Multi-specific antibodies targeting HPTP-beta (VE-PTP) and VEGF

Abstract

The present application relates to multispecific antibodies that target HPTP-beta (VE-PTP) and VEGF. The present disclosure provides compositions comprising multispecific compounds, including compounds that target phosphatase and receptor tyrosine kinase agonists. Also provided are methods of treating conditions associated with angiogenesis comprising administering a multispecific compound targeting a phosphatase and a receptor tyrosine kinase agonist.

Description

[0001] This application is a divisional application of the application with the filing date of September 23, 2019, application number 201980077336.9, and invention title "Multi - specific antibodies targeting HPTP - β (VE - PTP) and VEGF".

[0002] Cross-reference

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 735,331, filed on September 24, 2018, and U.S. Provisional Application No. 62 / 832,461, filed on April 11, 2019, each of which is incorporated herein by reference in its entirety. Background Art

[0004] Individual compounds having the ability to modulate different targets can be combined to generate multi - specific compounds. Such multi - specific compounds may have advantages over the parent compounds administered alone. These advantages can include, for example, a simpler dosing regimen, a longer half - life in a subject, or the ability to tightly bind to the target.

[0005] Incorporated by reference

[0006] Each patent, publication, and non - patent literature cited in this application is incorporated herein by reference in its entirety as if each were separately incorporated by reference. Summary of the Invention

[0007] In some embodiments, the present disclosure provides a compound comprising: (a) a first domain, wherein the first domain modulates a phosphatase, and the phosphatase modulates Tie2; and (b) a second domain that specifically binds to a receptor tyrosine kinase agonist.

[0008] In some embodiments, the present disclosure provides a method of treating a condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein. Brief Description of the Drawings

[0009] Figure 1 : Schematic diagram of a basic four - chain antibody unit. The light - chain sequence is represented by "SEQ A". The heavy - chain sequence is represented by "SEQ B". N and C represent the N - terminus and C - terminus, respectively. - S - S - represents a disulfide bond.

[0010] Figure 2: Schematic diagram of a tetravalent bispecific antibody with a sequence appended to the C-terminus of the heavy chain. The light chain sequence of the IgG isotype antibody unit is represented by "SEQ A". The heavy chain sequence of the IgG isotype antibody unit is represented by "SEQ B". The linker sequence is represented by "SEQ C". The appended antigen-binding domain sequence is represented by "SEQ D". N and C represent the N-terminus and C-terminus respectively. -S-S- represents a disulfide bond.

[0011] Figure 3 : Schematic diagram of a tetravalent bispecific antibody with a sequence appended to the C-terminus of the light chain. The light chain sequence of the IgG isotype antibody unit is represented by "SEQ A". The heavy chain sequence of the IgG isotype antibody unit is represented by "SEQ B". The linker sequence is represented by "SEQ C". The appended antigen-binding domain sequence is represented by "SEQ D". N and C represent the N-terminus and C-terminus respectively. -S-S- represents a disulfide bond.

[0012] Figure 4 : Schematic diagram of a hexavalent bispecific antibody with sequences appended to the C-termini of the heavy and light chains. The light chain sequence of the IgG isotype antibody unit is represented by "SEQ A". The heavy chain sequence of the IgG isotype antibody unit is represented by "SEQ B". The linker sequences are represented by "SEQ C" and "SEQ E". The appended antigen-binding domain sequence is represented by "SEQ D". N and C represent the N-terminus and C-terminus respectively. -S-S- represents a disulfide bond.

[0013] Figure 5 : Schematic diagram of a hexavalent trispecific antibody with sequences appended to the C-termini of the heavy and light chains. The light chain sequence of the IgG isotype antibody unit is represented by "SEQ A". The heavy chain sequence of the IgG isotype antibody unit is represented by "SEQ B". The linker sequences are represented by "SEQ C" and "SEQ E". The appended antigen-binding domain sequences are represented by "SEQ D" and "SEQ F". N and C represent the N-terminus and C-terminus respectively. -S-S- represents a disulfide bond.

[0014] Figure 6 : Schematic diagram of a bivalent bispecific antibody with two different heavy chain sequences and two different light chain sequences. The light chain sequences are represented by "SEQ A" and "SEQ D". The heavy chain sequences are represented by "SEQ B" and "SEQ C". N and C represent the N-terminus and C-terminus respectively. -S-S- represents a disulfide bond.

[0015] Figure 7: Schematic diagram of a trivalent trispecific antibody having two different heavy chain sequences, two different light chain sequences, and a sequence attached to the C-terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequence is represented by "SEQ E". The additional antigen-binding domain sequence is represented by "SEQ F". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0016] Figure 8 : Schematic diagram of a trivalent trispecific antibody having two different heavy chain sequences, two different light chain sequences, and a sequence attached to the C-terminus of one heavy chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequence is represented by "SEQ E". The additional antigen-binding domain sequence is represented by "SEQ F". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0017] Figure 9 : Schematic diagram of a tetravalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C-termini of two light chains. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen-binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0018] Figure 10 : Schematic diagram of a tetravalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C-termini of two heavy chains. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen-binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0019] Figure 11: Schematic diagram of a tetravalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences cis - attached to the C - terminus of one heavy chain and the C - terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen - binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N - terminus and C - terminus respectively. - S - S - represents a disulfide bond.

[0020] Figure 12 : Schematic diagram of a tetravalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences trans - attached to the C - terminus of one heavy chain and the C - terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen - binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N - terminus and C - terminus respectively. - S - S - represents a disulfide bond.

[0021] Figure 13 : Schematic diagram of a pentavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C - termini of two heavy chains and the C - terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQA" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E", "SEQ F", and "SEQ G". The additional antigen - binding domain sequences are represented by "SEQ H", "SEQ I", and "SEQ J". N and C represent the N - terminus and C - terminus respectively. - S - S - represents a disulfide bond.

[0022] Figure 14 : Schematic diagram of a pentavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C - terminus of one heavy chain and the C - termini of two light chains. The light chain sequences of the IgG isotype antibody units are represented by "SEQA" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E", "SEQ F", and "SEQ G". The additional antigen - binding domain sequences are represented by "SEQ H", "SEQ I", and "SEQ J". N and C represent the N - terminus and C - terminus respectively. - S - S - represents a disulfide bond.

[0023] Figure 15: Schematic diagram of a hexavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C-termini of the heavy and light chains. The light chain sequences of the IgG isotype antibody unit are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody unit are represented by "SEQ B" and "SEQ C". The connector sequences are represented by "SEQ E", "SEQF", "SEQ G" and "SEQ H". The attached antigen binding domain sequences are represented by "SEQ I", "SEQ J", "SEQ K" and "SEQL". N and C represent the N-terminus and C-terminus, respectively. -SS- represents a disulfide bond.

[0024] Figure 16 : ELISA data demonstrating binding of the tetravalent bispecific antibody HC2-AFL:LC1 to HPTP-β (upper panel) and VEGF (lower panel).

[0025] Figure 17 : ELISA data demonstrating binding of the tetravalent bispecific antibody HC2-BRO:LC1 to HPTP-β (upper panel) and VEGF (lower panel).

[0026] Figure 18 : ELISA data testing the binding of the tetravalent bispecific antibody HC2-RAN:LC1 to HPTP-β (upper panel) and VEGF (lower panel).

[0027] Figure 19 : Activation of Tie2 in HUVECs treated with tetravalent or hexavalent bispecific antibodies containing brolucizumab-derived or aflibercept-derived VEGF-binding domains. As demonstrated by immunoprecipitation and Western blotting, all bispecific antibodies tested increased basal Tie2 activation (in the absence of exogenous Ang1 or Ang2), while basal VEGFR2 phosphorylation (in the absence of exogenous VEGF) was unaffected.

[0028] Figure 20 : Tetravalent and hexavalent bispecific antibodies containing brolucizumab-derived or aflibercept-derived VEGF-binding domains enhanced Tie2 activation and inhibited VEGFR2 activation in HUVECs treated with Ang1 and VEGF, as demonstrated by immunoprecipitation and Western blotting.

[0029] Figure 21A , Figure 21B and Figure 21C: In HUVECs, including those treated with Ang1 and VEGF, tetravalent and hexavalent bispecific antibodies containing a VEGF-binding domain derived from abicipar or aflibercept enhance Tie2 activation and inhibit VEGFR2 activation. Figure 21A A Western blot is provided. Figure 21B Quantification of Tie2 activation is provided. Figure 21C Quantification of VEGFR2 phosphorylation is provided.

[0030] Figure 22A 、 Figure 22B and Figure 22C : In HUVECs, including those treated with Ang1 and VEGF, hexavalent bispecific antibodies containing a VEGF-binding domain derived from brolucizumab, aflibercept, or abicipar enhance Tie2 activation and inhibit VEGFR2 activation. Figure 22A A Western blot is provided. Figure 22B Quantification of Tie2 activation is provided. Figure 22C Quantification of VEGFR2 phosphorylation is provided.

[0031] Figure 23A and Figure 23B : As confirmed by quantification of electrochemiluminescence signals, in HUVECs treated with Ang1 and VEGF, tetravalent and hexavalent bispecific antibodies containing a VEGF-binding domain derived from brolucizumab, aflibercept, or abicipar enhance Tie2 activation and inhibit VEGFR2 activation. Figure 23A Quantification of Tie2 activation is provided. Figure 23B Quantification of VEGFR2 phosphorylation is provided.

[0032] Figure 24A and Figure 24B : As confirmed by quantification of electrochemiluminescence signals, in HUVECs treated with Ang1 and VEGF, hexavalent bispecific antibodies containing a VEGF-binding domain derived from brolucizumab, aflibercept, or abicipar enhance Tie2 activation and inhibit VEGFR2 activation. Figure 24A Quantification of Tie2 activation is provided. Figure 24B Quantification of VEGFR2 phosphorylation is provided.

[0033] Figure 25A and Figure 25B: As confirmed by electrochemiluminescence signal quantification, in HUVECs treated with Ang1 and VEGF, tetravalent and hexavalent bispecific antibodies containing the VEGF-binding domain derived from brolucizumab enhanced Tie2 activation and inhibited VEGFR2 activation. Figure 25A Quantification of Tie2 activation was provided. Figure 25B Quantification of VEGFR2 phosphorylation was provided.

[0034] Figure 26A and Figure 26B : As confirmed by electrochemiluminescence signal quantification, in HUVECs treated with Ang1 and VEGF, tetravalent and hexavalent bispecific antibodies containing the VEGF-binding domain derived from abicipar or aflibercept enhanced Tie2 activation and inhibited VEGFR2 activation. Figure 26A Quantification of Tie2 activation was provided. Figure 26B Quantification of VEGFR2 phosphorylation was provided.

[0035] Figure 27A and Figure 27B : As confirmed by electrochemiluminescence signal quantification, in HUVECs treated with Ang1 and VEGF, tetravalent and hexavalent bispecific antibodies containing the VEGF-binding domain derived from abicipar or aflibercept enhanced Tie2 activation and inhibited VEGFR2 activation. Figure 27A Quantification of Tie2 activation was provided. Figure 27B Quantification of VEGFR2 phosphorylation was provided. Detailed Description

[0036] The present disclosure provides compositions and methods for modulating phosphatases and kinases, such as receptor tyrosine kinases. Compositions and methods for modulating Tie2, such as to promote Tie2 phosphorylation, signal transduction, and / or activation, are provided. In some embodiments, the present disclosure provides compositions and methods for targeting phosphatases that modulate Tie2 signaling. In some embodiments, the phosphatase that modulates Tie2 signaling is human protein tyrosine phosphatase-β (HPTP-β).

[0037] Compositions and methods for modulating receptor tyrosine kinases, such as to reduce receptor tyrosine kinase phosphorylation, signal transduction, and / or activation, are provided. In some embodiments, the present disclosure provides compositions and methods for targeting receptor tyrosine kinase agonists, such as vascular endothelial growth factor (VEGF). In some embodiments, the present disclosure provides compositions and methods for targeting receptor tyrosine kinases, such as VEGF receptors.

[0038] In some embodiments, the present disclosure provides compositions and methods for targeting human protein tyrosine phosphatase-β (HPTP-β) or vascular endothelial protein tyrosine phosphatase (VE-PTP or VEPTP) and vascular endothelial growth factor (VEGF). In some embodiments, the present disclosure provides multispecific compounds, agents, antibodies, fragments or derivatives that target HPTP-β (VE-PTP) and VEGF.

[0039] The agents disclosed herein can be used to treat conditions characterized by, for example, vascular instability, angiogenesis, neovascularization, vascular leakage, and / or edema. The agents disclosed herein can be used to treat, for example, vascular disorders, ocular disorders, cancer, kidney disorders, and diabetic complications.

[0040] HPTP-β / VE-PTP, Tie2, and Vascular Stability

[0041] HPTP-β is a member of the receptor-like family of protein tyrosine phosphatases (PTPases). HPTP-β is a transmembrane protein found primarily in vascular endothelial cells that exhibits structural and functional similarities to cell adhesion molecules. Orthologs of HPTP-β have been found in a variety of species including, for example, zebrafish, chicken, dog, mouse, marmoset, and monkey. The orthologs are commonly referred to as vascular endothelial protein tyrosine phosphatase (VE-PTP). HPTP-β (VE-PTP) can affect vascular stability through its action on Tie2-mediated signaling.

[0042] Tie2 (tyrosine kinase 2 with immunoglobulin and epidermal growth factor homology domains) is a membrane receptor tyrosine kinase expressed primarily in vascular endothelial cells. Upstream factors can regulate Tie2 phosphorylation, affecting downstream signaling and vascular stabilization. Non-limiting examples of such factors include angiopoietin 1 (Ang1 / Angptl), angiopoietin 2 (Ang2 / Angpt2), and HPTP-β (VE-PTP).

[0043] Ang1 is an agonist of Tie2. The binding of Ang1 to Tie2 promotes receptor phosphorylation and downstream signaling to induce vascular stabilization by highly organized angiogenesis, strengthening endothelial cell junctions, enhancing endothelial viability, reducing endothelial inflammation, and improving endothelial function.

[0044] Ang2 functions as a context-dependent antagonist or agonist of Tie2. During angiogenesis, Ang2 acts as a negative regulator of Ang1-Tie2 signaling.

[0045] HPTP-β (VE-PTP) is a phosphatase that can regulate Tie2 signaling. HPTP-β (VE-PTP) can dephosphorylate the Tie2 receptor. Under physiological conditions, HPTP-β (VE-PTP) regulates the duration of Tie2 phosphorylation. Therefore, inhibition of HPTP-β (VE-PTP) can lead to increased Tie2 phosphorylation, increased Tie2-mediated signaling, and enhanced vascular stability. Inhibitors of HPTP-β (VE-PTP) are Tie2 activators. For example, compounds, inhibitors, antibodies, antibody fragments, variants, or derivatives that bind to HPTP-β (VE-PTP) can promote Tie2 phosphorylation, thereby activating Tie2 downstream signaling and promoting vascular stability.

[0046] Through the above process, HPTP-β (VE-PTP) activity can lead to conditions characterized by, for example, vascular instability, angiogenesis, neovascularization, vascular leakage, and / or edema. For example, HPTP-β (VE-PTP) activity can lead to vascular disorders, ocular disorders, cancer, kidney disorders, diabetic complications, and other conditions. Inhibition of HPTP-β (VE-PTP) activity can alleviate such conditions.

[0047] VEGF and Vascular Stability

[0048] Vascular endothelial growth factor (VEGF) is mainly present in endothelial cells and is associated with pathological neovascularization in many diseases. VEGF is a member of the cystine knot growth factor superfamily, the PDGF family, and the VEGF family. VEGF can act as an angiogenic factor. The VEGF family consists of VEGF-A, VEGF-B, VEGF-C, VEGF-D, and placental growth factor (PGF). There are nine VEGF-A isoforms: VEGF 121 、VEGF 145 、VEGF 148 、VEGF 162 、VEGF 165 、VEGF 165b 、VEGF 183 、VEGF 189 and VEGF 206 .

[0049] VEGF is a hypoxia-regulated gene, and VEGF levels increase in hypoxic or ischemic conditions. VEGF is an agonist of VEGF receptors (VEGFRs). VEGFRs are receptor tyrosine kinases; the binding of VEGF to VEGFRs can lead to phosphorylation of the receptors, which subsequently leads to phosphorylation of downstream signal transducers. VEGFR-mediated signal transduction can result in abnormal vasculogenesis, angiogenesis, and vascular permeability, leading to pathological vascular instability. Thus, inhibition of VEGF can lead to reduced VEGFR-mediated signal transduction and enhanced vascular stability. For example, an inhibitor, antibody, antibody fragment, variant, or derivative thereof that binds VEGF can reduce VEGFR ligation, thereby reducing VEGFR-mediated signal transduction and promoting vascular stability. Non-limiting examples of agents that bind VEGF include aflibercept — a recombinant protein that comprises the VEGF-binding portions of human VEGF receptors 1 and 2 fused to the Fc portion of human IgG1; brolucizumab — a humanized single-chain antibody fragment (scFv); RTH258 — a humanized single-chain antibody fragment (scFv); ranibizumab — a humanized monoclonal antibody fragment (Fab); bevacizumab — a humanized monoclonal antibody; conbercept — a recombinant fusion protein that comprises the extracellular domains from VEGF receptors 1 and 2 fused to the Fc portion of human IgG1; Abicipar — a designed ankyrin repeat protein (DARPin); MP0112 — a DARPin; MP0250 — a DARPin; CT-322 — an adnectin; and PRS-050 — an anticalin.

[0050] Through the above processes, VEGF can cause conditions characterized, for example, by vascular instability, angiogenesis, neovascularization, vascular leakage, and / or edema. For example, VEGF can cause vascular disorders, ocular disorders, cancer, kidney disorders, diabetic complications, and other conditions. For example, local ischemia in the eye can lead to increased VEGF production, which can result in vascular leakage and pathological neovascularization in the retina. Inhibition of VEGFR-mediated signal transduction can alleviate such conditions.

[0051] Receptor tyrosine kinases (RTKs) and receptor tyrosine kinase agonists

[0052] Receptor tyrosine kinases (RTKs) are cell surface receptors that are involved in the regulation of cell growth, differentiation, and survival. Binding of an agonist to an RTK can cause adjacent RTKs to associate with each other, forming a dimer. Dimerization can lead to cross-phosphorylation - each RTK in the dimer phosphorylates multiple tyrosine residues on the other RTK. Once cross-phosphorylated, the cytoplasmic tail of the RTK can initiate signal transduction pathways, for example, by acting as a docking platform for various intracellular proteins. RTK signaling can result in changes in gene transcription and expression in the cell.

[0053] Non-limiting examples of RTKs include AATK, AATYK, AATYK1, AATYK2, ACH, ALK, ARK, AXL, BDB, BDB1, BEK, BFGFR, BREK, Brt, CAK, CCK4, CD115, CD117, CD135, CD136, CD140a, CD140b, CD167, CD202b, CD220, CD221, CD246, CD309, CD331, CD332, CD333, CD334, CDHF12, CDHR16, CDw136, CEK, CEK2, CEK3, c-Eyk, CFD1, C-FMS, C-Kit, cprk, c-ros-1, CSF1R, CSFR, D3S3195, DDR1, DDR2, DFNB97, DKFZp761P1010, Dtk, ECT1, EDDR1, EGFR, EphA10, EphA1-8, EphB1, EphB2, EphB3, EphB4, EphB6, ErbB2, ErbB3, ErbB4, Etk-2, FGFR1, FGFR2, FGFR3, FGFR4, FLG, FLK1, FLK2, FLT, FLT1, FLT2, FLT3, FLT4, FMS, GAS9, H2, H3, H4, H5, HGFR, HSCR1, IGF1R, IGFIR, IGFR, INSR, INSRR, IRR, JKT5A, JTK11, JTK12, JTK13, JTK14, JTK2, JTK4, JTK5, JWS, KAL2, KDR, KGFR, KIAA0641, KIAA1079, KIAA1883, KIT, KPI2, K-SAM, LMR1, LMR2, LMR3, LMTK1, LMTK2, LMTK3, LTK, MCF3, MEN2A, MEN2B, Mer, MERTK, MET, MGC18216, MST1R, MTC, MTC1, MTRK1, MuSK, NEP, NOK, N-SAM, NTRK2, NTRK3, NTRK4, NTRKR1, NTRKR2, PBT, PCL, PDGFR, PDGFR1, PDGFR2, PDGFRA, PDGFR-alpha, PDGFRB, PDGFR-beta, PPP1R100, PPP1R101, PPP1R77, PTC, PTK3A, PTK7, PTK8, RCCP2, Rek, RET, RET51, RON, ROR1, ROR2, ROS, ROS1, RP38, RSE, RTK6, RYK, Ryk, RYK1, SCFR, Sky, STK, STYK1, SuRTK106, TEK, TIE1,TIE2, Tif, TK14, TK25, TKT, TRK, TrkA, TrkB, TrkC, TYK1, TYKLM3, TYRO10, Tyro12, Tyro3, Tyro7, UFO, VEGFR, VEGFR1, VEGFR2, VEGFR3, VMCM and VMCM1.

[0054] Non-limiting examples of RTK agonists include VEGF, Ang1, Ang2, BDNF, EGF, FGF, HGF, IGF, insulin, MSP, NGF, NT-3 and PDGF.

[0055] Antibodies and antigen-binding compounds

[0056] A basic four-chain antibody unit comprises two identical heavy chain (H) polypeptide sequences and two identical light chain (L) polypeptide sequences. Each heavy chain may comprise an N-terminal variable (V H ) region and three or four C-terminal constant (C H 1, C H 2, C H 3 and C H ) regions. Each light chain may comprise an N-terminal variable (V L ) region and a C-terminal constant (C L ) region. The light chain variable region is aligned with the heavy chain variable region, and the light chain constant region is aligned with the heavy chain constant region C H1 . The pairing of the heavy chain variable region and the light chain variable region together forms a single antigen-binding site. Each light chain is linked to the heavy chain by a covalent disulfide bond. Depending on the heavy chain isotype, the two heavy chains are linked to each other by one or more disulfide bonds. Each heavy chain and light chain also contain regularly spaced intra-chain disulfide bonds. The C-terminal constant region of the heavy chain contains the Fc region of the antibody, which mediates effector functions, for example, by interaction with Fc receptors or complement proteins. Figure 1 A simple representative schematic of the basic four-chain antibody unit is provided; the light chain sequence is denoted by "SEQ A". The heavy chain sequence is denoted by "SEQ B", -S-S- denotes a disulfide bond, and N and C denote the N-terminal and C-terminal, respectively.

[0057] Based on the amino acid sequence of the constant region, the light chain can be named κ or λ. Based on the amino acid sequence of the constant region, the heavy chain can be named α, δ, ε, γ, or μ. Based on the heavy chain, antibodies are divided into five immunoglobulin classes or isotypes. IgA contains α heavy chains, IgD contains δ heavy chains, IgE contains ε heavy chains, IgG contains γ heavy chains, and IgM contains μ heavy chains. Antibodies of the IgG, IgD, and IgE classes contain monomers (two heavy chains and two light chains) of the above four-chain unit, while antibodies of the IgM and IgA classes can contain multimers of the four-chain unit. Based on the differences in sequence and function of the heavy chain constant region, the α and γ classes are further divided into subclasses. The subclasses of IgA and IgG expressed in humans include IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0058] The constant region is minimally involved in antigen binding. Instead, the constant region can mediate various effector functions. For example, due to interactions with different Fc receptors and / or complement proteins, different IgG isotypes or subclasses can be associated with different effector functions or therapeutic properties. For example, an antibody containing an Fc region that engages an activating Fc receptor can participate in antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), induction of signal transduction via immunoreceptor tyrosine-based activation motifs (ITAMs), and induction of cytokine secretion. For example, an antibody containing an Fc region that engages an inhibitory Fc receptor can induce signal transduction via immunoreceptor tyrosine-based inhibitory motifs (ITIMs).

[0059] Different antibody subclasses have different abilities to initiate immune effector functions. For example, IgG1 and IgG3 can effectively recruit complement to activate CDC, and IgG2 elicits minimal ADCC. IgG4 has a lesser ability to trigger immune effector functions. Modification of the constant region can also affect antibody properties, such as enhancement or reduction of Fc receptor ligation, enhancement or reduction of ADCC, enhancement or reduction of ADCP, enhancement or reduction of CDC, enhancement or reduction of signal transduction via ITAMs, enhancement or reduction of cytokine induction, enhancement or reduction of signal transduction via ITIMs, extension or shortening of the half-life, or enhancement or reduction of co-engagement of antigen and Fc receptor. Modifications can include, for example, amino acid mutations, alteration of post-translational modifications (such as glycosylation), combination of domains from different isotypes or subclasses, or combinations thereof.

[0060] The compounds or antibodies of the present disclosure may comprise constant regions or Fc regions that are selected or modified to provide suitable antibody characteristics, e.g., suitable properties for treating the diseases or conditions disclosed herein. In some embodiments, IgG1 may be used, for example, to promote immune activating effector functions (e.g., ADCC, ADCP, CDC, ITAM signaling, cytokine induction, or combinations thereof, for treating cancer). In some embodiments, e.g., in situations where an antagonistic property of the antibody is desired without immune effector function (e.g., for treating ocular conditions), IgG4 may be used.

[0061] Non-limiting examples of antibody modifications and their effects are provided in Table 1.

[0062]

[0063] The variable (V) regions mediate antigen binding and define the specificity of a particular antibody for an antigen. The variable regions contain relatively invariant sequences called framework regions and hypervariable regions, which vary widely in sequence between antibodies with different binding specificities. The variable region of each antibody heavy or light chain contains four framework regions separated by three hypervariable regions. The variable regions of the heavy and light chains fold in such a way as to bring the hypervariable regions into close proximity to form an antigen-binding site. The four framework regions predominantly adopt a β-sheet conformation, while the three hypervariable regions form loops that connect the β-sheet structures and, in some cases, form part of the β-sheet structures.

[0064] There are amino acid residues within the hypervariable regions that primarily determine the binding specificity of the antibody. The sequences containing these residues are called complementarity-determining regions (CDRs). One antigen-binding site of an antibody contains six CDRs, three within the hypervariable region of the light chain and three within the hypervariable region of the heavy chain. The CDRs in the light chain are named L1, L2, and L3, while the CDRs in the heavy chain are named H1, H2, and H3. The CDRs may also be designated as LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3, respectively. The contribution of each CDR to antigen binding varies among antibodies. The lengths of the CDRs can vary. For example, the lengths of the CDRs are typically 5 to 14 residues, but there are CDRs as short as 0 residues or as long as 25 residues or longer.

[0065] There are several methods for predicting or assigning CDR sequences. These methods may use different numbering systems, e.g., because sequence insertions and deletions are numbered differently.

[0066] The Kabat method was developed by aligning a limited number of antibody sequences and determining the positions of the most variable residues. Based on the alignment, a numbering scheme was introduced for the residues in the variable regions. This numbering scheme can be used to determine the positions marking the start and end of each CDR. One iteration of the Kabat numbering system uses the following residue positions to determine the CDRs in the light chain variable region: LCDR1 around residues 24 - 34; LCDR2 around residues 50 - 56; and LCDR3 around residues 89 - 97. One iteration of the Kabat numbering system uses the following residue positions to determine the CDRs in the heavy chain variable region: HCDR1 around residues 31 - 35; HCDR2 around residues 50 - 65; and HCDR3 around residues 95 - 102.

[0067] The Chothia method was developed based on the analysis of three-dimensional antibody structures. The analysis determined that the hypervariable loops adopt a restricted set of conformations based on the presence of certain residues at key positions in the CDRs and flanking framework regions. This method uses a numbering scheme similar to the Kabat method, but numbers insertions and deletions differently. One iteration of the Chothia numbering system uses the following residue positions to determine the CDRs in the light chain variable region: LCDR1 around residues 24 - 34; LCDR2 around residues 50 - 56; and LCDR3 around residues 89 - 97. One iteration of the Chothia numbering system uses the following residue positions to determine the CDRs in the heavy chain variable region: HCDR1 around residues 26 - 34; HCDR2 around residues 52 - 56; and HCDR3 around residues 95 - 102.

[0068] The IMGT method (International ImMunoGeneTics database) was developed by integrating existing definitions of framework regions and CDRs, structural data, and data from alignments of antibody variable region sequences. This integration led to the identification of conserved residues in the framework regions, which can be used as reference points for identifying CDRs. Examples of conserved residues in the variable regions include cysteine at approximately residue 23 (in framework region 1), tryptophan at approximately residue 41 (in framework region 2), a hydrophobic amino acid at approximately residue 89 (in framework region 3), cysteine at approximately residue 104 (in framework region 3), and phenylalanine or tryptophan at approximately residue 118 (in framework region 4). By using an algorithm based on computational alignment, CDRs can be identified in the sequences encoding the variable regions of the target antibodies.

[0069] The IMGT numbering method always assigns the same number to conserved amino acids, but allows the lengths of the CDR and framework regions to vary. Thus, the IMGT numbering of residues is not necessarily continuous. The lengths of the CDRs determined by the IMGT method can vary. For example, LCDR1 or HCDR1 can be about 5 to about 12 amino acids, LCDR2 or HCDR2 can be about 0 to about 10 amino acids, and LCDR3 or HCDR3 can be about 5 to about 91 amino acids.

[0070] Based on multiple structural alignments of available antibody-antigen complexes, the Paratome method was developed. It was found that the structural positions involved in antigen binding were similar among the antibodies examined, and the antibody sequences in the dataset were annotated with antigen-binding regions (ABRs, analogous to CDRs). The ABRs in a query sequence can be determined using computational tools that first align the query sequence against antibodies with resolved antibody-antigen structures and then infer the positions of the ABRs based on that alignment. Antibodies with resolved structures can also determine the ABRs using structural alignment methods rather than sequence-based alignment methods.

[0071] A subset of residues within the CDR contacts the antigen. These residues that contact the antigen can be referred to as specific-determining residues (SDRs). However, other residues outside the SDRs can contribute to binding activity by helping to maintain the conformation of the binding site. The number of SDRs in an antibody can vary depending on the size and type of the antigen being recognized. For example, 0 - 14 SDRs can be found within the CDR. SDRs can be enriched in certain residues such as tyrosine, serine, tryptophan, and asparagine.

[0072] Monoclonal antibodies can be obtained from a substantially homogeneous antibody population, i.e., all the individual antibodies that make up the population are identical except for possibly minor amounts of naturally occurring mutations. In contrast to polyclonal antibody preparations that include different antibodies against different epitopes, each monoclonal antibody is directed against a single epitope.

[0073] The compounds of the present invention can be monoclonal antibodies, for example, chimeric antibodies in which portions of the heavy and / or light chains are identical or homologous to the corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remaining portions of the chains are identical or homologous to the corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass or antigen-binding fragments of such antibodies.

[0074] Antibody fragments or antigen-binding fragments can comprise a portion of an antibody, such as the antigen-binding region or variable region of a whole antibody. Non-limiting examples of antibody fragments include Fab, Fab', F(ab′) 2, dimers and trimers of Fab conjugates, Fv, scFv, minibodies, diabodies, triabodies and tetra-bodies, and linear antibodies. Fab and Fab′ are antigen-binding fragments that may comprise the V of the heavy chain linked to the light chain by a disulfide bond L and C L domains, and the V H and C H 1 domains. F(ab’) 2 may comprise two Fabs or Fab’s linked by a disulfide bond. Fv may comprise V H and V L domains held together by non-covalent interactions. scFv (single-chain variable fragment) is a fusion protein that may comprise V H and V L domains linked by a peptide linker. Manipulating the orientation of the V H and V L domains and the linker length can be used to create different forms of molecules that can be monomeric, dimeric (diabody), trimeric (triabody) or tetrameric (tetrabody). Minibodies are scFv-C H 3 fusion proteins that can be assembled into bivalent dimers.

[0075] Non-limiting examples of epitopes include amino acids, sugars, lipids, phosphoryl groups and sulfonyl groups. Epitopes may have specific three-dimensional structural properties and / or specific charge properties. Epitopes can be conformational or linear.

[0076] For human administration, monoclonal antibodies produced in non-human species can be further optimized through a humanization process to reduce the likelihood of immunogenicity while retaining target specificity. The humanization process involves integrating human DNA into the genetic sequence of the gene that produces the isolated antibody. The recombinant DNA is cloned and expressed in cells for large-scale production of the new humanized antibody.

[0077] An example of a humanized antibody is a modified chimeric antibody. As described above, chimeric antibodies can be produced. The chimeric antibody is further mutated outside the CDRs to replace the non-human sequences in the variable region with homologous human sequences. Another example of a humanized antibody is a CDR-grafted antibody, in which non-human CDR sequences are introduced into the human heavy and light chain variable sequences of a human antibody scaffold to replace the corresponding human CDR sequences.

[0078] Humanized antibodies can be produced in mammalian cells, bioreactors or transgenic animals such as mice, chickens, sheep, goats, pigs or marmosets. The transgenic animal can insert most of the genome that produces human antibodies into the genome of the animal.

[0079] In addition to antibodies and antibody fragments, other antigen-binding compounds can bind to target molecules. Non-limiting examples of non-antibody-derived antigen-binding compounds include ankyrins, ankyrin repeat proteins, designed ankyrin repeat proteins (DARPins), affibodies, avimers, adnectins, anticalins, Fynomers, Kunitz domains, knottins, β-hairpin mimetics, and their receptors and derivatives, e.g., VEGF receptors, or VEGF-binding portions of human VEGF receptors 1 and 2.

[0080] Designed ankyrin repeat proteins (DARPins) can be protein scaffolds based on ankyrin repeats. A DARPin can comprise one or more ankyrin repeats, which contain common sequence and / or structural motifs. When compared to each other, individual ankyrin repeats can contain common sequence and / or structural motifs, although containing mutations, substitutions, additions, and / or deletions. A DARPin can comprise, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more ankyrin repeats. A DARPin can comprise an N-terminal capping repeat, one or more internal ankyrin repeats, and a C-terminal capping repeat. Each ankyrin repeat can comprise framework residues and protein interaction residues. The framework residues can contribute to the structure or folding topology, e.g., the structure of the ankyrin repeat or the interaction with adjacent ankyrin repeats. The protein interaction residues can facilitate binding to the target molecule, e.g., by direct interaction with the target molecule or by stabilizing the direct interaction residues in a conformation that permits binding.

[0081] Compounds, antibodies, their fragments or derivatives, or other compounds that bind to a target molecule in the present disclosure can have a K of, for example, less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, less than about 900 pM, less than about 800 pM, less than about 700 pM, less than about 600 pM, less than about 500 pM, less than about 400 pM, less than about 300 pM, less than about 200 pM, less than about 100 pM, less than about 90 pM, less than about 80 pM, less than about 70 pM, less than about 60 pM, less than about 50 pM, less than about 40 pM, less than about 30 pM, less than about 20 pM, less than about 10 pM, less than about 9 pM, less than about 8 pM, less than about 7 pM, less than about 6 pM, less than about 5 pM, less than about 4 pM, less than about 3 pM, less than about 2 pM, less than about 1 pM, less than about 900 fM, less than about 800 fM, less than about 700 fM, less than about 600 fM, less than about 500 fM, less than about 400 fM, less than about 300 fM, less than about 200 fM, less than about 100 fM, less than about 90 fM, less than about 80 fM, less than about 70 fM, less than about 60 fM, less than about 50 fM, less than about 40 fM, less than about 30 fM, less than about 20 fM, or less than about 10 fM D Bind to the target.

[0082] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds that bind to a target molecule in the present disclosure can be, for example, from about 10 fM to about 500 nM, from about 30 fM to about 500 nM, from about 30 fM to about 400 nM, from about 30 fM to about 300 nM, from about 30 fM to about 200 nM, from about 30 fM to about 100 nM, from about 30 fM to about 90 nM, from about 30 fM to about 80 nM, from about 30 fM to about 70 nM, from about 30 fM to about 60 nM, from about 30 fM to about 50 nM, from about 30 fM to about 40 nM, from about 30 fM to about 30 nM, from about 30 fM to about 20 nM, from about 30 fM to about 10 nM, from about 30 fM to about 9 nM, from about 30 fM to about 8 nM, from about 30 fM to about 7 nM, from about 30 fM to about 6 nM, from about 30 fM to about 5 nM, from about 30 fM to about 4 nM, from about 30 fM to about 3 nM, from about 30 fM to about 2 nM, from about 30 fM to about 1 nM, from about 30 fM to about 900 pM, from about 30 fM to about 800 pM, from about 30 fM to about 700 pM, from about 30 fM to about 600 pM, from about 30 fM to about 500 pM, from about 30 fM to about 400 pM, from about 30 fM to about 300 pM, from about 30 fM to about 200 pM, from about 30 fM to about 100 pM, from about 30 fM to about 90 pM, from about 30 fM to about 80 pM, from about 30 fM to about 70 pM, from about 30 fM to about 60 pM, from about 30 fM to about 50 pM, from about 30 fM to about 40 pM, from about 30 fM to about 30 pM, from about 30 fM to about 20 pM, from about 30 fM to about 10 pM, from about 30 fM to about 1 pM, from about 30 fM to about 900 fM, from about 30 fM to about 800 fM, from about 30 fM to about 700 fM, from about 30 fM to about 600 fM, from about 30 fM to about 500 fM, from about 30 fM to about 400 fM, from about 30 fM to about 300 fM, from about 30 fM to about 200 fM, from about 30 fM to about 100 fM, from about 30 fM to about 500 nM, from about 30 fM to about 400 nM, from about 30 fM to about 300 nM, from about 30 fM to about 200 nM, from about 30 fM to about 100 nM, from about 30 fM to about 90 nM, from about 30 fM to about 80 nM, from about 30 fM to about 70 nM, from about 30 fM to about 60 nM, from about 30 fM to about 50 nM, from about 30 fM to about 40 nM, from about 30 fM to about 30 nM, from about 30 fM to about 20 nM, from about 30 fM to about 10 nM, from about 30 fM to about 9 nM, from about 30 fM to about 8 nM, from about 30 fM to about 7 nM, from about 30 fM to about 6 nM, from about 30 fM to about 5 nM, from about 30 fM to about 4 nM, from about 30 fM to about 3 nM, from about 30 fM to about 2 nM, from about 30 fM to about 1 nM, from about 30 fM to about 900 pM, from about 30 fM to about 800 pM, from about 30 fM to about 700 pMfrom about 30 fM to about 600 pM, from about 30 fM to about 500 pM, from about 30 fM to about 400 pM, from about 30 fM to about 300 pM, from about 30 fM to about 200 pM, from about 30 fM to about 100 pM, from about 30 fM to about 90 pM, from about 30 fM to about 80 pM, from about 30 fM to about 70 pM, from about 30 fM to about 60 pM, from about 30 fM to about 50 pM, from about 30 fM to about 40 pM, from about 30 fM to about 30 pM, from about 30 fM to about 20 pM, from about 30 fM to about 10 pM, from about 1 pM to about 500 nM, from about 1 pM to about 400 nM, from about 1 pM to about 300 nM, from about 1 pM to about 200 nM, from about 1 pM to about 100 nM, from about 1 pM to about 90 nM, from about 1 pM to about 80 nM, from about 1 pM to about 70 nM, from about 1 pM to about 60 nM, from about 1 pM to about 50 nM, from about 1 pM to about 40 nM, from about 1 pM to about 30 nM, from about 1 pM to about 20 nM, from about 1 pM to about 10 nM, from about 1 pM to about 9 nM, from about 1 pM to about 8 nM, from about 1 pM to about 7 nM, from about 1 pM to about 6 nM, from about 1 pM to about 5 nM, from about 1 pM to about 4 nM, from about 1 pM to about 3 nM, from about 1 pM to about 2 nM, from about 1 pM to about 1 nM, from about 1 pM to about 900 pM, from about 1 pM to about 800 pM, from about 1 pM to about 700 pM, from about 1 pM to about 600 pM, from about 1 pM to about 500 pM, from about 1 pM to about 400 pM, from about 1 pM to about 300 pM, from about 1 pM to about 200 pM, from about 1 pM to about 100 pM, from about 1 pM to about 90 pM, from about 1 pM to about 80 pM, from about 1 pM to about 70 pM, from about 1 pM to about 60 pM, from about 1 pM to about 50 pM, from about 1 pM to about 40 pM, from about 1 pM to about 30 pM, from about 1 pM to about 20 pM, from about 1 pM to about 10 pM, from about 100 pM to about 500 nM, from about 100 pM to about 400 nM, from about 100 pM to about 300 nM, from about 100 pM to about 200 nM, from about 100 pM to about 100 nM, from about 100 pM to about 90 nM, from about 100 pM to about 80 nM, from about 100 pM to about 70 nM, from about 100 pM to about 60 nM, from about 100 pM to about 50 nM, from about 100 pM to about 40 nM, from about 100 pM to about 30 nM, from about 100 pM to about 20 nM, from about 100 pM to about 10 nM, from about 100 pM to about 9 nM, from about 100 pM to about 8 nM, from about 100 pM to about 7 nM, from about 100 pM to about 6 nM, from about 100 pM to about 5 nM, from about 100 pM to about 4 nM, from about 100 pM to about 3 nM, from about 100 pM to about 2 nM, from about 100 pM to about 1 nM, from about 100 pM to about 900 pM, from about 100 pM to about 800 pM, from about 100 pM to about 700 pM, from about 100 pM to about 600 pMK of about 100 pM to about 500 pM, about 100 pM to about 400 pM, about 100 pM to about 300 pM, or about 100 pM to about 200 pM, D binds to the target.

[0083] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 500 fM to about 500 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 1 pM to about 500 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 60 pM to about 500 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 100 pM to about 500 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 1 pM to about 300 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 1 pM to about 200 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 1 pM to about 120 pM D and bind to HPTP-β (VE-PTP). In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 1 pM to about 70 pM D and bind to HPTP-β (VE-PTP).

[0084] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 30 fM to about 900 pM D and bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 30 fM to about 600 pM D and bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of about 30 fM to about 200 pM DBind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 30 fM to about 30 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 30 fM to about 40 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 30 fM to about 1 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 30 fM to about 200 fM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 1 pM to about 900 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 1 pM to about 600 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 1 pM to about 200 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 1 pM to about 30 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 1 pM to about 40 pM D Bind to VEGF.

[0085] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 30 fM to about 2 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 35 fM to about 200 fM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 100 fM to about 2 pM D Bind to VEGF.

[0086] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure can have a K of from about 20 pM to about 1 nM DBind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of from about 20 pM to about 800 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of from about 20 pM to about 350 pM D Bind to VEGF.

[0087] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of from about 30 fM to about 700 pM D Bind to VEGF.

[0088] In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of from about 40 fM to about 520 pM D Bind to VEGF. In some embodiments, the compounds, antibodies, fragments or derivatives thereof, or other compounds of the present disclosure may have a K of from about 40 fM to about 110 pM D Bind to VEGF.

[0089] Multivalent multispecific compounds

[0090] Antigen-binding compounds can be combined to generate multivalent and / or multispecific compounds. Such multivalent and / or multispecific compounds may have advantages over the parent compounds administered alone. These advantages may include, for example, a simpler dosing regimen, a longer half-life in the subject, and the ability to bind tightly to the target antigen.

[0091] Multispecific antibodies can be generated by a variety of methods. In one method, for example, monospecific antibodies or derivatives thereof can be chemically conjugated by chemically conjugating two IgG antibody units into a conjugate.

[0092] In another method, additional antigen-binding domains can be attached to a conventional IgG antibody or a derivative thereof using cloning techniques. The additional antigen-binding domains can be, for example, single variable domains (sVDs), single-chain variable fragments (scFvs), single-chain Fabs, peptides, ankyrins, ankyrin repeat proteins, designed ankyrin repeat proteins (DARPins), affibodies, avimers, adnectins, anticalins, Fynom ers, Kunitz domains, knottins, β-hairpin mimetics, tetrameric polyethylene glycol-clustered peptides, peptides derived from one or more receptors (such as VEGF receptors, or the VEGF-binding portions of human VEGF receptors 1 and 2), or derivatives thereof.

[0093] Additional antigen-binding domains (e.g., sVD, scFv, single-chain Fab, peptide, ankyrin, ankyrin repeat protein, DARPin, affibody, avimer, adnectin, anticalin, Fynomer, Kunitz domain, knottin, β-hairpin mimetic, tetravalent polyethylene glycol-clustered peptide, or a peptide derived from one or more receptors) can be attached, for example, via a peptide linker to the N- or C-terminus of the light and / or heavy chain of IgG or Fab. In some embodiments, an scFv can be attached to the C-terminus of the heavy chain of IgG to provide a tetravalent bispecific antibody. An scFv can be attached to the C-terminus of the light chain of IgG to provide a tetravalent bispecific antibody. An scFv can be attached to the C-terminus of the light chain and the C-terminus of the heavy chain of IgG to provide a hexavalent bispecific antibody. In some embodiments, a DARPin can be attached to the C-terminus of the heavy chain of IgG to provide a tetravalent bispecific antibody. A DARPin can be attached to the C-terminus of the light chain of IgG to provide a tetravalent bispecific antibody. A DARPin can be attached to the C-terminus of the light chain and the C-terminus of the heavy chain of IgG to provide a hexavalent bispecific antibody. Other examples of multispecific antibodies generated by cloning techniques include: (i) DVD-Ig TM (dual variable domain immunoglobulin, tandem ligation of a second V H and V L to the N-terminus of HC and LC, respectively), (ii) Tandemab (combinatorial tandem ligation of 2 V H -C H 1 with a common LC), (iii) DNL (natural association of two antibodies or antibody fragments anchored to the DDD (dimerization and docking domain) from PKA (protein kinase A) and the AD (anchoring domain) from A-kinase anchoring protein (AKAP), respectively), (iv) LUZ-Y (a leucine zipper tethered to the C-terminus of HC, followed by removal by proteolysis), (v) 2-in-1 IgG (identical LC and HC capable of dual recognition), and (vi) mAb 2 (engineered loop in the C H 3 domain of IgG to obtain a second specificity).

[0094] Another class of multispecific antibodies can be characterized by a structure having variable domains or scFv as structural units. Non-limiting examples of such multispecific antibodies include two V H domains joined in tandem, diabodies (heterodimers containing two polypeptide chains that encode V H -V L or V L -V H in the order of V L A-V H B and VH A-V L B, linker with 5 amino acids), dsDbs (interchain disulfide bond between the V L and V H ), DARTs (bispecific affinity retargeting, interchain disulfide bond between 2 Vs L ), scDbs (single-chain diabodies), tandAbs (diabody dimers through a flexible linker between them) and 2 scFvs tandemly linked by an adjustable linker.

[0095] Another class of multispecific antibodies may contain different antigen-binding fragments while retaining the basic IgG structure. Such antibodies may contain, for example, two different heavy chains and / or two different light chains. Various techniques can be used to promote the pairing of desired light and heavy chain combinations rather than random chain association. Non-limiting examples of such techniques include using a common light chain, orthogonal Fab interfaces (introducing complementary mutations at the LC and HC interfaces in one Fab without changes to the other Fab), CrossMab (where one Fab V H or C H 1 domain can be switched with a partner V L or C L domain while the other Fab remains unaffected), and replacing the Fab with a single-chain antigen-binding domain. Further examples include engineering strategies that can introduce mutations into the C H 3 domain to promote heterodimerization based on steric or electrostatic complementarity. The "knobs in holes" method may involve creating a "knob" by replacing the threonine at position 366 with a bulky tryptophan residue on one heavy chain and forming a corresponding "hole" through triple mutations (T366S, L368A, and Y407V) on the partner heavy chain. Another method may involve creating alternating human IgG and IgA fragments in C H 3 to provide a so-called SEEDbody (strand-exchange engineered domain) to direct heavy chain heterodimerization.

[0096] A non-limiting schematic of a multispecific antibody is provided in Figures 2 - 15 .

[0097] Figure 2 A schematic of a tetravalent bispecific antibody with a sequence appended to the C-terminus of the heavy chain is provided. The light chain sequence of the IgG isotype antibody unit is represented by "SEQ A". The heavy chain sequence of the IgG isotype antibody unit is represented by "SEQ B". The linker sequence is represented by "SEQ C". The appended antigen-binding domain sequence is represented by "SEQ D". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0098] Figure 3 Schematic diagrams of tetravalent bispecific antibodies with sequences appended to the C-terminus of the light chain are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B". The linker sequences are represented by "SEQ C". The appended antigen-binding domain sequences are represented by "SEQ D". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0099] Figure 4 Schematic diagrams of hexavalent bispecific antibodies with sequences appended to the C-terminus of the heavy and light chains are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B". The linker sequences are represented by "SEQ C" and "SEQ E". The appended antigen-binding domain sequences are represented by "SEQ D". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0100] Figure 5 Schematic diagrams of hexavalent trispecific antibodies with sequences appended to the C-terminus of the heavy and light chains are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B". The linker sequences are represented by "SEQ C" and "SEQ E". The appended antigen-binding domain sequences are represented by "SEQ D" and "SEQ F". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0101] Figure 6 Schematic diagrams of bivalent bispecific antibodies with two different heavy chain sequences and two different light chain sequences are provided. The light chain sequences are represented by "SEQ A" and "SEQ D". The heavy chain sequences are represented by "SEQ B" and "SEQ C". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0102] Figure 7 Schematic diagrams of trivalent trispecific antibodies with two different heavy chain sequences, two different light chain sequences, and a sequence appended to the C-terminus of one of the light chains are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequence is represented by "SEQ E". The appended antigen-binding domain sequence is represented by "SEQ F". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0103] Figure 8Schematic diagrams of trivalent trispecific antibodies having two different heavy chain sequences, two different light chain sequences, and a sequence attached to the C-terminus of one heavy chain are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequence is represented by "SEQ E". The additional antigen-binding domain sequence is represented by "SEQ F". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0104] Figure 9 Schematic diagrams of tetravalent antibodies having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C-termini of two light chains are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen-binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0105] Figure 10 Schematic diagrams of tetravalent antibodies having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C-termini of two heavy chains are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen-binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0106] Figure 11 Schematic diagrams of tetravalent antibodies having two different heavy chain sequences, two different light chain sequences, and sequences cis-attached to the C-terminus of one heavy chain and the C-terminus of one light chain are provided. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen-binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N-terminus and C-terminus, respectively. -S-S- represents a disulfide bond.

[0107] Figure 12Schematic diagram of a tetravalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences trans - attached to the C - terminus of one heavy chain and the C - terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E" and "SEQ F". The additional antigen - binding domain sequences are represented by "SEQ G" and "SEQ H". N and C represent the N - terminus and C - terminus, respectively. -S - S- represents a disulfide bond.

[0108] Figure 13 Schematic diagram of a pentavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C - termini of two heavy chains and the C - terminus of one light chain. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E", "SEQ F", and "SEQ G". The additional antigen - binding domain sequences are represented by "SEQ H", "SEQ I", and "SEQ J". N and C represent the N - terminus and C - terminus, respectively. -S - S- represents a disulfide bond.

[0109] Figure 14 Schematic diagram of a pentavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C - terminus of one heavy chain and the C - termini of two light chains. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQ D". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E", "SEQ F", and "SEQ G". The additional antigen - binding domain sequences are represented by "SEQ H", "SEQ I", and "SEQ J". N and C represent the N - terminus and C - terminus, respectively. -S - S- represents a disulfide bond.

[0110] Figure 15 Schematic diagram of a hexavalent antibody having two different heavy chain sequences, two different light chain sequences, and sequences attached to the C - termini of the heavy and light chains. The light chain sequences of the IgG isotype antibody units are represented by "SEQ A" and "SEQD". The heavy chain sequences of the IgG isotype antibody units are represented by "SEQ B" and "SEQ C". The linker sequences are represented by "SEQ E", "SEQ F", "SEQ G", and "SEQ H". The additional antigen - binding domain sequences are represented by "SEQ I", "SEQ J", "SEQ K", and "SEQ L". N and C represent the N - terminus and C - terminus, respectively. -S - S- represents a disulfide bond.

[0111] Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP) can be combined with antigen-binding compounds specific for, for example, RTK agonists to provide multivalent multispecific compounds that can bind both HPTP-β (VE-PTP) and RTK agonists. For example, antigen-binding compounds specific for HPTP-β (VE-PTP) can be combined with antigen-binding compounds specific for VEGF, Ang1, Ang2, BDNF, EGF, FGF, HGF, IGF, insulin, MSP, NGF, NT-3, PDGF, or any combination thereof, to provide multivalent multispecific compounds that can bind both HPTP-β (VE-PTP) and RTK agonists.

[0112] Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP) can be combined with antigen-binding compounds specific for, for example, VEGF to provide multivalent multispecific compounds that can bind both HPTP-β (VE-PTP) and VEGF. Compounds that bind both HPTP-β (VE-PTP) and VEGF can inhibit HPTP-β (VE-PTP), activate Tie2, inhibit the binding of VEGF to VEGFR, and inhibit VEGFR signaling.

[0113] Sequences derived from aflibercept (a recombinant protein containing the VEGF-binding portions of human VEGF receptors 1 and 2) can be combined with antigen-binding compounds specific for HPTP-β (VE-PTP). For example, sequences derived from aflibercept can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP)-specific antibody to provide a tetravalent bispecific antibody ( Figure 2 ). Sequences derived from aflibercept can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP)-specific antibody to provide a tetravalent bispecific antibody ( Figure 3 ). Sequences derived from aflibercept can be fused to the C-termini of the light and heavy chains of an HPTP-β (VE-PTP)-specific antibody to provide a hexavalent bispecific antibody ( Figure 4 ).

[0114] Sequences derived from brolucizumab (a humanized single-chain antibody fragment (scFv) inhibitor of VEGF that binds to the receptor-binding site of VEGF) can be combined with antigen-binding compounds specific for HPTP-β (VE-PTP). For example, sequences from brolucizumab can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP)-specific antibody to provide a tetravalent bispecific antibody ( Figure 2)。Sequences derived from brolucizumab can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。Sequences derived from brolucizumab can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0115] Sequences derived from ranibizumab (a humanized monoclonal antibody fragment (Fab) that binds and inhibits the activity of VEGF) can be combined with an antigen-binding compound specific for HPTP-β (VE-PTP). For example, the sequence from ranibizumab can be cloned into an scFv, and the ranibizumab-derived scFv can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 2 )。The ranibizumab-derived scFv can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。The ranibizumab-derived scFv can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0116] Sequences derived from bevacizumab (a humanized monoclonal antibody that binds and inhibits the activity of VEGF) can be combined with an antigen-binding compound specific for HPTP-β (VE-PTP). For example, the sequence from bevacizumab can be cloned into an scFv, and the bevacizumab-derived scFv can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 2 )。The bevacizumab-derived scFv can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。The bevacizumab-derived scFv can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0117] Sequences derived from conbercept (a recombinant protein containing the VEGF-binding portions of VEGF receptors 1 and 2) can be combined with an antigen-binding compound specific for HPTP-β (VE-PTP). For example, the sequence derived from conbercept can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody (Figure 2 )。The sequence derived from Conbercept can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。The sequence derived from Conbercept can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0118] The sequence derived from abicipar (a DARPin that binds VEGF) can be combined with an antigen-binding compound specific for HPTP-β (VE-PTP). For example, the sequence derived from abicipar can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 2 )。The sequence derived from abicipar can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。The sequence derived from abicipar can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0119] An antigen-binding compound specific for HPTP-β (VE-PTP) can be combined with a DARPin or its amino acid sequence, such as an amino acid sequence comprising any one of SEQ ID NOs: 158-217. An amino acid sequence comprising any one of SEQ ID NOs: 158-217 can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 2 )。An amino acid sequence comprising any one of SEQ ID NOs: 158-217 can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP) specific antibody to provide a tetravalent bispecific antibody ( Figure 3 )。An amino acid sequence comprising any one of SEQ ID NOs: 158-217 can be fused to the C-terminus of the light and heavy chains of an HPTP-β (VE-PTP) specific antibody to provide a hexavalent bispecific antibody ( Figure 4 )。

[0120] Multivalent VEGF-specific compounds can be combined with antigen-binding compounds specific for HPTP-β (VE-PTP). For example, one binding domain (e.g., an aflibercept-derived sequence) can be fused to the C-terminus of the heavy chain of an anti-HPTP-β (VE-PTP) antibody, while another binding domain (e.g., a brolucizumab-derived sequence) can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP)-specific antibody to provide a hexavalent trispecific antibody ( Figure 5 ).

[0121] If different heavy chains are used in a basic four-chain IgG antibody unit (e.g., using the "knobs-into-holes" method), antibodies can be generated that are divalent, trivalent, tetravalent, pentavalent or hexavalent, and monospecific, bispecific, trispecific, tetraspecific, pentaspecific or hexaspecific.

[0122] In one non-limiting example, one arm of the antibody unit can contain a CDR specific for HPTP-β (VE-PTP), while the other arm of the antibody unit can contain a CDR specific for VEGF ( Figure 6 ).

[0123] In one non-limiting example, both antigen-binding fragments of the basic antibody unit can be specific for HPTP-β (VE-PTP), and a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence or a bevacizumab-derived sequence) can be fused to one light chain to provide a trivalent bispecific antibody ( Figure 7 ).

[0124] In one non-limiting example, both antigen-binding fragments of the basic antibody unit can be specific for HPTP-β (VE-PTP), and a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence or a bevacizumab-derived sequence) can be fused to one heavy chain to provide a trivalent bispecific antibody ( Figure 8 ).

[0125] In one non-limiting example, both antigen-binding fragments of the basic antibody unit can be specific for HPTP-β (VE-PTP), and a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence or a bevacizumab-derived sequence) can be fused to one light chain, and a second different binding domain specific for VEGF can be fused to the other light chain to provide a tetravalent trispecific antibody ( Figure 9 ).

[0126] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, and a second different binding domain specific for VEGF can be fused to the other heavy chain to provide a tetravalent trispecific antibody ( Figure 10 ).

[0127] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, and a second different binding domain specific for VEGF can be fused in cis to one light chain to provide a tetravalent trispecific antibody ( Figure 11 ).

[0128] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, and a second different binding domain specific for VEGF can be fused in trans to one light chain to provide a tetravalent trispecific antibody ( Figure 12 ).

[0129] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, a second different binding domain specific for VEGF can be fused to a second heavy chain, and a third different binding domain specific for VEGF can be fused to one light chain to provide a pentavalent tetra-specific antibody ( Figure 13 ).

[0130] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, a second different binding domain specific for VEGF can be fused to one light chain, and a third different binding domain specific for VEGF can be fused to the other light chain to provide a pentavalent tetra-specific antibody ( Figure 14 ).

[0131] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), a binding domain specific for VEGF (e.g., an aflibercept-derived sequence, a brolucizumab-derived sequence, a ranibizumab-derived sequence, or a bevacizumab-derived sequence) can be fused to one heavy chain, a second different binding domain specific for VEGF can be fused to the other heavy chain, a third different binding domain specific for VEGF can be fused to one light chain, and a fourth binding domain specific for VEGF can be fused to the other light chain to provide a hexavalent penta-specific antibody ( Figure 15 ).

[0132] In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), one binding domain (e.g., an aflibercept-derived sequence) can be fused to the C-terminus of one heavy chain, and a second binding domain (e.g., a brolucizumab-derived sequence) is fused to the C-terminus of the other heavy chain to provide a tetravalent tri-specific antibody ( Figure 10 ). In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), an aflibercept-derived sequence can be fused to the C-terminus of one light chain, and a brolucizumab-derived sequence is fused to the C-terminus of the other light chain to provide a tetravalent tri-specific antibody ( Figure 9 ). In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), an aflibercept-derived sequence can be fused to the C-terminus of one heavy chain, and a brolucizumab-derived sequence is fused to the C-terminus of one light chain to provide a tetravalent tri-specific antibody ( Figure 11 、 Figure 12)。In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), the aflibercept-derived sequence can be fused to the C-terminus of one heavy chain, the brolucizumab-derived sequence can be fused to the C-terminus of the other heavy chain, and the ranibizumab-derived sequence can be fused to the C-terminus of both light chains to provide a hexavalent tetra-specific antibody( Figure 15 )。In a non-limiting example, two antigen-binding fragments of the basic antibody unit can both be specific for HPTP-β (VE-PTP), the aflibercept-derived sequence can be fused to the C-terminus of one heavy chain, the brolucizumab-derived sequence can be fused to the C-terminus of the other heavy chain, the ranibizumab-derived sequence can be fused to the C-terminus of one light chain, and the bevacizumab-derived sequence can be fused to the C-terminus of the other light chain to provide a hexavalent penta-specific antibody( Figure 15 )。

[0133] An antigen-binding compound specific for, for example, HPTP-β (VE-PTP) can be combined with other amino acid sequences to provide a multivalent multi-specific compound. For example, the compound enhances Tie2 activation, enhances Tie2 phosphorylation, enhances Tie2 signaling, reduces VEGFR activation, reduces VEGFR phosphorylation, reduces VEGFR signaling, or a combination thereof.

[0134] An antigen-binding compound specific for HPTP-β (VE-PTP) can be combined with a collagen IV-derived biomimetic peptide, for example, an amino acid sequence comprising SEQ ID NO:152 or SEQ ID NO:153. The amino acid sequence comprising SEQ ID NO:152 or SEQ ID NO:153 can be fused to the C-terminus of the heavy chain of an HPTP-β (VE-PTP)-specific antibody to provide a tetravalent bispecific antibody( Figure 2 )。The amino acid sequence comprising SEQ ID NO:152 or SEQ ID NO:153 can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP)-specific antibody to provide a tetravalent bispecific antibody( Figure 3 )。The amino acid sequence comprising SEQ ID NO:152 or SEQ ID NO:153 can be fused to the C-termini of the light and heavy chains of an HPTP-β (VE-PTP)-specific antibody to provide a hexavalent bispecific antibody( Figure 4 )。

[0135] Antigen-binding compounds specific for HPTP-β (VE-PTP) can be combined with Ang1 mimetics, such as vasculotide. Sequences derived from vasculotide can be fused to the C-terminus of the heavy chain of an antibody specific for HPTP-β (VE-PTP) to provide a tetravalent bispecific antibody ( Figure 2 ). Sequences derived from vasculotide can be fused to the C-terminus of the light chain of an antibody specific for HPTP-β (VE-PTP) to provide a tetravalent bispecific antibody ( Figure 3 ). Sequences derived from vasculotide can be fused to the C-terminus of both the light and heavy chains of an antibody specific for HPTP-β (VE-PTP) to provide a hexavalent bispecific antibody ( Figure 4 ).

[0136] Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP) can be combined with antigen-binding compounds specific for, for example, RTK to provide multivalent multispecific compounds that can bind both HPTP-β (VE-PTP) and RTK. For example, antigen-binding compounds specific for HPTP-β (VE-PTP) can be combined with antigen-binding compounds specific for VEGFR (VEGFR2) to provide multivalent multispecific compounds that can bind both HPTP-β (VE-PTP) and VEGFR.

[0137] Compounds that bind both HPTP-β (VE-PTP) and VEGFR can inhibit HPTP-β (VE-PTP), activate Tie2, inhibit the binding of VEGF to VEGFR, and inhibit VEGFR signaling.

[0138] Sequences derived from ramucirumab (a humanized monoclonal antibody that binds the extracellular domain of VEGFR2 and inhibits VEGFR2 signaling) can be combined with antigen-binding compounds specific for HPTP-β (VE-PTP). For example, sequences from ramucirumab can be cloned into an scFv, and the ramucirumab-derived scFv can be fused to the C-terminus of the heavy chain of an antibody specific for HPTP-β (VE-PTP) to provide a tetravalent bispecific antibody ( Figure 2 ). The ramucirumab-derived scFv can be fused to the C-terminus of the light chain of an antibody specific for HPTP-β (VE-PTP) to provide a tetravalent bispecific antibody ( Figure 3 ). The ramucirumab-derived scFv can be fused to the C-terminus of both the light and heavy chains of an antibody specific for HPTP-β (VE-PTP) to provide a hexavalent bispecific antibody ( Figure 4 ).

[0139] Multiple compounds that enhance Tie2 activation, enhance Tie2 phosphorylation, enhance Tie2 signaling, reduce VEGFR activation, reduce VEGFR phosphorylation, reduce VEGFR signaling, or combinations thereof can be combined with an antigen-binding compound specific for HPTP-β (VE-PTP). For example, one binding domain (e.g., a brolucizumab-derived sequence) can be fused to the C-terminus of the heavy chain of an anti-HPTP-β (VE-PTP) antibody, while another binding domain (e.g., a vasculotide-derived sequence) can be fused to the C-terminus of the light chain of an HPTP-β (VE-PTP)-specific antibody to provide a hexavalent trispecific antibody ( Figure 5 ).

[0140] The compounds, antibodies, or derivatives thereof disclosed herein can have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more domains. Each domain can modulate, bind to, antagonize, inhibit, or activate any target disclosed herein, such as a phosphatase, a phosphatase that regulates Tie2 signaling, a protein tyrosine phosphatase, a receptor-like protein tyrosine phosphatase, a Tie2 modulator, HPTP-β (VE-PTP), the extracellular domain of HPTP-β (VE-PTP), the first FN3 repeat of the extracellular domain of HPTP-β (VE-PTP), a kinase, a tyrosine kinase, a receptor tyrosine kinase, a receptor tyrosine kinase activator, a receptor tyrosine kinase agonist, a growth factor, a growth factor receptor activator, a growth factor receptor agonist, a cysteine knot growth factor superfamily member, an angiogenic factor, a PDGF family member, a VEGF receptor, a VEGF receptor agonist, a VEGF family member, VEGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, PGF, VEGF 121 , VEGF 145 , VEGF 148 , VEGF 162 , VEGF 165 , VEGF 165b , VEGF 183 , VEGF 189 or VEGF 206 .

[0141] Inhibitors, activators, modulators, and binding agents

[0142] The HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may include compounds, recombinant proteins, antibodies, antigen-binding fragments, variants or derivatives thereof, Tie2-mimetic peptides, tetraethylene glycol-clustered peptides, collagen IV biomimetic peptides, DARPins or derivatives thereof, affimers, avimers, adnectins, anticalins, Fynom ers, Kunitz domains, knottins, β-hairpin mimetics or peptides derived from one or more receptors, which are either alone or in combination with another amino acid sequence or multiple other amino acid sequences. The inhibitors, modulators or binders may be subject to modifications such as enzymatic cleavage or post-translational modifications.

[0143] In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may inhibit HPTP-β (VE-PTP) by interfering with the interaction between HPTP-β (VE-PTP) and Tie2. In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may inhibit HPTP-β (VE-PTP) by stabilizing the inactive conformation of HPTP-β (VE-PTP). In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may inhibit HPTP-β (VE-PTP) by promoting the internalization of HPTP-β (VE-PTP) (e.g., promoting endocytosis and degradation of HPTP-β (VE-PTP)). In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may inhibit HPTP-β (VE-PTP) by blocking the binding of ligands that activate HPTP-β (VE-PTP). In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may inhibit HPTP-β (VE-PTP) by regulating the oligomerization of HPTP-β (VE-PTP).

[0144] In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may bind to a dominant-negative isoform of HPTP-β (VE-PTP). The dominant-negative isoform may correspond to a form of HPTP-β (VE-PTP) lacking phosphatase activity, which may compete with endogenous HPTP-β (VE-PTP).

[0145] The HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may comprise multiple HPTP-β (VE-PTP) binding sites. In some embodiments, the HPTP-β (VE-PTP) inhibitor, modulator or binder may bind to two HPTP-β (VE-PTP) molecules simultaneously, bringing the two HPTP-β (VE-PTP) molecules into close proximity. In some embodiments, the HPTP-β (VE-PTP) inhibitor, modulator or binder may bind to three HPTP-β (VE-PTP) molecules simultaneously, bringing the three HPTP-β (VE-PTP) molecules into close proximity. In some embodiments, the HPTP-β (VE-PTP) inhibitor, modulator or binder may bind to four HPTP-β (VE-PTP) molecules simultaneously, bringing the four HPTP-β (VE-PTP) molecules into close proximity.

[0146] The HPTP-β (VE-PTP) inhibitors, modulators or binders of the present disclosure may be conjugated covalently or non-covalently to another moiety or vehicle. The moiety or vehicle may, for example, provide binding specificity for additional targets, inhibit degradation, extend the half-life, increase absorption, reduce toxicity, reduce immunogenicity, and / or increase the biological activity of the inhibitor, modulator or binder. Non-limiting examples of moieties that may be conjugated to the inhibitor, modulator or binder include the Fc domain of an immunoglobulin, a peptide, a lipid, a carbohydrate, a dendrimer, an oligosaccharide, a cholesterol group such as a steroid, and polymers such as polyethylene glycol (PEG), polylysine or dextran.

[0147] The compounds of the present disclosure can be used to target HPTP-β (VE-PTP) to restore Tie2 activity. The HPTP-β (VE-PTP) inhibitor, modulator or binder can thus be a Tie2 activator. In some embodiments, the compounds of the present disclosure can initiate or inhibit signal transduction cascades downstream of HPTP-β (VE-PTP) or Tie2, e.g., Akt / PI3-K signaling, Rac1 signaling, MAPK / Ras signaling or NF-κB signaling.

[0148] Inhibition of HPTP-β (VE-PTP) can lead to vascular stabilization, which may be beneficial for the treatment of conditions characterized, for example, by vascular instability, angiogenesis, neovascularization, vascular leakage, and / or edema. For example, inhibition of HPTP-β (VE-PTP) may be beneficial for the treatment of vascular disorders, ocular disorders, cancer, kidney disorders, diabetic complications, and other conditions. In some embodiments, the HPTP-β (VE-PTP) inhibitors, modulators, or binders of the present disclosure can be used to treat, for example, diabetic retinopathy, non-proliferative diabetic retinopathy (NPDR), glaucoma, intraocular pressure, ocular edema, ocular hemorrhage, ocular hypertension, ocular inflammation, ocular neovascularization, ocular vascular leakage, retinal perfusion, or retinopathy.

[0149] The Tie2 activators, modulators, or binders of the present disclosure can include, for example, compounds, recombinant proteins, peptides, antibodies, antigen-binding fragments, variants, or derivatives thereof, angiopoietin 1 recombinant protein, Ang1 mimetics, Tie2 agonists, HPTP-β (VE-PTP) phosphatase inhibitors, Tie2 peptidomimetics, tetraethylene glycol-clustered peptides, collagen IV biomimetic peptides, DARPins or derivatives thereof, affibodies, avimers, adnectins, anticalins, Fynom ers, Kunitz domains, knottins, β-hairpin mimetics, or peptides derived from one or more receptors, which are either alone or in combination with another amino acid sequence or multiple other amino acid sequences. The activator, modulator, or binder can undergo modifications such as enzymatic cleavage or post-translational modifications.

[0150] The Tie2 activators, modulators, or binders of the present disclosure can be conjugated covalently or non-covalently to another moiety or vehicle. The moiety or vehicle can, for example, provide binding specificity to an additional target, inhibit degradation, extend the half-life, increase absorption, reduce toxicity, reduce immunogenicity, and / or increase the biological activity of the activator, modulator, or binder. Non-limiting examples of moieties that can be conjugated to the activator, modulator, or binder include the Fc domain of an immunoglobulin, peptides, lipids, carbohydrates, dendrimers, oligosaccharides, cholesterol groups such as steroids, and polymers such as polyethylene glycol (PEG), polylysine, or dextran.

[0151] In some embodiments, the compounds of the present disclosure can initiate or inhibit signal transduction cascades downstream of Tie2, for example, Akt / PI3-K signal transduction, Rac1 signal transduction, MAPK / Ras signal transduction, or NF-κB signal transduction.

[0152] Activation of Tie2 can lead to vascular stabilization, which may be beneficial for the treatment of conditions characterized by, for example, vascular instability, angiogenesis, neovascularization, vascular leakage, and / or edema. For example, activation of Tie2 may be beneficial for the treatment of ocular conditions, cancer, kidney conditions, diabetic complications, and other conditions. In some embodiments, the Tie2 activators, modulators, or binders of the present disclosure can be used to treat diabetic retinopathy, non-proliferative diabetic retinopathy (NPDR), glaucoma, intraocular pressure, ocular edema, ocular hemorrhage, ocular hypertension, ocular inflammation, ocular neovascularization, ocular vascular leakage, retinal perfusion, or retinopathy.

[0153] The VEGF inhibitors, modulators, or binders of the present disclosure can include compounds, recombinant proteins, peptides, antibodies, antigen-binding fragments, variants or derivatives thereof, DARPins or derivatives thereof, affimers, avimers, adnectins, anticalins, Fynom ers, Kunitz domains, knottins, β-hairpin mimetics, tetravalent polyethylene glycol-clustered peptides, collagen IV biomimetic peptides, or peptides derived from one or more receptors (e.g., VEGF receptors, or the VEGF-binding portions of human VEGF receptors 1 and 2), either alone or in combination with another amino acid sequence or multiple other amino acid sequences. The inhibitors, modulators, or binders can undergo modifications, such as enzymatic cleavage or post-translational modifications.

[0154] The VEGF inhibitors, modulators, or binders of the present disclosure can contain multiple VEGF-binding sites. In some embodiments, the VEGF inhibitor, modulator, or binder can bind to two VEGF molecules simultaneously, bringing the two VEGF molecules into close proximity. In some embodiments, the VEGF inhibitor, modulator, or binder can bind to three VEGF molecules simultaneously, bringing the three VEGF molecules into close proximity. In some embodiments, the VEGF inhibitor, modulator, or binder can bind to four VEGF molecules simultaneously, bringing the four VEGF molecules into close proximity.

[0155] The VEGF inhibitors, modulators, or binders of the present disclosure can be conjugated covalently or non-covalently to another moiety or vehicle. The moiety or vehicle can, for example, provide binding specificity for an additional target, inhibit degradation, extend the half-life, increase absorption, reduce toxicity, reduce immunogenicity, and / or increase the biological activity of the inhibitor, modulator, or binder. Non-limiting examples of moieties that can be conjugated to the inhibitor, modulator, or binder include the Fc domain of an immunoglobulin, a peptide, a lipid, a carbohydrate, a dendrimer, an oligosaccharide, a cholesterol moiety such as a steroid, and polymers such as polyethylene glycol (PEG), polylysine, or dextran.

[0156] The VEGFR inhibitors, modulators or binders of the present disclosure may include compounds, recombinant proteins, antibodies, antigen-binding fragments, variants or derivatives thereof, tetraethylene glycol clustered peptides, collagen IV biomimetic peptides, DARPins or derivatives thereof, affimers, avimers, adnectins, anticalins, Fynom ers, Kunitz domains, knottins, β-hairpin mimetics or peptides derived from one or more receptors, which are either alone or in combination with another amino acid sequence or multiple other amino acid sequences. The inhibitor, modulator or binder may undergo modifications such as enzymatic cleavage or post-translational modifications.

[0157] The VEGFR inhibitors, modulators or binders of the present disclosure may comprise multiple VEGFR binding sites. In some embodiments, the VEGFR inhibitor, modulator or binder may bind to two VEGFR molecules simultaneously, bringing the two VEGFR molecules into close proximity. In some embodiments, the VEGFR inhibitor, modulator or binder may bind to three VEGFR molecules simultaneously, bringing the three VEGFR molecules into close proximity. In some embodiments, the VEGFR inhibitor, modulator or binder may bind to four VEGFR molecules simultaneously, bringing the four VEGFR molecules into close proximity.

[0158] The VEGFR inhibitors, modulators or binders of the present disclosure may be conjugated covalently or non-covalently to another moiety or vehicle. The moiety or vehicle may, for example, provide binding specificity for an additional target, inhibit degradation, extend the half-life, increase absorption, reduce toxicity, reduce immunogenicity, and / or increase the biological activity of the inhibitor, modulator or binder. Non-limiting examples of moieties that may be conjugated to the inhibitor, modulator or binder include the Fc domain of an immunoglobulin, peptides, lipids, carbohydrates, dendrimers, oligosaccharides, cholesterol groups such as steroids, and polymers such as polyethylene glycol (PEG), polylysine or dextran.

[0159] In some embodiments, the VEGFR inhibitors, modulators or binders of the present disclosure can inhibit VEGFR by stabilizing the inactive conformation of VEGFR. In some embodiments, the VEGFR inhibitors, modulators or binders of the present disclosure can inhibit VEGFR by promoting the internalization of VEGFR (e.g., promoting the endocytosis and degradation of HVEGFR). In some embodiments, the VEGFR inhibitors, modulators or binders of the present disclosure can inhibit VEGFR by blocking the binding of ligands that activate VEGFR (e.g., blocking the VEGF ligation of VEGFR). In some embodiments, the VEGFR inhibitors, modulators or binders of the present disclosure can inhibit VEGFR by modulating the oligomerization of VEGFR (e.g., preventing or reducing the likelihood of dimerization or oligomerization of VEGFR).

[0160] The compounds of the present disclosure can be used to interfere with the interaction between VEGF and VEGFR, thereby reducing the phosphorylation and downstream signaling of VEGFR. In some embodiments, the inhibition of VEGF can reduce abnormal angiogenesis, vasculogenesis or vascular permeability, thereby reducing pathological vascular instability. In some embodiments, the inhibition of VEGF may be beneficial for the treatment of conditions characterized by vascular instability, angiogenesis, neovascularization, vascular leakage and / or edema. For example, the inhibition of VEGF may be beneficial for the treatment of vascular disorders, ocular disorders, cancer, kidney disorders, diabetic complications and other disorders. In some embodiments, the VEGF inhibitors, modulators or binders of the present disclosure can be used to treat diabetic retinopathy, non-proliferative diabetic retinopathy (NPDR), glaucoma, intraocular pressure, ocular edema, ocular hemorrhage, ocular hypertension, ocular inflammation, ocular neovascularization, ocular vascular leakage, retinal perfusion or retinopathy. In some embodiments, the inhibition of VEGF can alleviate cancer.

[0161] Method

[0162] The promotion of Tie2 signaling and the inhibition of VEGFR signaling can lead to vascular stability, which may be beneficial for the treatment of conditions with a vascular component. The compounds disclosed herein can be used to treat, for example, diseases characterized by vascular system alterations, diseases characterized by reduced Tie2 activation, or diseases characterized by VEGF involvement in the pathogenesis, whether progressive or non-progressive, acute or chronic.

[0163] In some embodiments, the compounds disclosed herein can be used to treat ocular disorders. The compounds disclosed herein can be used to treat, for example, age-related macular degeneration (dry), age-related macular degeneration (wet), atopic keratitis, Bests disease, blepharitis, blurred vision, choroidal neovascularization, chronic retinal detachment, chronic uveitis / vitreitis, chorioretinitis, conjunctivitis, corneal contact lens overwear, corneal graft neovascularization, corneal graft rejection, corneal neovascularization, cystoid macular edema, diabetic macular edema, diplopia, diabetic retinopathy, diseases associated with redness (neovascularization of the cornea), diseases caused by abnormal proliferation of fibrovascular or fibrous tissue, including all forms of proliferative vitreoretinopathy, Eales disease, elevated intraocular pressure, epidemic keratoconjunctivitis, floaters, glaucoma, yellow hard exudates within 500 μm of the center of the fovea with adjacent retinal thickening, hyperviscosity syndrome, infections causing chorioretinitis, infections causing retinitis, iris neovascularization, ischemic retinopathy, decreased contrast, macular telangiectasia, mariginal keratolysis, multifocal chorioretinitis, myopia, neovascular glaucoma, non-proliferative diabetic retinopathy (NPDR), ocular edema, ocular hemorrhage, ocular histoplasmosis, ocular hypertension, ocular inflammation, ocular ischemia, ocular neovascularization, ocular trauma, ocular vascular leakage, fovea, optic disc edema, pars planitis, phylectenulosis, polypoidal choroidal vasculopathy, post-laser complications, proliferative diabetic retinopathy, pterygium keratitis, radial keratotomy, retinal angiomatous proliferation, retinal degeneration, retinal edema (including macular edema), retinal neovascularization, retinal perfusion, retinal thickening within 1 disc diameter of the center of the fovea, retinal thickening within 500 μm of the center of the fovea, retinal vein occlusion (central or branch), retinitis, retinitis pigmentosa, retinopathy, retinopathy of prematurity, scleritis, Stargarts disease, superior limbic keratitis, surgically induced edema, surgically induced neovascularization, Terrien marginal degeneration, trachoma, trauma, uveitis, vasculitis (such as central retinal vein occlusion) or other ophthalmic diseases in which the ocular disease or disorder is associated with ocular neovascularization, vascular leakage or retinal edema or a combination thereof.

[0164] In some embodiments, the compounds disclosed herein can be used to treat diabetic complications (e.g., comorbidities of diabetes). The compounds disclosed herein can be used to treat, for example, acute glomerulonephritis, acute myocardial infarction, amputation, amyotrophy, aneurysm, angina, aortic aneurysm, aortic dissection, atherosclerosis, atherosclerotic cardiovascular disease, atrial fibrillation, autonomic neuropathy, blindness, cardiovascular complications of diabetes, cerebrovascular complications of diabetes, Charcot joint disease, chronic glomerulonephritis, chronic renal failure, claudication, clinically significant macular edema, coronary artery disease, cranial nerve palsy, cystoid macular degeneration, cystoid macular edema, diabetic cardiomyopathy, diabetic hand joint disease, diabetic coma, diabetic encephalopathy, diabetic foot ulcer, diabetic hyperglycemia, diabetic hyperlipidemia, diabetic hyperosmolar syndrome, diabetic hypoglycemia, diabetic ketoacidosis, diabetic myonecrosis, diabetic nephropathy, diabetic neuropathy, diabetic ophthalmic diseases, diabetic peripheral vascular disease, diabetic retinopathy, diffuse idiopathic skeletal hyperostosis, Dupuytren's contracture, embolism, end-stage renal disease, erectile dysfunction, Forestier's disease, gangrene, gas gangrene, gastroparesis / diarrhea, heart failure, hyperglycemic crisis, hypertension, ischemic heart disease, ketoacidosis, lipohypertrophy, metabolic complications of diabetes, mononeuropathy, myocardial infarction, nephritis, nephropathy, nephrosis, nephrotic syndrome, neurogenic bladder, neuropathic arthropathy, neuropathy, orthostatic hypotension, osteoarthritis, osteoporosis, periodontal disease, peripheral vascular disease, polyneuropathy, proliferative retinopathy, renal failure, renal insufficiency, restrictive lung disease, retinal detachment, retinal edema, retinopathy, stroke, thrombosis, transient ischemic attack, ulceration, ventricular fibrillation, vitreous hemorrhage, or combinations thereof.

[0165] In some embodiments, the compounds disclosed herein can be used to treat kidney disorders. The compounds disclosed herein can be used to treat, for example, acute kidney injury, acute proliferative glomerulonephritis, adenine phosphoribosyltransferase deficiency, Alport syndrome, analgesic nephropathy, autosomal dominant polycystic kidney disease, autosomal recessive polycystic kidney disease, Balkan endemic nephropathy, benign nephrosclerosis, Bright's disease, cardiorenal syndrome, CFHR5 nephropathy, chronic kidney disease, chronic kidney disease - mineral and bone disorder, congenital nephrotic syndrome, conorenal syndrome, contrast-induced nephropathy, cystic kidney disease, Dent's disease, diabetic nephropathy, diffuse proliferative glomerulonephritis, distal renal tubular acidosis, diuresis, EAST syndrome, end-stage renal disease, epithelial-mesenchymal transition, Epstein syndrome, Fanconi syndrome, Fechtner syndrome, focal proliferative glomerulonephritis, focal segmental glomerulosclerosis, Fraley syndrome, Galloway Mowat syndrome, Gitelman syndrome, glomerular cystic kidney disease, glomerulopathy, Goodpasture syndrome, high anion gap metabolic acidosis, HIV-associated nephropathy, horseshoe kidney, hydronephrosis, hypertensive nephropathy, IgA nephropathy, interstitial nephritis, juvenile nephropathy, kidney cancer, nephropathy, nephrolithiasis, Lightwood–Albright syndrome, lupus nephritis, malaria nephropathy, medullary cystic kidney disease, medullary sponge kidney, membranous glomerulonephritis, Mesoamerican nephropathy, milk-alkali syndrome, minimal membranous glomerulonephritis, multicystic dysplastic kidney, nephritis, nephrocalcinosis, nephrogenic diabetes insipidus, renal hypertrophy, nephroptosis, nephrosis, nephrotic syndrome, nutcracker syndrome, papillary renal syndrome, phosphatic nephropathy, polycystic kidney disease, primary hyperoxaluria, proximal renal tubular acidosis, pyelonephritis, pyonephrosis, rapidly progressive glomerulonephritis, renal hypoplasia, renal colic, renal artery stenosis, renal cysts, renal ischemia, renal osteodystrophy, renal papillary necrosis, renal tubular acidosis, renal vein thrombosis, secondary hypertension, snake-bite fibular-polydactyly polycystic kidney syndrome, shunt nephritis, sickle cell nephropathy, thin basement membrane disease, transplant glomerulopathy, tubulointerstitial nephritis and uveitis, tubulopathy, uremia, uremic frost, Wunderlich syndrome, or a combination thereof.

[0166] In some embodiments, the compounds disclosed herein can be used to treat cancer. The compounds disclosed herein can be used to treat, for example, acute leukemia, astrocytoma, biliary tract cancer (cholangiocarcinoma), bone cancer, breast cancer, brainstem glioma, bronchioloalveolar carcinoma, adrenal cancer, anal area cancer, bladder cancer, endocrine system cancer, esophageal cancer, head or neck cancer, kidney cancer, parathyroid cancer, penile cancer, pleural / peritoneal cancer, salivary gland cancer, small intestine cancer, thyroid cancer, ureteral cancer, urethral cancer, cervical cancer, endometrial cancer, fallopian tube cancer, renal pelvis cancer, renal pelvis cancer, vaginal cancer, vulvar cancer, cervical cancer, chronic leukemia, colon cancer, colorectal cancer, cutaneous melanoma, ependymoma, epidermoid tumor, Ewing sarcoma, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, hematologic malignancies, hepatocellular (liver) cancer, hepatoma, Hodgkin's disease, intraocular melanoma, Kaposi sarcoma, lung cancer, lymphoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, muscle cancer, central nervous system tumors (CNS), neuronal cancer, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, pediatric malignancies, pituitary adenoma, prostate cancer, rectal cancer, renal cell cancer, soft tissue sarcoma, schwannoma, skin cancer, spinal axis tumors, squamous cell carcinoma, stomach cancer, synovial sarcoma, testicular cancer, uterine cancer or tumors and their metastases, including refractory forms of any of the above cancers, or combinations thereof.

[0167] In some embodiments, the compounds disclosed herein can be used to treat diseases characterized by alterations in the vasculature, diseases characterized by reduced Tie2 activation, or diseases characterized by VEGF involvement in pathogenesis. The compounds disclosed herein can be used to treat, for example, acne rosacea, acute lung injury, acute respiratory distress syndrome (ARDS), adhesion formation following abdominal surgery, obesity, albuminuria, allergic edema, allergy, angina, angiofibroma, arteriosclerosis, arterial occlusion, ascites, atheroma, atherosclerosis, asthma, avascular necrosis, bacterial ulcer, Bartonella bacilliformis infection, Behcet's disease, Buerger's disease (thromboangiitis obliterans), cardiac fibrosis, cardiac hypertrophy, cardiomyopathy, carotid obstructive disease, cerebral infarction, chemical burn, COPD, Crohn's disease, cytokine-induced vascular leakage, blood flow instability, diabetes (including non-insulin-dependent diabetes), dysfunctional uterine bleeding, endometriosis, Epstein-Barr virus infection, erectile dysfunction, hirsutism, follicular cyst, foot ulcer (e.g., diabetic foot ulcer), fungal ulcer, giant cell arteritis, glomerulosclerosis, Graves' disease, Hashimoto's autoimmune thyroiditis, hemangioma, hemangioendothelioma, hemophilic joint, hemorrhage, hepatitis C, hereditary hemorrhagic telangiectasia (HHT), herpes simplex infection, herpes zoster infection, hypertension, idiopathic thrombocytopenic purpura, poor wound healing, inflammatory and infectious processes (e.g., hepatitis, pneumonia, glomerulonephritis), interstitial fibrosis, ischemia, nephropathy, leishmaniasis, leukomalacia, lipidosis, liver regeneration, lupus nephritis, Lyme disease, lymphoproliferative disorders, malaria (Plasmodium infection), Mooren's ulcer, multiple sclerosis, mycobacterial infection, myocardial infarction, nasal polyps, nephropathy, neuronal inflammation, neuropathy, obesity, osteomyelitis, osteophyte, ovarian hyperstimulation, Paget's disease, pannus growth, peripheral arterial disease, peritoneal sclerosis, pemphigoid, polyarteritis, protozoal infection, pseudoxanthoma elasticum, psoriasis, pulmonary hypertension, pyogenic granuloma, renal fibrosis, respiratory distress, rheumatoid arthritis, rickettsial infection, keloid, sepsis, sickle cell anemia, Stevens-Johnson disease, stroke, synovitis, systemic lupus erythematosus, syphilis, goiter, thyroiditis, toxic shock syndrome, toxoplasmosis, trauma, ulcerative colitis, vascular leakage, vascular leak syndrome, vascular malformation (e.g., Osler-Weber syndrome), venous occlusion, viral hemorrhagic fever (e.g., dengue fever), vitamin A deficiency, warts, or Wegener's granulomatosis, or combinations thereof.

[0168] Sequence

[0169] As used herein, the abbreviations for L- and D-enantiomer amino acids are as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly); histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Val). In some embodiments, the amino acid is the L-enantiomer. In some embodiments, the amino acid is the D-enantiomer.

[0170] Table 2 shows the amino acid sequences of the humanized V H antibody regions that bind to HPTP-β (VE-PTP). SEQ ID NO:1 is V H1 , SEQ ID NO:2 is V H2 , SEQ ID NO:3 is V H3 , and SEQ ID NO:4 is V H4 .

[0171]

[0172] Table 3 shows the amino acid sequences of the humanized V L antibody regions that bind to HPTP-β (VE-PTP). SEQ ID NO:5 is V L1 , SEQ ID NO:6 is V L2 , SEQ ID NO:7 is V L3 , and SEQ ID NO:8 is V L4 .

[0173]

[0174] Each V H domain can be synthesized in-frame with a constant domain sequence such as a human IgG1, IgG2, IgG3, IgG4, IgE, IgA1, IgA2, IgM, or IgD sequence. The DNA sequence encoding the entire heavy chain sequence can be codon-optimized and verified.

[0175] Table 4 provides exemplary amino acid sequences of constant domain sequences. In some embodiments, the V HThe domain is synthesized in-frame with the human IgG1 constant domain sequence. The human IgG1 constant domain sequence may comprise SEQ ID NO:220. In some embodiments, the V H The domain is synthesized in-frame with the human IgG2 constant domain sequence. The human IgG2 constant domain sequence may comprise SEQ ID NO:221. In some embodiments, the V H The domain is synthesized in-frame with the human IgG3 constant domain sequence. The human IgG3 constant domain sequence may comprise SEQ ID NO:222. In some embodiments, the V H The domain is synthesized in-frame with the human IgG4 constant domain sequence. The human IgG4 isotype constant domain sequence can be mutated to proline instead of serine at position 228 to reduce Fab-arm exchange (stabilize the S228P mutation). The amino acid sequence of the IgG4 constant domain with the S228P mutation may comprise SEQ ID NO:9. In some embodiments, the V H The domain is synthesized in-frame with the human IgE constant domain sequence. The human IgE constant domain sequence may comprise SEQ ID NO:223. In some embodiments, the V H The domain is synthesized in-frame with the human IgA1 constant domain sequence. The human IgA1 constant domain sequence may comprise SEQ ID NO:224. In some embodiments, the V H The domain is synthesized in-frame with the human IgA2 constant domain sequence. The human IgA2 constant domain sequence may comprise SEQ ID NO:225. In some embodiments, the V H The domain is synthesized in-frame with the human IgM constant domain sequence. The human IgM constant domain sequence may comprise SEQ ID NO:226. In some embodiments, the V H The domain is synthesized in-frame with the human IgD constant domain sequence. The human IgD constant domain sequence may comprise SEQ ID NO:227.

[0176]

[0177]

[0178] Each V L The domain can be synthesized in-frame with a human light chain constant domain sequence, such as a kappa (IgK) or lambda (IgL) chain. The entire light chain sequence can then be codon-optimized and the DNA sequence can be verified. Table 5 provides exemplary light chain constant domain sequences.

[0179] In some embodiments, the V LThe domain is synthesized in-frame with the human IgK constant domain sequence. The human IgK constant domain sequence can include SEQ ID NO:10. In some embodiments, the V disclosed herein L The domain is synthesized in-frame with the human IgL constant domain sequence. The human IgL constant domain sequence can include SEQ ID NO:228.

[0180]

[0181] The signal peptide can result in higher protein expression and / or secretion in cells. The following signal peptides can be appended to all constructs disclosed herein.

[0182] Heavy chain signal peptide (SEQ ID NO:11): MGWTLVFLFLLSVTAGVHS

[0183] Light chain signal peptide (SEQ ID NO:12): MVSSAQFLGLLLLCFQGTRC

[0184] Signal peptidase can cleave the signal peptide from the protein, for example, during secretion, thereby generating a mature protein that does not contain the signal peptide sequence. In some embodiments, the signal peptide is excised from the compounds or antibodies of the present disclosure. In some embodiments, the mature compounds or antibodies of the present disclosure do not contain a signal peptide.

[0185] Tables 6 and 7 list the complete amino acid sequences of the humanized heavy and light chains that bind to HPTP-β (VE-PTP), respectively. HC1, HC2, HC3, and HC4 are the human IgG4 isotype constant domains conjugated to V H1 , V H2 , V H3 and V H4 respectively, with a stabilizing S228P mutation and appended with a signal peptide. Amino acids 1-19 of SEQ ID NO:13, 14, 15, and 16 are the heavy chain signal peptide (SEQ ID NO:11). HC1, HC2, HC3, and HC4 without the appended signal peptide (SEQ ID NO:246, 247, 248, and 249) are also shown. In some embodiments, the mature compounds or antibodies of the present disclosure do not contain a signal peptide.

[0186] LC1, LC2, LC3, and LC4 are the human IgL constant domains conjugated to V L1 , V L2 , V L3 and V L4Joined human IgK isotype constant domains, with a signal peptide appended. Amino acids 1-20 of SEQ ID NO:17, 18, 19, and 20 are the light chain signal peptide (SEQ ID NO:12). LC1, LC2, LC3, and LC4 (SEQ ID NO:250, 251, 252, and 253) without the appended signal peptide are also shown. In some embodiments, the mature compounds or antibodies of the present disclosure do not contain a signal peptide.

[0187]

[0188]

[0189]

[0190]

[0191]

[0192] Any V H region disclosed herein can be combined with any V L region disclosed herein, with or without additional sequences appended, to produce a compound that binds to anti-HPTP-β (VE-PTP). For example, the signal peptide and constant region sequences can be appended to the V H and V L regions shown in Tables 6 and 7, respectively, and the resulting heavy and light chains can be paired in any combination to form an antibody. Table 8 shows 16 possible pairings of HC1-4 and LC1-4 that bind to HPTP-β (VE-PTP). Transfection and expression of each anti-HPTP-β (VE-PTP) antibody in Table 8 can be explored.

[0193]

[0194]

[0195] An antibody or antigen-binding compound that is specific for HPTP-β (VE-PTP) can be combined with an antibody or compound that activates Tie2, inhibits VEGF, or inhibits VEGFR to form a multispecific compound.

[0196] Table 9 provides the sequences of collagen IV-derived biomimetic peptides. SEQ ID NO:152 is AXT-107, a collagen IV-derived biomimetic peptide that targets integrin and can inhibit VEGFR phosphorylation / activation / signal transduction and promote Tie2 phosphorylation / activation / signal transduction. SEQ ID NO:153 provides a consensus sequence, where X is any standard amino acid or non-genetically encoded amino acid. In some embodiments, X at position 7 is M, A, or G; X at position 9 is F, A, Y, or G; X at position 0 is M, A, G, D-alanine (dA), or norleucine (Nle); X at position 11 is F, A, Y, G, or 4-chlorophenylalanine (4-CiPhe); X at positions 12 and 18 are independently selected from 2-aminobutyric acid (Abu), G, S, A, V, T, I, L, or allylglycine (AllylGly).

[0197]

[0198] Table 10 provides sequences related to vasculotide, a synthetic Ang1 mimetic peptide that can act as a Tie2 agonist and activate Tie2 signal transduction. SEQ ID NO:229 provides the sequence of a synthetic heptamer that binds to the Tie2 receptor. SEQ ID NO:230 provides an octamer with a cysteine residue added at the N-terminus, allowing covalent tethering to a polyethylene glycol backbone to generate a tetrameric polyethylene oxide cluster form of the peptide.

[0199]

[0200] An antibody or antigen-binding compound specific for HPTP-β (VE-PTP) can be combined with an antibody or compound specific for VEGF or VEGFR to form a multispecific compound that binds HPTP-β (VE-PTP) and VEGF or VEGFR. Tables 11, 12, 13, 14, 15, and 17 provide example sequences of compounds specific for VEGF. Table 16 provides the sequences of antibodies that bind to VEGFR.

[0201] Table 11 provides sequences related to aflibercept, a recombinant protein that contains the VEGF-binding portions of human VEGF receptors 1 and 2 fused to the Fc portion of human IgG1. SEQ ID NO:21 is the complete amino acid sequence of aflibercept. SEQ ID NO:22 is a shortened sequence that contains the VEGF-binding portions of human VEGF receptors 1 and 2 and does not contain the Fc portion of IgG.

[0202]

[0203] Table 12 provides the sequence of brolucizumab (SEQ ID NO:23), a humanized single-chain antibody fragment (scFv) inhibitor of VEGF that binds to the receptor-binding site of VEGF, thereby interfering with the interaction of VEGF with VEGFR1 and VEGFR2.

[0204]

[0205] Table 13 provides the sequences related to ranibizumab, a humanized monoclonal antibody fragment (Fab) that binds to VEGF and inhibits its activity. SEQ ID NO:24 is the heavy-chain sequence of ranibizumab. SEQ ID NO:25 is the light-chain sequence of ranibizumab. SEQ ID NO:26 is a shortened sequence of the heavy chain of ranibizumab, which can be used to clone a single-chain antibody fragment (scFv). SEQ ID NO:27 is a shortened sequence of the light chain of ranibizumab, which can be used to clone a single-chain antibody fragment (scFv). SEQ ID NO:28 is a single-chain antibody fragment (scFv) containing SEQ ID NO:26, SEQ ID NO:27, and SEQ ID NO:33 (linker peptide, underlined).

[0206]

[0207] Table 14 provides the sequences related to bevacizumab, a humanized monoclonal antibody that binds to VEGF and inhibits its activity. SEQ ID NO:29 is the heavy-chain sequence of bevacizumab. SEQ ID NO:30 is the light-chain sequence of bevacizumab.

[0208]

[0209]

[0210] Table 15 provides the sequences related to conbercept, a recombinant protein that contains the extracellular domains of VEGF receptors 1 and 2 fused to the Fc portion of human IgG1. SEQ ID NO:154 is the complete amino acid sequence of conbercept. SEQ ID NO:155 is a shortened sequence containing the sequences of VEGF receptors 1 and 2 that does not contain the Fc portion of IgG.

[0211]

[0212] Table 16 provides sequences related to ramucirumab, a humanized monoclonal antibody that binds to the extracellular domain of VEGFR2 and inhibits VEGFR2 signaling. SEQ ID NO:156 is the heavy chain sequence of ramucirumab. SEQ ID NO:157 is the light chain sequence of ramucirumab.

[0213]

[0214] Table 17 provides DARPins that bind to VEGF and inhibit VEGFR signaling and

[0215] DARPin-derived amino acid sequences. SEQ ID NO:158 - 168 contain engineered ankyrin repeats with binding specificity for VEGF. SEQ ID NO:169 contains engineered ankyrin repeats with binding specificity for VEGF and engineered ankyrin repeats with binding specificity for serum albumin. SEQ ID NO:170 contains engineered ankyrin repeats with binding specificity for VEGF, engineered ankyrin repeats with binding specificity for hepatocyte growth factor, and engineered ankyrin repeats with binding specificity for serum albumin. SEQ ID NO:171 - 177 contain engineered ankyrin repeats with binding specificity for VEGF. SEQ ID NO:173 provides the sequence of abicipar, which contains engineered ankyrin repeats with binding specificity for VEGF. SEQ ID NO:178 - 190 provide individual engineered ankyrin repeat sequence motifs with binding specificity for VEGF, where X represents any amino acid. SEQ ID NO:191 - 217 contain engineered ankyrin repeats with binding specificity for VEGF.

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222] Antibodies or antigen-binding compounds specific for VEGF can be combined with antibodies or antigen-binding compounds specific for HPTP-β (VE-PTP) to form multispecific compounds, such as bispecific compounds that bind VEGF and HPTP-β (VE-PTP). Any compound specific for VEGF in the present disclosure can be combined with any compound specific for HPTP-β (VE-PTP) in the present disclosure. Any compound specific for VEGF or HPTP-β (VE-PTP) in the present disclosure can be modified, if necessary, to generate a multispecific compound. Non-limiting examples of modifications necessary to generate a multispecific compound include adding amino acid residues, removing amino acid residues, replacing amino acid residues, and using linkers. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more residues can be added to the N-terminus and / or C-terminus of the sequences disclosed herein, and the resulting sequences can be used to generate multispecific constructs. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more residues can be removed from the N-terminus and / or C-terminus of the sequences disclosed herein, and the remaining sequences can be used to generate multispecific constructs. For example, N- and / or C-terminal residues can be removed from SEQ ID NO:173 (e.g., to generate SEQ ID NO:244), and the truncated sequence can be used in a multispecific construct. In some embodiments, the sequences in any one of SEQ ID NOs: 13-20 or 246-253 can be modified. For example, one or more C-terminal residues can be removed (e.g., the C-terminal lysine can be removed from any one of SEQ ID NOs: 13-16 or 246-249, and the remaining residues (residues 1-467) can be used in a multispecific construct).

[0223] The compounds described herein can include linkers between different domains of the compound. The linker can be a chemical bond, such as a covalent or non-covalent bond. The linkers described herein can include flexible or rigid linkers.

[0224] The linker of the present disclosure may include a chemical linker. For example, two amino acid sequences of the present disclosure may be linked together by a chemical linker. Each chemical linker of the present disclosure may be an alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene or heteroarylene, any of which is optionally substituted. In some embodiments, the chemical linker of the present disclosure may be an ester, ether, amide, thioether or polyethylene glycol (PEG). In some embodiments, the linker may, for example, reverse the order of the amino acid sequences in a compound such that the amino acid sequences linked by the linker are head-to-head rather than head-to-tail. Non-limiting examples of such linkers include diesters of dicarboxylic acids such as oxalate diesters, malonate diesters, succinate diesters, glutarate diesters, adipate diesters, pimelate (pimetyl) diesters, fumarate diesters, maleate diesters, phthalate diesters, isophthalate diesters and terephthalate diesters. Non-limiting examples of such linkers include diamides of dicarboxylic acids such as oxalamide, malonamide, succinamide, glutaramide, adipamide, pimelamide (pimetylamide), fumaramide, maleamide, phthalamide, isophthalamide and terephthalamide. Non-limiting examples of such linkers include diamides of diamino linkers such as ethylenediamine, 1,2-bis(methylamino)ethane, 1,3-diaminopropane, 1,3-bis(methylamino)propane, 1,4-bis(methylamino)butane, 1,5-bis(methylamino)pentane, 1,6-bis(methylamino)hexane and piperazine.

[0225] Non-limiting examples of optional substituents include hydroxy groups, mercapto groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, carboxaldehyde groups, imine groups, alkyl groups, haloalkyl groups, alkenyl groups, haloalkenyl groups, alkynyl groups, haloalkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, aralkyloxy groups, heterocyclic groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ureido groups, epoxy groups and ester groups.

[0226] The linker can be a peptide. The linker may comprise a linker sequence, such as a linker peptide sequence. The length of the linker sequence can be, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 51, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70 amino acid residues.

[0227] The flexible linker may have a sequence comprising a stretch of glycine and serine residues. The small size of glycine and serine residues provides flexibility and allows mobility of the linked functional domains. Incorporation of serine or threonine can maintain the stability of the linker in aqueous solution by forming hydrogen bonds with water molecules, thereby reducing unfavorable interactions between the linker and the protein moiety.

[0228] The flexible linker may also comprise additional amino acids, such as threonine and alanine, to maintain flexibility, and polar amino acids, such as lysine and glutamine, to improve solubility.

[0229] The flexible linker may comprise repeats of SEQ ID NO:42 (GGGS), for example, SEQ ID NO:42 - 55. The flexible linker may comprise repeats of SEQ ID NO:31 (GGGGS), for example, SEQ ID NO:31 - 41. Several other types of flexible linkers can also be used, including SEQ ID NO:59 (KESGSVSSEQLAQFRSLD) and SEQ ID NO:60 (EGKSSGSGSESKST). The SEQ ID NO:61 (GSAGSAAGSGEF) linker can also be used, in which large hydrophobic residues are minimized to maintain good solubility in aqueous solution. The length of the flexible linker can be adjusted to allow proper folding or achieve optimal bioactivity of the fusion protein.

[0230] The rigid connector may have, for example, an alpha helical structure. An alpha-helical rigid connector may serve as a spacer between protein domains. The rigid connector may comprise a repetition of SEQ ID NO:62 (EAAAK), for example, SEQ ID NO:62-66. The rigid connector may comprise a repetition of SEQ ID NO:67 (EAAAR), for example, SEQ ID NO:67-72. The rigid connector may have a proline-rich sequence (XP) n, where X represents alanine, lysine, glutamine or any amino acid. The presence of proline in a non-helical connector may increase rigidity and allow effective separation of protein domains.

[0231] The linker may comprise any of the sequences disclosed in Table 18, which may be used to link any portion of a compound disclosed herein to any portion of another compound disclosed herein:

[0232]

[0233]

[0234] The VEGF-binding compounds and VEGFR-binding compounds described in Table 11, Table 12, Table 13, Table 14, Table 15, Table 16 and Table 17 can be combined with the HPTP-β (VE-PTP)-binding compounds described in Table 2, Table 3, Table 6, Table 7 and Table 8.

[0235] Any of the 16 antibodies described in Table 8 can be combined with aflibercept or aflibercept-related sequences to generate bispecific antibodies. Non-limiting illustrative examples are provided in Figure 2 , Figure 3 and Figure 4 For example, SEQ ID NO: 22 can be appended to SEQ ID NO: 14 or SEQ ID NO: 247 of antibody HC2: LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2: LC1 is appended with SEQ ID NO: 22 ( Figure 2 ). SEQ ID NO:22 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody in which the light chain of HC2:LC1 is appended with SEQ ID NO:22 ( Figure 3 ). SEQ ID NO:22 can be appended to SEQ ID NO:14 or SEQ ID NO:247, and SEQ ID NO:22 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody in which the heavy and light chains of HC2:LC1 are appended with SEQ ID NO:22 (Figure 4 )

[0236] Any one of the 16 antibodies described in Table 8 can be combined with brolucizumab or a brolucizumab-related sequence to generate a bispecific antibody. Non-limiting illustrative examples are presented in Figure 2 、 Figure 3 and Figure 4 . For example, SEQ ID NO:23 can be appended to SEQ ID NO:14 or SEQ ID NO:247 of the antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with SEQ ID NO:23 ( Figure 2 ). SEQ ID NO:23 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of the antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with SEQ ID NO:23 ( Figure 3 ). SEQ ID NO:23 can be appended to SEQ ID NO:14 or SEQ ID NO:247, and SEQ ID NO:23 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of the antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy chain and light chain of HC2:LC1 are appended with SEQ ID NO:23 ( Figure 4 )

[0237] Any one of the 16 antibodies described in Table 8 can be combined with ranibizumab or a ranibizumab-related sequence to generate a bispecific antibody. Non-limiting illustrative examples are presented in Figure 2 、 Figure 3 and Figure 4 . For example, SEQ IDNO:28 can be appended to SEQ ID NO:14 or SEQ ID NO:247 of the antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with SEQ ID NO:28 ( Figure 2 ). SEQ ID NO:28 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of the antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with SEQ ID NO:28 ( Figure 3) SEQ ID NO:28 can be appended to SEQ ID NO:14 or SEQ ID NO:247, and SEQ ID NO:28 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy and light chains of HC2:LC1 are appended with SEQ ID NO:28( Figure 4 )。

[0238] Any one of the 16 antibodies described in Table 8 can be combined with bevacizumab or a bevacizumab-related sequence to generate a bispecific antibody. Non-limiting illustrative examples are presented in Figure 2 、 Figure 3 and Figure 4 For example, the antigen-binding scFv of bevacizumab can be generated as shown for ranibizumab in Table 13. The bevacizumab-derived antigen-binding scFv can be appended to SEQ ID NO:14 or SEQ ID NO:247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with the bevacizumab-derived antigen-binding scFv( Figure 2 )。The bevacizumab-derived antigen-binding scFv can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with the bevacizumab-derived antigen-binding scFv( Figure 3 )。The bevacizumab-derived antigen-binding scFv can be appended to SEQ ID NO:14 or SEQ ID NO:247, and the bevacizumab-derived antigen-binding scFv can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy and light chains of HC2:LC1 are appended with the bevacizumab-derived antigen-binding scFv( Figure 4 )。

[0239] Any one of the 16 antibodies described in Table 8 can be combined with abicipar or an abicipar-related sequence to generate a bispecific antibody. Non-limiting illustrative examples are presented in Figure 2 、 Figure 3 and Figure 4presented. For example, SEQ ID NO: 173 or SEQ ID NO: 244 can be appended to SEQ ID NO: 14, SEQ ID NO: 247, residues 1 - 467 of SEQ ID NO: 14 or residues 1 - 467 of SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with SEQ ID NO: 173 or SEQ ID NO: 244( Figure 2 ). SEQ ID NO: 173 or SEQ ID NO: 244 can be appended to SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with SEQ ID NO: 173 or SEQ ID NO: 244( Figure 3 ). SEQ ID NO: 173 or SEQ ID NO: 244 can be appended to SEQ ID NO: 14, SEQ ID NO: 247, residues 1 - 467 of SEQ ID NO: 14 or residues 1 - 467 of SEQ ID NO: 247, and SEQ ID NO: 173 or SEQ ID NO: 244 can be appended to SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy and light chains of HC2:LC1 are appended with SEQ ID NO: 173 or SEQ ID NO: 244( Figure 4 ).

[0240] Any one of the 16 antibodies described in Table 8 can be combined with Conbercept or a Conbercept - related sequence to generate a bispecific antibody. Non - limiting illustrative examples are presented in Figure 2 , Figure 3 and Figure 4 . For example, SEQ ID NO: 155 can be appended to SEQ ID NO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with SEQ ID NO: 155( Figure 2 ). SEQ ID NO: 155 can be appended to SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with SEQ ID NO: 155( Figure 3) SEQ ID NO:155 can be appended to SEQ ID NO:14 or SEQ ID NO:247, and SEQ ID NO:155 can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy and light chains of HC2:LC1 are appended with SEQ ID NO:155( Figure 4 )

[0241] Any of the 16 antibodies described in Table 8 can be combined with ramucirumab or a ramucirumab-related sequence to generate a bispecific antibody. Non-limiting illustrative examples are presented in Figure 2 、 Figure 3 and Figure 4 . For example, the antigen-binding scFv of ramucirumab can be generated as shown for ranibizumab in Table 13. The ramucirumab-derived antigen-binding scFv can be appended to SEQ ID NO:14 or SEQ ID NO:247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with the ramucirumab-derived antigen-binding scFv( Figure 2 ) Figure 3 ) The ramucirumab-derived antigen-binding scFv can be appended to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with the ramucirumab-derived antigen-binding scFv( Figure 4 )

[0242] Any of the 16 antibodies described in Table 8 can be combined with a DARPin, DARPin repeat, or its sequence to generate a multispecific antibody. Non-limiting illustrative examples are in Figure 2 、 Figure 3 and Figure 4presented. For example, an amino acid sequence comprising any one of SEQ ID NOs: 158 - 217 can be appended to SEQ ID NO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with an amino acid sequence comprising any one of SEQ ID NOs: 158 - 217( Figure 2 ). An amino acid sequence comprising any one of SEQ ID NOs: 158 - 217 can be appended to SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with an amino acid sequence comprising any one of SEQ ID NOs: 158 - 217( Figure 3 ). An amino acid sequence comprising any one of SEQ ID NOs: 158 - 217 can be appended to SEQ ID NO: 14 or SEQ ID NO: 247 and SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalent bispecific antibody, wherein the heavy and light chains of HC2:LC1 are appended with an amino acid sequence comprising any one of SEQ ID NOs: 158 - 217( Figure 4 ).

[0243] The Tie2 activating compounds described in Tables 9 and 10 can be combined with the HPTP-β (VE-PTP) binding compounds described in Tables 2, 3, 6, 7, and 8.

[0244] Any one of the 16 antibodies described in Table 8 can be combined with a collagen IV-derived biomimetic peptide sequence to generate a multispecific antibody. Non-limiting illustrative examples are presented in Figure 2 , Figure 3 and Figure 4 . For example, an amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153 can be appended to SEQ ID NO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is appended with an amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153( Figure 2 ). An amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153 can be appended to SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is appended with an amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153(Figure 3 )。 The amino acid sequences containing SEQ ID NO:152 or SEQ ID NO:153 can be attached to SEQ ID NO:14 or SEQ ID NO:247 of antibody HC2:LC1 and SEQ ID NO:17 or SEQ ID NO:250 to generate a hexavalent bispecific antibody, wherein the heavy chain and light chain of HC2:LC1 are attached with the amino acid sequence containing SEQ ID NO:152 or SEQ ID NO:153( Figure 4 )。

[0245] Any one of the 16 antibodies described in Table 8 can be combined with an Ang2 mimic or its sequence to generate a multispecific antibody. Non-limiting schematic examples are presented in Figure 2 、 Figure 3 and Figure 4 For example, the amino acid sequence containing any one of SEQ ID NO:229 - 230 can be attached to SEQ ID NO:14 or SEQ ID NO:247 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the heavy chain of HC2:LC1 is attached with the amino acid sequence containing any one of SEQ ID NO:229 - 230( Figure 2 )。 The amino acid sequence containing any one of SEQ ID NO:229 - 230 can be attached to SEQ ID NO:17 or SEQ ID NO:250 of antibody HC2:LC1 to generate a tetravalent bispecific antibody, wherein the light chain of HC2:LC1 is attached with the amino acid sequence containing any one of SEQ ID NO:229 - 230( Figure 3 )。 The amino acid sequence containing any one of SEQ IDNO:229 - 230 can be attached to SEQ ID NO:14 or SEQ ID NO:247 of antibody HC2:LC1 and SEQ ID NO:17 or SEQ ID NO:250 to generate a hexavalent bispecific antibody, wherein the heavy chain and light chain of HC2:LC1 are attached with the amino acid sequence containing any one of SEQ ID NO:229 - 230( Figure 4 )。

[0246] The Tie2 activating compounds described in Table 9 and Table 10 can be combined with the VEGF binding and VEGFR binding compounds described in Table 11, Table 12, Table 13, Table 14, Table 15, Table 16 and Table 17.

[0247] Any compound in the present disclosure, such as the above-mentioned multispecific fusion construct, can be modified when necessary to promote the desired protein folding or biological activity.

[0248] CDR

[0249] The sequences in the present disclosure may include Complementary Determining Regions (CDRs). The CDRs can be identified by the Kabat method, the Chothia method, the IMGT method, or the Paratome method. The CDRs of the sequences herein can be, for example, of a length of 0 to 91 residues, 0 to 25 residues, 5 to 14 residues, about 0 residues, about 1 residue, about 2 residues, about 3 residues, about 4 residues, about 5 residues, about 6 residues, about 7 residues, about 8 residues, about 9 residues, about 10 residues, about 11 residues, about 12 residues, about 13 residues, about 14 residues, about 15 residues, about 16 residues, about 17 residues, about 18 residues, about 19 residues, about 20 residues, about 21 residues, about 22 residues, about 23 residues, about 24 residues, or about 25 residues.

[0250] The compounds of the present disclosure having binding specificity for HPTP-β (VE-PTP) or the ability to modulate HPTP-β (VE-PTP) may include any of the CDRs in, for example, Table 19, Table 20, Table 21, Table 22, Table 23, or Table 24.

[0251] Table 19 provides non-limiting examples of HCDR1 sequences specific for HPTP-β (VE-PTP).

[0252]

[0253] Table 20 provides non-limiting examples of HCDR2 sequences specific for HPTP-β (VE-PTP).

[0254]

[0255] Table 21 provides non-limiting examples of HCDR3 sequences specific for HPTP-β (VE-PTP).

[0256]

[0257]

[0258] Table 22 provides non-limiting examples of LCDR1 sequences specific for HPTP-β (VE-PTP).

[0259]

[0260] Table 23 provides non-limiting examples of LCDR2 sequences that are specific for HPTP-β (VE-PTP).

[0261]

[0262] Table 24 provides non-limiting examples of LCDR3 sequences that are specific for HPTP-β (VE-PTP).

[0263]

[0264] Compounds of the present disclosure having binding specificity for VEGF or the ability to modulate VEGF may comprise any of the CDRs in Tables 25, 26, 27, 28, 29, or 30.

[0265] Table 25 provides non-limiting examples of HCDR1 sequences that are specific for VEGF.

[0266]

[0267] Table 26 provides non-limiting examples of HCDR2 sequences that are specific for VEGF.

[0268]

[0269] Table 27 provides non-limiting examples of HCDR3 sequences that are specific for VEGF.

[0270]

[0271]

[0272] Table 28 provides non-limiting examples of LCDR1 sequences that are specific for VEGF.

[0273]

[0274] Table 29 provides non-limiting examples of LCDR2 sequences that are specific for VEGF.

[0275]

[0276] Table 30 provides non-limiting examples of LCDR3 sequences that are specific for VEGF.

[0277]

[0278] Table 31 provides the aflibercept-derived sequences corresponding to the D2 domain of human VEGF receptor 1 (SEQ ID NO: 147) and the D3 domain of human VEGF receptor 2 (SEQ ID NO: 148).

[0279]

[0280]

[0281] Table 32 provides the abicipar-derived sequences. SEQ ID NOs: 233 - 236 correspond to the ankyrin repeats within abicipar. SEQ ID NOs: 237 - 242 provide the consensus sequences of the VEGF-binding ankyrin repeats. In SEQ ID NO: 237, X1 is K, T, or Y; X2 is N or M; X3 is T or F; X4 is S or A; X5 is H or R; X6 is A, Y, H, or N; and X7 is A or T. In SEQ ID NO: 238, X1 is K, M, N, R, or V; X2 is Y, H, M, or V; X3 is F, L, M, or V; X4 is R, H, V, A, K, or N; X5 is F, D, H, T, Y, M, or K; and X6 is A, H, N, or Y. In SEQ ID NO: 239, X1 is L, S, or T; X2 is G, S, or C; X3 is S or A; X4 is Q, S, M, or N; X5 is L, M, or Q; and X6 is A, H, N, Y, or D. In SEQ ID NO: 240, X1 is K, S, I, N, T, or V; X2 is K, N, W, A, H, M, Q, or S; X3 is F, Q, L, H, or V; X4 is F or T; X5 is Q or H; X6 is Y or S; X7 is N, H, Y, or M; and X8 is A, H, N, or Y. In SEQ ID NO: 241, X1 is A, N, R, V, Y, E, H, I, K, L, Q, S, or T; X2 is S, A, N, R, D, F, L, P, T, or Y; X3 is T, V, S, A, L, or F; X4 is W, F, or H; X5 is P, I, A, L, S, T, V, or Y; X6 is W, F, I, L, T, or V; X7 is L or P; and X8 is A, H, N, or Y. In SEQ ID NO: 242, X1 is H, Q, A, K, R, D, I, L, M, N, V, or Y; X2 is Y, F, or H; X3 is Q, F, or T; X4 is W, M, G, H, N, or T; X5 is T, A, M, L, or V; X6 is I, L, V, D, or T; and X7 is A, H, N, or Y. SEQ ID NO: 244 provides the truncated sequence derived from abicipar, which contains the designed ankyrin repeats with binding specificity for VEGF.

[0282]

[0283]

[0284] Homology

[0285] The sequence of the present compound may have at least about 70% homology, at least about 71% homology, at least about 72% homology, at least about 73% homology, at least about 74% homology, at least about 75% homology, at least about 76% homology, at least about 77% homology, at least about 78% homology, at least about 79% homology, at least about 80% homology, at least about 81% homology, at least about 82% homology, at least about 83% homology, at least about 84% homology, at least about 85% homology, at least about 86% homology, at least about 87% homology, at least about 88% homology, at least about 89% homology, at least about 90% homology, at least about 91% homology, at least about 92% homology, at least about 93% homology, at least about 94% homology, at least about 95% homology, at least about 96% homology, at least about 97% homology, at least about 98% homology, at least about 99% homology, at least about 99.1% homology, at least about 99.2% homology, at least about 99.3% homology, at least about 99.4% homology, at least about 99.5% homology, at least about 99.6% homology, at least about 99.7% homology, at least about 99.8% homology, at least about 99.9% homology, at least about 99.91% homology, at least about 99.92% homology, at least about 99.93% homology, at least about 99.94% homology, at least about 99.95% homology, at least about 99.96% homology, at least about 99.97% homology, at least about 99.98% homology or at least about 99.99% homology with the amino acid sequences provided herein.

[0286] Homology between two or more sequences can be determined using a variety of methods and software programs, such as NCBI BLAST, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, or another suitable method or algorithm.

[0287] Pharmaceutical composition

[0288] The pharmaceutical compositions of the present disclosure can be any combination of the pharmaceutical compounds described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. The pharmaceutical composition facilitates the administration of the compound to an organism.

[0289] A pharmaceutical preparation for administration may include an aqueous solution of the active compound in a water-soluble form. A suspension of the active compound can be prepared as an oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. An aqueous injection suspension may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. The suspension may also contain suitable stabilizers or agents that increase the solubility of the compound to allow the preparation of highly concentrated solutions. The active ingredient may be in powder form for reconstitution with a suitable vehicle such as sterile pyrogen-free water before use.

[0290] When practicing the methods of treatment or use provided herein, a therapeutically effective amount of a compound described herein is administered to a subject having a disease or condition to be treated, in the form of a pharmaceutical composition. In some embodiments, the subject is a mammal, such as a human. The therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors.

[0291] The pharmaceutical composition can be formulated using one or more physiologically acceptable carriers, which include excipients and auxiliaries that facilitate processing the active compound into a pharmaceutically acceptable product. The formulation can be modified according to the chosen route of administration. A pharmaceutical composition containing a compound described herein can be prepared, for example, by mixing, dissolving, emulsifying, encapsulating, entrapping, or tabletting processes.

[0292] The pharmaceutical composition may comprise at least one pharmaceutically acceptable carrier, diluent, or excipient and the compound described herein in free base or pharmaceutically acceptable salt form. The pharmaceutical composition may contain solubilizers, stabilizers, tonicity enhancing agents, buffers, and preservatives.

[0293] Methods for preparing a composition containing a compound described herein include formulating the compound with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, and cachets. Liquid compositions include, for example, solutions in which the compound is dissolved, emulsions containing the compound, or solutions containing liposomes, micelles, or nanoparticles containing the compound disclosed herein. Semi-solid compositions include, for example, gels, suspensions, and creams. The composition can be a liquid solution or suspension, a solid form suitable for dissolving or suspending in a liquid before use, or an emulsion. These compositions may also contain small amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically acceptable additives.

[0294] Non-limiting examples of dosage forms suitable for the present disclosure include liquids, powders, gels, nano-suspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0295] Non-limiting examples of pharmaceutically acceptable excipients suitable for the present disclosure include binders, disintegrants, anti-adhesives, antistatic agents, surfactants, antioxidants, coating agents, colorants, plasticizers, preservatives, suspending agents, emulsifying agents, antimicrobial agents, spheronizing agents, and any combination thereof.

[0296] The compositions of the present disclosure can be, for example, in an immediate-release form or a controlled-release formulation. Immediate-release formulations can be formulated to make the compound act rapidly. Non-limiting examples of immediate-release formulations include readily soluble formulations. Controlled-release formulations can be pharmaceutical formulations that have been adjusted such that the release rate and release profile of the active agent can match physiological requirements and chronotherapeutic requirements, or have been formulated to achieve a programmed rate of release of the active agent. Non-limiting examples of controlled-release formulations include granules, delayed-release granules, hydrogels (e.g., of synthetic or natural origin), other gelling agents (e.g., gelling dietary fibers), matrix-based formulations (e.g., formulations containing polymeric materials with at least one active ingredient dispersed therein), particles within a matrix, polymeric mixtures, and granulated masses.

[0297] In some embodiments, the controlled-release formulation is in a delayed-release form. The delayed-release form can be formulated to delay the action of the compound for an extended period of time. The delayed-release form can be formulated to delay the release of an effective dose of one or more compounds, e.g., for about 4 hours, about 8 hours, about 12 hours, about 16 hours, or about 24 hours.

[0298] The controlled-release formulation can be in a sustained-release form. The sustained-release form can be formulated to maintain, for example, the action of the compound over an extended period of time. The sustained-release form can be formulated to provide an effective dose of any of the compounds described herein (e.g., provide a physiologically effective blood distribution) over about 4 hours, about 8 hours, about 12 hours, about 16 hours, or about 24 hours.

[0299] The disclosed compositions can optionally contain a pharmaceutically acceptable preservative.

[0300] Non-limiting examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins 1999), each of which is incorporated herein by reference in its entirety.

[0301] The compounds described herein can be conveniently formulated into pharmaceutical compositions consisting of one or more pharmaceutically acceptable carriers. See, for example, Remington’s Pharmaceutical Sciences, latest edition, E.W. Martin, Mack Pub. Co., Easton, PA, incorporated herein by reference in its entirety, which discloses typical carriers and conventional methods of preparing pharmaceutical compositions. Such carriers can be carriers for administering the composition to humans and non-humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH. The pharmaceutical composition can also contain one or more additional active ingredients, such as antimicrobial agents, anti-inflammatory agents, and anesthetics.

[0302] Non-limiting examples of pharmaceutically acceptable carriers include saline, Ringer's solution, and dextrose solutions. In some embodiments, the pH of the solution can be from about 5 to about 8, and can be from about 7 to about 7.5. Additional carriers include sustained release articles, such as semipermeable matrices of solid hydrophobic polymers containing the compound. The matrix can be in the form of a shaped article, such as a membrane, liposome, microparticle, or microcapsule.

[0303] Compositions suitable for topical administration can be used. In some embodiments, the compositions of the present disclosure can include a liquid containing an active agent in the form of a solution, a suspension, or both. The liquid composition can include a gel. The liquid composition can be, for example, aqueous. The composition is an in-situ gellable aqueous composition. In an alternative, the composition is an in-situ gellable aqueous solution. Such a composition can contain a gelling agent that effectively promotes gelling at a concentration when in contact with the eye or with the tear fluid outside the eye. The aqueous composition can have an eye-compatible pH and osmotic pressure. The composition can include an ophthalmic depot formulation that contains an active agent for subconjunctival administration. Particles containing the active agent can be encapsulated in a biocompatible pharmaceutically acceptable polymer or lipid encapsulant. The depot formulation can be suitable for releasing all or substantially all of the active substance over an extended period of time. The polymer or lipid matrix (if present) can be suitable for degrading sufficiently after releasing all or substantially all of the active agent to be transported from the site of administration. The depot formulation can be a liquid formulation containing a pharmaceutically acceptable polymer and the dissolved or dispersed active agent. Upon injection, the polymer forms a depot at the injection site, for example, by gelling or precipitating. The composition can include a solid article that can be inserted into a suitable location in the eye, such as between the eye and the eyelid or in the conjunctival sac, where the article releases the active agent. A solid article suitable for implantation in the eye in such a manner can contain a polymer and can be biodegradable or non-biodegradable.

[0304] In addition to the agents disclosed herein, the pharmaceutical formulation can further contain additional carriers, as well as thickening agents, diluents, buffers, preservatives, and surfactants.

[0305] The pH of the disclosed composition can range from about 3 to about 12. The pH of the composition can be, for example, about 3 to about 4, about 4 to about 5, about 5 to about 6, about 6 to about 7, about 7 to about 8, about 8 to about 9, about 9 to about 10, about 10 to about 11, or about 11 to about 12 pH units. The pH of the composition can be, for example, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, or about 12 pH units. The pH of the composition can be, for example, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or at least 12 pH units. The pH of the composition can be, for example, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, at most 10, at most 11, or at most 12 pH units. The pH of the pharmaceutical formulations disclosed herein can be from about 5.5 to about 6.5. For example, the pH of the formulations of the present disclosure can be about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, or about 6.5. In some embodiments, the pH is 6.2 ± 0.3, 6.2 ± 0.2, 6.2 ± 0.1, about 6.2, or 6.2.

[0306] If the pH is outside the range desired by the formulator, sufficient pharmaceutically acceptable acids and bases can be used to adjust the pH.

[0307] Depending on the intended mode of administration, the pharmaceutical composition can be in the form of a solid, semi - solid, or liquid dosage form, such as tablets, suppositories, pills, capsules, powders, liquids, suspensions, lotions, creams, or gels, e.g., in unit - dose form suitable for precise dosing in a single administration.

[0308] For solid compositions, non - toxic solid carriers include, for example, pharmaceutical - grade mannitol, lactose, starch, magnesium stearate, saccharin sodium, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0309] Non - limiting examples of pharmaceutically active agents suitable for combination with the compositions of the present disclosure include anti - infectives (i.e., aminoglycosides), antiviral agents, antimicrobials, anticholinergics / antispasmodics, antidiabetic agents, antihypertensive agents, antineoplastic agents, cardiovascular agents, central nervous system agents, coagulation regulators, hormones, immunomodulators, immunosuppressants, and ophthalmic products.

[0310] In some embodiments, the pharmaceutical compositions provided herein comprise a mixture of a therapeutically effective amount of a compound with a pharmaceutically acceptable carrier and / or excipient, such as saline, phosphate buffered saline, phosphate and amino acids, polymers, polyols, sugars, buffers, preservatives, and other proteins. Exemplary agents include octylphenoxypolyethoxyethanol compounds, polyethylene glycol monostearate compounds, polyoxyethylene sorbitan fatty acid esters, sucrose, fructose, dextrose, maltose, glucose, mannitol, dextran, sorbitol, inositol, galactitol, xylitol, lactose, trehalose, bovine or human serum albumin, citrate, acetate, Ringer's solution and Hank's solution, cysteine, arginine, carnitine, alanine, glycine, lysine, valine, leucine, polyvinylpyrrolidone, polyethylene, and diols.

[0311] In some embodiments, the pharmaceutical formulations disclosed herein may comprise: (i) a compound or antibody disclosed herein; (ii) a buffer solvate; (iii) a nonionic detergent; (iv) a tonicity agent; and (v) a stabilizer. In some embodiments, the pharmaceutical formulations disclosed herein are stable liquid pharmaceutical formulations.

[0312] In some embodiments, the ophthalmic formulations disclosed herein may comprise: (i) a compound or antibody disclosed herein; (ii) a buffer solvate; (iii) a nonionic detergent; (iv) a tonicity agent; and (v) a stabilizer. In some embodiments, the ophthalmic formulations disclosed herein are stable liquid pharmaceutical formulations or stable liquid ophthalmic formulations.

[0313] In some embodiments, the pharmaceutical formulations or ophthalmic formulations disclosed herein are liquid formulations that may comprise from about 5 mg / mL to about 150 mg / mL of an antibody or compound, from about 7.5 mg / mL to about 140 mg / mL of an antibody or compound, from about 10 mg / mL to about 130 mg / mL of an antibody or compound, from about 10 mg / mL to about 100 mg / mL of an antibody or compound, from about 20 mg / mL to about 80 mg / mL of an antibody or compound, or from about 30 mg / mL to about 70 mg / mL of an antibody or compound. For example, the formulations of the present disclosure may comprise about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, or about 150 mg / mL of a compound, antibody, or antigen-binding fragment thereof described herein.

[0314] In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise a buffer. In some embodiments, the buffer is used to maintain a stable pH and help stabilize the compounds or antibodies disclosed herein. In some embodiments, the buffer or buffer system comprises at least one buffer having a buffering range that completely or partially overlaps with the range of pH 5.5 - 7.4. In some embodiments, the pKa of the buffer is about 6.2 ± 0.5. In some embodiments, the buffer comprises a sodium phosphate buffer. In some embodiments, the sodium phosphate is present at a concentration of about 5 mM to about 15 mM, about 6 mM to about 14 mM, about 7 mM to about 13 mM, about 8 mM to about 12 mM, about 9 mM to about 11 mM, or about 10 mM. In certain embodiments, the buffer system comprises 10 mM of sodium phosphate with a pH of 6.2 ± 0.3 or 6.1 ± 0.3.

[0315] In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise a nonionic detergent. In some embodiments, the nonionic detergent is a nonionic polymer containing a polyoxyethylene moiety. In some embodiments, the nonionic detergent is any one or more of polysorbate 20, poloxamer 188, or polyethylene glycol 3350. In some embodiments, the nonionic detergent is polysorbate 20. In some embodiments, the nonionic detergent is polysorbate 80. In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise from about 0.01% to about 1% of the nonionic detergent. For example, the formulations of the present disclosure may comprise about 0.0085%, about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.1%, about 1.15%, about 1.2%, about 1.25%, about 1.3%, about 1.35%, about 1.4%, about 1.45%, about 1.5%, about 1.55%, about 1.6%, about 1.65%, about 1.7%, about 1.75%, about 1.8%, about 1.85%, about 1.9%, about 1.95%, or about 2% of polysorbate 20, polysorbate 80, or poloxamer 188.

[0316] In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise a tonicity agent. In some embodiments, the tonicity agent is sodium chloride or potassium chloride. In some embodiments, the tonicity agent is sodium chloride. In some embodiments, sodium chloride is present at a concentration of from about 5 mM to about 100 mM, from about 10 mM to about 50 mM, or about 40 mM.

[0317] In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise a stabilizer. In some embodiments, the stabilizer is a heat stabilizer that can stabilize the antibodies or compounds disclosed herein under heat stress conditions. In some embodiments, when a solution comprising a compound or antibody and a heat stabilizer is maintained at about 45 °C for up to about 28 days, the stabilizer maintains greater than about 93% of the compound or antibody in its native conformation. In some embodiments, when a solution comprising a compound or antibody and a heat stabilizer is maintained at about 45 °C for up to about 28 days, the stabilizer prevents aggregation of the compound or antibody and less than 4% of the compound or antibody aggregates. In some embodiments, when a solution comprising a compound or antibody and a heat stabilizer is maintained at about 37 °C for up to about 28 days, the stabilizer maintains more than about 96% of the compound or antibody in its native conformation. In some embodiments, when a solution comprising a compound or antibody and a heat stabilizer is maintained at about 37 °C for up to about 28 °C, the stabilizer prevents aggregation of the compound or antibody and less than about 2% of the compound or antibody aggregates.

[0318] In some embodiments, the heat stabilizer is a sugar or sugar alcohol, such as sucrose, sorbitol, glycerol, trehalose, or mannitol, or any combination thereof. In some embodiments, the stabilizer is a sugar. In some embodiments, the sugar is sucrose, mannitol, or trehalose. In some embodiments, the stabilizer is sucrose. In some embodiments, the pharmaceutical or ophthalmic formulations disclosed herein may comprise from about 1% to about 20% sugar or sugar alcohol, from about 2% to about 18% sugar or sugar alcohol, from about 3% to about 15% sugar or sugar alcohol, from about 4% to about 10% sugar or sugar alcohol, or about 5% sugar or sugar alcohol. For example, the pharmaceutical or ophthalmic formulations of the present disclosure may comprise about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14% sugar or sugar alcohol (such as sucrose, trehalose, or mannitol). In some embodiments, the concentration of the stabilizer is from about 1% w / v to about 20% w / v. In some embodiments, the stabilizer is sucrose at a concentration of from about 1% w / v to about 15% w / v or from about 1% w / v to about 10% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 5% w / v or about 5% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 7.5% w / v or about 7.5% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 10% w / v or about 10% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 12.5% w / v or about 12.5% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 15% w / v or about 15% w / v. In some embodiments, the stabilizer is sucrose at a concentration of 20% w / v or about 20% w / v.

[0319] Administration of the pharmaceutical composition

[0320] The pharmaceutical compositions disclosed herein may be administered in a therapeutically effective amount by various forms and routes, including, for example, oral, topical, parenteral, intravenous injection, intravenous infusion, subcutaneous injection, subcutaneous infusion, intramuscular injection, intramuscular infusion, intradermal injection, intradermal infusion, intraperitoneal injection, intraperitoneal infusion, intracerebral injection, intracerebral infusion, subarachnoid injection, subarachnoid infusion, intraocular injection, intraspinal injection, intrasternal injection, ophthalmic administration, endothelial administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, intraarterial administration, intrathecal administration, inhalation, intralesional administration, intradermal administration, epidural administration, absorption through epithelial or mucocutaneous linings (such as oral mucosa, rectal, and intestinal mucosa), intracapsular administration, subcapsular administration, intracardiac administration, transtracheal administration, subepidermal administration, subarachnoid administration, subcapsular administration, intraspinal canal administration, or intrasternal administration.

[0321] The pharmaceutical composition can be administered locally, for example, by directly injecting the compound into an organ, optionally in the form of a depot formulation or a sustained release formulation or an implant. The pharmaceutical composition can be provided in the form of an immediate release formulation, an extended release formulation, or an intermediate release formulation. The immediate release form can provide immediate release. The extended release formulation can provide controlled release or sustained delayed release.

[0322] In some embodiments, a pump can be used to deliver the pharmaceutical composition. In some embodiments, a pen delivery device can be used to deliver, for example, subcutaneously, the compositions of the present disclosure. Such a pen delivery device can be reusable or disposable. The reusable pen delivery device can use a replaceable cartridge containing the pharmaceutical composition disclosed herein. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. The disposable pen does not have a replaceable cartridge. Instead, the disposable pen delivery device is pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the pharmaceutical composition in the reservoir is emptied, the entire device is discarded.

[0323] The pharmaceutical compositions disclosed herein can be administered by any suitable form or route, such as to the eye, including, for example, topical, oral, systemic, intravitreal, intracameral, intracranial, subconjunctival, sub-Tenon's, retrobulbar, intraocular, intrastromal, intracorneal, posterior juxtascleral, periocular, subretinal, or suprachoroidal administration. Delivery methods can include invasive methods for directly delivering the composition to eye cells. In some embodiments, a liquid pharmaceutical composition comprising an antibody or a compound can be administered by subretinal injection, intravitreal injection (e.g., anterior, middle, or posterior vitreous injection), intravitreal implant, intraorbital injection, intraorbital administration, subcutaneous injection, intracameral injection, intracranial injection, subconjunctival injection, subconjunctival implant, trans-temporal intracameral injection, intrastromal injection, intracorneal injection, aqueous humor injection, sub-Tenon's injection, or sub-Tenon's implant. The composition can be administered by injecting the formulation into any part of the eye, including the anterior chamber, posterior chamber, vitreous chamber (intravitreal), native retina, and / or subretinal space.

[0324] The pharmaceutical compositions disclosed herein can be delivered by non-invasive methods. Examples of non-invasive modes of administering the formulation can include the use of a needleless injection device, as well as topical administration, such as eye drops to the cornea. A variety of administration routes can be employed to effectively deliver the pharmaceutical composition. In some embodiments, the composition is delivered by multiple administration routes, such as subretinal and intravitreal, to increase the efficiency of antibody delivery. In some embodiments, a vitrectomy is performed prior to subretinal and / or intravitreal injection.

[0325] In some embodiments, a liquid formulation comprising an antibody or compound at 10 mg / mL to 120 mg / mL is in a pre-filled syringe and is administered intravitreally in a volume of up to about 500 μL. In some embodiments, a liquid formulation comprising an antibody or compound at 10 mg / mL to 120 mg / mL is in a pre-filled syringe and is administered intravitreally in a volume of up to about 100 μL. In some embodiments, a liquid formulation comprising an antibody or compound at 10 mg / mL to 120 mg / mL is in a pre-filled syringe and is administered intravitreally in a volume of about 50 μL.

[0326] The pharmaceutical compositions disclosed herein can be targeted to any suitable eye cells, including, for example, endothelial cells such as vascular endothelial cells, retinal cells such as retinal pigment epithelium (RPE), corneal cells, fibroblasts, astrocytes, glial cells, pericytes, iris epithelial cells, neurogenic cells, ciliary epithelial cells, Müller cells, muscle cells around and attached to the eye, such as cells of the lateral rectus muscle, orbital adipocytes, cells of the sclera and episclera, cells of the trabecular meshwork, or connective tissue cells.

[0327] Administration

[0328] The compounds, antibodies or therapeutic agents described herein can be administered before, during or after the onset of a disease or condition, and the timing of administration of a composition containing the compound, antibody or therapeutic agent can be varied. For example, the composition can be used as a prophylactic agent and can be administered continuously to a subject susceptible to a condition or disease to reduce the likelihood of occurrence of the disease or condition. The composition can be administered to a subject already suffering from the disease or condition in an amount sufficient to cure or at least partially arrest the symptoms of the disease or condition or to cure, treat, improve or alleviate the condition. The composition can be administered to the subject as soon as possible during or after the onset of symptoms. Administration of the compound, antibody or therapeutic agent can be initiated within 48 hours before the onset of symptoms, within 24 hours before the onset of symptoms, within 6 hours before the onset of symptoms or within 3 hours of the onset of symptoms. Any of the formulations described herein can be used and can be initially administered by any practical route, such as by any of the routes described herein. The compound, antibody or therapeutic agent can be administered as soon as practicable after detection or suspicion of the onset of a disease or condition and for the length of time necessary to treat the disease, for example, from about 1 month to about 3 months. The length of treatment can vary for each subject. The amount effective for such use can vary based on the severity and course of the disease or condition, previous treatment, the health status, weight and response to the drug of the subject, and the judgment of the attending physician. Improvement of clinical symptoms can be monitored, for example, by indirect ophthalmoscopy, fundus photography, fluorescein angiopathy, electroretinography, external eye examination, slit lamp biomicroscopy, applanation tonometry, thickness measurement, optical coherence tomography or autorefaction.

[0329] The pharmaceutical compositions described herein can be in the form of unit dosage forms suitable for the precise administration of a single dose. In a unit dosage form, the formulation can be divided into unit doses containing a suitable amount of one or more compounds, antibodies or therapeutic agents. The unit dose can be in the form of a package containing a discrete amount of the formulation. Non-limiting examples are packaged injectables, vials and ampoules. The aqueous suspension compositions disclosed herein can be packaged in non-reclosable single-dose containers. Reclosable multi-dose containers can be used, for example, with or without a preservative. The injectable formulations disclosed herein can be present in unit dosage form, for example, in ampoules or multi-dose containers with a preservative.

[0330] The various compounds, antibodies, or therapeutic agents disclosed herein can be administered in any order or simultaneously. If administered simultaneously, the various compounds, antibodies, or therapeutic agents can be provided in a single, unified form or in multiple forms, such as as multiple separate injections or infusions. The compounds, antibodies, or therapeutic agents can be packaged together in one package or separately in multiple packages. One or all of the compounds, antibodies, or therapeutic agents can be administered in multiple doses. If not administered simultaneously, the timing between multiple administrations can vary by up to about one month.

[0331] Intraocular injections can be performed at any time interval to increase delivery efficiency and / or minimize or avoid damage to surrounding tissues. The time interval between two or more intraocular injections can be, for example, about 1 minute to about 60 minutes, about 1 minute to about 5 minutes, about 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 15 minutes to about 20 minutes, about 20 minutes to about 25 minutes, about 25 minutes to about 30 minutes, about 30 minutes to about 35 minutes, about 35 minutes to about 40 minutes, about 40 minutes to about 45 minutes, about 45 minutes to about 50 minutes, about 50 minutes to about 55 minutes, or about 55 minutes to about 60 minutes.

[0332] Intraocular injections can be performed at any rate. The rate of intraocular injection can be, for example, about 1 μL / sec to about 500 μL / sec, about 1 μL / sec to about 10 μL / sec, about 10 μL / sec to about 20 μL / sec, about 20 μL / sec to about 30 μL / sec, about 30 μL / sec to about 40 μL / sec, about 40 μL / sec to about 50 μL / sec, about 50 μL / sec to about 60 μL / sec, about 60 μL / sec to about 70 μL / sec, about 70 μL / sec to about 80 μL / sec, about 80 μL / sec to about 90 μL / sec, about 90 μL / sec to about 100 μL / sec, about 100 μL / sec to about 110 μL / sec, about 110 μL / sec to about 120 μL / sec, about 120 μL / sec to about 130 μL / sec, about 130 μL / sec to about 140 μL / sec, about 140 μL / sec to about 150 μL / sec, about 150 μL / sec to about 160 μL / sec, about 160 μL / sec to about 170 μL / sec, about 170 μL / sec to about 180 μL / sec, about 180 μL / sec to about 190 μL / sec, about 190 μL / sec to about 200 μL / sec, about 200 μL / sec to about 300 μL / sec, about 300 μL / sec to about 400 μL / sec, or about 400 μL / sec to about 500 μL / sec.

[0333] The compounds, antibodies or therapeutic agents disclosed herein can be administered at a dose of 0.0001 mg / kg to about 1000 mg / kg, about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 20 mg / kg, about 0.02 mg / kg to about 7 mg / kg, about 0.03 mg / kg to about 5 mg / kg, about 0.05 mg / kg to about 3 mg / kg, about 0.1 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.2 mg / kg to about 0.6 mg / kg, about 0.3 mg / kg to about 0.7 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.1 mg / kg to about 0.9 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 10 mg / kg, about 5 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg or about 3 mg / kg to about 7 mg / kg of the subject's mass.

[0334] The compounds, antibodies or therapeutic agents described herein can be administered at any desired interval. The administration of the compound, antibody or therapeutic agent can have a regular or irregular dosing schedule to suit the person to whom the compound, antibody or therapeutic agent is administered or the subject receiving the compound, antibody or therapeutic agent. For example, the compound, antibody or therapeutic agent can be administered twice a day, once a day, five times a week, four times a week, three times a week, twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every five weeks, once every six weeks, once every eight weeks, once every two months, once every twelve weeks, once every three months, once every four months, once every six months, once a year or at a less frequent rate. In some embodiments, the administration is once every other week.

[0335] The amount administered can be the same amount in each dose, or the dose can vary between doses. For example, a first amount can be administered in the morning and a second amount can be administered in the evening.

[0336] The compounds, antibodies or therapeutic agents described herein can be administered in any amount necessary or convenient. For example, the compounds described herein can be administered by any route of administration in an amount of about 0.05 mg to about 300 mg, about 0.1 mg to about 300 mg, about 0.1 mg to about 200 mg, about 0.1 mg to about 100 mg, about 0.05 mg to about 1.5 mg, about 0.1 mg to about 1.5 mg, about 0.05 mg to about 1 mg, about 1 mg to about 1.5 mg, about 0.5 mg to about 6 mg, about 1 mg to about 4 mg, about 2 mg to about 10 mg, about 10 mg to about 30 mg, about 30 mg to about 50 mg, about 50 mg to about 70 mg, about 70 mg to about 100 mg or about 0.1 mg to about 1 mg, about 0.05 mg, about 0.06 mg, about 0.07 mg, about 0.08 mg, about 0.09 mg, about 0.1 mg, about 0.11 mg, about 0.12 mg, about 0.13 mg, about 0.14 mg, about 0.15 mg, about 0.16 mg, about 0.17 mg, about 0.18 mg, about 0.19 mg, about 0.2 mg, about 0.21 mg, about 0.22 mg, about 0.23 mg, about 0.24 mg, about 0.25 mg, about 0.26 mg, about 0.27 mg, about 0.28 mg, about 0.29 mg, about 0.3 mg, about 0.31 mg, about 0.32 mg, about 0.33 mg, about 0.34 mg, about 0.35 mg, about 0.36 mg, about 0.37 mg, about 0.38 mg, about 0.39 mg, about 0.4 mg, about 0.41 mg, about 0.42 mg, about 0.43 mg, about 0.44 mg, about 0.45 mg, about 0.46 mg, about 0.47 mg, about 0.48 mg, about 0.49 mg, about 0.5 mg, about 0.51 mg, about 0.52 mg, about 0.53 mg, about 0.54 mg, about 0.55 mg, about 0.56 mg, about 0.57 mg, about 0.58 mg, about 0.59 mg, about 0.6 mg, about 0.61 mg, about 0.62 mg, about 0.63 mg, about 0.64 mg, about 0.65 mg, about 0.66 mg, about 0.67 mg, about 0.68 mg, about 0.69 mg, about 0.7 mg, about 0.71 mg, about 0.72 mg, about 0.73 mg, about 0.74 mg, about 0.75 mg, about 0.76 mg, about 0.77 mg, about 0.78 mg, about 0.79 mg, about 0.8 mg, about 0.81 mg, about 0.82 mg, about 0.83 mg, about 0.84 mg, about 0.85 mg, about 0.86 mg, about 0.87 mg, about 0.88 mg, about 0.89 mg, about 0.9 mg, about 0.91 mg, about 0.92 mg, about 0.93 mg, about 0.94 mg, about 0.95 mg, about 0.96 mg, about 0.administered to a subject in an amount of about 97 mg, about 0.98 mg, about 0.99 mg, about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg or about 300 mg.

[0337] Combination therapy

[0338] The pharmaceutical compositions provided herein can be administered in combination with other therapies, such as chemotherapy, radiotherapy, surgery, anti-inflammatory agents or vitamins. The other agents can be administered before, after or concomitantly with the pharmaceutical composition.

[0339] In some embodiments, the compounds or antibodies described herein can be used alone or in combination with one or more therapeutic agents as components of a mixture.

[0340] In some embodiments, the present disclosure provides co-administration of a multispecific compound or antibody that targets, for example, HPTP-β (VE-PTP) and VEGF, with one or more additional anti-VEGF agents, which can stabilize the vasculature to prevent neovascularization. In some embodiments, co-administration of the multispecific compound or antibody with one or more additional anti-VEGF agents can stabilize the vasculature to prevent leakage. The anti-VEGF agent can be a compound, recombinant protein, antibody, antigen-binding fragment, variant or derivative thereof (e.g., scFv), a protein comprising one or more engineered ankyrin repeats, a designed ankyrin repeat protein (DARPin), ankyrin, ankyrin repeat protein, affibody, avimer, adnectin, anticalin, Fynomer, Kunitz domain, knottin, β-hairpin mimetic, or a peptide derived from one or more receptors such as a VEGF receptor, or the VEGF-binding portions of human VEGF receptors 1 and 2.

[0341] Non-limiting examples of anti-VEGF agents include bevacizumab ranibizumab aflibercept Conbercept, brolucizumab, RTH258, VEGF receptor tyrosine kinase inhibitors such as sorafenib, sunitinib, axitinib, pazopanib, vandetinib, cabozantinib, regorafenib, and 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474), VEGF variants, soluble VEGF receptor fragments or traps, aptamers capable of blocking VEGF or VEGFR (e.g., pegaptanib), neutralizing anti-VEGFR antibodies or fragments thereof (e.g., ramucirumab, p1C11, 1121, 1121B), anti-KDR antibodies, anti-flt1 antibodies, low molecular weight inhibitors of VEGFR tyrosine kinase, DARPins that bind VEGF (e.g., abicipar, MP0112, MP0250), proteins containing one or more designed ankyrin repeats that bind VEGF, adnectins (e.g., CT-322), anticalins (e.g., PRS-050), collagen IV-derived biomimetic peptides (e.g., AXT-107).

[0342] Other non-limiting examples of VEGF modulators include anti-inflammatory agents such as dexamethasone, fluocinolone, and triamcinolone. Multispecific compounds or antibodies that target, for example, HPTP-β (VE-PTP) and VEGF can be co-administered with any additional anti-VEGF agent in any combination, such as at the start of treatment, at any time during treatment, or at any time after treatment with the additional anti-VEGF agent has ended. Additionally, the dose of the multispecific compound or antibody can be adjusted during treatment. Additionally, the dose of the additional anti-VEGF agent can be adjusted during treatment. The multispecific compound or antibody can be administered, for example, once a month, once every three months, once every six months, or once a year, where the additional anti-VEGF agent is administered at any frequency between treatments. Methods of treating the diseases or conditions disclosed herein are also disclosed. The method includes administering to a subject:

[0343] a) a therapeutically effective amount of a multispecific compound or antibody that targets, for example, HPTP-β (VE-PTP) and VEGF; and

[0344] b) a therapeutically effective amount of an additional anti-VEGF agent;

[0345] wherein the administration of the multispecific compound or antibody and the additional anti-VEGF agent can be carried out as described herein.

[0346] In some embodiments, the present disclosure provides for the co - administration of a multispecific compound or antibody that targets, for example, HPTP - β (VE - PTP) and VEGF with one or more additional anti - HPTP - β (VE - PTP) agents, which can stabilize the vasculature to prevent neovascularization. In some embodiments, the co - administration of the multispecific compound or antibody with one or more additional anti - HPTP - β (VE - PTP) agents can stabilize the vasculature to prevent leakage. The anti - HPTP - β (VE - PTP) agent can be a compound, recombinant protein, antibody, antigen - binding fragment, variant or derivative thereof (e.g., scFv), a protein comprising one or more engineered ankyrin repeats, a designed ankyrin repeat protein (DARPin), ankyrin, ankyrin repeat protein, affibody, avimer, adnectin, anticalin, Fynomer, Kunitz domain, knottin, β - hairpin mimetic, or a peptide derived from one or more receptors. In some embodiments, the additional anti - HPTP - β (VE - PTP) agent can activate Tie2 signaling by promoting protein phosphorylation such as phosphorylation of the Tie2 protein. In some embodiments, the additional anti - HPTP - β (VE - PTP) agent can bind to HPTP - β (VE - PTP).

[0347] A multispecific compound or antibody that targets, for example, HPTP - β (VE - PTP) and VEGF can be co - administered with any additional anti - HPTP - β (VE - PTP) agent in any combination, for example, at the start of treatment, at any time during treatment, or at any time after treatment with the additional anti - HPTP - β (VE - PTP) agent has ended. The dose of the multispecific compound or antibody can be adjusted during treatment. The dose of the additional anti - HPTP - β (VE - PTP) agent can be adjusted during treatment. For example, the multispecific compound or antibody can be administered monthly, quarterly, semi - annually, or annually, with the additional anti - HPTP - β (VE - PTP) agent being administered at any frequency between treatments. Also disclosed herein is a method of treating a disease or condition disclosed herein. The method comprises administering to a subject:

[0348] a) a therapeutically effective amount of a multispecific compound or antibody that targets, for example, HPTP - β (VE - PTP) and VEGF; and

[0349] b) a therapeutically effective amount of an additional anti - HPTP - β (VE - PTP) agent;

[0350] wherein the administration of the multispecific compound or antibody and the additional anti - HPTP - β (VE - PTP) agent can be carried out as described herein.

[0351] In some embodiments, the present disclosure provides for the co - administration of a multi - specific compound or antibody that targets, for example, HPTP - β (VE - PTP) and VEGF with one or more additional Tie2 receptor - activating compounds. The additional Tie2 receptor - activating compounds can be, for example, angiopoietin - 1 recombinant protein, Ang1 mimetic, Tie2 agonist, peptide, HPTP - β (VE - PTP) phosphatase inhibitor, Tie2 - peptidomimetic, tetra - polyethylene glycol - clustered peptide, collagen IV biomimetic peptide, compound, recombinant protein, antibody, antigen - binding fragment, variants or derivatives thereof (e.g., scFv), affibody, avimer, adnectin, protein comprising one or more engineered ankyrin repeats, designed ankyrin repeat protein (DARPin), ankyrin, ankyrin repeat protein, affibody, avimer, adnectin, anticalin, Fynomer, Kunitz domain, knottin, β - hairpin mimetic, or a peptide derived from one or more receptors. In some embodiments, the one or more additional Tie2 receptor - activating compounds are small molecules. In some embodiments, the one or more additional Tie2 receptor - activating compounds improve drainage through the ocular lymphatics, Schlemm's canal, or limbal lymphatics. In some embodiments, the one or more additional Tie2 receptor - activating compounds are administered in the form of eye drops. In some embodiments, the one or more additional Tie2 receptor - activating compounds are administered to treat primary open - angle glaucoma, age - related macular degeneration, cardiovascular disease, or cystic kidney disease. In some embodiments, the one or more additional Tie2 receptor - activating compounds can be, for example, MAN - 01, AXT - 107, or vasculotide. In some embodiments, the compounds disclosed herein can be co - administered with, for example, MAN - 01, AXT - 107, or vasculotide.

[0352] The multi - specific compound or antibody that targets HPTP - β (VE - PTP) and VEGF can be co - administered with any additional Tie2 receptor - activating compound in any combination, for example, at the start of treatment, at any time during treatment, or at any time after treatment with the additional Tie2 receptor - activating compound has ended. The dose of the multi - specific compound or antibody can be adjusted during treatment. The dose of the additional Tie2 receptor - activating compound can be adjusted during treatment. For example, the multi - specific compound or antibody can be administered monthly, quarterly, semi - annually, or annually, wherein the additional Tie2 receptor - activating compound is administered at any frequency between the multi - specific compound or antibody treatments. Also disclosed herein are methods of treating the diseases or conditions disclosed herein. The method comprises administering to a subject:

[0353] a) A multispecific compound or antibody that is therapeutically effective and targets, for example, HPTP-β (VE-PTP) and VEGF; and

[0354] b) A therapeutically effective Tie2 activator;

[0355] wherein the administration of the multispecific compound or antibody and the additional Tie2 receptor activating compound can be carried out as described herein.

[0356] Mouse model

[0357] Oxygen-induced ischemic retinopathy model

[0358] The oxygen-induced ischemic retinopathy model can be considered to mimic various aspects of proliferative retinopathy neovascularization and proliferative diabetic retinopathy. One-week-old mice can be placed in a sealed chamber and exposed to high oxygen (75 ± 3% oxygen) for five days, thereby causing oxygen-induced neovascularization between days 17 and 21 after birth, for example, at the junction between the vascularized and avascular retina. The compound of interest, such as the antibody or compound of the present disclosure, can be administered to the mice to determine its effect on neovascularization and / or vascular leakage. Neovascularization and / or vascular leakage can be evaluated as described below.

[0359] Rho / VEGF mouse model

[0360] The Rho / VEGF mouse model can mimic various aspects of neovascular age-related macular degeneration. Transgenic mice with vascular endothelial growth factor (VEGF) expression driven by the rhodopsin promoter (rho / VEGF mice) can develop retinal neovascularization, retinal angiomatous proliferation, and retinal vascular leakage. In rho / VEGF mice, VEGF expression in photoreceptors can begin between days 5 and 10 after birth, during which time the deep capillary bed develops. Neovascularization can originate from the deep capillary bed of the retina and grow into the subretinal space. The compound of interest, such as the antibody or compound of the present disclosure, can be administered to the mice to determine its effect on neovascularization and / or vascular leakage. Neovascularization and / or vascular leakage can be evaluated as described below.

[0361] Tet / opsin / VEGF mouse model

[0362] Mice (Tet / opsin / VEGF mice) with VEGF under the control of an inverted tetracycline transactivator (rtTA)-inducible promoter coupled to the rhodopsin promoter can be used as an induction model for neovascularization, retinal vascular leakage, and retinal detachment. In these mice, VEGF transgene expression in the retina can be induced by administration of doxycycline. Three to four days after VEGF induction, neovascularization may become apparent. The neovascularization may become more extensive and may lead to extraretinal folds, which then lead to complete retinal detachment within about five days. A compound of interest, such as an antibody or compound of the present disclosure, can be administered to the mice to determine the effect on neovascularization, vascular leakage, and / or retinal detachment. Neovascularization and / or vascular leakage can be evaluated as described below. To evaluate retinal detachment, the eye can be frozen in a cryostat embedding solution, a ten-micron section can be cut through the entire eye, and the section can be stained with Hoechst. The section can be examined by light microscopy, the average length of retinal detachment for each section can be measured by image analysis, and the percentage of retinal detachment can be calculated.

[0363] Tet / opsin / Ang2 mouse model

[0364] Tet / opsin / Ang2 mice have inducible Ang2 expression in the retina. These mice can be used to study the effects of Ang2 expression under various experimental conditions. For example, Tet / opsin / Ang2 mice can be used to determine the effect of Ang2 expression on neovascularization when VEGF levels are high and low, or to determine the effect of Ang2 expression in an oxygen-induced ischemic retinopathy model. A compound of interest, such as an antibody or compound of the present disclosure, can be administered to the mice to determine the effect of Ang2 expression on therapeutic efficacy.

[0365] Laser-induced choroidal neovascularization model

[0366] The laser-induced choroidal neovascularization model can be considered to mimic aspects of neovascular age-related macular degeneration. Anesthetized mice can be dilated, and burns can be delivered to the retina using a krypton laser, for example, through a slit lamp system and a coverslip used as a contact lens. Multiple burns can be created in one eye, for example, three burns at three locations in each eye. The burns can cause rupture of Bruch's membrane. Choroidal neovascularization can be evaluated at multiple time points after laser treatment, for example, one week, two weeks, or four weeks after laser treatment. A compound of interest, such as an antibody or compound of the present disclosure, can be administered to the mice to determine the effect on neovascularization. The eyecup can be stained with FITC-labeled GSA, the choroid can be flat-mounted and sealed, and the area of choroidal neovascularization at each site of Bruch's membrane rupture can be measured by fluorescence microscopy and image analysis. Neovascularization and / or vascular leakage can also be evaluated as described below.

[0367] Assessment of neovascularization

[0368] Multiple techniques can be used to assess neovascularization.

[0369] Fluorescein angiography can be accomplished by taking serial fundus photographs after injection of a dye to visualize blood vessels, allowing determination of the presence, location, and size of neovascular complexes. For example, an intraperitoneal injection of 0.3 mL of 1% sodium fluorescein can be performed, serial fundus photographs taken, and the area of choroidal neovascularization, total lesion area, and leakage area can be measured.

[0370] The eye can be processed for evaluation by fluorescence, light, or electron microscopy. For example, mice can be perfused with fluorescently labeled dextran, or an antibody targeting GSA or PECAM can be injected into the eye. The eye can be processed for observation under a fluorescence microscope, and the degree of neovascularization can be quantified, for example, by quantifying the area of neovascularization per retina, by quantifying the area of neovascularization per site of Bruch's membrane rupture, or by quantifying the number of nuclei of neovessels extending from the retina into the vitreous.

[0371] The area of retinal neovascularization can be determined, for example, using FITC-labeled GSA lectin and fluorescence microscopy. The eye can be fixed in 10% formalin, dissected intact, washed with PBS, blocked in 8% porcine serum, and stained with FITC-labeled GSA lectin for a period of time (e.g., 40 - 50 minutes) suitable for staining retinal neovascularization and vitreous vessels but not normal retina. The retina can be flat-mounted and sealed, digital images can be obtained using a fluorescence microscope, and merged into a single image of the entire retina. Software can be used to measure the area of retinal neovascularization per retina.

[0372] The area of subretinal neovascularization can be determined, for example, using FITC-labeled GSA lectin and fluorescence microscopy. The eye can be fixed in 10% formalin, the retina can be dissected, blocked in 5% porcine serum, stained with FITC-conjugated GSA for 2 hours to stain vascular cells, and flat-mounted with the photoreceptor side up. The area of subretinal neovascularization can be measured by fluorescence microscopy and image analysis.

[0373] Assessment of retinal vascular leakage

[0374] Retinal vascular leakage can be evaluated by measuring extravasated serum albumin using immunofluorescence techniques. For example, the eye can be fixed in 10% formalin, the retina can be dissected, washed, blocked, and stained with anti-albumin antibody and a fluorescent-conjugated secondary antibody. The blood vessels can be labeled by counterstaining with GSA lectin, and the retina can be mounted and sealed. The retina can be examined by fluorescence microscopy, and the albumin staining area can be determined by image analysis. Retinal vascular leakage is associated with, for example, diabetic macular edema and macular edema due to retinal vein occlusion.

[0375] Miles assay for vascular leakage

[0376] The Miles assay can be used to evaluate vascular leakage of dermal subcutaneous blood vessels. Evans blue is a dye that binds to albumin. Under physiological conditions, the endothelium is impermeable to albumin, so Evans blue-bound albumin is restricted within the blood vessels. Under pathological conditions that promote increased vascular permeability, the endothelial cells partially lose their tight junctions. The endothelium becomes permeable to small proteins such as albumin. 1% Evans blue dye in PBS can be injected intravenously into mice, and VEGF can be injected intradermally. Thirty minutes after intradermal injection, the tissue at the intradermal injection site can be excised and extracted with formamide to evaluate the extravasation of Evans blue dye. Vascular leakage can be quantified by measuring the dye incorporated per milligram of tissue, for example, using densitometry and a standard curve. A test compound, such as an antibody or compound of the present disclosure, can be administered to the mice to determine its effect on vascular leakage.

[0377] Cancer models

[0378] The compounds or antibodies of the present disclosure can be tested for their efficacy as cancer treatments in a variety of cancer models. In some cancer models, cancer cells from a cell line can be implanted into recipient animals, such as mice. Non-limiting examples of suitable mouse strains include C57BL6, BALB / C, C3H, FVB / N, and FVB / N-Tg(MMTV-PyVT)634Mul. Non-limiting examples of suitable cancer cell lines include 4T1, E0771, and P0008. For example, cells of the 4T1 or E0771 cell line can be implanted into the mammary pad as a model of breast cancer. 4T1 or E0771 cells can be implanted into, for example, the third mammary fat pad of female nude mice.

[0379] The size of solid tumors can be measured with calipers, and the tumor volume can be calculated. The tumors can be processed for histopathological evaluation or fluorescence microscopy to assess, for example, tumor area, tumor grade, metastasis count, metastasis area, number of cell nuclei per tumor focus, intratumoral blood vessel diameter, intratumoral blood vessel density, tumor blood vessel maturity, pericyte coverage, proximity between pericytes and endothelial cells, or to grade tumor foci as intravascular or extravascular.

[0380] The spontaneous metastasis model can be used to study the effect of the compounds or antibodies of the present disclosure on metastasis. After implanting the tumor, the primary tumor can be resected (e.g., when it reaches 5 mm in size), and then the metastasis in the animal can be evaluated macroscopically or by histopathological evaluation, for example, to determine the number and size of metastases in the lung, liver, lymph nodes, and bone.

[0381] To evaluate the effect of vascular stability on metastasis, a model in which tumor cells can be injected intravenously can be used, and then intravascular metastasis and extravasation metastasis in the animal can be evaluated. For example, 4T-1 cells can be injected intravenously, and the lung can be processed for histopathological evaluation to quantify intravascular metastasis and extravascular metastasis.

[0382] Metastases can be counted and measured macroscopically, for example, by immersing the lung in Bouin's solution and then examining it with a stereomicroscope.

[0383] To measure tumor vascular permeability in vivo, in vivo multiphoton microscopy can be used. For example, at pre-treatment and post-treatment time points, animals can be injected with fluorescently labeled bovine serum albumin. At each time point, two different regions within the tumor can be selected, and 200-μm image stacks of BSA fluorescence can be recorded by taking pictures through each region every ten minutes for one hour using a multiphoton laser scanning microscope. This analysis method can involve three-dimensional vascular tracing to create vascular metrics, and three-dimensional maps of voxel intensity over time relative to the distance to the nearest blood vessel. Using three-dimensional image registration, the images can be corrected for sample movement over time. The normalized transvascular flux can be calculated.

[0384] Exemplary CDR combinations

[0385] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR3.

[0386] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1 and HCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1 and HCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1 and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1 and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1 and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2 and HCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2 and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2 and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2 and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3 and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3 and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3 and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR1 and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR1 and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR2 and LCDR3.

[0387] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, and HCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise LCDR1, LCDR2, and LCDR3.

[0388] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, and LCDR1. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, LCDR1, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, LCDR1, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR3, LCDR1, LCDR2, and LCDR3.

[0389] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, LCDR1, and LCDR2. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, LCDR1, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, LCDR1, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR3, LCDR1, LCDR2, and LCDR3. In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0390] In some embodiments, the antibodies or compounds of the present disclosure comprise HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0391] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 81-87. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 88-90. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 91-93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 97-98.

[0392] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80 and any one of SEQ ID NOs: 81 - 87. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80 and any one of SEQ ID NOs: 88 - 90. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80 and any one of SEQ ID NOs: 91 - 93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80 and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80 and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81 - 87 and any one of SEQ ID NOs: 88 - 90. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81 - 87 and any one of SEQ ID NOs: 91 - 93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81 - 87 and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81 - 87 and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88 - 90 and any one of SEQ ID NOs: 91 - 93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88 - 90 and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88 - 90 and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 91 - 93 and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 91 - 93 and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 94 - 96 and any one of SEQ ID NOs: 97 - 98.

[0393] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 81-87, and any one of SEQ ID NO: 88-90. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 81-87, and any one of SEQ ID NO: 91-93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 81-87, and any one of SEQ ID NO: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 81-87, and any one of SEQ ID NO: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 88-90, and any one of SEQ ID NO: 91-93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 88-90, and any one of SEQ ID NO: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 88-90, and any one of SEQ ID NO: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 91-93, and any one of SEQ ID NO: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 91-93, and any one of SEQ ID NO: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76-80, any one of SEQ ID NO: 94-96, and any one of SEQ ID NO: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 81-87, any one of SEQ ID NO: 88-90, and any one of SEQ ID NO: 91-93.In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81-87, any one of SEQ ID NOs: 88-90, and any one of SEQ ID NOs: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81-87, any one of SEQ ID NOs: 88-90, and any one of SEQ ID NOs: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81-87, any one of SEQ ID NOs: 91-93, and any one of SEQ ID NOs: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81-87, any one of SEQ ID NOs: 91-93, and any one of SEQ ID NOs: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 81-87, any one of SEQ ID NOs: 94-96, and any one of SEQ ID NOs: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88-90, any one of SEQ ID NOs: 91-93, and any one of SEQ ID NOs: 94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88-90, any one of SEQ ID NOs: 91-93, and any one of SEQ ID NOs: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 88-90, any one of SEQ ID NOs: 94-96, and any one of SEQ ID NOs: 97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 91-93, any one of SEQ ID NOs: 94-96, and any one of SEQ ID NOs: 97-98.

[0394] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 88 - 90, and any one of SEQ ID NOs: 91 - 93. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 88 - 90, and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 88 - 90, and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 91 - 93, and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 91 - 93, and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 81 - 87, any one of SEQ ID NOs: 94 - 96, and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 88 - 90, any one of SEQ ID NOs: 91 - 93, and any one of SEQ ID NOs: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 88 - 90, any one of SEQ ID NOs: 91 - 93, and any one of SEQ ID NOs: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 76 - 80, any one of SEQ ID NOs: 88 - 90, any one of SEQ ID NOs: 94 - 96, and any one of SEQ ID NOs: 97 - 98.In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:76-80, any one of SEQ ID NO:91-93, any one of SEQ ID NO:94-96, and any one of SEQ ID NO:97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:81-87, any one of SEQ ID NO:88-90, any one of SEQ ID NO:91-93, and any one of SEQ ID NO:94-96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:81-87, any one of SEQ ID NO:88-90, any one of SEQ ID NO:91-93, and any one of SEQ ID NO:97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:81-87, any one of SEQ ID NO:88-90, any one of SEQ ID NO:94-96, and any one of SEQ ID NO:97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:81-87, any one of SEQ ID NO:91-93, any one of SEQ ID NO:94-96, and any one of SEQ ID NO:97-98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:88-90, any one of SEQ ID NO:91-93, any one of SEQ ID NO:94-96, and any one of SEQ ID NO:97-98.

[0395] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 91 - 93, and any one of SEQ ID NO: 94 - 96. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 91 - 93, and any one of SEQ ID NO: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 94 - 96, and any one of SEQ ID NO: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 91 - 93, any one of SEQ ID NO: 94 - 96, and any one of SEQ ID NO: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 91 - 93, any one of SEQ ID NO: 94 - 96, and any one of SEQ ID NO: 97 - 98. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 91 - 93, any one of SEQ ID NO: 94 - 96, and any one of SEQ ID NO: 97 - 98.

[0396] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO: 76 - 80, any one of SEQ ID NO: 81 - 87, any one of SEQ ID NO: 88 - 90, any one of SEQ ID NO: 91 - 93, any one of SEQ ID NO: 94 - 96, and any one of SEQ ID NO: 97 - 98.

[0397] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82.

[0398] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76 and SEQ ID NO:81. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76 and SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81 and SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:94 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77 and SEQ ID NO:82. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77 and SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82 and SEQ ID NO:88.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82 and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82 and SEQ ID NO:97.

[0399] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, and SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:94, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, and SEQ ID NO:88. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:94.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:94, and SEQ ID NO:97.

[0400] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97.

[0401] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:94. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97.

[0402] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:76, SEQ ID NO:81, SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:91, SEQ ID NO:94, and SEQ ID NO:97.

[0403] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:97.

[0404] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78 and SEQ ID NO:83. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78 and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78 and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78 and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83 and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83 and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83 and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:92 and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:92 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:95 and SEQ ID NO:97.

[0405] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:95, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97.

[0406] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:92. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:95, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97.

[0407] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:95. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:89, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97.

[0408] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:78, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:92, SEQ ID NO:95, and SEQ ID NO:97.

[0409] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86.

[0410] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:84. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84 and SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84 and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84 and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90 and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90 and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:93 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:93 and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:96 and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79 and SEQ ID NO:86. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86 and SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86 and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86 and SEQ ID NO:98.

[0411] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, and SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:96, and SEQ ID NO:98.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, and SEQ ID NO:90. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:96, and SEQ ID NO:98.

[0412] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:93. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98.

[0413] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98.

[0414] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:79, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:93, SEQ ID NO:96, and SEQ ID NO:98.

[0415] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87.

[0416] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:85. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85 and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:96 and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80 and SEQ ID NO:87. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87 and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87 and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87 and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87 and SEQ ID NO:97.

[0417] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:96, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, and SEQ ID NO:89. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:96, and SEQ ID NO:97.

[0418] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:91. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97.

[0419] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:96. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97.

[0420] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, and SEQ ID NO:97.

[0421] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:99 - 113. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:114 - 123. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:124 - 131. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NO:144 - 146.

[0422] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113 and any one of SEQ ID NOs: 114 - 123. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113 and any one of SEQ ID NOs: 124 - 131. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113 and any one of SEQ ID NOs: 132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113 and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113 and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114 - 123 and any one of SEQ ID NOs: 124 - 131. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114 - 123 and any one of SEQ ID NOs: 132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114 - 123 and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114 - 123 and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124 - 131 and any one of SEQ ID NOs: 132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124 - 131 and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124 - 131 and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 132 - 137 and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 132 - 137 and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 138 - 143 and any one of SEQ ID NOs: 144 - 146.

[0423] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 114 - 123, and any one of SEQ ID NOs: 124 - 131. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 114 - 123, and any one of SEQ ID NOs: 132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 114 - 123, and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 114 - 123, and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 124 - 131, and any one of SEQ ID NOs: 132 - 137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 124 - 131, and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 124 - 131, and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 132 - 137, and any one of SEQ ID NOs: 138 - 143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 132 - 137, and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99 - 113, any one of SEQ ID NOs: 138 - 143, and any one of SEQ ID NOs: 144 - 146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114 - 123, any one of SEQ ID NOs: 124 - 131, and any one of SEQ ID NOs: 132 - 137.In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146.

[0424] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, and any one of SEQ ID NOs: 132-137. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146.In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146.

[0425] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 138-143. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146. In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146.

[0426] In some embodiments, the antibodies or compounds of the present disclosure comprise any one of SEQ ID NOs: 99-113, any one of SEQ ID NOs: 114-123, any one of SEQ ID NOs: 124-131, any one of SEQ ID NOs: 132-137, any one of SEQ ID NOs: 138-143, and any one of SEQ ID NOs: 144-146.

[0427] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:103, SEQ ID NO:118, SEQ ID NO:125, SEQ ID NO:132, SEQ ID NO:140, and SEQ ID NO:144. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:108, SEQ ID NO:123, SEQ ID NO:127, SEQ ID NO:135, SEQ ID NO:143, and SEQ ID NO:145. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:113, SEQ ID NO:123, SEQ ID NO:130, SEQ ID NO:135, SEQ ID NO:143, and SEQ ID NO:145.

[0428] In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 103, SEQ ID NO:118, SEQ ID NO:125, SEQ ID NO:132, SEQ ID NO:140, and SEQ ID NO:144. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 103, SEQ ID NO:118, SEQ ID NO:125, SEQ IDNO:132, SEQ ID NO:140, and SEQ ID NO:144. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 108, SEQ ID NO:123, SEQ ID NO:127, SEQ ID NO:135, SEQ ID NO:143, and SEQID NO:145. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 108, SEQ ID NO:123, SEQ ID NO:127, SEQ ID NO:135, SEQ ID NO:143, and SEQ ID NO:145. In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:85, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 113, SEQ ID NO:123, SEQ ID NO:130, SEQ IDNO:135, SEQ ID NO:143, and SEQ ID NO:145.In some embodiments, the antibodies or compounds of the present disclosure comprise SEQ ID NO:80, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO 113, SEQ ID NO:123, SEQ ID NO:130, SEQ ID NO:135, SEQ ID NO:143, and SEQ ID NO:145.

[0429] Example

[0430] Example 1: A tetravalent bispecific antibody comprising antibody HC2:LC1 and an aflibercept-derived sequence.

[0431] To generate a tetravalent bispecific antibody that fuses the heavy chain of antibody HC2:LC1 to an aflibercept-derived VEGF-binding domain, an amino acid sequence was generated that contains the following additional amino acid sequences from the N-terminus to the C-terminus:

[0432] 1) SEQ ID NO:14 (heavy chain of antibody HC2:LC1);

[0433] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0434] 3) SEQ ID NO:22 (aflibercept-derived sequence).

[0435] The resulting polypeptide (SEQ ID NO:149) was co-expressed with SEQ ID NO:17 to provide the tetravalent bispecific antibody HC2-AFL:LC1, which contains the sequences shown in Table 33. Amino acids 1-19 of SEQ ID NO:149 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-AFL:LC1 does not contain a signal peptide. For example, the mature HC2-AFL:LC1 of the present disclosure may comprise SEQ ID NO:254 and SEQ ID NO:250.

[0436]

[0437]

[0438] Example 2: A tetravalent bispecific antibody comprising antibody HC2:LC1 and a brolucizumab-derived sequence.

[0439] To generate a tetravalent bispecific antibody that fuses the heavy chain of antibody HC2:LC1 to the VEGF-binding domain derived from brolucizumab, an amino acid sequence was generated that contains the following additional amino acid sequences from the N-terminus to the C-terminus:

[0440] 1) SEQ ID NO:14 (heavy chain of antibody HC2:LC1);

[0441] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0442] 3) SEQ ID NO:23 (sequence derived from brolucizumab).

[0443] The resulting polypeptide (SEQ ID NO:150) was co-expressed with SEQ ID NO:17 to provide the tetravalent bispecific antibody HC2-BRO:LC1, which contains the sequences shown in Table 34. Amino acids 1-19 of SEQ ID NO:150 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-BRO:LC1 does not contain a signal peptide. For example, the mature HC2-BRO:LC1 of the present disclosure may contain SEQ ID NO:255 and SEQ ID NO:250.

[0444]

[0445]

[0446] Example 3: Tetravalent bispecific antibody comprising antibody HC2:LC1 and a sequence derived from ranibizumab.

[0447] To generate a tetravalent bispecific antibody that fuses the heavy chain of antibody HC2:LC1 to the VEGF-binding domain derived from ranibizumab, an amino acid sequence was generated that contains the following additional amino acid sequences from the N-terminus to the C-terminus:

[0448] 1) SEQ ID NO:14 (heavy chain of antibody HC2:LC1);

[0449] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0450] 3) SEQ ID NO:28 (sequence derived from ranibizumab).

[0451] The resulting polypeptide (SEQ ID NO:151) was co-expressed with SEQ ID NO:17 to provide the tetravalent bispecific antibody HC2-RAN:LC1, which comprises the sequences shown in Table 35. Amino acids 1-19 of SEQ ID NO:151 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-RAN:LC1 does not comprise a signal peptide. For example, the mature HC2-RAN:LC1 of the present disclosure may comprise SEQ ID NO:256 and SEQ ID NO:250.

[0452]

[0453]

[0454] Example 4: Characterization of bispecific compounds.

[0455] The following multispecific antibodies of the present disclosure were generated: (i) HC2-RAN:LC1, (ii) HC2-AFL:LC1, (iii) HC2-BRO:LC1, (iv) HC2-ABI:LC1, (v) HC2:LC1-AFL, (vi) HC2:LC1-BRO, (vii) HC2:LC1-ABI, (viii) HC2-AFL:LC1-AFL, (ix) HC2-BRO:LC1-BRO, and (x) HC2-ABI:LC1-ABI. Enzyme-linked immunosorbent assays (ELISAs) were performed to determine the binding of these antibodies to VEGF and HPTP-β (VE-PTP). Binding to HPTP-β (VE-PTP) was confirmed for all constructs, and binding to VEGF was confirmed for all constructs except HC2-RAN:LC1.

[0456] As shown in Table 36, Figure 16 , Figure 17 and Figure 18 the tetravalent bispecific antibodies described in Examples 1-3 were generated and characterized.

[0457] Table 36 provides the results of small-scale production and bispecific candidate characterization.

[0458]

[0459] **m / c = measured / calculated

[0460] Figure 16ELISA data demonstrating the binding of the tetravalent bispecific antibody HC2-AFL:LC1 to HPTP-β (upper panel) and VEGF (lower panel) are provided. The binding is compared to controls (HC2:LC1 in the upper panel and VEGF-R2-Fc chimera in the lower panel).

[0461] Figure 17 ELISA data demonstrating the binding of the tetravalent bispecific antibody HC2-BRO:LC1 to HPTP-β (upper panel) and VEGF (lower panel) are provided. The binding is compared to controls (HC2:LC1 in the upper panel and VEGF-R2-Fc chimera in the lower panel).

[0462] Figure 18 ELISA data demonstrating the binding of the tetravalent bispecific antibody HC2-RAN:LC1 to HPTP-β (upper panel) and VEGF (lower panel) are provided. The binding is compared to controls (HC2:LC1 in the upper panel and VEGF-R2-Fc chimera in the lower panel).

[0463] Example 5: Tetravalent bispecific antibodies comprising the antibody HC2:LC1 and abicipar-derived sequences.

[0464] To generate a tetravalent bispecific antibody that fuses the heavy chain of the antibody HC2:LC1 to an abicipar-derived VEGF-binding domain, an amino acid sequence was generated that contains the following additional amino acid sequences from the N-terminus to the C-terminus:

[0465] 1) Residues 1-467 of SEQ ID NO:14 (heavy chain of the antibody HC2:LC1);

[0466] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0467] 3) SEQ ID NO:244 (abicipar-derived sequence).

[0468] The resulting polypeptide (SEQ ID NO:231) was co-expressed with SEQ ID NO:17 to provide the tetravalent bispecific antibody HC2-ABI:LC1, which contains the sequences shown in Table 37. Amino acids 1-19 of SEQ ID NO:231 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-ABI:LC1 does not contain a signal peptide. For example, the mature HC2-ABI:LC1 of the present disclosure may contain SEQ ID NO:257 and SEQ ID NO:250.

[0469]

[0470]

[0471] Example 6: Hexavalent bispecific antibody comprising antibody HC2:LC1 and brolucizumab-derived sequences.

[0472] To generate a heavy chain with a brolucizumab-derived VEGF-binding domain added at the C-terminus, an amino acid sequence was generated that contained the following additional amino acid sequences from the N-terminus to the C-terminus:

[0473] 1) SEQ ID NO:14 (heavy chain of antibody HC2:LC1);

[0474] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0475] 3) SEQ ID NO:23 (brolucizumab-derived sequence).

[0476] To generate a light chain with a brolucizumab-derived VEGF-binding domain added at the C-terminus, an amino acid sequence was generated that contained the following additional amino acid sequences from the N-terminus to the C-terminus:

[0477] 1) SEQ ID NO:17 (light chain of antibody HC2:LC1);

[0478] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0479] 3) SEQ ID NO:23 (brolucizumab-derived sequence).

[0480] The resulting polypeptides SEQ ID NO:150 and SEQ ID NO:218 were co-expressed to provide the hexavalent bispecific antibody HC2-BRO:LC1-BRO shown in Table 38. Amino acids 1-19 of SEQ ID NO:150 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:218 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-BRO:LC1-BRO does not contain a signal peptide. For example, the mature HC2-BRO:LC1-BRO of the present disclosure may comprise SEQ ID NO:255 and SEQ ID NO:258.

[0481]

[0482] Example 7: Hexavalent bispecific antibody comprising antibody HC2:LC1 and aflibercept-derived sequence.

[0483] To generate a heavy chain with an aflibercept-derived VEGF-binding domain added at the C-terminus, an amino acid sequence was generated that contained the following additional amino acid sequences from the N-terminus to the C-terminus:

[0484] 1) SEQ ID NO:14 (heavy chain of antibody HC2:LC1);

[0485] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0486] 3) SEQ ID NO:22 (aflibercept-derived sequence).

[0487] To generate a light chain with an aflibercept-derived VEGF-binding domain added at the C-terminus, an amino acid sequence was generated that contained the following additional amino acid sequences from the N-terminus to the C-terminus:

[0488] 1) SEQ ID NO:17 (light chain of antibody HC2:LC1);

[0489] 2) SEQ ID NO:31 (linker peptide, underlined); and

[0490] 3) SEQ ID NO:22 (aflibercept-derived sequence).

[0491] The resulting polypeptides SEQ ID NO:149 and SEQ ID NO:219 were co-expressed to provide the hexavalent bispecific antibody HC2-AFL:LC1-AFL shown in Table 39. Amino acids 1-19 of SEQ ID NO:149 are the heavy chain signal peptide (SEQ ID NO:11). Amino acids 1-20 of SEQ ID NO:219 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, the mature HC2-AFL:LC1-AFL does not contain a signal peptide. For example, the mature HC2-AFL:LC1-AFL of the present disclosure may comprise SEQ ID NO:254 and SEQ ID N...

Claims

1. An antibody comprising: (a) a first domain that binds to HPTP-β; and (b) a second domain that binds to VEGF; wherein the second domain comprises a sequence that is at least 80% identical to any one of SEQ ID NO:147, SEQ ID NO:148, or SEQ ID NO:233-242.

2. The antibody according to claim 1, wherein the first domain comprises: (i) a heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NO:76-80, the amino acid sequence of any one of SEQ ID NO:81-87, and the amino acid sequence of any one of SEQ ID NO:88-90; and (ii) a light chain variable region comprising the amino acid sequence of any one of SEQ ID NO:91-93, the amino acid sequence of any one of SEQ ID NO:94-96, and the amino acid sequence of any one of SEQ ID NO:97-98.

3. The antibody according to claim 2, wherein: (i) the heavy chain variable region of the first domain comprises the amino acid sequence of SEQ ID NO:76, the amino acid sequence of SEQ ID NO:81, and the amino acid sequence of SEQ ID NO:88; and (ii) the light chain variable region of the first domain comprises the amino acid sequence of SEQ ID NO:91, the amino acid sequence of SEQ ID NO:94, and the amino acid sequence of SEQ ID NO:

97.

4. The antibody according to claim 1, wherein the antibody comprises: (i) a heavy chain amino acid sequence comprising the heavy chain variable region of the first domain and having a sequence that is at least 80% identical to SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, or SEQ ID NO:257; and (ii) a light chain amino acid sequence comprising the light chain variable region of the first domain and having a sequence that is at least 80% identical to SEQ ID NO:250, SEQ ID NO:251, SEQ ID NO:252, or SEQ ID NO:

253.

5. The antibody according to claim 4, wherein the antibody comprises: (i) a heavy chain amino acid sequence comprising the heavy chain variable region of the first domain and having a sequence of SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, or SEQ ID NO:257; and (ii) a light chain amino acid sequence comprising the light chain variable region of the first domain and having a sequence of SEQ ID NO:

250.

6. The antibody according to claim 5, wherein the antibody comprises: (i) a heavy chain amino acid sequence comprising the heavy chain variable region of the first domain and having a sequence of SEQ ID NO:254; and (ii) The amino acid sequence of the light chain comprising the variable region of the light chain of the first domain, and its sequence is SEQ ID NO:

250.

7. The antibody according to claim 1, wherein the antibody comprises: (i) The amino acid sequence of the heavy chain comprising the variable region of the heavy chain of the first domain, and its sequence is at least 80% identical to any one of SEQ ID NO:262 or SEQ ID NO:246 - 249; and (ii) The amino acid sequence of the light chain comprising the variable region of the light chain of the first domain, and its sequence is at least 80% identical to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260 or SEQ ID NO:

261.

8. The antibody according to claim 7, wherein the antibody comprises: (i) The amino acid sequence of the heavy chain comprising the variable region of the heavy chain of the first domain, and its sequence is SEQ ID NO:247 or SEQ ID NO:262; and (ii) The amino acid sequence of the light chain comprising the variable region of the light chain of the first domain, and its sequence is SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260 or SEQ ID NO:

261.

9. The antibody according to claim 8, wherein the antibody comprises: (i) The amino acid sequence of the heavy chain comprising the variable region of the heavy chain of the first domain, and its sequence is SEQ ID NO:247; and (ii) The amino acid sequence of the light chain comprising the variable region of the light chain of the first domain, and its sequence is SEQ ID NO:

259.

10. The antibody according to claim 1, wherein the antibody comprises: (i) The amino acid sequence of the heavy chain comprising the variable region of the heavy chain of the first domain, and its sequence is at least 80% identical to any one of SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256 or SEQ ID NO:257; and (ii) The amino acid sequence of the light chain comprising the variable region of the light chain of the first domain, and its sequence is at least 80% identical to SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260 or SEQ ID NO:261.

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