e1) to (e21) below:
(e1) [FFB0185] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, HCDR3 comprising the amino acid sequence of SEQ ID NO: 13, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, LCDR3 comprising the amino acid sequence of SEQ ID NO: 16;
(e2) [FFB0219] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24;
(e3) [FFB0261] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32;
(e4) [FFB0294] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40;
(e5) [FFB0299] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48;
(e6) [FFB0335] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56;
(e7) [FFB0365] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64;
(e8) [FFB0368] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72;
(e9) [FFB0374] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80;
(e10) [FFB0453] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88;
(e11) [FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(e12) [FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(e13) [FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(e14) [FFB0065] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120;
(e15) [FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(e16) [FFB0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136;
(e17) [FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(e18) [FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(e19) [FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(e20) [FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, LCDR3 comprising the amino acid sequence of SEQ ID NO: 168; and
(e21) [FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, LCDR3 comprising the amino acid sequence of SEQ ID NO: 176.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 compete for binding to FGFR2 with an antigen-binding domain comprising any one of (f1) to (f116) below, or as described in Tables 1 to 4, or each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (f1) to (f116) below:
(f1) [FFB0185 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f2) [FFB0185 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f3) [FFB0185 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f4) [FFB0185 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f5) [FFB0185 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f6) [FFB0185 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f7) [FFB0185 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f8) [FFB0185 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f9) [FFB0185 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 11, HCDR2 comprising the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 13 and LCDR1 comprising the amino acid sequence of SEQ ID NO: 14, LCDR2 comprising the amino acid sequence of SEQ ID NO: 15, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 16, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f10) [FFB0219 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f11) [FFB0219 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f12) [FFB0219 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f13) [FFB0219 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f14) [FFB0219 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f15) [FFB0219 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f16) [FFB0219 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f17) [FFB0219 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f18) [FFB0219 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 19, HCDR2 comprising the amino acid sequence of SEQ ID NO: 20, HCDR3 comprising the amino acid sequence of SEQ ID NO: 21, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 22, LCDR2 comprising the amino acid sequence of SEQ ID NO: 23, LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f19) [FFB0261 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f20) [FFB0261// FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f21) [FFB0261// FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f22) [FFB0261// FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f23) [FFB0261// FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f24) [FFB0261// FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f25) [FFB0261// FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f26) [FFB0261// FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f27) [FFB0261// FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 27, HCDR2 comprising the amino acid sequence of SEQ ID NO: 28, HCDR3 comprising the amino acid sequence of SEQ ID NO: 29, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 30, LCDR2 comprising the amino acid sequence of SEQ ID NO: 31, LCDR3 comprising the amino acid sequence of SEQ ID NO: 32, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f28) [FFB0294 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f29) [FFB0294 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f30) [FFB0294 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f31) [FFB0294 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f32) [FFB0294 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f33) [FFB0294 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f34) [FFB0294 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f35) [FFB0294 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f36) [FFB0294 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 35, HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, HCDR3 comprising the amino acid sequence of SEQ ID NO: 37, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 38, LCDR2 comprising the amino acid sequence of SEQ ID NO: 39, LCDR3 comprising the amino acid sequence of SEQ ID NO: 40, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f37) [FFB0299 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f38) [FFB0299// FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f39) [FFB0299// FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f40) [FFB0299// FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f41) [FFB0299// FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f42) [FFB0299// FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f43) [FFB0299// FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f44) [FFB0299// FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f45) [FFB0299// FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 43, HCDR2 comprising the amino acid sequence of SEQ ID NO: 44, HCDR3 comprising the amino acid sequence of SEQ ID NO: 45, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 46, LCDR2 comprising the amino acid sequence of SEQ ID NO: 47, LCDR3 comprising the amino acid sequence of SEQ ID NO: 48, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f46) [FFB0335 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f47) [FFB0335 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f48) [FFB0335 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f49) [FFB0335 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f50) [FFB0335 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f51) [FFB0335 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f52) [FFB0335 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f53) [FFB0335 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f54) [FFB0335 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 51, HCDR2 comprising the amino acid sequence of SEQ ID NO: 52, HCDR3 comprising the amino acid sequence of SEQ ID NO: 53, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 54, LCDR2 comprising the amino acid sequence of SEQ ID NO: 55, LCDR3 comprising the amino acid sequence of SEQ ID NO: 56, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f55) [FFB0365 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f56) [FFB0365 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f57) [FFB0365 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f58) [FFB0365 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f59) [FFB0365 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f60) [FFB0365 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f61) [FFB0365 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f62) [FFB0365 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f63) [FFB0365 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 59, HCDR2 comprising the amino acid sequence of SEQ ID NO: 60, HCDR3 comprising the amino acid sequence of SEQ ID NO: 61, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 62, LCDR2 comprising the amino acid sequence of SEQ ID NO: 63, LCDR3 comprising the amino acid sequence of SEQ ID NO: 64, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f64) [FFB0368 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f65) [FFB0368 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f66) [FFB0368 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f67) [FFB0368 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f68) [FFB0368 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f69) [FFB0368 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f70) [FFB0368 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f71) [FFB0368 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f72) [FFB0368 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 67, HCDR2 comprising the amino acid sequence of SEQ ID NO: 68, HCDR3 comprising the amino acid sequence of SEQ ID NO: 69, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 70, LCDR2 comprising the amino acid sequence of SEQ ID NO: 71, LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f73) [FFB0374 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f74) [FFB0374 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f75) [FFB0374 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f76) [FFB0374 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f77) [FFB0374 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f78) [FFB0374 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f79) [FFB0374 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f80) [FFB0374 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f81) [FFB0374 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 75, HCDR2 comprising the amino acid sequence of SEQ ID NO: 76, HCDR3 comprising the amino acid sequence of SEQ ID NO: 77, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 78, LCDR2 comprising the amino acid sequence of SEQ ID NO: 79, LCDR3 comprising the amino acid sequence of SEQ ID NO: 80, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f82) [FFB0453 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f83) [FFB0453 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f84) [FFB0453 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f85) [FFB0453 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f86) [FFB0453 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f87) [FFB0453 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f88) [FFB0453 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f89) [FFB0453 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f90) [FFB0453 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 83, HCDR2 comprising the amino acid sequence of SEQ ID NO: 84, HCDR3 comprising the amino acid sequence of SEQ ID NO: 85, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 86, LCDR2 comprising the amino acid sequence of SEQ ID NO: 87, LCDR3 comprising the amino acid sequence of SEQ ID NO: 88, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f91) [FFB0065 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f92) [FFB0065 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f93) [FFB0065 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f94) [FFB0065 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f95) [FFB0065 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f96) [FFB0065 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f97) [FFB0065 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f98) [FFB0065 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f99) [FFB0065 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 115, HCDR2 comprising the amino acid sequence of SEQ ID NO: 116, HCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 118, LCDR2 comprising the amino acid sequence of SEQ ID NO: 119, LCDR3 comprising the amino acid sequence of SEQ ID NO: 120, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128;
(f100) [FFB0080 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f101) [FFB0080 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f102) [FFB0080 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f103) [FFB0080 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f104) [FFB0080 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f105) [FFB0080 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f106) [FFB0080 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f107) [FFB0080 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, LCDR3 comprising the amino acid sequence of SEQ ID NO: 128, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176;
(f108) [FFB0088 // FFB0341] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 91, HCDR2 comprising the amino acid sequence of SEQ ID NO: 92, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 93, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 94, LCDR2 comprising the amino acid sequence of SEQ ID NO: 95, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 96;
(f109) [FFB0088 // FFB0396] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 99, HCDR2 comprising the amino acid sequence of SEQ ID NO: 100, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 101, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 102, LCDR2 comprising the amino acid sequence of SEQ ID NO: 103, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 104;
(f110) [FFB0088 // FFB0464] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 107, HCDR2 comprising the amino acid sequence of SEQ ID NO: 108, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 109, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 110, LCDR2 comprising the amino acid sequence of SEQ ID NO: 111, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 112;
(f111) [FFB0088 // FFB0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 139, HCDR2 comprising the amino acid sequence of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 141, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 142, LCDR2 comprising the amino acid sequence of SEQ ID NO: 143, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 144;
(f112) [FFB0088 // FFB0035] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 147, HCDR2 comprising the amino acid sequence of SEQ ID NO: 148, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 149, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 150, LCDR2 comprising the amino acid sequence of SEQ ID NO: 151, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 152;
(f113) [FFB0088 // FFB0094] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 155, HCDR2 comprising the amino acid sequence of SEQ ID NO: 156, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 157, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 158, LCDR2 comprising the amino acid sequence of SEQ ID NO: 159, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 160;
(f114) [FFB0088 // FFB0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 163, HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 166, LCDR2 comprising the amino acid sequence of SEQ ID NO: 167, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 168;
(f115) [FFB0088 // FFB0057] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR1 comprising the amino acid sequence of SEQ ID NO: 171, HCDR2 comprising the amino acid sequence of SEQ ID NO: 172, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 173, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 174, LCDR2 comprising the amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 176; and
(f116) [FFB0088 // FFB0080] HCDR1 comprising the amino acid sequence of SEQ ID NO: 131, HCDR2 comprising the amino acid sequence of SEQ ID NO: 132, HCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 134, LCDR2 comprising the amino acid sequence of SEQ ID NO: 135, LCDR3 comprising the amino acid sequence of SEQ ID NO: 136; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 123, HCDR2 comprising the amino acid sequence of SEQ ID NO: 124, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 125, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 126, LCDR2 comprising the amino acid sequence of SEQ ID NO: 127, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 128.

According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule, or antibody fragment thereof, of the present invention, the antigen-binding molecule comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 212, and a light chain comprising an amino acid sequence of SEQ ID NO: 213, and one of the following combinations selected from (g1) to (g5) below:
(g1) a heavy chain comprising an amino acid sequence of SEQ ID NO: 210, and a light chain comprising an amino acid sequence of SEQ ID NO: 211,
(g2) a heavy chain comprising an amino acid sequence of SEQ ID NO: 214, and a light chain comprising an amino acid sequence of SEQ ID NO: 215,
(g3) a heavy chain comprising an amino acid sequence of SEQ ID NO: 216, and a light chain comprising an amino acid sequence of SEQ ID NO: 217,
(g4) a heavy chain comprising an amino acid sequence of SEQ ID NO: 218, and a light chain comprising an amino acid sequence of SEQ ID NO: 219, and
(g5) a heavy chain comprising an amino acid sequence of SEQ ID NO: 220, and a light chain comprising an amino acid sequence of SEQ ID NO: 221.

According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule, or antibody fragment thereof, of the present invention, the antigen-binding molecule comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 388, and a light chain comprising an amino acid sequence of SEQ ID NO: 389, and one of the following combinations selected from (g6) to (g7) below:
(g6) a heavy chain comprising an amino acid sequence of SEQ ID NO: 390, and a light chain comprising an amino acid sequence of SEQ ID NO: 391, and
(g7) a heavy chain comprising an amino acid sequence of SEQ ID NO: 386, and a light chain comprising an amino acid sequence of SEQ ID NO: 387.

According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule, or antibody fragment thereof, of the present invention, the antigen-binding molecule comprises one of the following combinations selected from (g8) to (g13) below:
(g8) a heavy chain comprising an amino acid sequence of SEQ ID NO: 392, and a light chain comprising an amino acid sequence of SEQ ID NO: 393;
(g9) a heavy chain comprising an amino acid sequence of SEQ ID NO: 396, and a light chain comprising an amino acid sequence of SEQ ID NO: 397;
(g10) a heavy chain comprising an amino acid sequence of SEQ ID NO: 400, and a light chain comprising an amino acid sequence of SEQ ID NO: 401;
(g11) a heavy chain comprising an amino acid sequence of SEQ ID NO: 402, and a light chain comprising an amino acid sequence of SEQ ID NO: 403;
(g12) a heavy chain comprising an amino acid sequence of SEQ ID NO: 404, and a light chain comprising an amino acid sequence of SEQ ID NO: 405; and
(g13) a heavy chain comprising an amino acid sequence of SEQ ID NO: 408, and a light chain comprising an amino acid sequence of SEQ ID NO: 409; and
one of the following combinations selected from (g14) to (g16) below:
(g14) a heavy chain comprising an amino acid sequence of SEQ ID NO: 394, and a light chain comprising an amino acid sequence of SEQ ID NO: 395;
(g15) a heavy chain comprising an amino acid sequence of SEQ ID NO: 398, and a light chain comprising an amino acid sequence of SEQ ID NO: 399; and
(g16) a heavy chain comprising an amino acid sequence of SEQ ID NO: 406, and a light chain comprising an amino acid sequence of SEQ ID NO: 407.

According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 9, 17, 25, 33, 41, 49, 57, 65, 73, 81, 89, 97, 105, 113, 121, 129, 137, 145, 153, 161, and 169, or any one of VH described in Tables 1 to 4. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a heavy chain (HC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 208, 210, 212, 214, 216, 218, 220, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, and 408, or any one of HC described in Tables 1 to 4 and Tables 7 to 8.

According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 10, 18, 26, 34, 42, 50, 58, 66, 74, 82, 90, 98, 106, 114, 122, 130, 138, 146, 154, 162, and 170, or any one of VL described in Tables 1 to 4. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a light chain (LC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 209, 211, 213, 215, 217, 219, 221, 222, 223, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, and 409, or any one of LC described in Tables 1 to 4 and Tables 7 to 8.

According to one aspect of agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a VH described in Tables 1 to 4, or as described in [0173], and a VL described in Tables 1 to 3, or as described in [0174]. According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a HC described in Tables 1 to 4 and Tables 7 to 8, or as described in [0173], and a LC described in Tables 1 to 4 and Tables 7 to 8, or as described in [0174].

According to one aspect of biparatopic agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of VH described in Tables 1 to 4, or as described in [0173], and a pair of VL described in Tables 1 to 4, or as described in [0174]. According to one aspect of the agonistic anti-FGFR2 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of HC described in Tables 1 to 4 and Tables 7 to 8, or as described in [0173], and a pair of LC described in Tables 1 to 4 and Tables 7 to 8, or as described in [0174].

According to one aspect of the agonistic antigen-binding molecule of the present invention, binding of the first antigen-binding domain and the second antigen-binding domain to FGFR2b protein leads to agonistic activity as follows:
(i) binding of the molecule to FGFR2 protein via a cell-surface expressed FGFR2b in a target cell activates signaling in the target cell, thereby exhibiting agonistic activity, and the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR2b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR2b protein in the target cell, in particular 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) binding of the molecule to FGFR2 protein via a cell-surface expressed FGFR2b in a target cell induces proliferation of the target cell, thereby exhibiting agonistic activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of agonistic activity; or
(iv) phosphorylation of the FGFR2b protein is detected in the presence of the molecule in an in vitro cell assay, wherein FGFR2b protein phosphorylation is indicative of agonistic activity.

anti-FGFR1 agonistic antigen-binding molecules
FGFR1 refers to a tyrosine protein kinase that acts as a cell-surface receptor for fibroblast growth factors. The protein is also known as basic fibroblast growth factor receptor 1(BFGFR1) and CD331. Structurally, a FGFR1 amino acid sequence is a receptor tyrosine kinase protein having a signal peptide, at least one or more immunoglobulin (Ig)-like domains, an acidic box, a transmembrane domain, and a split tyrosine kinase domain and has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of NCBI accession No. NP_ NP_001167536.1, NP_ NP_001167534.1. Structurally, a FGFR1 nucleic acid sequence has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the nucleic acid sequence of NCBI accession No. NM_001174065, NM_001174063. The FGFR1 signal peptide may be retained or cleaved off.

FGFR1 plays an essential role in the regulation of cell proliferation, differentiation, migration and apoptosis, and in the regulation of embryonic development. Activation of FGFR1 results in phosphorylation of PLCG1, FRS2, and PAK4 and activation of several signaling cascades. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. Phosphorylation of FRS2 triggers recruitment of GRB2, GAB1, PIK3R1 and SOS1, and mediates activation of RAS, MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling pathway, as well as of the AKT1 signaling pathway. FGFR1 signaling is down-regulated by ubiquitination, internalization and degradation. Mutations that lead to constitutive kinase activation or impair normal FGFR1 maturation, internalization and degradation lead to aberrant signaling. Likewise, over-expressed FGFR1 promotes activation of STAT1. In some of these cases, such as in some cancers, FGFR1 aberrations result in the formation of FGFR1 fusion proteins, or FGFR1 oncogenic isoforms, such as FGFR1-ZNF198 (NPL 19).

Fibroblast growth factor (FGF) 1 (FGF1), FGF2 are agonists for IIIb FGFR1 (FGFR1b) by binding to IgIII domain of FGFR1b (NPL 13). FGF7 and FGF10 are also weak agonists for FGFR1b (NPL 9). Various types of FGF ligands are agonists for the other IIIc FGFR1 (FGFR1c), including FGF1, FGF2, FGF4, FGF8, FGF9, FGF19, FGF21, FGF23, and FGF19, FGF21, FGF23 act on β-Klotho in complex with FGFR1c as a heterodimerization (NPL 10, NPL 11). As well as FGFR2, binding of FGF ligands to FGFR1b with heparin or heparan sulfate proteoglycans (HSPGs) is thought to be important for agonistic FGF signaling activation as receptor homodimerization (NPL 3). Although there are several reports for agonisms of FGFR1, including homodimerization-based agonist for both FGFR1b and FGFR1c (NPL 12, NPL 13), strong FGFR1b specific agonist that could show better agonistic activity than FGF ligands have not been identified yet.

Without wishing to be bound by theory, the present inventors hypothesized that the antigen-binding molecules described herein in part mimic the binding of FGF ligand such as FGF2 with FGFR1b, wherein binding of FGF ligands to FGFR1b has been elucidated to involve binding to more than one domain of FGFR1b. The aspects and embodiments described herein are based, at least in part, on the discovery that antigen-binding molecules that bind FGFR1b on different domains or bind to FGFR1b on different, or different (i.e. unique and non-overlapping) epitopes on the same domain is capable of activating FGFR1b. In a preferred embodiment, an anti-FGFR1b antigen-binding molecule that binds FGFR1b on different domains or binds to FGFR1b on different, or different (i.e. unique and non-overlapping) epitopes on the same domain, wherein the antigen-binding molecule is an agonist, is provided.

In aspects of the present invention, an anti-FGFR1b antigen-binding molecule described herein does not substantially bind to mutated FGFR1, such as FGFR1 fusion proteins, or FGFR1 oncogenic isoforms.

In yet further embodiments, the RTK protein is FGFR1b.

The terms "FGFR1b" and "FGFR1IIIb", as used herein, refer to the amino acid sequence as set forth in NCBI accession No. NP_ NP_001167536.1, and is encoded by the FGFR1 gene residing on chromosome 8. The epithelial splicing regulatory protein 1 (ESRP1) isoform is responsible for the FGFR1 splicing and consequent expression of the epithelial FGFR1b isoform.

The terms "FGFR1c" and "FGFR1IIIc", as used herein, refer to the epithelial FGFR1c isoform, and refer to the amino acid sequence as set forth in NCBI accession No. NP_001167534.1.

In the present disclosure, an anti-FGFR1b antigen binding molecule comprising a first antigen-binding domain and a second antigen-binding domain, and wherein the molecule is an agonist is provided.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human FGFR1 protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first FGFR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein, wherein the first epitope and the second epitope are different from each other.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human FGFR1 protein, wherein the molecule comprises a first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR1 protein and a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein, wherein the first epitope and the second epitope are different from each other.

According to one aspect, the first antigen-binding domain or the first means binds a first FGFR1 protein and the second antigen-binding domain or the second means binds a second FGFR1 protein.

According to one aspect, the human FGFR1 protein is human FGFR1b protein.

According to one aspect, each of the extracellular domains is independently selected from an Immunoglobulin (Ig)-like domain of the FGFR1 protein.

According to one aspect of the agonistic antigen-binding molecule of the present invention, wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR1 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR1 protein, the first domain and the second domain is the same.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first epitope in a first domain of the FGFR1 protein and the second antigen-binding domain or the second means binds to a second epitope in a second domain of the FGFR1 protein, and the first domain and the second domain is the same, but the first epitope and the second epitopes are different. In a further aspect, the first epitope and the second epitope are unique, non-overlapping epitopes.

According to another aspect of the agonistic antigen-binding molecule of the present invention, wherein the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first domain of the FGFR1 protein and the second antigen-binding domain or the second means binds to a second domain of the FGFR1 protein, and the second domain is different from the first domain.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind an Ig-like domain of the FGFR1 protein, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgI domain of FGFR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgII domain of FGFR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIII domain of FGFR1;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIb domain of FGFR1;
(v) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgI and IgIIIc domain of FGFR1;
(vi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgII domain of FGFR1;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIII domain of FGFR1;
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIb domain of FGFR1;
(ix) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgII and IgIIIc domain of FGFR1;
(x) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIII domain of FGFR1;
(xi) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIb domain of FGFR1;
(xii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIII and IgIIIc domain of FGFR1;
(xiii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR1; and
(xiv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIc domain of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds an Ig-like domain of the FGFR1 in the following combinations selected from the group consisting of:
(i) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR1;
(ii) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgIII domain of FGFR1;
(iii) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgII domain of FGFR1; and
(iv) the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the IgIIIb and IgI domain of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, (i) the first antigen-binding domain or the first means binds the IgIIIb domain of FGFR1, or (ii) the first antigen-binding domain or the first means binds specifically to IgIIIb of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the FGFR1 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds an Ig-like domain of FGFR1, selected from the group consisting of IgI, IgII, IgIII, IgIIIb, and IgIIIc of FGFR1.
In one aspect, the second antigen binding domain or the second means binds to IgI of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgI of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgII and IgIII of FGFR1.
In one aspect, the second antigen binding domain or the second means binds to IgII of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgII of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgIII of FGFR1.
In one aspect, the second antigen binding domain or the second means binds to IgIII of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIII of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
In one aspect, the second antigen binding domain or the second means binds to IgIIIb of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIb of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.
In one aspect, the second antigen binding domain or the second means binds to IgIIIc of FGFR1, or (ii) the second antigen-binding domain or the second means binds specifically to IgIIIc of FGFR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIb of FGFR1, and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain, or the first epitope of the first antigen-binding domain and the second epitope of the second antigen-binding domain are non-overlapping. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means does not compete for binding to the same or different domain of FGFR1, in particular the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgIII of FGFR1. According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgIIIb of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIIIb and IgI of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain or the first means binds a first epitope in the IgIII domain of FGFR1, and the second antigen-binding domain or the second means binds a second epitope in the IgIII domain of FGFR1, and wherein the first and the second epitope are different. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgIII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgIII and IgI of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to IgI and IgII of FGFR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain is selected from the group consisting of FFA0029, and FFA0048.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain is selected from the group consisting of FFA0020, FFA0026, FFA0032, FFA0043, FFA0047, FFA0055, FFA0064, and FFA0088.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain is the same, and is selected from the group consisting of FFA0020, FFA0026, FFA0029, FFA0032, FFA0043, FFA0047, FFA0048, FFA0055, FFA0064, and FFA0088.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain are different, and are each independently selected from the group consisting of FFA0020, FFA0026, FFA0029, FFA0032, FFA0043, FFA0047, FFA0048, FFA0055, FFA0064, and FFA0088.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain complementarity determining regions (HCDR1, HCDR2, HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, LCDR3) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 9, the amino acid sequence identifiers (SEQ ID NOs) of the variable region of the heavy chain (VH), and the variable region of the light chain (VL) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 9.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain variable regions (VH), light chain variable regions (VL), heavy chains (HC), and light chains (LC) of the biparatopic anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 9 and 14.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to FGFR1 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (h1) to (h2) below, or as described in Tables 9 and 14, or comprises any one of (h1) to (h2) below:
(h1) [FFA0029] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249; and
(h2) [FFA0048] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to FGFR1 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (i1) to (i2) below, or as described in Tables 9 and 14, or comprises any one of (i1) to (i2) below:
(i1) [FFA0029] LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and
(i2) [FFA0048] LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to FGFR1 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (j1) to (j8) below, or as described in Tables 9 and 14, or wherein the second antigen-binding domain comprises any one of (j1) to (j8) below:
(j1) [FFA0020] HCDR1 comprising the amino acid sequence of SEQ ID NO: 231, HCDR2 comprising the amino acid sequence of SEQ ID NO: 232, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 233;
(j2) [FFA0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 239, HCDR2 comprising the amino acid sequence of SEQ ID NO: 240, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 241;
(j3) [FFA0032] HCDR1 comprising the amino acid sequence of SEQ ID NO: 255, HCDR2 comprising the amino acid sequence of SEQ ID NO: 256, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 257;
(j4) [FFA0043] HCDR1 comprising the amino acid sequence of SEQ ID NO: 263, HCDR2 comprising the amino acid sequence of SEQ ID NO: 264, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 265;
(j5) [FFA0047] HCDR1 comprising the amino acid sequence of SEQ ID NO: 271, HCDR2 comprising the amino acid sequence of SEQ ID NO: 272, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 273;
(j6) [FFA0055] HCDR1 comprising the amino acid sequence of SEQ ID NO: 287, HCDR2 comprising the amino acid sequence of SEQ ID NO: 288, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 289;
(j7) [FFA0064] HCDR1 comprising the amino acid sequence of SEQ ID NO: 295, HCDR2 comprising the amino acid sequence of SEQ ID NO: 296, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 297; and
(j8) [FFA0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 303, HCDR2 comprising the amino acid sequence of SEQ ID NO: 304, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 305.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to FGFR1 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (k1) to (k8) below, or as described in Tables 9 and 14, or wherein the second antigen-binding domain comprises any one of (k1) to (k8) below:
(k1) [FFA0020] LCDR1 comprising the amino acid sequence of SEQ ID NO: 234, LCDR2 comprising the amino acid sequence of SEQ ID NO: 235, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 236;
(k2) [FFA0026] LCDR1 comprising the amino acid sequence of SEQ ID NO: 242, LCDR2 comprising the amino acid sequence of SEQ ID NO: 243, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 244;
(k3) [FFA0032] LCDR1 comprising the amino acid sequence of SEQ ID NO: 258, LCDR2 comprising the amino acid sequence of SEQ ID NO: 259, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 260;
(k4) [FFA0043] LCDR1 comprising the amino acid sequence of SEQ ID NO: 266, LCDR2 comprising the amino acid sequence of SEQ ID NO: 267, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 268;
(k5) [FFA0047] LCDR1 comprising the amino acid sequence of SEQ ID NO: 274, LCDR2 comprising the amino acid sequence of SEQ ID NO: 275, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 276;
(k6) [FFA0055] LCDR1 comprising the amino acid sequence of SEQ ID NO: 290, LCDR2 comprising the amino acid sequence of SEQ ID NO: 291, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 292;
(k7) [FFA0064] LCDR1 comprising the amino acid sequence of SEQ ID NO: 298, LCDR2 comprising the amino acid sequence of SEQ ID NO: 299, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 300; and
(k8) [FFA0088] LCDR1 comprising the amino acid sequence of SEQ ID NO: 306, LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 308.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to FGFR1 with an antigen-binding domain comprising any one of (l1) to (l10) below, or as described in Tables 9 and 14, or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (l1) to (l10) below:
(l1) [FFA0020] HCDR1 comprising the amino acid sequence of SEQ ID NO: 231, HCDR2 comprising the amino acid sequence of SEQ ID NO: 232, HCDR3 comprising the amino acid sequence of SEQ ID NO: 233, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 234, LCDR2 comprising the amino acid sequence of SEQ ID NO: 235, LCDR3 comprising the amino acid sequence of SEQ ID NO: 236;
(l2) [FFA0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 239, HCDR2 comprising the amino acid sequence of SEQ ID NO: 240, HCDR3 comprising the amino acid sequence of SEQ ID NO: 241, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 242, LCDR2 comprising the amino acid sequence of SEQ ID NO: 243, LCDR3 comprising the amino acid sequence of SEQ ID NO: 244;
(l3) [FFA0029] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, LCDR3 comprising the amino acid sequence of SEQ ID NO: 252;
(l4) [FFA0032] HCDR1 comprising the amino acid sequence of SEQ ID NO: 255, HCDR2 comprising the amino acid sequence of SEQ ID NO: 256, HCDR3 comprising the amino acid sequence of SEQ ID NO: 257, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 258, LCDR2 comprising the amino acid sequence of SEQ ID NO: 259, LCDR3 comprising the amino acid sequence of SEQ ID NO: 260;
(l5) [FFA0043] HCDR1 comprising the amino acid sequence of SEQ ID NO: 263, HCDR2 comprising the amino acid sequence of SEQ ID NO: 264, HCDR3 comprising the amino acid sequence of SEQ ID NO: 265, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 266, LCDR2 comprising the amino acid sequence of SEQ ID NO: 267, LCDR3 comprising the amino acid sequence of SEQ ID NO: 268;
(l6) [FFA0047] HCDR1 comprising the amino acid sequence of SEQ ID NO: 271, HCDR2 comprising the amino acid sequence of SEQ ID NO: 272, HCDR3 comprising the amino acid sequence of SEQ ID NO: 273, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 274, LCDR2 comprising the amino acid sequence of SEQ ID NO: 275, LCDR3 comprising the amino acid sequence of SEQ ID NO: 276;
(l7) [FFA0048] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, LCDR3 comprising the amino acid sequence of SEQ ID NO: 284;
(l8) [FFA0055] HCDR1 comprising the amino acid sequence of SEQ ID NO: 287, HCDR2 comprising the amino acid sequence of SEQ ID NO: 288, HCDR3 comprising the amino acid sequence of SEQ ID NO: 289, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 290, LCDR2 comprising the amino acid sequence of SEQ ID NO: 291, LCDR3 comprising the amino acid sequence of SEQ ID NO: 292;
(l9) [FFA0064] HCDR1 comprising the amino acid sequence of SEQ ID NO: 295, HCDR2 comprising the amino acid sequence of SEQ ID NO: 296, HCDR3 comprising the amino acid sequence of SEQ ID NO: 297, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 298, LCDR2 comprising the amino acid sequence of SEQ ID NO: 299, LCDR3 comprising the amino acid sequence of SEQ ID NO: 300; and
(l10) [FFA0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 303, HCDR2 comprising the amino acid sequence of SEQ ID NO: 304, HCDR3 comprising the amino acid sequence of SEQ ID NO: 305, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 306, LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, LCDR3 comprising the amino acid sequence of SEQ ID NO: 308.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 compete for binding to FGFR1 with an antigen-binding domain comprising any one of (m1) to (m16) below, or as described in Tables 9 and 14, or each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (m1) to (m16) below:
(m1) [FFA0029 // FFA0020] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 231, HCDR2 comprising the amino acid sequence of SEQ ID NO: 232, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 233, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 234, LCDR2 comprising the amino acid sequence of SEQ ID NO: 235, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 236;
(m2) [FFA0029 // FFA0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 239, HCDR2 comprising the amino acid sequence of SEQ ID NO: 240, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 241, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 242, LCDR2 comprising the amino acid sequence of SEQ ID NO: 243, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 244;
(m3) [FFA0029 // FFA0032] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 255, HCDR2 comprising the amino acid sequence of SEQ ID NO: 256, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 257, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 258, LCDR2 comprising the amino acid sequence of SEQ ID NO: 259, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 260;
(m4) [FFA0029 // FFA0043] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 263, HCDR2 comprising the amino acid sequence of SEQ ID NO: 264, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 265, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 266, LCDR2 comprising the amino acid sequence of SEQ ID NO: 267, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 268;
(m5) [FFA0029 // FFA0047] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 271, HCDR2 comprising the amino acid sequence of SEQ ID NO: 272, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 273, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 274, LCDR2 comprising the amino acid sequence of SEQ ID NO: 275, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 276;
(m6) [FFA0029 // FFA0055] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO:287, HCDR2 comprising the amino acid sequence of SEQ ID NO: 288, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 289, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 290, LCDR2 comprising the amino acid sequence of SEQ ID NO: 291, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 292;
(m7) [FFA0029 // FFA0064] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 295, HCDR2 comprising the amino acid sequence of SEQ ID NO: 296, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 297, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 298, LCDR2 comprising the amino acid sequence of SEQ ID NO: 299, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 300;
(m8) [FFA0029 // FFA0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 247, HCDR2 comprising the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 249, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 250, LCDR2 comprising the amino acid sequence of SEQ ID NO: 251, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 252; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 303, HCDR2 comprising the amino acid sequence of SEQ ID NO: 304, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 305, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 306, LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 308;
(m9) [FFA0048 // FFA0020] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 231, HCDR2 comprising the amino acid sequence of SEQ ID NO: 232, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 233, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 234, LCDR2 comprising the amino acid sequence of SEQ ID NO: 235, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 236;
(m10) [FFA0048 // FFA0026] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 239, HCDR2 comprising the amino acid sequence of SEQ ID NO: 240, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 241, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 242, LCDR2 comprising the amino acid sequence of SEQ ID NO: 243, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 244;
(m11) [FFA0048 // FFA0032] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 255, HCDR2 comprising the amino acid sequence of SEQ ID NO: 256, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 257, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 258, LCDR2 comprising the amino acid sequence of SEQ ID NO: 259, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 260;
(m12) [FFA0048 // FFA0043] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 263, HCDR2 comprising the amino acid sequence of SEQ ID NO: 264, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 265, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 266, LCDR2 comprising the amino acid sequence of SEQ ID NO: 267, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 268;
(m13) [FFA0048 // FFA0047] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 271, HCDR2 comprising the amino acid sequence of SEQ ID NO: 272, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 273, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 274, LCDR2 comprising the amino acid sequence of SEQ ID NO: 275, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 276;
(m14) [FFA0048 // FFA0055] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 287, HCDR2 comprising the amino acid sequence of SEQ ID NO: 288, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 289, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 290, LCDR2 comprising the amino acid sequence of SEQ ID NO: 291, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 292;
(m15) [FFA0048 // FFA0064] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 295, HCDR2 comprising the amino acid sequence of SEQ ID NO: 296, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 297, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 298, LCDR2 comprising the amino acid sequence of SEQ ID NO: 299, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 300; and
(m16) [FFA0048 // FFA0088] HCDR1 comprising the amino acid sequence of SEQ ID NO: 279, HCDR2 comprising the amino acid sequence of SEQ ID NO: 280, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 281, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 282, LCDR2 comprising the amino acid sequence of SEQ ID NO: 283, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 284; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 303, HCDR2 comprising the amino acid sequence of SEQ ID NO: 304, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 305, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 306, LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 308.

According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule, or antibody fragment thereof, of the present invention, the antigen-binding molecule comprises one of the following combinations selected from (n1) to (n2) below:
(n1) a heavy chain comprising an amino acid sequence of SEQ ID NO: 364, and a light chain comprising an amino acid sequence of SEQ ID NO: 365, and
(n2) a heavy chain comprising an amino acid sequence of SEQ ID NO: 372, and a light chain comprising an amino acid sequence of SEQ ID NO: 373; and
one of the following combinations selected from (n3) to (n10) below:
(n3) a heavy chain comprising an amino acid sequence of SEQ ID NO: 362, and a light chain comprising an amino acid sequence of SEQ ID NO: 363,
(n4) a heavy chain comprising an amino acid sequence of SEQ ID NO: 366, and a light chain comprising an amino acid sequence of SEQ ID NO: 367,
(n5) a heavy chain comprising an amino acid sequence of SEQ ID NO: 368, and a light chain comprising an amino acid sequence of SEQ ID NO: 369,
(n6) a heavy chain comprising an amino acid sequence of SEQ ID NO: 370, and a light chain comprising an amino acid sequence of SEQ ID NO: 371,
(n7) a heavy chain comprising an amino acid sequence of SEQ ID NO: 374, and a light chain comprising an amino acid sequence of SEQ ID NO: 375,
(n8) a heavy chain comprising an amino acid sequence of SEQ ID NO: 376, and a light chain comprising an amino acid sequence of SEQ ID NO: 377,
(n9) a heavy chain comprising an amino acid sequence of SEQ ID NO: 378, and a light chain comprising an amino acid sequence of SEQ ID NO: 379, and
(n10) a heavy chain comprising an amino acid sequence of SEQ ID NO: 380, and a light chain comprising an amino acid sequence of SEQ ID NO: 381.

According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 229, 237, 245, 253, 261, 269, 277, 285, 293, and 301, or any one of VH described in Table 9. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a heavy chain (HC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 362, 364, 366, 368, 370, 372, 374, 376, 378, and 380, or any one of HC described in Table 14.

According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 230, 238, 246, 254, 262, 270, 278, 286, 294, and 302, or any one of VL described in Table 9. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a light chain (LC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 363, 365, 367, 369, 371, 373, 375, 377, 379, and 381, or any one of LC described in Table 14.

According to one aspect of agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a VH described in Table 9, or as described in [0226], and a VL described in Table 9, or as described in [0227]. According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a HC described in Table 14, or as described in [0226], and a LC described in Table 14, or as described in [0227].

According to one aspect of biparatopic agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of VH described in Table 9, or as described in [0226], and a pair of VL described in Table 9, or as described in [0227]. According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of HC described in Table 14, or as described in [0226], and a pair of LC described in Table 14, or as described in [0227].

According to one aspect of the agonistic antigen-binding molecule of the present invention, binding of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means to FGFR1b protein leads to agonistic activity as follows:
(i) binding of the molecule to FGFR1 protein via a cell-surface expressed FGFR1b in a target cell activates signaling in the target cell, thereby exhibiting agonistic activity, and the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR1b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR1b protein in the target cell, in particular 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
(ii) binding of the molecule to FGFR1 protein via a cell-surface expressed FGFR1b in a target cell induces proliferation of the target cell, thereby exhibiting agonistic activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of agonistic activity; or
(iv) phosphorylation of the FGFR1b protein is detected in the presence of the molecule in an in vitro cell assay, wherein FGFR1b protein phosphorylation is indicative of agonistic activity.

anti-DDR1 agonistic antigen-binding molecules
DDR1 refers to a tyrosine protein kinase that acts as a cell-surface receptor for collagen. The protein is also known as CD167. Structurally, a DDR1 amino acid sequence is a receptor tyrosine kinase protein having a signal peptide, at least one or more immunoglobulin (Ig)-like domains, an acidic box, a transmembrane domain, and a split tyrosine kinase domain and has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of NCBI accession No. NP_ 001945.3, NP_054699.2, NP_054700.2, NP_001189450.1, NP_001189451.1. Structurally, a DDR1 nucleic acid sequence has over its full length at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the nucleic acid sequence of NCBI accession No. NM_001954.5, NM_013993, NM_013994, NM_001202521, NM_001202522. The DDR1 signal peptide may be retained or cleaved off.

DDR1 play an important role in regulation of cell proliferation, survival migration, adhesion, aggregation, and ECM remodeling. Src kinases is thought to be required for phosphorylation of specific tyrosine residues in the KD of the DDR1, and the phosphorylation of these tyrosine residues recruits the binding of various proteins containing Src homology-2 (SH2) and phosphotyrosine binding domains. Furthermore, these phosphorylated residues of KD lead to autophosphorylation of several other tyrosine residues present in IJM domain, which serves as binding sites of numerous adaptor proteins including SHC transforming protein 1 or SH2 domain containing transforming protein 1, Nck2, tyrosine phosphatase protein or SHP-2, C-terminal Src kinase, non-muscle myosin heavy chain-IIA, Pyk2 of focal adhesion kinase (FAK) superfamily, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), cell division control protein-42 (Cdc42), extracellular signal regulated kinase - mitogen-activated protein kinase (MAPK1), activator protein 1 (AP1), and also signal transducers and activators of protein (STAT) family of intracellular transcription factors (STAT1a/b, STAT3, and STAT5). DDR1 also activates other major pathways including Ras/Raf/ERK and PI3K/AKT pathways (NPL 15).

DDR1 is activated upon collagen binding at the DS domain, and then initiates downstream signaling pathways including receptor phosphorylation (NPL 15). DDR1 forms constitutive dimers, and collagen induced lateral association of DDR1 dimers and phosphorylation between dimers are suggested, however, mechanisms of agonistic signal activation of DDR1 are not completely understood yet (NPL 20). There is a report about mutation in transmembrane region of DDR1 (DDR1-TM, NPL 21). DDR1-TM shows defective transmembrane helix association with impaired signal activation, suggesting that appropriate two DDR1 association would be important for its agonistic signal activation.

Without wishing to be bound by theory, the present inventors hypothesized that the antigen-binding molecules described herein in part mimic DDR1 activation, wherein binding of collagen to DDR1 has been elucidated to involve binding or association to more than one domain of DDR1. The aspects and embodiments described herein are based, at least in part, on the discovery that antigen-binding molecules that bind DDR1 on different domains or bind to DDR1 on different, or different (i.e. unique and non-overlapping) epitopes on the same domain is capable of activating DDR1. In a preferred embodiment, an anti-DDR1 antigen-binding molecule that binds DDR1 on different domains or binds to DDR1 on different, or different (i.e. unique and non-overlapping) epitopes on the same domain, wherein the antigen-binding molecule is an agonist, is provided.

In aspects of the present invention, an anti- DDR1 antigen-binding molecule described herein does not substantially bind to mutated DDR1, such as DDR1 fusion proteins, or DDR1 oncogenic isoforms.

In yet further embodiments, the RTK protein is DDR1.

The term "DDR1", as used herein, refers to the amino acid sequence as set forth in NCBI accession No. NP_ 001945.3, NP_054699.2, NP_054700.2, NP_001189450.1, NP_001189451.1, and is encoded by the DDR1 gene residing on chromosome 6.

In the present disclosure, an anti-DDR1 antigen binding molecule comprising a first antigen-binding domain and a second antigen-binding domain, and wherein the molecule is an agonist is provided.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human DDR1 protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first DDR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein, wherein the first epitope and the second epitope are different from each other.

The present invention therefore provides an agonistic antigen-binding molecule that binds to a human DDR1 protein, wherein the molecule comprises a first means for binding a first epitope of an extracellular domain (ECD) of a first DDR1 protein and a second means for binding a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein, wherein the first epitope and the second epitope are different from each other.

According to one aspect, the first antigen-binding domain or the first means binds a first DDR1 protein and the second antigen-binding domain or the second means binds a second DDR1 protein.

According to one aspect, the human DDR1 protein is human DDR1a protein or human DDR1b protein.

According to one aspect, each of the extracellular domains is independently selected from a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 protein.

According to one aspect of the agonistic antigen-binding molecule of the present invention, wherein the first antigen-binding domain or the first means binds to a first domain of the DDR1 protein and the second antigen-binding domain or the second means binds to a second domain of the DDR1 protein, the first domain and the second domain is the same.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first epitope in a first domain of the DDR1 protein and the second antigen-binding domain or the second means binds to a second epitope in a second domain of the DDR1 protein, and the first domain and the second domain is the same, but the first epitope and the second epitopes are different. In a further aspect, the first epitope and the second epitope are unique, non-overlapping epitopes.

According to another aspect of the agonistic antigen-binding molecule of the present invention, wherein the agonistic antigen-binding molecule is biparatopic, and wherein the first antigen-binding domain or the first means binds to a first domain of the DDR1 protein and the second antigen-binding domain or the second means binds to a second domain of the DDR1 protein, and the second domain is different from the first domain.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 protein.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS domain of DDR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS and DS-like domain of DDR1;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and EJXM region of DDR1;
(iv) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and DS-like domain of DDR1;
(vii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS-like domain and EJXM region of DDR1; and
(viii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the EJXM region and EJXM region of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of the DDR1 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and the DS domain of DDR1;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and DS-like domain of DDR1; and
(iii) wherein the first antigen-binding domain and the second antigen-binding domain or the first means and the second means, independently of each other, bind the DS domain and EJXM region of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, (i) the first antigen-binding domain or the first means binds the DS domain of DDR1, or (ii) the first antigen-binding domain binds or the first means specifically to the DS domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS-like domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the EJXM region of DDR1.

According to one aspect of the agostic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds a second epitope, and wherein the second epitope resides in the same as or different domain of the DDR1 from the first epitope, or wherein the second antigen-binding domain or the second means binds a second epitope of the ECD of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain or the second means binds a discoidin (DS) domain, a DS-like domain, or an extracellular juxtamembrane (EJXM) region of DDR1.
In one aspect, (i) the second antigen binding domain or the second means binds to the DS domain of DDR1, or (ii) the second antigen-binding domain or the second means binds specifically to the DS domain of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS-like domain and/or the EJXM region domain of DDR1.
In one aspect, (i) the second antigen binding domain or the second means binds to the DS-like domain and/or the EJXM region domain of DDR1, or (ii) the second antigen-binding domain or the second means binds specifically to the DS-like domain and/or the EJXM region of DDR1 and shows less than 10%, or less than 5%, or 0% cross-reactivity with the DS domain of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first epitope of the first antigen-binding domain is different from the second epitope of the second antigen-binding domain, or the first epitope of the first antigen-binding domain and the second epitope of the second antigen-binding domain are non-overlapping. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same or different domain of DDR1, in particular the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to the DS domain and the DS domain of DDR1. According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to the DS domain and the DS-like domain and/or the EJXM region of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule binds biparatopically to the DS-like domain and/or the EJXM region and the DS-like domain and/or the EJXM region domain of DDR1.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain binds a first epitope in the DS domain of DDR1, and the second antigen-binding domain binds a second epitope in the DS domain of DDR1, and wherein the first and the second epitope are different. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain binds a first epitope in the DS domain of DDR1, and the second antigen-binding domain binds a second epitope in the DS-like domain and/or the EJXM region domain of DDR1, and wherein the first and the second epitope are different. According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain or the first means and the second means do not compete for binding to the same domain of FGFR2.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain is selected from the group consisting of DAA0011 and DAA0045.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain is selected from the group consisting of DAA0007, DAA0012, and DAA0024.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain is the same, and is selected from the group consisting of DAA0011, DAA0045, DAA0007, DAA0012, and DAA0024.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first and the second antigen-binding domain are different, and are each independently selected from the group consisting of DAA0011, DAA0045, DAA0007, DAA0012, and DAA0024.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain complementarity determining regions (HCDR1, HCDR2, HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, LCDR3) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 10, the amino acid sequence identifiers (SEQ ID NOs) of the variable region of the heavy chain (VH), and the variable region of the light chain (VL) of the agonistic antigen-binding molecule, monoparatopic and biparatopic anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Table 10.

The amino acid sequence identifiers (SEQ ID NOs) of the heavy chain variable regions (VH), light chain variable regions (VL), heavy chains (HC), and light chains (LC) of the biparatopic anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention are presented in Tables 10 and 13.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to DDR1 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (o1) to (o2) below, or as described in Table 10, or comprises any one of (o1) to (o2) below:
(o1) [DAA0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 343, HCDR2 comprising the amino acid sequence of SEQ ID NO: 344, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 345; and
(o2) [DAA0011] HCDR1 comprising the amino acid sequence of SEQ ID NO: 319, HCDR2 comprising the amino acid sequence of SEQ ID NO: 320, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 321.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain competes for binding to DDR1 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (p1) to (p2) below, or as described in Table 10, or comprises any one of (p1) to (p2) below:
(p1) [DAA0045] LCDR1 comprising the amino acid sequence of SEQ ID NO: 346, LCDR2 comprising the amino acid sequence of SEQ ID NO: 347, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 348; and
(p2) [DAA0011] LCDR1 comprising the amino acid sequence of SEQ ID NO: 322, LCDR2 comprising the amino acid sequence of SEQ ID NO: 323, and LCDR3 comprising the amino acid sequence of SEQ ID NO:324.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to DDR1 with an antigen-binding domain comprising any one of HCDR1, HCDR2 and HCDR3 of an antigen-binding domain comprising any one of (q1) to (q3) below, or as described in Table 10, or wherein the second antigen-binding domain comprises any one of (q1) to (q3) below:
(q1) [DAA0007] HCDR1 comprising the amino acid sequence of SEQ ID NO: 311, HCDR2 comprising the amino acid sequence of SEQ ID NO: 312, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 313;
(q2) [DAA0024] HCDR1 comprising the amino acid sequence of SEQ ID NO: 335, HCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 337; and
(q3) [DAA0012] HCDR1 comprising the amino acid sequence of SEQ ID NO: 327, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 329.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the second antigen-binding domain competes for binding to DDR1 with an antigen-binding domain comprising any one of LCDR1, LCDR2 and LCDR3 of an antigen-binding domain comprising any one of (r1) to (r3) below, or as described in Table 10, or wherein the second antigen-binding domain comprises any one of (r1) to (r3) below:
(r1) [DAA0007] LCDR1 comprising the amino acid sequence of SEQ ID NO: 314, LCDR2 comprising the amino acid sequence of SEQ ID NO: 315, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 316;
(r2) [DAA0024] LCDR1 comprising the amino acid sequence of SEQ ID NO: 338, LCDR2 comprising the amino acid sequence of SEQ ID NO: 339, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 340; and
(r3) [DAA0012] LCDR1 comprising the amino acid sequence of SEQ ID NO: 330, LCDR2 comprising the amino acid sequence of SEQ ID NO: 331, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 332.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain bind the same epitope and wherein the first antigen-binding domain and the second antigen-binding domain compete for binding to DDR1 with an antigen-binding domain comprising any one of (s1) to (s5) below, or as described in Table 10, or wherein the first antigen-binding domain and the second antigen-binding domain comprises any one of (s1) to (s5) below:
(s1) [DAA0011] HCDR1 comprising the amino acid sequence of SEQ ID NO: 319, HCDR2 comprising the amino acid sequence of SEQ ID NO: 320, HCDR3 comprising the amino acid sequence of SEQ ID NO: 321, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 322, LCDR2 comprising the amino acid sequence of SEQ ID NO: 323, LCDR3 comprising the amino acid sequence of SEQ ID NO: 324;
(s2) [DAA0045] HCDR1 comprising the amino acid sequence of SEQ ID NO: 343, HCDR2 comprising the amino acid sequence of SEQ ID NO: 344, HCDR3 comprising the amino acid sequence of SEQ ID NO: 345, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 346, LCDR2 comprising the amino acid sequence of SEQ ID NO: 347, LCDR3 comprising the amino acid sequence of SEQ ID NO: 348;
(s3) [DAA0007] HCDR1 comprising the amino acid sequence of SEQ ID NO: 311, HCDR2 comprising the amino acid sequence of SEQ ID NO: 312, HCDR3 comprising the amino acid sequence of SEQ ID NO: 313, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 314, LCDR2 comprising the amino acid sequence of SEQ ID NO: 315, LCDR3 comprising the amino acid sequence of SEQ ID NO: 316;
(s4) [DAA0012] HCDR1 comprising the amino acid sequence of SEQ ID NO: 327, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, HCDR3 comprising the amino acid sequence of SEQ ID NO: 329, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 330, LCDR2 comprising the amino acid sequence of SEQ ID NO: 331, LCDR3 comprising the amino acid sequence of SEQ ID NO: 332; and
(s5) [DAA0024] HCDR1 comprising the amino acid sequence of SEQ ID NO: 335, HCDR2 comprising the amino acid sequence of SEQ ID NO: 336, HCDR3 comprising the amino acid sequence of SEQ ID NO: 337, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 338, LCDR2 comprising the amino acid sequence of SEQ ID NO: 339, LCDR3 comprising the amino acid sequence of SEQ ID NO: 340.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 compete for binding to DDR1 with an antigen-binding domain comprising any one of (t1) to (t6) below, or as described in Tables 10 and 13, or each of the first antigen-binding domain and the second antigen-binding domain comprises any one of (t1) to (t6) below:
(t1) [DAA0045 // DAA0007] HCDR1 comprising the amino acid sequence of SEQ ID NO: 343, HCDR2 comprising the amino acid sequence of SEQ ID NO: 344, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 345, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 346, LCDR2 comprising the amino acid sequence of SEQ ID NO: 347, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 348; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 311, HCDR2 comprising the amino acid sequence of SEQ ID NO: 312, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 313, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 314, LCDR2 comprising the amino acid sequence of SEQ ID NO: 315, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 316;
(t2) [DAA0045 // DAA0024] HCDR1 comprising the amino acid sequence of SEQ ID NO: 343, HCDR2 comprising the amino acid sequence of SEQ ID NO: 344, HCDR3 comprising the amino acid sequence of SEQ ID NO: 345, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 346, LCDR2 comprising the amino acid sequence of SEQ ID NO: 347, LCDR3 comprising the amino acid sequence of SEQ ID NO: 348; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 335, HCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 337, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 338, LCDR2 comprising the amino acid sequence of SEQ ID NO: 339, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 340;
(t3) [DAA0045 // DAA0012] HCDR1 comprising the amino acid sequence of SEQ ID NO: 343, HCDR2 comprising the amino acid sequence of SEQ ID NO: 344, HCDR3 comprising the amino acid sequence of SEQ ID NO: 345, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 346, LCDR2 comprising the amino acid sequence of SEQ ID NO: 347, LCDR3 comprising the amino acid sequence of SEQ ID NO: 348; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 327, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 329, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 330, LCDR2 comprising the amino acid sequence of SEQ ID NO: 331, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 332;
(t4) [DAA0011 // DAA0007] HCDR1 comprising the amino acid sequence of SEQ ID NO: 319, HCDR2 comprising the amino acid sequence of SEQ ID NO: 320, HCDR3 comprising the amino acid sequence of SEQ ID NO: 321, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 322, LCDR2 comprising the amino acid sequence of SEQ ID NO: 323, LCDR3 comprising the amino acid sequence of SEQ ID NO: 324; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 311, HCDR2 comprising the amino acid sequence of SEQ ID NO: 312, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 313, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 314, LCDR2 comprising the amino acid sequence of SEQ ID NO: 315, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 316;
(t5) [DAA0011 // DAA0024] HCDR1 comprising the amino acid sequence of SEQ ID NO: 319, HCDR2 comprising the amino acid sequence of SEQ ID NO: 320, HCDR3 comprising the amino acid sequence of SEQ ID NO: 321, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 322, LCDR2 comprising the amino acid sequence of SEQ ID NO: 323, LCDR3 comprising the amino acid sequence of SEQ ID NO: 324; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 335, HCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 337, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 338, LCDR2 comprising the amino acid sequence of SEQ ID NO: 339, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 340; and
(t6) [DAA0011 // DAA0012] HCDR1 comprising the amino acid sequence of SEQ ID NO: 319, HCDR2 comprising the amino acid sequence of SEQ ID NO: 320, HCDR3 comprising the amino acid sequence of SEQ ID NO: 321, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 322, LCDR2 comprising the amino acid sequence of SEQ ID NO: 323, LCDR3 comprising the amino acid sequence of SEQ ID NO: 324; and HCDR1 comprising the amino acid sequence of SEQ ID NO: 327, HCDR2 comprising the amino acid sequence of SEQ ID NO: 328, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 329, and LCDR1 comprising the amino acid sequence of SEQ ID NO: 330, LCDR2 comprising the amino acid sequence of SEQ ID NO: 331, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 332.

According to one aspect of the agonistic anti-DDR1 antigen-binding molecule, or antibody fragment thereof, of the present invention, the antigen-binding molecule comprises one of the following combinations selected from (u1) to (u2) below:
(u1) a heavy chain comprising an amino acid sequence of SEQ ID NO: 354, and a light chain comprising an amino acid sequence of SEQ ID NO: 355, and
(u2) a heavy chain comprising an amino acid sequence of SEQ ID NO: 358, and a light chain comprising an amino acid sequence of SEQ ID NO: 359; and
one of the following combinations selected from (u3) to (u5) below:
(u3) a heavy chain comprising an amino acid sequence of SEQ ID NO: 352, and a light chain comprising an amino acid sequence of SEQ ID NO: 353,
(u4) a heavy chain comprising an amino acid sequence of SEQ ID NO: 356, and a light chain comprising an amino acid sequence of SEQ ID NO: 357, and
(u5) a heavy chain comprising an amino acid sequence of SEQ ID NO: 360, and a light chain comprising an amino acid sequence of SEQ ID NO: 361.

According to one aspect of the agonistic anti-DDR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 309, 317, 325, 333, and 341, or any one of VH described in Table 10. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a heavy chain (HC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 352, 354, 356, 358, and 360, or any one of HC described in Table 13.

According to one aspect of the agonistic anti- DDR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises a light chain variable domain (VL) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 310, 318, 326, 334, and 342, or any one of VL described in Table 10. According to one aspect of the agonistic antigen-binding molecule of the present invention, the antigen-binding molecule comprises a light chain (LC) comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 353, 355, 357, 359, and 361, or any one of LC described in Table 13.

According to one aspect of agonistic anti-DDR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a VH described in Table 10, or as described in [0274], and a VL described in Table 10, or as described in [0275]. According to one aspect of the agonistic anti-FGFR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a HC described in Table 13, or as described in [0274], and a LC described in Table 13, or as described in [0275].

According to one aspect of biparatopic agonistic anti-DDR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of VH described in Table 10, or as described in [0274], and a pair of VL described in Table 10, or as described in [0275]. According to one aspect of the agonistic anti-DDR1 antigen-binding molecule or antibody or fragment thereof, of the present invention, the antigen-binding molecule comprises any combination of a pair of HC described in Table 13, or as described in [0274], and a pair of LC described in Table 13, or as described in [0275].

According to one aspect of the agonistic antigen-binding molecule of the present invention, binding of the first antigen-binding domain and the second antigen-binding domain or the first means and the second means to DDR1 protein leads to agonistic activity as follows:
(i) binding of the molecule to DDR1 protein via a cell-surface expressed DDR1a in a target cell activates signaling in the target cell, thereby exhibiting agonistic activity, and the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of DDR1a by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed DDR1a protein in the target cell, in particular wherein the intracellular protein downstream of DDR1a is selected from the group consisting of: SHP-2, FAK, NF-kappa B, Cdc42, MAPK1, AP1, Ras/Raf/ERK, PI3K/AKT, STAT1/3/5; or
(ii) binding of the molecule to DDR1 protein via a cell-surface expressed DDR1a in a target cell induces proliferation of the target cell, thereby exhibiting agonistic activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of agonistic activity; or
(iv) phosphorylation of the DDR1a protein is detected in the presence of the molecule in an in vitro cell assay, wherein DDR1a protein phosphorylation is indicative of agonistic activity.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the first antigen-binding domain and the second antigen-binding domain are each independently selected from a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, optionally wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain, in particular wherein said at least one disulfide bond is formed between the group selected from the following:
(i) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
(ii) an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iii) an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
(iv) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
(v) an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
(vi) an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a 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 originate is present at a position selected from the group consisting of positions 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, according to EU numbering, in the CH1 region.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, wherein said one disulfide bond is formed between an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL 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 originate is present at a position selected from the group consisting of positions 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 numbering in the CL region.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, wherein said one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of the second antigen-binding domain, and the amino acid residue in the CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197 according to EU numbering, and the amino acid residue in the CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128 according to Kabat numbering.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, wherein said one disulfide bond is formed between an amino acid residue in a VH region of a first antigen-binding domain and an amino acid residue in a VH 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 originate is present at a position selected from the group consisting of positions 6, 8, 16, 20, 25, 26, 28, 74, and 82b according to Kabat numbering in the VH region.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, wherein said one disulfide bond is formed between an amino acid residue in a VHH region of a first antigen-binding domain and an amino acid residue in a VHH 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 originate is present at a position selected from the group consisting of positions 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 numbering in the VHH region.

According to one aspect of the agonistic antigen-binding molecule of the present invention, 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 which are not in a hinge region, the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, wherein said one disulfide bond is formed between an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of the second antigen-binding domain, and wherein when the VL is of a subclass kappa, the amino acid residue from which the bond between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 21, 27, 58, 77, 100, 105, and 107 according to Kabat numbering in the VL region (subclass kappa); or
wherein when the VL is of a subclass lambda, the amino acid residue from which the bonds between the first and second antigen-binding domains originate is present at a position selected from the group consisting of positions 6, 19, 33, and 34 according to Kabat numbering in the VL region (subclass lambda).

According to one aspect of the agonistic antigen-binding molecule of the present invention, the at least one disulfide bond is formed between an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain. In one aspect, at least one disulfide bond is formed between the amino acid residue at position 191 in a CH1 region of a first antigen-binding domain (according to Kabat numbering) and the amino acid residue at position 126 in a CL region of a second antigen-binding domain (according to Kabat numbering), preferably the at least one disulfide bond is formed between S191C in the CH1 region of a first antigen-binding domain (according to Kabat numbering) and K126C in a CL region of a second antigen-binding domain (according to Kabat numbering).

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule comprises an antibody constant region, in particular (i) the antibody constant region is a human antibody constant region, or (ii) the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule is characterized by two antigen-binding domains, or the molecule is a bivalent molecule, or the first antigen-binding domain and second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.

According to one aspect of the agonistic antigen-binding molecule of the present invention, the molecule comprises more than two antigen-binding domains, or the molecule is tri-valent, tetra-valent, or multivalent. Optionally, the molecule comprises a third antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain.

According to one aspect of the agonistic antigen-binding molecule of the present invention, wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain. Optionally, wherein the molecule comprises a third and a fourth antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain.

The first antigen-binding domains of the agonistic antigen-binding molecules of the present invention are encompassed by the term "first means for binding a first epitope of a first RTK protein". In particular, the first antigen-binding domains of the anti-FGFR2b agonistic antigen-binding molecules, the anti-FGFR1b agonistic antigen-binding molecules, and the anti-DDR1 agonistic antigen-binding molecules of the present invention are encompassed by the term "first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR2 protein", "first means for binding a first epitope of an extracellular domain (ECD) of a first FGFR1 protein", and "first means for binding a first epitope of an extracellular domain (ECD) of a first DDR1 protein", respectively.

The second antigen-binding domains of the agonistic antigen-binding molecules of the present invention are encompassed by the term "a second means for binding a second epitope of the first or a second RTK protein". In particular, the second antigen-binding domains of the anti-FGFR2b agonistic antigen-binding molecules, the anti-FGFR1b agonistic antigen-binding molecules, and the anti-DDR1 agonistic antigen-binding molecules of the present invention are encompassed by the term "second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein", "second means for binding a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein", and "second means for binding a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein", respectively.

Assays
The agonistic antigen binding molecule of the present invention, provided herein may be identified, screened for, or characterized for their physical/chemical properties and/or biological activities by various assays known in the art.

In one aspect, the agonistic antigen binding molecule of the invention is tested for its antigen binding activity, e.g. by known methods such as ELISA, Western blot, Luciferase Reporter Assay, etc.

In one aspect, competition assays may be used to identify antigen binding molecules that compete with the agonistic antigen binding molecule of the invention for binding to the same antigen. In certain embodiments, such a competing antigen binding molecule binds to the same epitope (e.g. a linear or a conformational epitope) that is bound by the agonistic antigen binding molecule of the present invention. Detailed exemplary methods for mapping an epitope to which an antigen-binding molecule, antibody or fragments thereof, binds are provided in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

In an exemplary competition assay, immobilized antigen (i.e. FGFR2 and isotype variants thereof) is/are incubated in a solution comprising a first labeled antigen-binding molecule or antibody that binds to antigen (e.g., any one of the antigen-binding molecule or antibody described herein) and a second unlabeled antigen-binding molecule or antibody that is being tested for its ability to compete with the first antigen-binding molecule or antibody for binding to antigen. The second antigen-binding molecule or antibody may be present in a hybridoma supernatant. As a control, immobilized antigen is incubated in a solution comprising the first labeled antigen-binding molecule or antibody but not the second unlabeled antigen-binding molecule or antibody. After incubation under conditions permissive for binding of the first antigen-binding molecule or antibody to antigen, excess unbound antigen-binding molecule or antibody is removed, and the amount of label associated with immobilized antigen is measured. If the amount of label associated with immobilized antigen is substantially reduced in the test sample relative to the control sample, then that indicates that the second antigen-binding molecule or antibody is competing with the first antigen-binding molecule or antibody for binding to antigen. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

In one aspect, assays are provided for identifying agonistic antigen binding molecules, agonistic anti-FGFR2b molecules and fragments thereof, having biological activity, in particular agonistic activity. Biological activity, in particular agonistic activity, may include an increase or enhancement in FGFR2b signaling, an increase in expression of an intracellular protein downstream of FGFR2b, an increase in the proliferation of lung epithelial cells, an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells, or an induction in proliferation of lung epithelial cells or AT2 cells measured by an increase in ATP expression or BrdU expression that is directly proportional to the concentration of viable cells, detection of or an increase in FGFR2b dimerization and phosphorylation of FGFR2b. Antigen binding molecules having such agonistic activity in vivo and/or in vivo are also provided.

In certain embodiments, an agonistic antigen binding molecule or an agonistic anti-FGFR2b antigen binding molecule is tested for agonistic activity.

Methods for assaying agonistic activity
Increase or enhancement in FGFR2b, FGFR1b, or DDR1 signaling
An increase or enhancement in FGFR2b, FGFR1b, or DDR1 signaling introduced by the binding of the agonistic antigen binding molecule of the present invention may be assayed by in vitro measurement of an increase in presence or phosphorylation of a downstream protein of FGFR2b, FGFR1b, or DDR1 signaling, wherein such downstream proteins of FGFR2b or FGFR1b include for example FRS, PLCG1, PAK4, GRB2, GAB1, PIK3R1, SOS1, MAPK1/ERK2, MAPK3/ERK1, and AKT1 and such downstream proteins of DDR1 include for example SHP-2, FAK, NF-kappa B, Cdc42, MAPK1, AP1, Ras/Raf/ERK, PI3K/AKT, STAT1/3/5. The increase can be assayed using various protein quantification methods known in the art, such as for example, Bradford assay, Bovine Serum Albumin (BSA) assay, enzyme-linked immunosorbent assay (ELISA), gel electrophoresis, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, immunoblotting, silver staining and coomassie blue staining.

In one embodiment, the increase corresponds to at least or equivalent to 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 15%, or 20%, or 25%, or 30%, or 35%, or 40%, or 45%, or 50%, or 55%, or 60%, or 65%, or 70%, or 75%, or 80%, or 85%, or 90%, or 95% or 100% increase when comparing the expression of the downstream protein after binding of the agonistic antigen binding molecule of the present invention to antigen compared to prior to binding of the agonistic antigen binding molecule of the present invention to antigen. The increase may be expressed in comparative units, e.g. fold-changes, or as suitably employed by the skilled person.

Increase in the proliferation of target cells, i.e. lung epithelial cells, an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells
An increase in proliferation of target cells may be assayed by in vitro measurement of an increase in ATP or BrdU or other proliferation related biomarkers or proteins wherein such increase is directly proportional to the concentration of viable target cells, in cell or organoid assays, or immunohistochemical staining of cell proliferation markers, such as MTT, WST-1, PCNA, Ki67 (Mki67), Cdc20, BrdU etc, or other cell proliferation assays well known in the art.

In one embodiment, efficacy of the agonistic antigen binding molecules of the present invention may be evaluated by measurement of alveolar type II (AT2) cell marker such as surfactant protein C (Sftpc) or measurement of fibrotic markers such as fibronectin 1 (Fn1) or collagen type I alpha 1 chain (Col1a1) in target cells or organoids, or in fibrosis model such as bleomycin induced mouse lung fibrosis model. An increase of alveolar type II (AT2) cell marker and/or reduction of fibrotic marker refer to efficacy of the agonistic antigen binding molecules of the present invention in the treatment and/or prevention fibrosis or inflammation.

Cell proliferation assays well known in the art measure cell viability. One such assay is the CellTiter-Glo (registered trademark) Luminescent Cell Viability Assay, which is commercially available from Promega (Madison, WI). That assay determines the number of viable cells in culture based on quantitation of ATP present, which is an indication of metabolically active cells. See Crouch et al. (1993) J. Immunol. Meth. 160: 81-88 US Pat. No. 6602677. The assay may be conducted in 96- or 384-well format, making it amenable to automated high-throughput screening (HTS). See Cree et al. (1995) AntiCancer Drugs 6:398-404. The assay procedure involves adding a single reagent (CellTiter-Glo (registered trademark) Reagent) directly to cultured cells. This results in cell lysis and generation of a luminescent signal produced by a luciferase reaction. The luminescent signal is proportional to the amount of ATP present, which is directly proportional to the number of viable cells present in culture. Data can be recorded by luminometer or CCD camera imaging device. The luminescence output is expressed as relative light units (RLU). Another assay for cell proliferation is the "MTT" assay, a colorimetric assay that measures the oxidation of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to formazan by mitochondrial reductase. Like the CellTiter-Glo (registered trademark) assay, this assay indicates the number of metabolically active cells present in a cell culture. See, e.g., Mosmann (1983) J. Immunol. Meth. 65:55-63, and Zhang et al. (2005) Cancer Res. 65:3877-3882. Another assay for cell proliferation is the "BrdU" assay, an immunoassay that measures the amount of incorporated BrdU. The Bromodeoxyuridine / 5-bromo-2’-deoxyuridine (BrdU) is an analog of the nucleoside thymidine which is incorporated into replicating DNA during cell proliferation. This assay indicates the number of replicating cells present in a cell culture. See e.g., Messele et al. (2000) Clin. Diagn. Lab. Immunol., 7: 687-692.

In one embodiment, an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the agonistic antigen binding molecule of the present invention compared to in the target cell in the absence of the agonistic antigen binding molecule of the present invention indicates an increase in target cell proliferation. The increase may be expressed in comparative units, e.g. percentages, or as suitably employed by the skilled person.

Reduction or inhibition of fibrosis
Reduction or inhibition of fibrosis may be assayed by measurement of reduction of fibrotic biomarker, such as fibronectin 1 (Fn1) or collagen type I alpha 1 chain (Col1a1) in cell or organoid assays, or immunohistochemical staining of the fibrotic markers, or other fibrosis assays well known in the art.

The present invention is also concerned with a pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention and a pharmaceutically acceptable carrier.

The present invention is also concerned with an immunoconjugate comprising the agonistic antigen-binding molecule of the present invention. An "immunoconjugate" is an antibody conjugated to one or more heterologous molecule(s) as defined herein.

The present invention is also concerned with the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention, for use in the treatment or prevention of tissue-degenerative diseases, including tissue fibrosis.

Fibrosis is an essential and vital process in the wound healing process. As used herein, the term "fibrosis" comprises neoplastic fibrosis, neoplastic fibrosis, and fibrosis induced by chemicals or external stimuli. Excessive fibrosis is common in a variety of rare and common diseases and is largely associated with the pathogenic mechanism of the disease. Diseases characterized by excessive fibrosis include systemic sclerosis, scleroderma, hypertrophic cardiomyopathy, dilated cardiomyopathy (DCM), atrial fibrillation, ventricular fibrillation, myocarditis, liver cirrhosis, renal disease, etc. As described herein, the fibrosis is fibrosis of a respiratory organ, a circulatory system organ, a digestive system organ, a urinary system organ, a nervous system organ and / or a musculoskeletal system organ. As described herein, the fibrosis refers to fibrosis of lung, heart, blood vessel, liver, small intestine, large intestine, pancreas, kidney, eye, brain, skin, bone marrow and / or muscle tissue. In one embodiment, the fibrosis is fibrosis of the lung, wherein the diseases characterized by lung fibrosis include, but are not limited to pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, obstructive bronchitis, asbestosis, silicosis, chronic pulmonary hypertension, AIDS associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma.

The present invention is also concerned with the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention, for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.

In one aspect of the present invention, the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.

In one aspect of the present invention, the treatment of any disease, disorder, or condition is susceptible of being improved or prevented by one of by the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b, FGFR1b, or DDR1 signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

The present invention is also concerned with the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention, for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.

In one aspect of the present invention, the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.

In one aspect of the present invention,
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.

The present invention is also concerned with the use of the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention, in the manufacture of a medicament for the treatment or prevention of a disease or disorder, or a condition related to injury of lung epithelial cells, in an individual in need thereof.

The present invention is also concerned with a method of treating a disease or disorder, or a condition related to injury of lung epithelial cells, in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention.

In the present invention, the condition related to the injury of lung epithelial cells in an individual can be selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.

In the present invention, the condition related to the injury of lung epithelial cells in an individual can be one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b, FGFR1b, or DDR1 signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.

The present invention is also concerned with the use of the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.

The present invention is also concerned with a method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the agonistic antigen-binding molecule of the present invention, or the pharmaceutical composition comprising the agonistic antigen-binding molecule of the present invention, or the immunoconjugate comprising the agonistic antigen-binding molecule of the present invention.

In the present invention, the fibrosis or inflammation can be selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
In one aspect of the present invention,
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.

The present invention is also concerned with an isolated nucleic acid encoding the agonistic antigen-binding molecule of the present invention, or a plurality of isolated nucleic acids encoding agonistic antigen-binding molecule of the present invention.

The present invention is also concerned with a host cell comprising the nucleic acid encoding the agonistic antigen-binding molecule of the present invention or the plurality of nucleic acids encoding the agonistic antigen-binding molecule of the present invention.

The present invention is also concerned with a method of producing the agonistic antigen-binding molecule of the present invention, or a process for producing the agonistic antigen-binding molecule of the present invention comprising the following steps: (a) culturing the host cell of the present invention, and (b) recovering the agonistic antigen-binding molecule of the present invention that is produced in step (a).

The present invention is also concerned with a kit for use in a method of the present disclosure, which contain the agonistic antigen-binding molecule of the present invention or an agonistic antigen-binding molecule produced by a method of the present invention. The kit may be packaged with an additional pharmaceutically acceptable carrier or medium, or instruction manual describing how to use the kits, etc.

In another aspect of the invention, an article of manufacture containing materials useful for the treatment, prevention and/or diagnosis of the disorders described above is provided. The article of manufacture comprises a container and a label on or a package insert associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is by itself or combined with another composition effective for treating, preventing and/or diagnosing the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active ingredient in the composition is the agonistic antigen-binding molecule of the present invention. The label or package insert indicates that the composition is used for treating the condition of choice. Moreover, the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises the agonistic antigen-binding molecule of the present invention; and (b) a second container with a composition contained therein, wherein the composition comprises a further cytotoxic or otherwise therapeutic agent. The article of manufacture in this embodiment of the invention may further comprise a package insert indicating that the compositions can be used to treat a particular condition. Alternatively, or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.As shown above in detail, according to one aspect, anti-FGFR2b antigen-binding molecules, wherein the molecules are agonists are provided. Exemplary anti-FGFR2b antigen-binding molecules, wherein the molecules are agonists according to this aspect are listed in Tables 1 to 4 herein.

Table 1 sets forth the amino acid sequence identifiers of the heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), heavy chain variable regions (VH) and light chain variable regions (VL) of the monoparatopic anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

Table 2 sets forth the amino acid sequence identifiers of the heavy chain (HC), and the light chain (LC) of the biparatopic anti-FGFR2b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

Table 3 sets forth the amino acid sequence identifiers of the heavy chains (HC), and light chains (LC) of the biparatopic LINC-Ig anti-FGFR2b antibodies, or antigen-binding fragments thereof, disclosed herein.

Table 4 sets forth exemplary biparatopic anti-FGFR2 antigen binding molecules, antibodies, or antigen-binding fragments thereof, of the present invention comprising combinations of a first antigen-binding domain and a second antigen-binding domain selected from Tables 1 to 3.

As shown above in detail, according to one aspect, anti-FGFR1b antigen-binding molecules, wherein the molecules are agonists are provided. Exemplary anti-FGFR1b antigen-binding molecules, wherein the molecules are agonists according to this aspect are listed in Tables 9 and 14 herein.

Table 9 sets forth the amino acid sequence identifiers of the heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), heavy chain variable regions (VH) and light chain variable regions (VL) of the monoparatopic anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

Table 14 sets forth the amino acid sequence identifiers of the heavy chain (HC), and the light chain (LC) of the biparatopic anti-FGFR1b antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

As shown above in detail, according to one aspect, anti-DDR1 antigen-binding molecules, wherein the molecules are agonists are provided. Exemplary anti-DDR1 antigen-binding molecules, wherein the molecules are agonists according to this aspect are listed in Tables 10 and 13 herein.

Table 10 sets forth the amino acid sequence identifiers of the heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3), and the light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), heavy chain variable regions (VH) and light chain variable regions (VL) of the monoparatopic anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

Table 13 sets forth the amino acid sequence identifiers of the heavy chain (HC), and the light chain (LC) of the biparatopic anti-DDR1 antigen-binding molecules, antibodies, or antigen-binding fragments thereof, disclosed herein.

Antigen binding molecules may be produced using recombinant methods and compositions, e.g., as described in U.S. Patent No. 4,816,567. In one embodiment, isolated nucleic acid encoding an anti-FGFR2b antigen-binding molecule, an anti-FGFR1b antigen-binding molecule, or an anti-DDR1 antigen-binding molecule described herein is provided. Such nucleic acid may encode an amino acid sequence comprising the VL and/or an amino acid sequence comprising the VH of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule (e.g., the light and/or heavy chains of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule). In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acid are provided. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule and an amino acid sequence comprising the VH of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule. In one embodiment, the host cell is eukaryotic, e.g., a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., Y0, NS0, Sp2/0 cell). In one embodiment, a method of making an anti-FGFR2b antigen-binding molecule, an anti-FGFR1b antigen-binding molecule, or an anti-DDR1 antigen-binding molecule is provided, wherein the method comprises culturing a host cell comprising a nucleic acid encoding the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule, as provided above, under conditions suitable for expression of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule, and optionally recovering the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule from the host cell (or host cell culture medium).

For recombinant production of an anti-FGFR2b antigen-binding molecule, an anti-FGFR1b antigen-binding molecule, or an anti-DDR1 antigen-binding molecule, nucleic acid encoding an anti-FGFR2b antigen-binding molecule, an anti-FGFR1b antigen-binding molecule, or an anti-DDR1 antigen-binding molecule, e.g., as described above, is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule).

Suitable host cells for cloning or expression of antigen-binding molecule-encoding vectors include prokaryotic or eukaryotic cells described herein. For example, antigen-binding molecules may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antigen-binding molecule fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, describing expression of antigen-binding molecule fragments in E. coli.) After expression, the anti-FGFR2b antigen-binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antigen-binding molecule-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been "humanized," resulting in the production of an antigen-binding molecule with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).

Suitable host cells for the expression of glycosylated antigen-binding molecules are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

Plant cell cultures can also be utilized as hosts. See, e.g., US Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES^TM technology for producing antigen-binding molecules in transgenic plants).

Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR^- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2/0. For a review of certain mammalian host cell lines suitable for antigen-binding molecule production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

In certain embodiments, the invention provides agonistic antigen-binding molecules, and anti-FGFR2b antigen binding molecules, anti-FGFR1b antigen binding molecules, or anti-DDR1 antigen binding molecules obtainable by a method set out above.

Any one of the antigen-binding molecules, and anti-FGFR2b antigen-binding molecules, anti-FGFR1b antigen binding molecules, or anti-DDR1 antigen binding molecules provided herein may be used in therapeutic methods.

It is understood that any one of the above formulations or therapeutic methods may be carried out using an immunoconjugate of the invention in place of or in addition to an anti-FGFR2 antigen binding molecule, an anti-FGFR1b antigen-binding molecule, or an anti-DDR1 antigen-binding molecule.

The anti-FGFR2 antigen binding molecules, the anti-FGFR1b antigen binding molecules, and the anti-DDR1 antigen binding molecules herein can be administered by a method in which a nucleic acid encoding the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, and the anti-DDR1 antigen-binding molecule, respectively, is administered or incorporated into a body using a vector or the like, and the antigen binding molecule is directly expressed in a body, but may be administered without using a vector. As vectors, viral vectors, plasmid vectors are exemplified, and even adenoviral vectors are exemplified. The nucleic acid encoding the an anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule may be administered directly to the body, or cells into which the nucleic acid encoding the an anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule are introduced may be administered to the body. For example, the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, and the anti-DDR1 antigen-binding molecule may be administered by a method in which mRNA encoding the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, and the anti-DDR1 antigen-binding molecule, respectively, may be subjected to chemical modifications to enhance the stability of mRNA in vivo, and mRNA may be administered directly to humans and the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, and the anti-DDR1 antigen-binding molecule, respectively, may be expressed in vivo (see EP2101823B, WO2013/120629). Also, B cells into which the nucleic acid encoding the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule is introduced may be administered (Sci Immunol. (2019), 4(35), eaax0644). Bacteria into which the nucleic acid encoding the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule is introduced may also be administered (Nature Reviews Cancer (2018) 18, 727-743).

As a non-limiting example of techniques that can be combined with an anti-FGFR2 antigen binding molecules herein, the generation of T cells secreting T cell redirecting antibodies utilizing the anti-FGFR2 antigen binding molecule, the anti-FGFR1b antigen-binding molecule, or the anti-DDR1 antigen-binding molecule is exemplified (Trends Immunol.(2019) 40(3) 243-257). One of the non-limiting techniques for the generation is the introduction of a nucleic acid encoding a bispecific antibody, comprising a binding domain for any one of the subunits forming a T-cell receptor (TCR) complex on T cells, in particular a binding domain for CD3 epsilon chain among CD3, and a binding domain for FGFR2, FGFR1, or DDR1, into an effector cell such as a T cell by genetic modification techniques.

In certain embodiments, any one of the anti-FGFR2 antigen binding molecules, provided herein is useful for detecting the presence of FGFR2 in a biological sample. The term "detecting" as used herein encompasses quantitative or qualitative detection. In certain embodiments, a biological samples comprises a cell or tissue, such as lung or skin tissue. In one embodiment, an anti-FGFR2 antigen binding molecule for use in a method of diagnosis or detection is provided. In a further aspect, a method of detecting the presence of FGFR2 in a biological sample is provided. In certain embodiments, the method comprises contacting the biological sample with an anti-FGFR2 antigen binding molecule as described herein under conditions permissive for binding of the anti-FGFR2 antigen binding molecule to FGFR2 and detecting whether a complex is formed between the anti-FGFR2 antigen binding molecule and FGFR2. Such method may be an in vitro or in vivo method. In one embodiment, an anti-FGFR2 antigen binding molecule is used to select subjects eligible for therapy with an anti-FGFR2 antigen binding molecule, e.g., where FGFR2 is a biomarker for selection of patients.

In certain embodiments, any one of the anti-FGFR1 antigen binding molecules, provided herein is useful for detecting the presence of FGFR1 in a biological sample. The term "detecting" as used herein encompasses quantitative or qualitative detection. In certain embodiments, a biological samples comprises a cell or tissue, such as lung or skin tissue. In one embodiment, an anti-FGFR1 antigen binding molecule for use in a method of diagnosis or detection is provided. In a further aspect, a method of detecting the presence of FGFR1 in a biological sample is provided. In certain embodiments, the method comprises contacting the biological sample with an anti-FGFR1 antigen binding molecule as described herein under conditions permissive for binding of the anti-FGFR1 antigen binding molecule to FGFR1 and detecting whether a complex is formed between the anti-FGFR1 antigen binding molecule and FGFR1. Such method may be an in vitro or in vivo method. In one embodiment, an anti-FGFR1 antigen binding molecule is used to select subjects eligible for therapy with an anti-FGFR1 antigen binding molecule, e.g., where FGFR1 is a biomarker for selection of patients.

In certain embodiments, any one of the anti-DDR1 antigen binding molecules, provided herein is useful for detecting the presence of DDR1 in a biological sample. The term "detecting" as used herein encompasses quantitative or qualitative detection. In certain embodiments, a biological samples comprises a cell or tissue, such as lung or skin tissue. In one embodiment, an anti-DDR1 antigen binding molecule for use in a method of diagnosis or detection is provided. In a further aspect, a method of detecting the presence of DDR1 in a biological sample is provided. In certain embodiments, the method comprises contacting the biological sample with an anti-DDR1 antigen binding molecule as described herein under conditions permissive for binding of the anti-DDR1 antigen binding molecule to DDR1 and detecting whether a complex is formed between the anti-DDR1 antigen binding molecule and DDR1. Such method may be an in vitro or in vivo method. In one embodiment, an anti-DDR1 antigen binding molecule is used to select subjects eligible for therapy with an anti-FGFR1 antigen binding molecule, e.g., where DDR1 is a biomarker for selection of patients.

In certain embodiments, labeled anti-FGFR2 antigen binding molecules, labeled anti-FGFR1 antigen binding molecules, or labeled anti-DDR1 antigen binding molecules are provided. Labels include, but are not limited to, labels or moieties that are detected directly (such as fluorescent, chromophoric, electron-dense, chemiluminescent, and radioactive labels), as well as moieties, such as enzymes or ligands, that are detected indirectly, e.g., through an enzymatic reaction or molecular interaction. Exemplary labels include, but are not limited to, the radioisotopes ³²P, ¹⁴C, ¹²⁵I, ³H, and ¹³¹I, fluorophores such as rare earth chelates or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, luceriferases, e.g., firefly luciferase and bacterial luciferase (U.S. Patent No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, horseradish peroxidase (HRP), alkaline phosphatase, beta-galactosidase, glucoamylase, lysozyme, saccharide oxidases, e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase, heterocyclic oxidases such as uricase and xanthine oxidase, those coupled with an enzyme that employs hydrogen peroxide to oxidize a dye precursor such as HRP, lactoperoxidase, or microperoxidase, biotin/avidin, spin labels, bacteriophage labels, stable free radicals, and the like. In one embodiment, the labeled anti-FGFR2 antigen binding molecules are provided as diagnostic reagents.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the invention.

The following are examples of methods and compositions of the invention. It is understood that various other embodiments may be practiced, given the general description provided above.

Example 1: Generation of bivalent monoparatopic and bivalent biparatopic anti-FGFR2 antibodies
Anti-FGFR2 antibodies were prepared, selected and assayed in two immunization campaigns as follows:

Example 1.1 Immunization using mixture of mouse FGFR2IIIb and FGFGR2IIIc variant
Two NZW rabbits were immunized with mixture of mouse FGFR2IIIb (FGFR2b) variant (SEQ ID NO: 1) plasmid and mouse FGFR2IIIc (FGFR2c) variant (SEQ ID NO: 2) plasmid coated gold particles (1micro g DNA per shot for total 40 shots) by Helios Gene Gun system (Bio-Rad) according to the manufacturer’s instruction (Yuxin Chen et al. J. Virol. (2013) 87(18):10232). In addition, combination of intradermal administration (200 micro g DNA per site for total 2 sites at shoulders) and intramuscular administration (400 micro g DNA per site for total 2 sites at two legs) with mixture of mouse FGFR2b plasmid and mouse FGFR2c plasmid were carried out. The immunization was repeated weekly for 8 times followed by blood and spleen collection. For B cell selection, IgG positive and mouse FGFR2 binding B cells, or IgG positive and mouse FGFR2IIIb and/or human FGFR2IIIb binding B cells were collected. B cells were collected and sorted using a cell sorter and then plated and cultured according to the procedure described in WO2016098356A1. After cultivation, B cell culture supernatants were collected for further analysis and B cell pellets were cryopreserved.

Specific binding to mouse FGFR2 was checked by flow cytometry using selected B cell supernatant with mouse FGFR2IIIb variant stably expressing FreeStyle^TM 293-F cell line, mouse FGFR2IIIc variant stably expressing FreeStyle^TM 293-F cell line and human FGFR2IIIb variant (UniProt Reference Sequence: P21802-3, SEQ ID NO: 5) stably expressing Ba/F3 cell line. B cells with good binding capability to mouse FGFR2 were preferred and selected for gene cloning.

Example 1.2 Immunization using mixture of mouse and human FGFR2IIIb variant
Four JW rabbits were immunized with recombinant human FGFR2IIIb protein (ACROBiosystems, FGB-H5256) and mouse FGFR2IIIb protein (ACROBiosystems, FGB-M52H5) protein. Six alternative doses (3 times for each antigen) were administered over a period of 2 months, followed by blood and spleen collection. For B cell selection, IgG positive and mouse FGFR2IIIb binding -and/or human FGFR2IIIb-binding B cells were collected. B cells were collected and sorted using a cell sorter and then plated and cultured according to the procedure described in WO2016098356A1. After cultivation, B cell culture supernatants were collected for further analysis and B cell pellets were cryopreserved.

Specific binding to mouse FGFR2 and/or human FGFR2 was checked by flow cytometry using selected B cell supernatant with mouse FGFR2IIIb variant stably expressing FreeStyle^TM 293-F cell line, mouse FGFR2IIIc variant stably expressing FreeStyle^TM 293-F cell line and human FGFR2IIIb variant stably expressing Ba/F3 cell line.
B cells with good binding capability to mouse FGFR2 and/or human FGFR2 were preferred and selected for gene cloning.

Example 1.3 Recombinant expression and purification of bivalent monoparatopic anti-FGFR2 antibodies
The RNAs of selected B cell lines were purified from the cryopreserved cell pellets using the ZR-96 Quick-RNA kits (ZYMO RESEARCH, Cat No. R1053). The DNAs encoding antigen-binding domains comprising antigen-binding heavy chain variable region, VH, and light chain variable region, VL, and their corresponding HCDRs and LCDRs respectively, in the selected B cell lines were amplified by reverse transcription PCR. The antigen-binding domains were named FFB0001 to FFB0470, of which, a selected representative number of antigen-binding domains are shown in Table 1 below. Heavy chain expression vectors were produced that encode a heavy chain variable region selected from Table 1 in combination with a heavy chain constant region SG1 (SEQ ID NO: 7). Light chain expression vectors were produced that encode a light chain variable region selected from Table 1 in combination with a light chain constant region SK2 (SEQ ID NO: 8). Recombinant antibodies (bivalent monoparatopic antibodies) were expressed in Expi293^TM Expression System (Thermo Fisher Scientific, Carlsbad, CA, USA), and antibody purification was carried out using Protein A affinity chromatography and gel filtration.

Table 1 shows the names and SED ID NOs of antigen-binding domains comprising variable regions (VH and VL), and their corresponding HCDRs and LCDRs of selected antibodies for further evaluation.

Example 1.4 Generation of biparatopic bivalent anti-FGFR2 antibodies by FORCE technology
Biparatopic anti-FGFR2b antibodies with both arms targeting different epitopes of FGFR2b were generated by Format Chain Exchange (FORCE) process as described in WO2019077092A1 and Dengl. S. et al., 2020, and referring to Figure 1. In brief, the FORCE process is based on CH3-interface driven exchange reactions of initial molecules representing different binder/format combinations, e.g. knob-into-hole heterodimers. The initial molecules comprise two monoparatopic monovalent antibodies ("Fc-dummy containing monoparatopic monovalent antibody"), wherein the constant region comprises a normal Fc half and a dummy Fc half. Upon preferential knob-into-hole heterodimerization, biparatopic bivalent antibodies and dummy Fc heterodimers were produced. The dummy Fc heterodimers were removed after affinity tag purification.

Fc-dummy containing monoparatopic monovalent anti-FGFR2b antibodies were generated by co-transfection of expression vectors encoding heavy chain comprising either hole or knob mutations, respective light chains, and matching dummy chains, either FEdumkHis001 (SEQ ID No. 224) or FEdumhHis001 (SEQ ID No. 225) into Expi293^TM cells (Thermo Fisher, Carlsbad, CA, USA). Antibody purification of monoparatopic monovalent antibodies was carried out using Protein A affinity chromatography and gel filtration.

The exchange reaction was initiated by incubating equal molar amounts of two Fc-dummy containing monospecific monovalent antibodies with Tris (2-carboxyethyl)phosphine (TCEP) to temporary separate the interchain disulfide bonds on the hinge region according to method described in Dengl, Stefan, et al. ("Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices." Nature Communications 11.1 (2020): 4974). Chain rearrangement occurred spontaneously thereafter resulting in the production of biparatopic bivalent antibodies with preferential matching heterodimers and dummy chain heterodimers. Biparatopic bivalent antibodies were purified using IMAC PhyTip columns (Biotage/PhyNexus Inc., Sweden, Cat. No. PTB-91-20-03) according to manufacturer’s instructions. Table 2 and Figure 2 shows the names and SEQ ID NOs of biparatopic antibodies comprising the first antigen-binding domains and second antigen-binding domains, and their corresponding heavy chains, and light chains. Biparatopic anti-FGFR2b antibodies used in animal study were generated by a method known in the art.

Example 1.5 Generation of biparatopic bivalent anti-FGFR2 antibodies with LINC-Ig technology
Anti-FGFR2b biparatopic LINC-Ig antibodies were generated by utilizing LINC-Ig technology as described in PTL 5 and a method known in the art. In brief, LINC-Ig introduces one or more engineered disulfide bonds between the two Fabs of an antibody which reduces the flexibility of the two Fab arms. In the present example, one disulfide bond was introduced into each pair of heavy and light chain of selected bivalent biparatopic antibodies of Table 2 and the resultant biparatopic LINC-Ig antibodies are shown in Table 3. The disulfide bond pair comprises of amino acid substitutions S191C (according to Kabat numbering) on CH1 of constant heavy chain and K126C (according to Kabat numbering) on constant light chain, CL.

All monoparatopic bivalent antibodies with or without LINC-Ig mutation were expressed transiently using Expi293^TM Expression System (Thermo Fisher, Carlsbad, CA, USA). Antibody purification was carried out using Protein A affinity chromatography and gel filtration. Biparatopic bivalent antibody preparation was conducted according to a method known in the art. The anti-FGFR2b biparatopic LINC-Ig antibodies used in animal study were further treated with a reducing agent, Cysteine (15mM Cysteine, 50mM Tris-HCL, pH8.0) to de-cap the cysteines residues and efficiently promote the engineered disulfide bond pairs (PTL 5). The assembly of biparatopic bivalent antibodies with or without LINC-Ig mutation is shown in Figure 3.

Table 3 and Figure 4 shows the names and SEQ ID NOs of biparatopic LINC-Ig antibodies comprising the first antigen-binding domains and second antigen-binding domains, and their corresponding heavy chains, and light chains.

Example 2: Characterisation of anti-FGFR2b antibodies
Example 2.1 Binding analysis of anti-FGFR2b monoclonal antibody using FACS
To determine if antibody binds to mouse FGFR2, flow cytometry analysis was carried out by using mouse FGFR2IIIb stably expressing 293-F cell line described above. 2 microgram/mL of the anti-FGFR2b antibody were incubated on ice for 30 minutes with the cell line and washed with FACS buffer (0.1% BSA in PBS). Goat F(ab’)2 anti-Human IgG, Mouse ads-PE (Southern Biotech, Cat. 2043-09) was then added and incubated for 30 minutes on ice. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR2b antibodies was measured and the result is shown in Figure 5. Data acquisition was performed on LSRFortessa X-20 (Becton Dickinson), followed by analysis using the FlowJo software (Tree Star) and Excel software (Microsoft). Binding capability of the anti-FGFR2b antibodies to another mouse FGFR2b variant (GenBank ABL89196.1) was also confirmed.

To map more detailed binding information of the anti-FGFR2b antibodies, IgI domain (Pro38-Arg152) deleted mouse FGFR2IIIb variant (mouse FGFR2IIIb delta IgI, SEQ ID NO: 3) stably expressing FreeStyle^TM 293-F cell line was prepared. 2 microgram/mL of the anti-FGFR2 antibody were incubated on ice for 30 minutes with the cell line and washed with FACS buffer (0.1% BSA in PBS). Goat F(ab’)2 anti-Human IgG, Mouse ads-PE (Southern Biotech, Cat. 2043-09) was then added and incubated for 30 minutes on ice. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR2b antibodies was measured and the result is shown in Figure 6. In addition, IgIII domain (Ser252-Gln362) deleted mouse FGFR2 variant (mouse FGFR2 delta IgIII, SEQ ID NO: 4) transiently expressing FreeStyle^TM 293-F cell line was prepared. 2 microgram/mL of the anti-FGFR2b antibody were incubated on ice for 30 minutes with the cell line and washed with FACS buffer (0.1% BSA in PBS). Goat F(ab’)2 anti-Human IgG, Mouse ads-PE (Southern Biotech, Cat. 2043-09) was then added and incubated for 30 minutes on ice. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR2b antibodies was measured and the result is shown in Figure 7. Data acquisition was performed on LSRFortessa X-20 (Becton Dickinson), followed by analysis using the FlowJo software (Tree Star) and Excel software (Microsoft).

Antibodies that bind to full length mouse FGFR2IIIb and mouse FGFR2 delta IgIII but not to mouse FGFR2IIIb delta IgI are identified as binders to the IgI region, antibodies that bind to full length mouse FGFR2IIIb and mouse FGFR2IIIb delta IgI but not mouse FGFR2 delta IgIII are identified as binders to the IgIII region, while antibodies that bind full length mouse FGFR2IIIb, mouse FGFR2IIIb delta IgI and mouse FGFR2 delta IgIII are identified as binders to the IgII region. The binding results and estimated epitope location from this study were summarized in Table 5. In Table 5, "Yes" means that the tested antibody can bind to the cell line and "No" means that the tested antibody substantially cannot bind to the cell line.

Example 2.2 Binding analysis of anti-FGFR2b monoclonal antibodies to mouse FGFR2IIIc using FACS
To determine if antibody binds to mouse FGFR2IIIc, flow cytometry analysis was carried out by using mouse FGFR2IIIc stably expressing 293-F cell line described above. 2 microgram/mL of the anti-FGFR2b antibody were incubated on ice for 30 minutes with the cell line and washed with FACS buffer (0.1% BSA in PBS). Goat F(ab’)2 anti-Human IgG, Mouse ads-PE (Southern Biotech, Cat. 2043-09) was then added and incubated for 30 minutes on ice. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR2b antibodies was measured and the result is shown in Figure 8A and 8B. Data acquisition was performed on LSRFortessa X-20 (Becton Dickinson), followed by analysis using the FlowJo software (Tree Star) and Excel software (Microsoft).

Example 2.3 Binding analysis of anti-FGFR2b monoclonal antibodies to human FGFR2IIIb using FACS
To determine if antibody binds to human FGFR2IIIb, flow cytometry analysis was carried out by using human FGFR2IIIb stably expressing Ba/F3 cell line described above. 2 microgram/mL of the anti-FGFR2 antibody were incubated on ice for 30 minutes with the cell line and washed with FACS buffer (0.1% BSA in PBS). Goat F(ab’)2 anti-Human IgG, Mouse ads-PE (Southern Biotech, Cat. 2043-09) was then added and incubated for 30 minutes on ice. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR2b antibodies was measured and the result is shown in Figure 9. Data acquisition was performed on LSRFortessa X-20 (Becton Dickinson), followed by analysis using the FlowJo software (Tree Star) and Excel software (Microsoft). Figure 9 shows that FFB0026, FFB0035, FFB0057, FFB0065, FFB0185, FFB0219, FFB0261, FFB0294, FFB0299, FFB0335, FFB0341, FFB0365, FFB0368, FFB0374, FFB0396, FFB0453 and FFB0464 have binding activity to human FGFR2IIIb.

Example 2.4 Competition binding analysis of anti-FGFR2IIIb monoparatopic antibodies
Competition assay was conducted to evaluate if the first and the second antigen-binding domains making up a biparatopic antibody compete with each other for binding to FGFR2IIIb. Combinations of monoparatopic antibodies comprising the first antigen-binding domain and the second antigen-binding domain were allowed to compete with each other in binding to Histidine-tagged mouse FGFR2IIIb protein (ACROBiosystems, FGB-M52H5) by Surface Plasmon Resonance (SPR) using Biacore^TM T200 (Cytiva). An appropriate amount of FGFR2IIIb was captured on Series S Sensor Chip NTA (Cytiva) by chelating histidine tag to allow antibodies to interact. The running buffer used was 20 mM ACES, 150 mM NaCl, 0.05% (v/v) Tween20, 0.005% (w/v) NaN₃, 50 micro M EDTA, pH7.4.

To allow competitive interaction against mouse FGFR2IIIb captured on the sensor chip, first antibody or second antibody in the combination was injected at 500 nM at a flow rate of 10 micro L/min for 60 seconds, immediately followed by second injection using dual injection function of Biacore^TM T200 at 500 nM at a flow rate of 10 micro L/min for 60 seconds. In the second injection to eliminate an effect of antibody dissociation, mixture of first and second antibodies was injected. Next, the running buffer was injected at a flow rate of 10 micro L/min for 60 seconds. Then, 0.35 M EDTA was injected twice at a flow rate of 30 micro L/min for 60 seconds to regenerate the sensor chip. The measurement was done at 25 degrees C. Binding response at each reported point was obtained by processing with Biacore^TM T200 Evaluation Software (Cytiva).

Theoretical binding response for non-competitive combinations is calculated by a sum of the binding responses of first and second antibodies in the first injection. Considering assay variation, combinations of antibodies are determined as non-competitive combinations when additional binding responses of the combinations in the second injection show over greater than 80% of theoretical binding response, while combinations showing less than 80% of theoretical binding response are determined as competitive combinations. For the calculation above, we corrected binding response by capture level of mouse FGFR2IIIb on the sensor chip. The results of determined competition from this study were summarized in Table 6, where "Yes" means that the tested combination of antibodies is competitive to each other and "No" means that the tested combination of antibodies is not competitive to each other.

Example 2 demonstrates an exemplary number of anti-FGFR2 monoparatopic antibodies and anti-FGFR2 biparatopic antibodies were obtained. Amongst the anti-FGFR2 biparatopic antibodies, biparatopic antibodies comprising a first antigen-binding domain and a second antigen-binding domain that bind different Ig domains of FGFR were obtained. Biparatopic antibodies comprising a first antigen-binding domain and a second antigen-binding domain that bind the same Ig domain of FGFR were also obtained. Competition binding assay was then conducted, and the antigen binding domains were concluded as non-competitive in binding to FGFR2IIIb.

Example 3: Evaluation of agonistic activity of anti-FGFR2 monoparatopic and biparatopic antibodies in lung organoid culture
To evaluate the effect of the anti-FGFR2 monoparatopic and biparatopic antibodies of the present invention on proliferation of lung epithelial cells, lung organoid culture was established as described below and was subjected to the anti-FGFR2 antibodies prepared in Examples 1.3 and 1.4 or FGF7 (positive control). FGF7 is an epithelial FGFR2IIIb splicing variant selective agonist and is a known mitogen that induces proliferation of FGFR2IIIb-expressing lung epithelial cell.

Example 3.1 Establishment of mouse lung organoid culture
Lung organoids were established from C57BL/6 NTac male mice. Specific pathogen-free C57BL/6NTac male mice of 6-7 weeks of age were purchased from Invivos Pte Ltd (Singapore) and were acclimated for 1 week before the start of harvesting. Animals were maintained at 20 - 26 degrees C with a 12:12 hour light/dark cycle and fed with a commercial standard diet (5P75; PMI Nutrition INT'L (LabDiet), Missouri, United States) and tap water ad libitum.

Mice were euthanized under isoflurane anesthesia, and their lungs were collected. The lungs were digested in physiological saline containing 2 mg/mL Collagenase/Dispase (Roche) and 10 micro M Y-27632 (Sigma) with DNase I (Roche) by gentleMACS^TM (Miltenyi Biotec) using 37C_m_LDK_1 program. The collected lung cells were reacted in the ACK lysing buffer (Gibco) for 2 min to lyse the contained erythrocytes. After washing with 1% BSA/DMEM-F12 (Stem cell Technologies), the cell mass was removed with a 70 micro m cell strainer (Corning). The final fraction was centrifuged at 300 g for 5 min at 4 degrees C. The cell pellet was then dispersed in Matrigel^TM (Corning) and seeded on a 24 well plate. Matrigel^TM was polymerized at 37degrees C for 10 to 30 minutes and then cultured in 100 ng/mL FGF7 (R & D systems) and/or 100 ng/mL FGF10 (R&D systems) containing lung organoid maintenance medium, wherein the medium is composed of IntestiCult^TM Organoid Growth Medium (Stem cell Technologies) supplemented with 100 ng/mL Noggin (R & D systems), 1 mM Nicotinamide (Sigma), 3 micro M CHIR99021 (Sigma), and 1/100 Penicillin/Streptomycin (Gibco). Once lung organoids were established, Matrigel^TM was removed with Accumax^TM or Accutase^TM or Cell Dissociation Buffer (Stem Cell Technologies) and passaging were conducted. For the first day of culture after passaging, lung organoids were cultured with 10 micro M Y-27632 (Sigma) and maintained at 37 degrees C in a 5% CO₂ incubator.

Example 3.2 In vitro lung organoid proliferation by anti-FGFR2 monoparatopic and biparatopic antibodies
To conduct the lung organoid proliferation assay, lung organoids removed from Matrigel^TM were subcultured in lung organoid maintenance medium, and growth of the organoids was observed for the duration of 1 week in the lung organoid maintenance medium containing recombinant mouse FGF7 (Genscript; positive control) or anti-FGFR2 monoparatopic or biparatopic antibodies. Proliferative activity was assessed by CellTiter-Glo (registered trademark) 3D Cell Viability Assay (Promega). The organoids were imaged using CellVoyager CV 8000 High-Content Screening System (Yokogawa Denki). As shown in Figures 10A to 10K, each biparatopic antibody showed stronger proliferative activity than monoparatopic antibody. Also, each biparatopic antibody exhibited concentration-dependent proliferation of lung organoids. Furthermore, higher proliferative activity was also observed for these antibodies compared to reported anti-FGFR2IIIb antibody "N12433" (WO2021/247718A1).

Notably, most of the biparatopic antibodies induced maximum proliferation to the same extent as that induced by FGF7 (positive control). The induction of proliferative activity of the lung epithelial cells by anti-FGFR2 monoparatopic antibodies did not reach the same extent as that of FGF7 (positive control). This suggests that biparatopic binding to the FGFR2 receptor enhances the agonistic activity of the activated receptor.

Example 3.3 Comparison of agonistic activity of anti-FGFR2 monoparatopic antibodies with other anti-FGFR2 biparatopic antibodies
Agonistic activity of anti-FGFR2 monoparatopic and biparatopic antibodies derived from the combination of other antigen-binding domains were evaluated. As in Example 3.2, lung organoids removed from Matrigel^TM were subcultured in lung organoid maintenance medium, and growth of the organoids was observed for the duration of 15 days in the lung organoid maintenance medium containing 1 micro g/mL anti-FGFR2 monoparatopic or biparatopic antibodies. As shown in Figure 11A and 11B, the agonistic activity of the biparatopic format is greater than the monoparatopic format for the tested combination of IgI and IgII antigen-binding domains, and IgII and IgIII antigen-binding domains, respectively.

Example 4: Evaluation of agonistic activity of anti-FGFR2 monoparatopic and biparatopic antibodies in keratinocyte culture
To evaluate the effect of the anti-FGFR2 monoparatopic and biparatopic antibodies of the present invention on proliferation of skin keratinocyte, keratinocyte culture was established as described below and subjected to the anti-FGFR2 antibodies prepared in Examples 1.3 and 1.4 or FGF7 (positive control). FGF7 is an epithelial FGFR2IIIb splicing variant selective agonist and is a known mitogen that also induces proliferation of FGFR2IIIb-expressing keratinocyte.

Example 4.1 In vitro keratinocyte proliferation by anti-FGFR2 monoparatopic and biparatopic antibodies
MPEK-BL6 cells (CELLnTEC) were used as mouse primary keratinocytes. Keratinocytes were cultured and maintained in CnT-07 Epithelial Proliferation Medium (CELLnTEC) at 37 degrees C in a 5% CO₂ incubator. Keratinocytes were passaged and starved overnight in 0.03% BSA/DMEM-F12 medium (Stem cell Technologies). Keratinocytes were then cultured in the same medium containing recombinant mouse FGF7 (Genscript; positive control) supplemented with Heparin (Sigma) or anti-FGFR2 monoparatopic or biparatopic antibodies for 72 hours. Proliferative activity was assessed by Cell Proliferation ELISA BrdU (Roche). As shown in Figures 12A to 12H, both anti-FGFR2 monoparatopic and biparatopic antibodies exhibited concentration-dependent keratinocyte proliferative activity. However, while the agonistic activity of most of the biparatopic antibodies reached the same maximum extent as FGF7, the agonistic activity of the monoparatopic antibodies did not. This corroborates the observation in lung organoid proliferation in Example 3.

Example 5: Evaluation of agonistic activity of anti-FGFR2 monoparatopic and biparatopic antibodies in NIH3T3 cultures
Example 5.1 In vitro NIH3T3 proliferation by anti-FGFR2 monoparatopic and biparatopic antibodies
Since the IgIII domain of FGFR2 comprises the epithelial IIIb or mesenchymal IIIc isoform, the anti-FGFR2 monoparatopic or biparatopic antibodies of the present invention may activate FGFR2 signaling via binding to epithelial IIIb or mesenchymal IIIc of FGFR2. To evaluate subtype specificity of the antibodies, each of the antibodies in Working Example 3, and FGF7 were added to NIH3T3 cultures, which are thought to express the FGFR2IIIc splicing variant and not the FGFR2IIIb splicing variant (Kohl et al. 2002). NIH3T3 cells were cultured and passaged in DMEM high glucose medium (Gibco) containing 10% FBS (Sigma) at 37 degrees C in a 5% CO₂ incubator. Prior to the proliferation assay, NIH3T3 cells were cultured in the starvation medium comprising of 0.03% BSA/DMEM-F12 medium (Gibco) overnight. Then, the NIH3T3 cells were cultured in the starvation medium containing recombinant mouse FGF2 (R&D systems) or FGF18 (Novus Biologicals) or FGF7 (Genscript) supplemented with Heparin (Sigma) or the anti-FGFR2 monoparatopic or biparatopic antibodies of Example 3 for 72 hours. Proliferative activity was assessed by Cell Proliferation ELISA BrdU (Roche). As shown in Figures 13A to 13D, unlike FGF2 or FGF18 which has an agonistic activity on mesenchymal IIIc subtype and acts as positive control in the assay, the anti-FGFR2 monoparatopic and biparatopic antibodies and FGF7 did not exhibit any induction of proliferation of NIH3T3 cells.

In sum, Examples 1-5 taken together, demonstrate the anti-FGFR2b antibodies of the present invention showed proliferation of FGFR2IIIb-expressing lung epithelial cells and keratinocytes, and not FGFR2IIIc-expressing NIH3T3 cells. The biparatopic agonist anti-FGFR2b antibodies described herein selectively activate FGFR2IIIb signaling and enhance the agonistic activity compared to the monoparatopic agonist anti-FGFR2b antibodies.

Example 6: Evaluation of agonistic activity of anti-FGFR2b biparatopic LINC-Ig antibodies and Un-LINC-Ig antibodies
Example 6.1 In vitro lung organoid proliferation by anti-FGFR2b biparatopic LINC-Ig antibodies
Accordingly in the art, it was previously exemplified that a mutation in FGFR that promotes the change in receptor formation primed for dimerization shows strong signal activation that is observed in cancer associated mutation of FGFR3. To evaluate whether enhanced receptor dimerization by increasing the proximity of two different antigen-binding domains would result in stronger agonistic signal activation, anti-FGFR2b antibodies ("anti-FGFR2b biparatopic LINC-Ig antibodies") were developed, wherein these antibodies were engineered such that their antigen-binding domains are brought in closer proximity than their non-engineered counterparts ("anti-FGFR2b biparatopic Un-LINC-Ig antibodies"). Similar to Example 3, lung organoids were passaged and cultured for 2 weeks in the lung organoid maintenance medium containing recombinant mouse FGF7 (Genscript) or anti-FGFR2b biparatopic Un-LINC-Ig antibodies or anti-FGFR2b biparatopic LINC-Ig antibodies. Proliferative activity was assessed by CellTiter-Glo (registered trademark) 3D Cell Viability Assay (Promega). The organoids were imaged using CellVoyager CV 8000 High-Content Screening System (Yokogawa Denki).

As shown in Figure 14, anti-FGFR2b biparatopic LINC-Ig antibodies showed enhanced lung organoid growth compared to anti-FGFR2b biparatopic Un-LINC-Ig antibodies. Heavy chain-Light chain LINC-Ig (HL-LINC), and Light chain-Light chain LINC-Ig (LL-LINC) formats were evaluated. In HL-LINC, FFB0065//FFB0045 and FFB0065//FFB0057 biparatopic LINC-Ig showed slightly improved agonistic activity in lung organoid growth (ranging from 1.9 to 2.3 fold improvement) compared to the biparatopic Un-LINC-Ig counterparts. In LL-LINC, FFB0065//FFB0045, FFB0065//FFB0035 and FFB0065//FFB0057 showed improved agonistic activity in lung organoid growth (ranging from 3.2 - 8.2 fold improvement) compared to the biparatopic Un-LINC-Ig counterparts. Furthermore, as shown in Figure 15, lung organoids were grown in conditions comprising the anti-FGFR2b biparatopic LINC-Ig antibodies without exogenous FGF7.

Example 6.2 In vivo proliferation marker induction by anti-FGFR2b biparatopic LINC-Ig antibodies
In addition to in vitro lung organoid and keratinocyte proliferation, in vivo lung and skin tissue cell proliferation by anti-FGFR2b biparatopic LINC-Ig antibodies were evaluated.

As in Example 3.1, specific pathogen-free C57BL/6NTac male mice of 5-6 weeks of age were purchased from Invivos Pte Ltd (Singapore) and used for in vivo proliferation marker evaluation. The anti-FGFR2b biparatopic LINC-Ig antibodies were administered 5 mg/kg by intravenous injection every day from Day 0 to Day 2. Vehicle (PBS) and 250 micro g/kg mouse FGF7 (Genscript) were used as a control in this study. The animals were killed by exsanguination under isoflurane anaesthesia on Day 3. The lung and skin tissue were quickly removed from mice and each tissue was snap-frozen in liquid nitrogen or on dry ice for molecular analysis.

The anti-FGFR2b biparatopic LINC-Ig antibodies used in this study were prepared as described in Example 1.5, and each antibody information is summarized in Table 7.

Total RNA was extracted from lung and ear skin tissues using QIAzol Lysis Reagent and RNeasy Mini Kit (Qiagen). Mouse Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as the endogenous reference for each sample. Mki67 and Cell division cycle 20 (Cdc20) were used as proliferation markers. Relative mRNA expression values were calculated using double delta Ct analysis.

As shown in Figures 16A to 16D, LL-LINC_FFB0065//FFB0035 and LL-LINC_FFB0065//FFB0045 showed significant induction of proliferation markers in both lung and skin compared to Vehicle. Statistical analysis was performed using Dunnett’s multiple comparison test compared to Vehicle. When P values were of <0.05, differences were considered statistically significant (Statistical significance: ***P<0.001, ****P<0.0001).

Example 7: Evaluation of efficacy of anti-FGFR2b biparatopic antibodies
Example 7.1 Efficacy of anti-FGFR2b biparatopic LINC-Ig antibodies in bleomycin model
In addition to in vivo lung and skin tissue cell proliferation by anti-FGFR2b biparatopic LINC-Ig antibodies, efficacy of anti-FGFR2b biparatopic LINC-Ig antibodies in a bleomycin induced mouse lung fibrosis model (BLM model) was evaluated. The model is the best characterized and most widely used preclinical model, which reproduces many aspects of idiopathic pulmonary fibrosis (NPL 5, NPL 6, NPL 7).

As in Example 3.1, specific pathogen-free C57BL/6NTac male mice of 5-6 weeks of age were purchased from Invivos Pte Ltd (Singapore) and used for BLM model study. Bleomycin sulfate (Gold Bio, st. Louis, U.S. Pat. No. B-910-50) or saline is administered by osmotic minipump (Alzet, cupertino, CA, USA, model 1007D). On day 0, the micropump was loaded with bleomycin (50 mg/kg) dissolved in 90 microliters (micro l) of saline. Saline was administered to the unaffected control group. The micropump was incubated at 37 degrees C for 3-4 hours prior to implantation. The pump was implanted subcutaneously (s.c.) in the back through an incision under isoflurane anesthesia. On day 10, micropumps were removed from mice under isoflurane anesthesia. From day 14, all antibodies were administered by intravenous injection at 5mg/kg three times per week. The present study used the Sham operation (Sham) with saline injection (QD, s.c.) as a positive control and the bleomycin administration (BLM) with saline injection (QD, s.c.) as a negative control. Animals were monitored by their body weight during study and sacrificed by exsanguination under isoflurane anesthesia on day 28. The lungs were removed quickly and weighed. Portions of lung tissue were snap frozen on liquid nitrogen or dry ice for molecular analysis. The anti-FGFR2b biparatopic LINC-Ig antibodies used in this study were prepared as described in Example 1.5, and each antibody information is summarized in Table 7, and the detailed protocol is summarized in Figure 17.

Total RNA was extracted from lung tissue using QIAzol Lysis Reagent and RNeasy Mini Kit (Qiagen). Mouse Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as the endogenous reference for each sample. Mouse Surfactant protein C (Sftpc) was used as alveolar type II (AT2) cells marker, and Mouse fibronectin 1 (Fn1) and Collagen type I alpha 1 chain (Col1a1) were used as fibrotic markers. Relative mRNA expression values were calculated using double delta Ct analysis.

As shown in Figures 18A to 18C, LL-LINC_FFB0065//FFB0035 showed significant induction of alveolar type II (AT2) cells marker and reduction of fibrotic markers in lung compared to negative control. Statistical analysis was performed using Dunnett’s multiple comparison test compared to BLM. When P values were of <0.05, differences were considered statistically significant (Statistical significance: **P<0.01, ****P<0.0001).

Example 7.2 Efficacy of anti-FGFR2b biparatopic antibodies in bleomycin model
Additionally, efficacy of anti-FGFR2b biparatopic antibodies in a bleomycin induced mouse lung fibrosis model (BLM model) was also evaluated. As in Example 6.2, in vivo proliferation marker induction by anti-FGFR2b biparatopic antibodies was evaluated. The anti-FGFR2b biparatopic antibodies used in this study were prepared as described in Example 1.4, and each antibody information is summarized in Table 8.

As shown in Figures 19A and 19B, FFB0219//FFB0396, FFB0261//FFB0464, FFB0335//FFB0464, FFB0365//FFB0464, FFB0368//FFB0341, and FFB374//FFB0464 showed significant induction of proliferation markers in lung compared to Vehicle. When P values were of <0.05, differences were considered statistically significant (Statistical significance: ***P<0.001, ****P<0.0001).

As in Example 7.1, efficacy of those anti-FGFR2b biparatopic antibodies in a bleomycin induced mouse lung fibrosis model (BLM model) was evaluated. The anti-FGFR2b biparatopic antibodies used in this study were prepared as described in Example 1.4, and anti-KLH monoparatopic antibody IC17dK (KLH) was used as a negative control. Each anti-FGFR2b biparatopic antibody information is summarized in Table 8. All antibodies were administered by intravenous injection at 5mg/kg three times per week. The present study used the Sham operation (Sham) with anti-KLH monoparatopic antibody IC17dK injection (Sham + KLH) as a positive control and the bleomycin administration (BLM) with anti-KLH monoparatopic antibody IC17dK injection (BLM + KLH) injection as a negative control.

As shown in Figures 20A to 20C and 21A to 21C, all of anti-FGFR2b biparatopic antibodies showed significant induction of alveolar type II (AT2) cells marker and FFB0219//FFB0396, FFB0261//FFB0464 showed reduction of fibrotic markers in lung compared to negative control. Statistical analysis was performed using Dunnett’s multiple comparison test compared to BLM + KLH. When P values were of <0.05, differences were considered statistically significant (Statistical significance: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).

In addition to marker gene expression analysis, histopathological analysis for alveolar type II (AT2) cell proliferation was evaluated. As shown in Figures 22A and 22B, all of anti-FGFR2b biparatopic antibodies showed statistically significant increase in the number of alveolar type II (AT2) cells. Statistical analysis was performed using Tukey’s multiple comparisons test compared to BLM + KLH group. When P values were of <0.05, differences were considered statistically significant (Statistical significance: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).

Overall data demonstrated that anti-FGFR2b biparatopic antibodies regenerate alveolar type II (AT2) cells in bleomycin induced mouse lung fibrosis model.

Example 8: Generation of anti-DDR1 and FGFR1 antibodies
Example 8.1: Generation of anti-FGFR1 antibodies
Anti-FGFR1 antibodies were prepared, selected and assayed as follows:Two NZW rabbits were immunized with mixture of exon3 including mouse FGFR1IIIb (FGFR1b) variant (SEQ ID NO: 226) plasmid and mouse FGFR1IIIc (FGFR1c) variant (SEQ ID NO:227) plasmid coated gold particles (1 micro g DNA per shot for total 40 shots) by Helios Gene Gun system (Bio-Rad) according to the manufacturer’s instruction (Yuxin Chen et al. J. Virol. (2013) 87(18):10232). In addition, combination of intradermal administration (200 micro g DNA per site for total 2 sites at shoulders) and intramuscular administration (400 micro g DNA per site for total 2 sites at two legs) with mixture of mouse FGFR1IIIb variant plasmid and mouse FGFR1IIIc variant plasmid were carried out. The immunization was repeated weekly for 8 times followed by blood and spleen collection. For B cell selection, IgG positive and mouse FGFR1 binding B cells were collected. B cells were sorted using a cell sorter and then plated and cultured according to the procedure described in WO2016098356A1. After cultivation, B cell culture supernatants were collected for further analysis and B cell pellets were cryopreserved.

Specific binding to mouse FGFR1 was checked by flow cytometry using selected B cell supernatant with mouse FGFR1IIIb variant stably expressing Ba/F3 cell line, mouse FGFR1IIIc variant stably expressing Ba/F3 cell line and human FGFR1IIIb variant (Reference Sequence: NP_001167534) stably expressing FreeStyle^TM 293-F cell line.
B cells with good binding capability to mouse FGFR1IIIb variant and/or mouse FGFR1IIIc variant were preferred and selected for gene cloning.

The RNAs of selected B cell lines were purified from the cryopreserved cell pellets using the ZR-96 Quick-RNA kits (ZYMO RESEARCH, Cat No. R1053). The DNAs encoding antibody heavy-chain variable regions in the selected lines were amplified by reverse transcription PCR and recombined with DNA encoding SG181 heavy chain constant region (SEQ ID NO:228). The DNAs encoding antibody light-chain variable regions were also amplified by reverse transcription PCR and recombined with DNA encoding the k0MC light-chain constant region. Cloned antibodies were expressed in Expi293^TM Expression System (Thermo Fisher Scientific) and purified from culture supernatants.

Table 9 shows the names and SED ID NOs of variable regions (including VH, VL, and HVR/CDR) of selected identified antibodies for further evaluation.

Example 8.2: Generation of anti-DDR1 antibodies
Anti-DDR1 antibodies were prepared, selected and assayed as follows:
Three NZW rabbits were immunized with mixture of mouse DDR1 (SEQ ID NO:382) plasmid and mouse DDR2 (SEQ ID NO:383) plasmid coated gold particles (1 micro g DNA per shot for total 40 shots) by Helios Gene Gun system (Bio-Rad) according to the manufacturer’s instruction (Yuxin Chen et al. J. Virol. (2013) 87(18):10232). In addition, combination of intradermal administration (200 micro g DNA per site for total 2 sites at shoulders) and intramuscular administration (400 micro g DNA per site for total 2 sites at two legs) with mixture of mouse DDR1 plasmid and mouse DDR2 plasmid were carried out. The immunization was repeated weekly for 8 times followed by blood and spleen collection. For B cell selection, IgG positive and mouse DDR1 binding B cells were collected. B cells were sorted using a cell sorter and then plated and cultured according to the procedure described in WO2016098356A1. After cultivation, B cell culture supernatants were collected for further analysis and B cell pellets were cryopreserved.

Specific binding to mouse DDR1 was checked by flow cytometry using selected B cell supernatant with mouse DDR1 stably expressing Ba/F3 cell line and mouse DDR2 stably expressing Ba/F3 cell line.

The RNAs of selected B cell lines were purified from the cryopreserved cell pellets using the ZR-96 Quick-RNA kits (ZYMO RESEARCH, Cat No. R1053). DNAs encoding antibody heavy-chain variable regions in the selected lines were amplified by reverse transcription PCR and recombined with DNA encoding SG181 heavy chain constant region. DNAs encoding antibody light-chain variable regions were also amplified by reverse transcription PCR and recombined with DNA encoding the k0MC light-chain constant region. Cloned antibodies were expressed in Expi293^TM Expression System (Thermo Fisher Scientific) and purified from culture supernatants.

Table 10 shows the names and SED ID NOs of variable regions (including VH, VL, and HVR/CDR) of selected identified antibody for further evaluation.

Example 9: Characterization of anti-DDR1 and FGFR1b antibodies
Example 9.1 : Epitope analysis of anti-FGFR1 monoclonal antibodies
To determine antibody binding to mouse FGFR1, flow cytometry analysis was carried out by using mouse FGFR1IIIb variant stably expressing Ba/F3 cell line and mouse FGFR1IIIc variant stably expressing Ba/F3 cell line described above. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR1 antibodies was measured. Data acquisition was performed on iQue Screener (IntelliCyt), followed by analysis using the ForeCyt software (IntelliCyt) and Excel software (Microsoft).

To map more detail binding epitope of anti-FGFR1 antibodies, Ig1 domain deleted mouse FGFR1IIIb variant (mouse FGFR1IIIb delta Ig1, SEQ ID NO:349) stably expressing Ba/F3 cell line, Ig1 domain deleted mouse FGFR1IIIc variant (mouse FGFR1IIIc delta Ig1, SEQ ID NO:350) stably expressing Ba/F3 cell line and Ig3 domain deleted mouse FGFR1 variant (mouse FGFR1 delta Ig3, SEQ ID NO:351) stably expressing Ba/F3 cell line were prepared. Geometric Mean Fluorescent Intensity (MFI) of anti-FGFR1 antibodies was measured. Data acquisition was performed on iQue Screener (IntelliCyt), followed by analysis using the ForeCyt software (IntelliCyt) and Excel software (Microsoft).

Epitope of antibody which can bind to exon3 including mouse FGFR1IIIb and/or FGFR1IIIc and mouse FGFR1 delta Ig3 but not to mouse FGFR1IIIb delta Ig1 and/or mouse FGFR1IIIc delta Ig1 would be Ig1 region. Epitope of antibody which can bind to exon3 including mouse FGFR1IIIb and/or FGFR1IIIc and mouse FGFR1IIIb delta Ig1 and/or mouse FGFR1IIIc delta Ig1 but not to mouse FGFR1 delta Ig3 would be Ig3 region. While epitope of antibody which can bind both exon3 including mouse FGFR1IIIb and/or FGFR1IIIc, mouse FGFR1IIIb delta Ig1 and/or mouse FGFR1IIIc delta Ig1 and mouse FGFR1 delta Ig3 would be Ig2 region. The estimated epitope location from this study were summarized in Table 11.

Example 9.2: Epitope analysis of anti-DDR1 monoclonal antibodies
To determine antibody binding to mouse DDR1, flow cytometry analysis was carried out by using mouse DDR1 stably expressing Ba/F3 cell line. One-arm format antibodies were prepared to evaluate the binding by affinity mode. Geometric Mean Fluorescent Intensity (MFI) of anti-DDR1 antibodies was measured.

To map more detail binding epitope of anti-DDR1 antibodies, discoidin domain (Cys32-Cys186) deleted mouse DDR1 (mouse DDR1 delta DS, SEQ ID NO: 410) stably expressing Ba/F3 cell line was prepared. Binding to this cell line was measured by flow cytometry and MFI value was acquired. Epitope of antibody which can bind to full length mouse DDR1 but not to mouse DDR1 delta DS would be DS domain. While epitope of antibody which can bind both full length mouse DDR1 and mouse DDR1 delta DS would be discoidin-like (DSL) domain and extracellular juxtamembrane (EJXM) region (DSL-EJXM). The binding results and estimated epitope location from this study were summarized in Table12. In Table12, "Yes" means that antibody can bind to the cell line and "No" means that antibody cannot bind to the cell line.

Example 10: Evaluation of agonistic activity of biparatopic antibodies for other tyrosine kinase receptors including DDR1 and FGFR1b
Example 10.1 In vitro Ba/F3-DDR1 proliferation by anti-DDR1 biparatopic antibodies
To evaluate the effect of the anti-DDR1 monoparatopic and biparatopic antibodies of the present invention on proliferation of mouse DDR1 stably expressing Ba/F3 cell line (described in Example 8.2; Ba/F3-DDR1), Ba/F3-DDR1 culture was established as described below and subjected to the anti-DDR1 antibodies prepared in Examples 8 or collagen (Nitta Gelatin, molecular weight was used based on data sheet) as a positive control.

Ba/F3-DDR1 was cultured and maintained in RPMI1640 (Gibco) containing 10% FBS (Sigma), penicillin-streptomycin (Gibco), Geneticin (Gibco), and mouse IL-3 (R&D systems) at 37 degrees C in a 5% CO₂ incubator. Ba/F3-DDR1 was then passaged with the same culture medium without Geneticin and IL-3 containing various concentration of anti-DDR1 monoparatopic or biparatopic antibodies or collagen control for 72 hours. Proliferative activity was assessed by CellTiter-Glo (registered trademark) 3D Cell Viability Assay (Promega).

As shown in Figures 23A to 23C, enhanced proliferative activity by anti-DDR1 biparatopic antibodies compared to monoparatopic antibodies were observed. Especially, DAA0011//DAA0024 exhibited stronger activity than collagen control.

Example 10.2 Phosphorylation of DDR1 by anti-DDR1 biparatopic antibodies in Ba/F3-DDR1
In addition to proliferative activity, phosphorylation of DDR1 induced by anti-DDR1 monoparatopic and biparatopic antibodies was evaluated. As in Example 10.1, Ba/F3-DDR1 was cultured and maintained. Ba/F3-DDR1 was then passaged with RPMI1640 (Gibco) containing 0.1% FBS (Sigma), penicillin-streptomycin (Gibco) with 100nM of anti-DDR1 monoparatopic antibodies or 50nM of anti-DDR1 biparatopic antibodies for 60min. Phosphorylation of DDR1 was assessed by PathScan (registered trademark) Phospho-DDR1 (panTyr) Sandwich ELISA Kit (Cell Signaling Technology).

As shown in Figures 24A and 24B, enhanced Phosphorylation of DDR1 by anti-DDR1 biparatopic antibodies compared to monoparatopic antibodies were observed. The result above indicates that anti-DDR1 biparatopic antibodies have strong agonist activity on DDR1 with phosphorylation of the receptor.

Example 10.3 In vitro BaF3-FGFR1b cell proliferation by anti-FGFR1b biparatopic antibodies
Moreover, effects of anti-FGFR1b biparatopic antibodies were evaluated by using mouse FGFR1IIIb variant stably expressing Ba/F3 cell line (described in Example 8.1; Ba/F3-FGFR1b). Ba/F3-FGFR1b culture was established as described below and subjected to the anti-FGFR1b antibodies prepared in Examples 8 or FGF2 (mouse, R&D systems) as a positive control.

Ba/F3-FGFR1b was cultured and maintained in RPMI1640 (Gibco) containing 10% FBS (Sigma), penicillin-streptomycin (Gibco), Geneticin (Gibco), and mouse IL-3 (R&D systems) at 37 degrees C in a 5% CO₂ incubator. Ba/F3-FGFR1b was then passaged with the same culture medium without Geneticin and IL-3 containing various concentration of anti-FGFR1b monoparatopic or biparatopic antibodies or FGF2 control for 72 hours. Proliferative activity was assessed by CellTiter-Glo (registered trademark) 3D Cell Viability Assay (Promega).

As shown in Figures 25A to 25H, concentration-dependent proliferative activity by anti-FGFR1b biparatopic antibodies was observed. In particular, maximum activity of anti-FGFR1b biparatopic antibodies was stronger than that of monoparatopic antibodies and FGF2. These data demonstrated that biparatopic antibody could enhance agonist activity on not only FGFR2b, but also DDR1 and FGFR1b, and potentially, other tyrosine kinase receptors.

Claims

An agonistic antigen-binding molecule that binds to a receptor tyrosine kinase (RTK) protein, and wherein the molecule is an agonist of the RTK protein, 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 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.
The agonistic antigen-binding molecule of claim 1, wherein the RTK protein isa human Fibroblast growth factor receptor 2 (FGFR2) protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first FGFR2 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR2 protein, wherein the first epitope and the second epitope are different from each other.
The molecule of claim 2, wherein the first antigen-binding domain binds a first FGFR2 protein and the second antigen-binding domain binds a second FGFR2 protein.
The molecule of claims 2 or 3, which is a biparatopic antigen-binding molecule.
The molecule of any one of claims 2 to 4, wherein the human FGFR2 protein is human FGFR2b protein.
The molecule of any one of claims 2 to 5, wherein each of the extracellular domains is independently selected from an Immunoglobulin (Ig)-like domain of the FGFR2 protein.
The molecule of 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 is the same.
The molecule of 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.
The molecule of any one of claims 2 to 8, wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, bind an Ig-like domain of the FGFR2 protein, selected from the group consisting of IgI, IgII, IgIII, IgIIIb and IgIIIc of FGFR2.
The molecule of any one of claims 6 to 9, wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, binds an Ig-like domain of the FGFR2 in the following combinations selected from the group consisting of:
(i) wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, bind the IgIIIb and IgIIIb domain of FGFR2;
(ii) wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, bind the IgIIIb and IgIII domain of FGFR2;
(iii) wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, bind the IgIIIb and IgII domain of FGFR2; and
(iv) wherein the first antigen-binding domain and the second antigen-binding domain, independently of each other, bind the IgIIIb and IgI domain of FGFR2.
The molecule of any one of claims 6 to 10, wherein (i) the first antigen-binding domain binds the IgIIIb domain of FGFR2, or wherein (ii) the first antigen-binding domain binds specifically to IgIIIb of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgIIIc of FGFR2 and shows less than 10%, or less than 5%, or 0% cross-reactivity with IgI and IgII of FGFR2.
The molecule of any one of claims 4 to 11, wherein binding of the first antigen-binding domain and the second antigen-binding domain to FGFR2b protein leads to agonist activity as follows:
(i) wherein binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell activates signaling in the target cell, thereby exhibiting agonist activity, and wherein the activated signal in the target cell is measured by an increase in expression of an intracellular protein downstream of FGFR2b by at least 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or more compared to the expression of said intracellular protein prior to binding of the molecule to the cell-surface expressed FGFR2b protein in the target cell, in particular 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 binding of the molecule to FGFR2b protein via a cell-surface expressed FGFR2b in a target cell induces proliferation of the target cell, thereby exhibiting agonist activity, wherein the induction in proliferation of the target cell is measured by an increase in ATP expression by at least 1.5-fold, or 2-fold, or 2.5-fold or more, in the target cell in the presence of the molecule compared to in the target cell in the absence of the molecule, wherein the ATP expression is directly proportional to the concentration of viable target cells,
in particular wherein the target cell is a lung epithelial cell, optionally wherein the lung epithelial cell is selected from alveolar type II (AT2) cells; or
(iii) wherein receptor dimerization is detected in the presence of the molecule in an in vitro cell assay, wherein receptor dimerization is indicative of 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 FGFR2b protein phosphorylation is indicative of agonist activity.
The agonistic 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 a first epitope of an extracellular domain (ECD) of a first FGFR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second FGFR1 protein, wherein the first epitope and the second epitope are different from each other.
The agonistic antigen-binding molecule of claim 1, wherein the RTK protein is a human Discoidin Domain receptor 1 (DDR1) protein, wherein the molecule comprises a first antigen-binding domain that binds a first epitope of an extracellular domain (ECD) of a first DDR1 protein and a second antigen-binding domain that binds a second epitope of an extracellular domain (ECD) of the first or a second DDR1 protein, wherein the first epitope and the second epitope are different from each other.
The molecule of any one of claims 1 to 14, wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody, or wherein the first antigen-binding domain and the second antigen-binding domain are each a Fab, Fab’, F(ab)’2, scFab, scFv, Fv, VHH or single-domain antibody which are linked by at least one disulfide bond.
The molecule of 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 which are not in a hinge region.
The molecule of claim 16, wherein the at least one disulfide bond is an engineered disulfide bond which is not present in a wild type IgG, optionally wherein said at least one disulfide bond is formed between a CH1 region, a CL region, a VH or VHH region, or a VL region of the first antigen-binding domain and a CH1 region, a CL region, a VH or VHH region, or a VL region of the second antigen-binding domain, in particular wherein said at least one disulfide bond is formed between the group selected from the following:
[i] an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CH1 region of a second antigen-binding domain;
[ii] an amino acid residue in a CL region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
[iii] an amino acid residue in a CH1 region of a first antigen-binding domain and an amino acid residue in a CL region of a second antigen-binding domain;
[iv] an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VH or VHH region of a second antigen-binding domain;
[v] an amino acid residue in a VL region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain; and
[vi] an amino acid residue in a VH or VHH region of a first antigen-binding domain and an amino acid residue in a VL region of a second antigen-binding domain.
The molecule of any one of claims 1 to 17, wherein the molecule comprises an antibody constant region, in particular (i) wherein the antibody constant region is a human antibody constant region, or (ii) wherein the antibody constant region is selected from a human IgG1, IgG2, IgG3 or IgG4 constant regions.
The molecule of any one of claim 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 second antigen-binding domain are each a monovalent Fab or a monovalent VHH, or are each a monovalent scFab.
The molecule of any one of claims 1 to 19, wherein the molecule comprises more than two antigen-binding domains, or wherein the molecule is tri-valent, tetra-valent, or multivalent molecule, optionally wherein the molecule comprises a third antigen-binding domain, the third antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain.
The molecule of claim 20, wherein the molecule comprises a fourth antigen-binding domain, the fourth antigen-binding domain binds the same epitope on the same domain as the first or the second antigen-binding domain but not the same epitope as the third antigen-binding domain, optionally wherein the molecule comprises a third and a fourth antigen-binding domain, wherein the third antigen-binding domain binds the same epitope on the same domain as the first antigen-binding domain, and the fourth antigen-binding domain binds the same epitope on the same domain as the second antigen-binding domain.
A pharmaceutical composition comprising the molecule of any one of claims 1 to 21, and a pharmaceutically acceptable carrier.
An immunoconjugate protein comprising the molecule of any one of claims 1 to 21.
The molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23 for use in the treatment or prevention of a disease, disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
The molecule or pharmaceutical composition or immunoconjugate for the use of claim 24, wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
The molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23 for use in the treatment of any disease, disorder, or condition susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
The molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23 for use in the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
The molecule or pharmaceutical composition or immunoconjugate for the use of claim 27, wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
The molecule or pharmaceutical composition or immunoconjugate for the use of claim 28, wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
Use of molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23, in the manufacture of a medicament for the treatment or prevention of a disease, or disorder, or condition related to injury of lung epithelial cells in an individual in need thereof.
A method of treating a disease, or disorder, or condition related to injury of lung epithelial cells in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of claims 1 to 21, or the pharmaceutical composition of claim 22, or the immunoconjugate of claim 23.
The use of claim 30, or the method of claim 31, wherein the condition related to the injury of lung epithelial cells in an individual is selected from the group consisting of fibrosis of the lungs, inflammation of the lungs, senescence-related fibrosis of the lungs, asthma, a condition associated with exposure to environmental toxins, a condition associated with exposure to bacteria, a condition associated with exposure to a virus, cystic fibrosis, a pneumonectomy, a condition associated with exposure to radiation, a condition associated with exposure to tobacco smoke or chemicals, and bleomycin mediated epithelial injury.
The use of claim 30, or the method of claim 31, wherein the condition related to the injury of lung epithelial cells in an individual is one susceptible of being improved or prevented by one of the following selected from the group consisting of:
(i) an increase or enhancement in FGFR2b signaling;
(ii) an increase in the proliferation of lung epithelial cells; and
(iii) an increase in the proliferation of alveolar type II (AT2) progenitor cells or alveolar type II (AT2) cells.
Use of the molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23 in the manufacture of a medicament for the treatment or prevention of fibrosis or inflammation in an individual in need thereof.
A method of treating fibrosis or inflammation in an individual in need thereof, comprising administering to said individual a therapeutically effective amount of the molecule of any one of claims 1 to 21 or the pharmaceutical composition of claim 22 or the immunoconjugate of claim 23.
The use of claim 34, or the method of claim 35, wherein the fibrosis or inflammation is selected from fibrosis or inflammation of lung, heart, blood vessel, liver, bile duct, small intestine, large intestine, pancreas, kidney, eye, brain, skin, oral mucosa, thymus, bone marrow, or muscle tissue.
The use of claim 34, or the method of claim 35, wherein
(i) the fibrosis or inflammation of lung 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 associated pulmonary hypertension, sarcoidosis, interstitial lung tumor, and asthma;
(ii) the fibrosis or inflammation of liver or bile duct is selected from the group consisting of non-alcoholic steatohepatitis, or alcoholic steatohepatitis, or cholangitis, or Alagille syndrome;
(iii) the fibrosis or inflammation of pancreas is pancreatitis;
(iv) the fibrosis or inflammation of kidney is selected from the group consisting of chronic kidney disease or acute kidney disease;
(v) the fibrosis or inflammation of the oral mucosal is oral mucositis;
(vi) the fibrosis or inflammation of the skin is diabetic foot ulcer or epidermolysis bullosa;
(vii) the 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 an inflammatory bowel disease (IBD), optionally wherein the IBD is ulcerative colitis (UC) or Crohn’s disease.
An isolated nucleic acid encoding the molecule of any one of claims 1 to 21 or a plurality of isolated nucleic acids encoding the molecule of any one of claims 1 to 21.
A host cell comprising the nucleic acid or the plurality of nucleic acids of claim 38.
A method of producing the molecule, or a process for producing the molecule comprising the following steps (a) culturing the host cell of claim 39, and (b) recovering the molecule that is produced in step (a).