Humanized monoclonal antibodies targeting ve-PTP (HPTP-β)

Abstract

To provide humanized monoclonal antibodies that target VE-PTP (HPTP-β).SOLUTION: The disclosure provides a composition and a method for treating ocular conditions associated with angiogenesis, which comprises administering an antibody that targets a tyrosine phosphatase inhibitor in a subject. In one embodiment, a compound comprising a sequence that is at least 80% identical to SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12 is provided. In another embodiment, the sequence is at least 85% identical to SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12.SELECTED DRAWING: None

Description

[Technical field]

[0001] cross reference This application claims the benefit of U.S. Provisional Application No. 62 / 364,381, filed July 20, 2016, and U.S. Provisional Application No. 62 / 377,072, filed August 19, 2016, each of which is incorporated by reference in its entirety. [Background technology]

[0002] Monoclonal antibodies generated from non-human organisms may induce an immune response when administered to humans. Segments of these non-human monoclonal antibodies can be replaced with humanized sequences through a humanization process to reduce the possibility of drug immunogenicity while preserving target specificity.

[0003] Incorporation by Reference Each patent, publication, and non-patent literature cited in this application is incorporated herein by reference in its entirety, as if each was individually incorporated by reference. Summary of the Invention [Means for solving the problem]

[0004] In some embodiments, the invention provides compounds comprising a sequence that is at least 80% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12.

[0005] In some embodiments, the invention provides compounds comprising a sequence that is at least 80% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, or SEQ ID NO:23.

[0006] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:34.

[0007] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:29; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:35.

[0008] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:37. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows a sequence alignment of the murine VH sequence (truncated VH0; SEQ ID NO:47) with the four humanized variants SEQ ID NO:9 (VH1), SEQ ID NO:10 (VH2), SEQ ID NO:11 (VH3) and SEQ ID NO:12 (VH4).

[0010] [Diagram 2] FIG. 2 shows a sequence alignment of the murine VL sequence (truncated VL0; SEQ ID NO:48) with the four humanized VL variants SEQ ID NO:20 (VL1), SEQ ID NO:21 (VL2), SEQ ID NO:22 (VL3) and SEQ ID NO:23 (VL4).

[0011] [Diagram 3] FIG. 3 summarizes the hybridoma technology used to generate monoclonal antibodies against the HPTP-β extracellular domain (human VE-PTP / HPTP-β ECD).

[0012] [Figure 4] FIG. 4 shows immunoprecipitation Western blot (left panel) and flow cytometry results (right panel) of R15E6 bound to endogenous human VE-PTP (HPTP-β) in human endothelial cells.

[0013] [Diagram 5]FIG. 5 shows the binding specificity of R15E6 to recombinant, 6-His tagged human VE-PTP (HPTP-β), cynomolgus monkey PTP-β, and human PTP-η extracellular domain proteins by Western blot.

[0014] [Figure 6] FIG. 6 shows Western blot (Panel A) and quantification of concentration-dependent phosphorylation of Tie2 (Panel B) and enhanced endothelial cell viability by R15E6 (Panel C).

[0015] [Figure 7] FIG. 7 shows the effect of rat anti-mouse VE-PTP monoclonal antibody 109.1 on Tie2 (upper left panel), mouse VE-PTP expression (lower left panel), and Akt (right panel) in mouse endothelial cells as determined by Western blot.

[0016] [Figure 8] FIG. 8 shows Tie2 activation by polyclonal antibodies against mouse VE-PTP ECD after 1 hour (Panel A) and 3 minutes (Panel B) incubation in mouse endothelial cells.

[0017] [Figure 9] FIG. 9 shows in vitro inhibition of thrombin- and VEGF-induced endothelial cell permeability by mouse VE-PTP ECD polyclonal antibody.

[0018] [Figure 10] FIG. 10 shows in vivo inhibition of VEGF-induced skin vascular permeability by intravenous administration of monoclonal antibody 109.1 (panel A) and mouse VE-PTP ECD polyclonal antibody (PTP1-8) (panel B), as well as immunoprecipitation of Tie2 from lung lysates of mice injected with control IgG (-) or anti-VE-PTP antibody (+) (panel C).

[0019] [Figure 11] FIG. 11 shows the effect of anti-mouse VE-PTP monoclonal antibody 109.1 on retinal neovascularization (panels A and B) and choroidal neovascularization (panel C) following intraocular administration.

[0020] [Figure 12] FIG. 12 shows quantification of the mean area of ​​retinal neovascularization (Panels A and B) and choroidal neovascularization (Panel C) following intraocular administration of anti-mouse VE-PTP monoclonal antibody 109.1.

[0021] [Figure 13] FIG. 13 shows reducing (Panel A) and non-reducing (Panel B) SDS-PAGE of anti-human VE-PTP (anti-HPTP-β) humanized variant of R15E6.

[0022] [Figure 14] FIG. 14 shows the binding of HC1 variants of R15E6 and HC0LC0 to HPTP-β.

[0023] [Figure 15] FIG. 15 shows the binding of HC2 variants of R15E6 and HC0LC0 to HPTP-β.

[0024] [Figure 16] FIG. 16 shows the binding of HC3 variants of R15E6 and HC0LC0 to HPTP-β.

[0025] [Figure 17] FIG. 17 shows the binding of HC4 variants of R15E6 and HC0LC0 to HPTP-β.

[0026] [Figure 18] FIG. 18 shows Tie2 phosphorylation (left panel) and Akt activation (right panel) in the absence of Ang1 by various humanized antibodies as determined by Western blot.

[0027] [Figure 19] FIG. 19 shows concentration-dependent Tie2 activation (upper panel) and Akt activation (lower panel) by HC2LC4 and HC2LC1 in the absence of ligand as determined by Western blot.

[0028] [Figure 20] FIG. 20 shows Tie2 phosphorylation (left panel) and Akt activation (right panel) by HC2LC4 and HC2LC1 in the presence of Ang1 and Ang2 as determined by Western blot.

[0029] [Figure 21] FIG. 21 shows Tie2 phosphorylation (Panel A), human VE-PTP (HPTP-β) expression (Panel B), and Akt activation (Panel C) by HC2LC1 and HC2LC4 in the presence of Ang1 and / or Ang2 as determined by Western blot.

[0030] [Figure 22] FIG. 22 shows Akt activation by HC2LC1 alone and with excess recombinant human VE-PTP extracellular domain protein (βECD 6His or βECD Fc) as determined by Western blot.

[0031] [Figure 23] FIG. 23 shows the enhancement of endothelial cell viability by HC2LC1 and HC0LC0 compared to a parenteral antibody (R15E6) and a mouse IgG1 control.

[0032] [Figure 24] FIG. 24 shows the improved efficacy of anti-mouse VE-PTP antibody / aflibercept combination therapy for the treatment of retinal detachment in a mouse model.

[0033] [Diagram 25]FIG. 25 shows that subcutaneous administration of monoclonal antibody 109.1 reduced ischemic neovascularization in mice compared to vehicle. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] The present disclosure provides compositions and methods for targeting vascular endothelial protein tyrosine phosphatase (VE-PTP or VEPTP) or human protein tyrosine phosphatase-beta (HPTP-β), for example, to treat ocular disorders characterized by vascular instability, angiogenesis, neovascularization, vascular leakage, and edema. The compositions disclosed herein can activate Tie2 signaling by promoting protein phosphorylation, such as the phosphorylation of Tie2 protein.

[0035] VE-PTP is a member of the receptor-like family of protein tyrosine phosphatases (PTPases). VE-PTP is a transmembrane protein found primarily in vascular endothelial cells that shows structural and functional similarity to cell adhesion molecules. VE-PTP is found in a variety of species, including, for example, zebrafish, chicken, dog, mouse, marmoset, monkey, and human. The human orthologue of VE-PTP is HPTP-β.

[0036] Tie2 (tyrosine kinase with immunoglobulin and epidermal growth factor homology domains 2) is a membrane receptor tyrosine kinase expressed primarily in vascular endothelial cells throughout development. The major regulators of Tie2 phosphorylation are angiopoietin 1 (Ang1) and angiopoietin 2 (Ang2). Ang1 is an agonist of Tie2, and binding of Ang1 to Tie2 promotes receptor phosphorylation. Ang2 acts as a context-dependent antagonist or agonist of Tie2. Binding of Ang1 to Tie2 increases the level of endogenous Tie2 receptor phosphorylation and initiates downstream signaling to induce vascular stabilization through highly organized angiogenesis, tightening of endothelial cell-cell junctions (endothelial cell proximity), enhanced endothelial viability, reduced endothelial inflammation, and improved endothelial function. During angiogenesis, Ang2 acts as a negative regulator of Ang1-Tie2 signaling.

[0037] Under physiological conditions, the duration of Tie2 phosphorylation is regulated by VE-PTP (HPTP-β), which removes phosphate from Tie2 receptor. By inhibiting VE-PTP (HPTP-β), the level of Tie2 phosphorylation is substantially increased, restoring vascular stability. VE-PTP (HPTP-β) inhibitors, such as antibodies that bind to VE-PTP (HPTP-β), can activate Tie2 downstream signaling by inhibiting VE-PTP (HPTP-β). Inhibition of VE-PTP (HPTP-β) by inhibitors can result in vascular stability in subjects with ocular disorders as described herein.

[0038] The VE-PTP (HPTP-β) inhibitors of the present disclosure can include murine monoclonal antibody R15E6 produced by hybridoma cell line ATCC No. PTA-7580, which is immunoreactive with the extracellular domain (SEQ ID NO: 45) of human VE-PTP (HPTP-β) and immunoreactive with the first FN3 repeat (SEQ ID NO: 46) of human VE-PTP (HPTP-β). The VE-PTP (HPTP-β) inhibitors can include antibodies having the same or substantially the same biological properties as R15E6, antibody fragments of R15E6, where the fragments include one or both of the heavy and light chain variable regions, dimers or trimers of F(ab')2, Fab, Fv, scFv of R15E6, and diabodies, triabodies, or tetrabodies derived from R15E6. In some embodiments, fragments of the compounds described herein can be assayed for any biological activity described herein. In some embodiments, fragments of the compounds described herein are capable of activating Tie2 or providing the biological activity of the corresponding intact antibody at equipotent, increased, or decreased levels. [ka] [ka]

[0039] The VE-PTP (HPTP-β) inhibitors of the present disclosure can include antibodies, or antibody fragments, variants, or derivatives thereof, alone or in combination with other amino acid sequences. The inhibitors may be subject to modifications, such as enzymatic cleavage and post-translational modifications.

[0040] The VE-PTP (HPTP-β) inhibitors of the present disclosure can bind to a dominant-negative isoform of VE-PTP (HPTP-β). In some embodiments, the dominant-negative isoform can correspond to a form of VE-PTP (HPTP-β) that lacks phosphatase activity that can compete with endogenous VE-PTP (HPTP-β). Functional evaluation of the dominant-negative VE-PTP (HPTP-β) can be performed by delivery of a transgene and determining the effect on Tie2 phosphorylation.

[0041] The VE-PTP(HPTP-β) inhibitors of the present disclosure can include multiple VE-PTP(HPTP-β) binding sites. In some embodiments, the VE-PTP(HPTP-β) inhibitors can bind to two VE-PTP(HPTP-β) molecules simultaneously, thereby bringing the two VE-PTP(HPTP-β) molecules into close proximity. The VE-PTP(HPTP-β) inhibitors can bind to three VE-PTP(HPTP-β) molecules simultaneously, thereby bringing the three VE-PTP(HPTP-β) molecules into close proximity.

[0042] The VE-PTP (HPTP-β) inhibitors of the present disclosure may be conjugated to another moiety or vehicle by covalent or non-covalent bonding. The moiety or vehicle may, for example, inhibit degradation of the inhibitor, increase half-life, increase absorption, reduce toxicity, reduce immunogenicity, and / or increase biological activity. Non-limiting examples of moieties include Fc domains of immunoglobulins, polymers such as polyethylene glycol (PEG), polylysine, and dextran, lipids, cholesterol groups such as steroids, carbohydrates, dendrimers, oligosaccharides, and peptides.

[0043] The compounds of the present invention can be used to target VE-PTP (HPTP-β) to restore Tie2 activity and initiate downstream signaling cascades, including, for example, Akt / PI3-K signaling, Rac1 signaling, MAPK / Ras signaling. In some embodiments, the compounds of the present invention can inhibit NF-κB signaling. Activation of Tie2 can result in vascular stabilization, which can be beneficial for treating diabetes-related conditions and ocular conditions, including, for example, retinopathy, diabetic retinopathy, retinal perfusion, ocular neovascularization, ocular vascular leakage, ocular edema, intraocular pressure, ocular hypertension, ocular inflammation, glaucoma, and ocular bleeding. The compounds can also be effective for treating peripheral arterial disease, including, for example, wound healing, unstable blood flow, cardiac fibrosis, erectile dysfunction, cardiomyopathy, ischemic injury, cardiac hypertrophy, interstitial fibrosis, nephropathy, albuminuria, glomerulosclerosis, renal fibrosis, neuropathy, and neuronal inflammation.

[0044] antibody Antibodies consist of two identical heavy (H) chain polypeptide sequences and two identical light (L) chain polypeptide sequences. Each of the heavy chains contains one N-terminal variable (V H ) region and three C-terminal constant (C H 1. C H 2, and C H Each light chain contains one N-terminal variable (V L ) region and one C-terminal constant (C L 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 heavy and light chain variable regions align with each other. The pairing of the heavy and light chain variable regions together forms a single antigen-binding site. Each light chain is linked to a heavy chain by one covalent disulfide bond. The two heavy chains are linked to each other by one or more disulfide bonds depending on the heavy chain isotype. Each heavy and light chain also contains regularly spaced intrachain disulfide bridges.

[0045] Light chains from any vertebrate species can be designated kappa or lambda based on the amino acid sequence of the constant region. Depending on the amino acid sequence of the heavy chain constant region, immunoglobulins can be classified into five classes of immunoglobulins (IgA, IgD, IgE, IgG, and IgM), each with heavy chains designated alpha, delta, epsilon, gamma, and mu, respectively. The alpha and gamma classes are further divided into subclasses based on differences in the sequence and function of the heavy chain constant region. Subclasses of IgA and IgG expressed by humans include IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0046] The variable (V) region contains segments that can vary widely in sequence between antibodies. The variable region mediates antigen binding and defines the specificity of a particular antibody for its antigen. However, variability is not uniformly distributed across the range of the variable region. Instead, the variable region consists of relatively invariant stretches called framework regions (FR) of 15-30 amino acids separated by shorter regions of extreme variability called hypervariable regions, each of which is 9-12 amino acids long. Each of the variable regions of naturally occurring heavy and light chains adopts a predominantly f3 sheet configuration and contains four framework regions connected by three hypervariable regions, which form loops that connect, and in some cases form part of, the f3 sheet structure. The hypervariable regions in each chain are held together in close proximity by the framework regions and, together with the hypervariable regions from the other chain, contribute to the formation of the antigen-binding site of the antibody. The constant domains are not directly involved in binding the antibody to an antigen, but exhibit various effector functions, such as the participation of the antibody in antibody-dependent cellular cytotoxicity (ADCC).

[0047] The hypervariable region can include amino acid residues from a complementarity determining region (CDR), e.g., approximately residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in a light chain variable region, approximately residues 1-35 (H1), 50-65 (H2), and 95-102 (H3) in a heavy chain variable region, and / or residues from a hypervariable loop.

[0048] Monoclonal antibodies can be obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies that make up the population are identical except for possible naturally occurring mutations that may be present in small amounts. In contrast to polyclonal antibody preparations, which contain different antibodies against different epitopes, each monoclonal antibody is directed against a single epitope. In addition to specificity, monoclonal antibodies are beneficial in that each can be synthesized uncontaminated by other antibodies.

[0049] As used herein, a monoclonal antibody may be, for example, a chimeric antibody in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as antigen-binding fragments of such antibodies.

[0050] Antibody fragments can include portions of a multimeric antibody, such as the antigen-binding or variable regions of an intact 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 tetrabodies, and linear antibodies.

[0051] Non-limiting examples of epitopes include amino acids, sugars, lipids, phosphoryl, and sulfonyl groups. Epitopes can have specific three-dimensional structural characteristics and / or specific charge characteristics. Epitopes can be conformational or linear.

[0052] Humanized Monoclonal Antibodies Suitable antibodies that target VE-PTP can be identified using a variety of techniques. For example, candidate agents may be screened for binding to VE-PTP. Agents that bind to VE-PTP may be screened for activity, such as inhibition of VE-PTP-mediated dephosphorylation of Tie2. In some embodiments, candidate agents are first screened for activity in vivo.

[0053] The selection of suitable assays to use for identifying specific inhibitors depends on the nature of the candidate agent to be screened. For example, if the candidate is an antibody or peptibody that contains an Fc portion, fluorescence-activated cell sorting (FACS) analysis allows the candidate agent to be selected based on its ability to bind to cells expressing VE-PTP. The cells can endogenously express VE-PTP or can be genetically engineered to express VE-PTP. For other candidate agents, such as aptamers, other techniques can be utilized. For example, aptamers that specifically bind to VE-PTP can be selected using Systematic Evolution of Ligands by Exponential Enrichment (SELEX), which selects for specific aptamers through repeated rounds of in vitro selection.

[0054] VE-PTP inhibitors may be screened for inhibition of VE-PTP-mediated activity, such as Tie2 dephosphorylation. In one suitable Western blotting-based assay, human umbilical vein endothelial cells (HUVEC) are cultured in serum-free medium in the presence or absence of various concentrations of candidate drugs, and cell lysates are prepared, immunoprecipitated with Tie2 antibody, separated by polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to polyvinylidene difluoride (PVDF) membrane. The membrane-bound immunoprecipitated proteins are then Western blotted sequentially with anti-phosphotyrosine antibody to quantify Tie2 phosphorylation, followed by Tie2 antibody to quantify total Tie2. Tie2 phosphorylation is expressed as the ratio of anti-phosphotyrosine signal to total Tie2 signal. The greater the level of anti-phosphotyrosine signal, the greater the VE-PTP inhibition by the candidate drug.

[0055] Hybridoma technology is a method for producing large numbers of monoclonal antibodies that target a specific antigen. Hybridoma development begins by injecting a mammalian host, such as a mouse (rat) or rabbit, with an antigen that elicits an immune response. In response to the antigen, B lymphocytes (B cells) in the host produce antibodies that bind to the antigen. The B cells are then taken from the host and fused with immortal B cell cancer cells (myeloma cells) to produce a hybrid cell line known as a hybridoma. Hybridomas retain both the antibody-producing capacity of B cells and the increased longevity and reproducibility of myelomas. Hybridomas can be grown in culture starting with one surviving hybridoma cell to produce new cultures consisting of genetically identical hybridomas, which then produce one antibody (monoclonal) or a mixture of different antibodies (polyclonal) per culture. The myeloma cell lines used in this process are selected for their ability to grow in tissue culture and their inability to synthesize antibodies. After culturing, a primary screening process is carried out to identify and select the hybridoma that produces the most specific antibody. Non-limiting examples of antibody screening techniques include ELISA and immunocytochemical screening. The lead hybridoma identified from screening can then be characterized for reactivity, binding affinity, specificity, and cross-reactivity.

[0056] In some embodiments, the binding affinity (K D ) is as strong as about 70 pM to about 70 nM, 1 nM to about 70 nM, or at least about 1 nM. In some embodiments, the binding affinity (K D ) is about 4 nM to about 70 nM.

[0057] Other recombinant antibody engineering techniques include the use of non-mammalian viruses or yeast. These techniques rely on the rapid cloning of immunoglobulin gene segments to create antibody libraries with slightly different amino acid sequences from which antibodies with the desired specificity can be selected. Antibody libraries can be used to enhance specificity for target antigens, stability in various environmental conditions, therapeutic efficacy, and detectability in diagnostic applications.

[0058] For human administration, monoclonal antibodies generated from non-human species can be further optimized by humanization processes to reduce the possibility of immunogenicity while preserving target specificity. The humanization process involves the incorporation of human DNA into the genetic sequence of the gene that produces the isolated antibody. The recombinant DNA is then cloned and expressed in cells for large-scale production of the newly humanized antibody.

[0059] An example of humanized antibody is modified chimeric antibody. Chimeric antibody is generated as described above. Chimeric antibody is further mutated outside CDR to replace non-human sequence in variable region with homologous human sequence. Another example of humanized antibody is CDR-grafted antibody, where non-human CDR sequence is introduced into human heavy and light chain variable sequence of human antibody scaffold to replace corresponding human CDR sequence.

[0060] Humanized antibodies can be produced in mammalian cells, bioreactors, or transgenic animals such as mice, chickens, sheep, goats, pigs, and marmosets. Transgenic animals can have a substantial portion of the human antibody-producing genome inserted into the animal's genome.

[0061] A fully human monoclonal antibody corresponds to an antibody whose antigen-binding residues are entirely derived from a human immunoglobulin sequence or fragment thereof that is subject to selection. In some embodiments, the selection can be performed using phage display technology, in which a set of variable antibody domains are expressed on a filamentous phage coat protein and enriched for binding to a target antigen. In some embodiments, the selection can be performed using a transgenic animal, such as a mouse, rat, or rabbit, in which a full set of endogenous immunoglobulin genes is replaced with a full set of human immunoglobulin genes. In some embodiments, a full set of human immunoglobulin genes can be introduced into the genome of an animal in which endogenous antibody production renders the production of antibodies insufficient.

[0062] A humanized monoclonal antibody targeting VE-PTP (HPTP-β) The humanization process of the murine monoclonal antibody targeting VE-PTP (HPTP-β) described herein utilizes a combination of CDR grafting technology in conjunction with the antibody structure and a database of mature human IgG sequences. Human framework sequences were used as acceptor frameworks for the CDR sequences. All of these acceptor sequences were derived from mature human IgG from a human source and were not derived from phage display.

[0063] Four humanized variants were designed for the heavy and light chain variable domains of a murine monoclonal antibody raised against the recombinant extracellular domain of human VE-PTP (HPTP-β). The design was based on factors including homology, T cell epitopes, critical residues, and predicted structure.

[0064] The sequences disclosed herein were identified using the antibody numbering systems from IMGT and Kabat. These two numbering systems identify different residues in murine antibodies as belonging to the CDRs, and the combined IMGT / Kabat CDR sequences were used for optimal retention of CDR loop conformations.

[0065] Heavy chain sequence SEQ ID NO:1 is the murine monoclonal antibody R15E6 (V H0 )V H SEQ ID NO: 47 is the V domain of R15E6 without the murine signal peptide sequence (underlined). H SEQ ID NO:2 is the truncation sequence of the murine V domain of R15E6. H The closest human germline gene V region to the domain is Homo sapiens IGHV3-73. SEQ ID NO:3 and SEQ ID NO:4 are Homo sapiens IGHV3-72 and Homo sapiens IGHV3-48, respectively, which are further germline V regions similar to SEQ ID NO:2. The peptide sequences of SEQ ID NO:1 to SEQ ID NO:4 are shown in Table 1. [Table 1]

[0066] An online database of human IgG sequences was searched using the BLAST algorithm to identify murine V H The domains were searched for comparison and candidate human variable domains were selected from the top 200 BLAST results. The equivalent human variable domains were reduced to four candidates based on a combination of framework homology, preserving framework residues and canonical loop structures. Table 2 lists the four selected acceptor frameworks, SEQ ID NOs: 5-8. SEQ ID NO: 5 is AEX29600, SEQ ID NO: 6 is AAC51024, SEQ ID NO: 7 is AEX29289, and SEQ ID NO: 8 is ABA26204. [Table 2]

[0067] Mouse V H When the CDRs of the above were grafted onto the acceptor framework (SEQ ID NOs: 5 to 8), these sequences were converted into the humanized variants shown in Table 3. SEQ ID NO: 9 is V H1 and SEQ ID NO:10 is VH2 and SEQ ID NO:11 is V H3 and SEQ ID NO:12 is V H4 It is. [Table 3]

[0068] FIG. 1 shows the four humanized variants, SEQ ID NO:9 (V H1 ), SEQ ID NO:10(V H2 ), SEQ ID NO:11(V H3 ), and SEQ ID NO: 12 (V H4 ) with Mouse V H Sequence (Cut V H0 ; SEQ ID NO: 47). H / V K Residues important to the interface and canonical loop structures were conserved.

[0069] Mouse V H The percentage of homology of the humanized variants with V is shown in Table 4. The homology ranking of the humanized variants is as follows: H2 >V H1 >V H3 >V H4 It is. [Table 4]

[0070] Light chain sequence In Table 5 below, SEQ ID NO:13 is the murine V sequence of R15E6, including the murine signal peptide sequence (underlined). L Domain (V L0 SEQ ID NO: 48 is the V of R15E6 without the murine signal peptide sequence. L SEQ ID NO: 14 is the truncation sequence of the murine V domain of R15E6. L The closest human germline gene V region to the domain is Homo sapiens IGKV1-16. SEQ ID NO: 15 is IGKV4-1, which is highly similar to IGKV1-16. [Table 5]

[0071] An online database of human IgK sequences was searched for comparison with the murine VL domains using the BLAST search algorithm, and candidate human variable domains were selected from the top 200 BLAST results. These equivalent human variable domains were reduced to four candidates based on a combination of framework homology, preserving framework residues and canonical loop structures. Table 6 lists the four selected acceptor frameworks. SEQ ID NO: 16 is AF234256_1, SEQ ID NO: 17 is AAD03722, SEQ ID NO: 18 is AAY33352, and SEQ ID NO: 19 is AAZ09113. [Table 6]

[0072] Mouse V L Grafting the CDRs of V into these acceptor frameworks converted these sequences into the humanized variants shown in Table 7. SEQ ID NO: 20 represents V L1 and SEQ ID NO:21 is V L2 and SEQ ID NO:22 is V L3 and SEQ ID NO:23 is V L4 It is. [Table 7]

[0073] Figure 2 shows the four humanized V L SEQ ID NO: 20 (V L1 ), SEQ ID NO:21(V L2 ), SEQ ID NO:22(V L3 ), and SEQ ID NO: 23 (V L4 ) with Mouse V L Sequence (Cut V L0 ; SEQ ID NO: 48) is shown. L / V K Residues important to the interface and canonical loop structures were conserved.

[0074] Mouse V L The percentage of homology of the humanized variants with V is shown in Table 8. The homology ranking of the humanized variants is as follows: L3 >V L1 >V L2 >V L4 It is. [Table 8]

[0075] Designing humanized antibodies: The four Vs H and four V's L Chain combination The mouse variable chains were humanized by grafting the murine CDR sequences into the appropriate human antibody donor sequences as described above in the context of the IgG4 heavy chain for expression of the intact antibody protein. The panel of full-length humanized antibodies was then codon-optimized for expression in a Chinese Hamster Ovary (CHO) cell line. Four variants were designed for each variable chain using different human donor sequences, producing a matrix of 16 human antibodies for expression, whose heavy and light chain pairings are shown in Table 9. The complete mouse variable domain (V) grafted into a human IgG4 constant domain (IgG4) was expressed as a 2D mouse antibody with a 2D mouse CDR sequence. H0- V L0 ) was generated as a positive control in binding and functional assays. [Table 9]

[0076] T cell epitope screening Heavy chain. Presentation of peptide sequences in the groove of MHC class II molecules leads to activation and immunogenic responses of CD8+ T cells. To reduce this response, therapeutic proteins can be designed to avoid the incorporation of T cell epitopes that can activate T cells by reducing the affinity of binding to MHC class II molecules.

[0077] Murine Antibody V H and VL The humanized variant sequences were screened for MHC II binding peptides to determine that the humanization process had removed peptide sequences with high affinity using an in silico algorithm. Table 10 shows the results of the screening, with the high affinity T cell epitope core in bold (IC 50 <50 nM). Human germline sequences ICHV3-73, ICHV3-72, and ICHV3-48 were also analyzed for comparison. Any potential T cell epitopes present in the germline sequences and matched in the humanized variants are shown in italics. CDRs are underlined. [Table 10]

[0078] Light chain sequence Table 11 shows the results of the screening, with the high affinity T cell epitope core in bold (IC 50 <50 nM). The human germline sequences IGKV1-16 and IGKV4-1 were also analyzed for comparison, and any potential T cell epitopes present in the germline sequences and matched in the humanized variants are shown in italics. CDRs are underlined. [Table 11]

[0079] Mouse V L0 contains a T-cell epitope within the CDR3 / framework 4 region of the light chain that is absent in three of the humanized variants.

[0080] Post-translational modifications Fv glycosylation. The N-linked glycosylation motif is NXS / T, where X is any amino acid except proline. This motif is R15E6 V H or V L was not present in the murine or humanized variants of

[0081] Deamidation. The amino acid motifs SNG, ENN, LNG, and LNN may be prone to deamidation of asparagine to give aspartic acid. H or V L was not present in the murine or humanized variants of

[0082] Signal peptide. Murine antibody signal peptides can result in higher levels of expression in CHO cells. The following signal peptides can be incorporated into the humanized variants: Heavy chain signal peptide (SEQ ID NO:24): MGWTLVFLFLLSVTAGVHS Light chain signal peptide (SEQ ID NO: 25): MVSSAQFLGLLLLCFQGTRC

[0083] V H Each of the domains can be synthesized in frame with a human IgG4 isotype constant domain sequence with a stabilizing S228P mutation. The entire heavy chain sequence can be codon optimized and the DNA sequence verified. The amino acid sequence of the IgG4 constant domain with the S228P mutation (SEQ ID NO:26) is as follows: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

[0084] V LEach of the domains can be synthesized in frame with the human IgK isotype constant domain sequence. The entire light chain sequence can then be codon optimized and the DNA sequence verified. The amino acid sequence of the IgK constant domain (SEQ ID NO:27) is as follows: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0085] Each variant chain can be verified by DNA sequence analysis. Then the transient transfection and expression of each humanized antibody can be tracked. One chimeric antibody can be expressed to be used as a positive control and can have a murine variable domain, a human Ig constant domain. The 16 humanized variants have only a humanized variable domain and a human Ig constant domain. Table 12 shows the pairing of heavy and light chains that produces the 16 humanized variants with a humanized variable domain and a human Ig constant domain. [Table 12]

[0086] Tables 13 and 14 list the complete amino acid sequences of each humanized heavy and light chain. [Table 13-1] [Table 13-2] [Table 14]

[0087] The target sequence may have 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 100% homology, at least about 105% homology, at least about 106% homology, at least about 107% homology, at least about 108% homology, at least about 109% homology, at least about 110% homology, at least about 111% homology, at least about 112% homology, at least about 113% homology, at least about 114% homology, at least about 115% homology, at least about 116% homology, at least about 117% homology, at least about 118% homology, at least about 119% homology, at least about 120% homology, at least about 121% homology, at least about 122% homology, at least about 123% homology, at least about 124% homology, at least about 125% homology, at least about 126% homology, at least about 127% homology, at least about 128% homology, at least about 129% homology, at least about 130% homology, at least about 131% homolog The sequence may have 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.

[0088] Various methods and software programs can be used to determine homology between two or more peptides or nucleic acids, such as NCBI BLAST, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, or another suitable method or algorithm.

[0089] Pharmaceutical Compositions The pharmaceutical composition of the present invention may be a combination of any pharmaceutical compound described herein with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. The pharmaceutical composition may be administered in a therapeutically effective amount as a pharmaceutical composition by various forms and routes, including, for example, intravenous, intramuscular, oral, parenteral, ophthalmic, and topical administration.

[0090] The pharmaceutical composition may be administered to the eye via any suitable form or route, including, for example, topical, oral, systemic, intravitreal, intracameral, subconjunctival, subtenon, retrobulbar, intraocular, posterior juxtascleral, periocular, subretinal, and suprachoroidal administration. The composition may be administered by injecting the formulation into any part of the eye, including the anterior chamber, posterior chamber, vitreous cavity (intravitreal), retina proper, and / or subretinal space. The composition may be delivered by non-invasive methods. Non-invasive modes of administering the formulation may include using a needleless injection device. Multiple administration routes may be utilized for efficient delivery of the pharmaceutical composition.

[0091] The pharmaceutical composition may be, for example, a cell comprising endothelial cells such as vascular endothelial cells, retinal pigment epithelium, Any suitable ocular cells can be targeted, including cells of the retina, such as the retinal pigment epilthelium (RPE), corneal cells, fibroblasts, astrocytes, glial cells, pericytes, iris epithelial cells, cells of neural origin, ciliary epithelial cells, Müller cells, muscle cells that surround and attach to the eye, such as cells of the lateral rectus muscle, orbital fat cells, cells of the sclera and episclera, cells of the trabecular meshwork, and connective tissue cells.

[0092] The pharmaceutical composition can be administered locally, for example, via injection of the compound directly into the organ, optionally in a depot or sustained release formulation or implant.The pharmaceutical composition can be provided in the form of a rapid release formulation, an extended release formulation, or an intermediate release formulation.The rapid release form can provide immediate release.The extended release formulation can provide controlled release or sustained delayed release.

[0093] Pharmaceutical preparations for administration can include aqueous solutions of the active compounds in water-soluble form. Suspensions of the active compounds can be prepared as oily injection suspensions. 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. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Suspensions can also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient can be in powder form for constitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.

[0094] In carrying out the treatment or method of use provided herein, a therapeutically effective amount of the compound described herein is administered in a pharmaceutical composition to a subject having a disease or condition to be treated.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.

[0095] In some embodiments, the compounds described herein can be used alone or in combination with one or more therapeutic agents as a component of a mixture. For example, the VE-PTP (HPTP-β) inhibitors of the present disclosure can be co-formulated or co-administered with an antibody, such as an anti-VEGF agent. The anti-VEGF agent can be a compound, an antibody, or an antibody fragment, variant, or derivative thereof. Non-limiting examples of anti-VEGF agents include bevacizumab (Avastin®), ranibizumab (Lucentis®), and aflibercept (Eylea®). In some embodiments, the compounds described herein can be used before, during, or after treatment with an anti-VEGF agent.

[0096] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers, including excipients and auxiliaries, which facilitate the processing of active compounds into medicament-usable preparations.Formulations can be modified according to the route of administration selected.The pharmaceutical compositions comprising the compounds described herein can be prepared, for example, by mixing, dissolving, emulsifying, encapsulating, encapsulating or compressing processes.

[0097] The pharmaceutical composition may comprise at least one pharma- ceutically acceptable carrier, diluent, or excipient and a compound described herein in free base or pharma- ceutically acceptable salt form.The pharmaceutical composition may contain solubilizers, stabilizers, tonicity enhancers, buffers, and preservatives.

[0098] The method of preparing a composition comprising the compounds described herein includes formulating the compounds with one or more inert pharma- ceutically 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 compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, for example, gels, suspensions, and creams. The compositions may be in liquid solution or suspension, solid form suitable for dissolving or suspending in liquid before use, or as emulsions. These compositions may also contain small amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharma- ceutical acceptable additives.

[0099] Non-limiting examples of dosage forms suitable for use in the present invention include liquids, powders, gels, nanosuspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0100] Non-limiting examples of pharma- ceutically acceptable excipients suitable for use in the present invention include binders, disintegrants, anti-adherents, antistatic agents, surfactants, antioxidants, coating agents, colorants, plasticizers, preservatives, suspending agents, emulsifiers, antibacterial agents, spheronization agents, and any combination thereof.

[0101] The composition of the present invention may be, for example, an immediate release form or a controlled release formulation. An immediate release formulation may be formulated to allow the compound to act rapidly. Non-limiting examples of immediate release formulations include formulations that are easily soluble. A controlled release formulation may be a pharmaceutical formulation in which the release rate and release profile of the active agent are adapted to match physiological and chronotherapeutic requirements, or alternatively, formulated to provide the release of the active agent at a programmed rate. Non-limiting examples of controlled release formulations include granules, delayed release granules, hydrogels (e.g., synthetic or natural origin), other gelling agents (e.g., gel-forming dietary fibers), matrix-based formulations (e.g., formulations that include polymeric materials in which at least one active ingredient is dispersed), granules in a matrix, polymer mixtures, and granular masses.

[0102] In some cases, the controlled release formulation is a delayed release form.Delayed release form can be formulated to delay the action of compound for a long period of time.Delayed release form can be formulated to delay the release of effective dose of one or more compounds for, for example, about 4, about 8, about 12, about 16 or about 24 hours.

[0103] Controlled release formulations may be sustained release forms.Sustained release forms can be formulated, for example, to sustain the action of compound over a long period of time.Sustained release forms can be formulated to provide any of the effective doses of compounds described herein (for example, provide a physiologically effective blood profile) for about 4, about 8, about 12, about 16, or about 24 hours.

[0104] The disclosed compositions can optionally include a pharma- ceutically acceptable preservative.

[0105] Non-limiting examples of pharma- ceutically acceptable excipients can be found, for example, in Remington: 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, HA and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins 1999), each of which is incorporated by reference in its entirety.

[0106] The compounds described herein can be conveniently formulated into pharmaceutical compositions consisting of one or more pharma- ceutically acceptable carriers.See, for example, Remington's Pharmaceutical Sciences, latest edition, by EW Martin Mack Pub. Co., Easton, PA, which is incorporated by reference in its entirety. It discloses typical carriers and conventional methods for preparing pharmaceutical compositions. Such carriers may be carriers for administering the compositions to humans and non-humans, including solutions such as sterile water, saline, and buffer solutions at physiological pH. Pharmaceutical compositions can also include one or more additional active ingredients, such as antibacterial agents, anti-inflammatory agents, and anesthetics.

[0107] Non-limiting examples of pharma- ceutically acceptable carriers include saline, Ringer's solution, and dextrose solution. In some embodiments, the pH of the solution may be about 5 to about 8, or about 7 to about 7.5. Further carriers include sustained-release preparations such as semipermeable matrices of solid hydrophobic polymers containing the compound. The matrices may be in the form of shaped articles, e.g., films, liposomes, microparticles, or microcapsules.

[0108] The disclosed method relates to administering an antibody targeting VE-PTP (HPTP-β) as part of a pharmaceutical composition. A composition suitable for topical administration may be used. In some embodiments, the compositions of the present invention may include a liquid that includes an active agent in a solution, suspension, or both. The liquid composition may include a gel. The liquid composition may be, for example, aqueous. The composition is an aqueous composition capable of gelling in situ. To reiterate, the composition is an aqueous solution capable of gelling in situ. Such a composition may include a gelling agent at a concentration effective to promote gelling upon contact with the eye or the tear fluid outside the eye. The aqueous composition may have an ophthalmically-compatible pH and osmolality. The composition may include an ophthalmic depot formulation including an active agent for subconjunctival administration. The microparticles including the active agent may be embedded in a biocompatible pharma- ceutically acceptable polymer or lipid encapsulant. The depot formulation may be adapted to release all or substantially all of the active agent over an extended period of time. The polymer or lipid matrix, if present, may be adapted to degrade sufficiently to be transported away from the administration site after the release of all or substantially all of the active agent. The depot formulation may be a liquid formulation comprising a pharma- ceutically acceptable polymer and a dissolved or dispersed active agent. Upon injection, the polymer forms a depot at the injection site, for example, by gelling or precipitation. The composition may comprise a solid article that can be inserted into the eye at a suitable location, such as between the eye and the eyelid or in the conjunctival sac, where the article releases the active agent. The solid article suitable for implantation into the eye in such a manner may comprise a polymer and may be bioerodible or non-bioerodible.

[0109] Pharmaceutical formulations can include, in addition to the agents disclosed herein, additional carriers, as well as thickeners, diluents, buffers, preservatives, and surfactants.

[0110] The pH of the disclosed compositions may range from about 3 to about 12. The pH of the composition may 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 may 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 may 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 may 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. If the pH is outside the range desired by the formulator, the pH may be adjusted by using sufficient pharma- ceutical acceptable acids and bases.

[0111] Depending on the intended mode of administration, the pharmaceutical composition may be in the form of a solid, semi-solid or liquid dosage form, such as, for example, a tablet, suppository, pill, capsule, powder, liquid, suspension, lotion, cream, or gel, e.g., a unit dosage form suitable for single administration of a precise dosage amount.

[0112] For solid compositions, non-toxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0113] Non-limiting examples of pharma- ceutically active agents suitable for combination with the compositions of the present disclosure include anti-infective agents, i.e., aminoglycosides, antiviral agents, antibacterial agents, anti-cholinergic / anti-spasmotic agents, antidiabetic agents, antihypertensive agents, anti-neoplastic agents, cardiovascular agents, central nervous system agents, coagulation modifiers, hormones, immunological agents, immunosuppressants, and ophthalmic preparations.

[0114] In some embodiments, the pharmaceutical compositions provided herein include a therapeutically effective amount of the compound mixed with a pharma- ceutically acceptable carrier and / or excipient, such as saline, phosphate buffered saline, phosphates 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 and Hanks' solution, cysteine, arginine, carnitine, alanine, glycine, lysine, valine, leucine, polyvinylpyrrolidone, polyethylene, and glycols.

[0115] Methods of Administration and Treatment The pharmaceutical compositions described herein may be administered for preventive and / or therapeutic treatment. In therapeutic applications, the compositions may be administered to a subject already suffering from a disease or condition in an amount sufficient to treat or at least partially arrest the symptoms of the disease or condition, or to treat, cure, improve, or ameliorate the condition. The compositions may also be administered to reduce the likelihood of developing, contracting, or worsening the condition. Amounts effective for this use may vary based on the severity and course of the disease or condition, previous therapy, the subject's health, weight, and response to the drug, and the judgment of the treating physician.

[0116] Multiple therapeutic agents can be administered in any order or simultaneously.When simultaneously, multiple therapeutic agents can be provided in a single unified form, or in multiple forms, for example, as multiple separate pills.The agents can be packaged together or separately, in a single package or multiple packages.One or all of the therapeutic agents can be given in multiple doses.When not simultaneously, the timing between multiple doses can vary up to about one month.

[0117] The therapeutic agents described herein may be administered before, during, or after the onset of a disease or condition, and the timing of administration of the composition containing the therapeutic agent may vary. For example, the composition may be used as a prophylactic and may be administered continuously to a subject with a tendency to a condition or disease to reduce the likelihood of the onset of the disease or condition. The composition may be administered to the subject during or as soon as possible after the onset of symptoms. Administration of the therapeutic agent may begin within the first 48 hours of the onset of symptoms, within the first 24 hours of the onset of symptoms, within the first 6 hours of the onset of symptoms, or within 3 hours of the onset of symptoms. The initial administration may be via any practical route, for example, by any route described herein using any formulation described herein. The therapeutic agent may be administered as soon as practicable after the onset of a disease or condition is detected or suspected, and for the period of time required to treat the disease, such as, for example, from about 1 month to about 3 months. The length of treatment may vary from subject to subject.

[0118] The pharmaceutical compositions described herein may be in unit dosage form suitable for single administration of precise dosage. In unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. The unit dosage may be in the form of a package containing individual amounts of the formulation. Non-limiting examples are packaged injections, vials, or ampoules. Aqueous suspension compositions may be packaged in single-dose non-reclosable containers. Multi-dose reclosable containers may be used, for example, with or without preservatives. Injectable formulations may be presented in unit dosage form, for example, ampoules, or multi-dose containers with preservatives.

[0119] The pharmaceutical compositions provided herein may be administered in conjunction with other therapies, such as chemotherapy, radiation, surgery, anti-inflammatory agents, and selected vitamins. The other agents may be administered before, after, or simultaneously with the pharmaceutical composition.

[0120] dosage VE-PTP (HPTP-β) antibodies may be administered at a dosage of 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 based on the subject's body weight.

[0121] VE-PTP (HPTP-β) antibodies may be administered at any desired interval. The administration of the compound may have an irregular dosing schedule to suit either the person administering the compound or the subject receiving the compound. For example, the compound may be administered once a week, twice a week, three times a week, four times a week, six times a week, seven times a week, eight times a week, nine times a week, or ten times a week. The interval between daily doses may be any time interval, for example, every hour, every two hours, every three hours, every four hours, every five hours, every six hours, every seven hours, every eight hours, every nine hours, every ten hours, every eleven hours, or every twelve hours. Dosing schedules for administration of VE-PTP (HPTP-β) antibodies include, but are not limited to, once daily, three times weekly, twice weekly, once weekly, three times monthly, twice monthly, once every other month, once yearly, twice yearly, three times yearly, or four times yearly. In some embodiments, the VE-PTP (HPTP-β) antibody may be administered every other week.

[0122] Furthermore, the amount of VE-PTP (HPTP-β) antibody administered may be the same amount in each dose, or the dosage may vary between doses. For example, a first amount is administered in the morning and a second amount is administered in the evening. The dosage for administration may vary depending on the schedule of anti-VEGF administration.

[0123] The anti-VE-PTP antibody may be administered in combination with any anti-VEGF agent in any combination, for example, at the beginning of treatment, any time during treatment, or any time after treatment with the anti-VEGF agent has ended. Additionally, the dosage of the VE-PTP (HPTP-β) inhibitor may be adjusted during treatment. The amount of the anti-VEGF agent may also be adjusted during treatment.

[0124] Further non-limiting examples of VEGF modulating agents include non-inflammatory agents, such as dexamethasone, fluocinolone, and triamcinolone. Additionally, the disclosed methods may include an implant that delivers an anti-VEGF agent. For example, a VE-PTP (HPTP-β) inhibitor may be co-administered either before, during, or after the implant is provided to a subject suffering from a disease or condition described herein.

[0125] The anti-VEGF treatment may be administered, for example, monthly, once every three months, once every six months, or yearly, with the VE-PTP (HPTP-β) inhibitor being administered any frequency during treatment.

[0126] Also disclosed herein is a method of treating a disease or condition disclosed herein, the method comprising: a) a therapeutically effective amount of a VE-PTP (HPTP-β) inhibitor, and b) a therapeutically effective amount of an anti-VEGF agent to a subject, Administration of the VE-PTP (HPTP-β) inhibitor and the anti-VEGF agent may be in any manner desired by the administrator, for example, as further described herein.

[0127] VE-PTP (HPTP-β) binding agents or anti-VEGF agents may be administered in any amount necessary or convenient. For example, the compounds described herein may be administered to a subject by any route of administration at a dose of 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, 0.1 mg to about 1.5 mg, about 0.05 mg to about 1 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 0.17mg, about 0.18mg, about 0.19mg, about 0.2mg, about 0.21mg, about 0.22mg, about 0.23mg, about 0.24mg, about 0.25mg, about 0.26mg, about 0.27mg, about 0.28mg, about 0.29mg, about 0.3mg, about 0.31mg, About 0.32mg, about 0.33mg, about 0.34mg, about 0.35mg, about 0.36mg, about 0.37mg, about 0.38mg, about 0.39mg, about 0.4mg, about 0.41mg, about 0.42mg, about 0.43mg, about 0.44mg, about 0.45mg, about 0.46mg, 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 0.77mg, about 0.78mg, about 0.79mg, about 0.8mg, about 0.81mg, about 0.82mg, about 0.83mg, about 0.84mg, about 0.85mg, about 0.86mg, about 0.87mg, about 0.88mg, about 0.89mg, about 0.9mg, about 0.91mg, about 0.92mg, about 0.93mg, about 0.94mg, about 0.95mg, about 0.96mg, about 0.97mg, about 0.98mg, about 0.99mg, about 1mg, about 1.5mg, about 2mg, about 2.5mg, about 3mg, about 3.5mg, about 4mg, about 4.5mg, about 5mg, about 5.5mg, about 6mg, about 6.5mg, about 7mg, about 7.5mg, about 8mg, about 8.5mg, about 9mg, about 9.5mg, about 10mg, about 15mg, about 20mg, about 25mg, about 30m g, 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, It may be administered in an amount of 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.

[0128] intraocular delivery Disclosed herein are methods for intraocular delivery of compositions of the present invention to a subject having a disease or condition disclosed herein. The delivery method can include an invasive method for directly delivering the composition to ocular cells. In some embodiments, the liquid pharmaceutical composition comprising the antibody is delivered by subretinal injection. In some embodiments, the liquid pharmaceutical composition comprising the antibody is delivered by intravitreal or subcutaneous injection. In some embodiments, the liquid pharmaceutical composition comprising the antibody is delivered by intracavitary injection. In some embodiments, the composition is delivered by multiple routes of administration, e.g., subretinal and / or intravitreal, to increase the efficiency of antibody delivery. In some embodiments, the subretinal and / or intravitreal injection is preceded by a vitrectomy.

[0129] Intraocular injections can be performed over any time interval to improve efficiency of delivery and / or minimize or avoid damage to surrounding tissues. The time interval for 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.

[0130] Intraocular injection can be performed at any speed. The speed of intraocular injection can be, for example, about 1 μL / min to about 200 μL / min, about 1 μL / min to about 10 μL / min, about 10 μL / min to about 20 μL / min, about 20 μL / min to about 30 μL / min, about 30 μL / min to about 40 μL / min, about 40 μL / min to about 50 μL / min, about 50 μL / min to about 60 μL / min, about 60 μL / min to about 70 μL / min, about 70 μL / min to about 80 μL / min, about 80 μL / min to about 90 μL / min, about 90 μL / min to about 100 μL / min, The flow rate may be about 100 μL / min to about 110 μL / min, about 110 μL / min to about 120 μL / min, about 120 μL / min to about 130 μL / min, about 130 μL / min to about 140 μL / min, about 140 μL / min to about 150 μL / min, about 150 μL / min to about 160 μL / min, about 160 μL / min to about 170 μL / min, about 170 μL / min to about 180 μL / min, about 180 μL / min to about 190 μL / min, or about 190 μL / min to about 200 μL / min.

[0131] kit The present disclosure further relates to a kit containing the composition of the present disclosure. The kit comprises: A) a composition comprising an antibody targeting VE-PTP (HPTP-β), and B) A carrier for delivering the composition to a subject may include.

[0132] The kit may be modified to accommodate the dosing regimen prescribed for the subject to be treated. The following are non-limiting examples of kits for use with subjects receiving the compositions of the present disclosure by intraocular injection: A) an aqueous composition comprising: a) an antibody targeting the VE-PTP (HPTP-β) extracellular domain, and b) a carrier system comprising: i) a tonicity agent, and ii) water Including, The tonicity agent is present in an amount such that the reconstituted formulation comprises about 0.5% to about 10% by weight of the tonicity agent by volume of the carrier system. and B) Components for delivering the aqueous composition.

[0133] In some embodiments, the kit of the invention comprises: A) a composition for delivering an anti-VE-PTP antibody, and B) Compositions for delivering anti-VEGF agents Includes. The kit may be modified to suit the dosage regimen prescribed for the subject to be treated.The following is a non-limiting example of a kit for use with a patient receiving an intravenously delivered composition comprising the disclosed compound and an anti-VEGF agent administered intravitreally.This example provides for twice daily dosing of the disclosed compound for three months, and an injection of ranibizumab at the 12th week. A. Three packages, each package containing four vials, each vial containing a sufficient amount of a VE-PTP (HPTP-β) inhibitor to administer 5 mg of the disclosed compound twice daily for seven days; and B. A vial of ranibizumab for injection at the end of week 12 providing 0.5 mg of ranibizumab. The kit may include any combination of elements. Additionally, where the anti-VE-PTP antibodies disclosed herein are provided orally, a single container having a sufficient dose of the disclosed compound may be supplied with the kit. The kit may also provide written instructions for using and disposing of the delivered composition. The instructions may vary from kit to kit to reflect the prescribed dosing regimen. The instructions may describe any of the therapies, compounds, excipients, or methods of administration described herein.

[0134] The disclosed compositions may, for example, comprise about 1.5% to about 90% by weight based on the volume of the carrier system. Non-limiting examples of tonicity agents include dextrose, mannitol, and glycerin. Formulators may utilize more than one tonicity agent.

[0135] The kit may further include devices for administration, such as syringes, filter needles, extension tubing, cannulas, and subretinal syringes. Non-limiting examples of administration routes include intraocular, parenteral, and topical. Intraocular administration routes may include, for example, intravitreal, intracavitary, subconjunctival, subtenon, retrobulbar, intraocular, posterior juxtascleral, periocular, subretinal, and suprachoroidal. Delivery may be, for example, by syringe, needle, infusion pump, or injector. Syringes and injectors may be, for example, single dose, multiple dose, fixed dose, or variable dose. Non-limiting examples of injectors include pen injectors, autoinjectors, and electronic patch injector systems.

[0136] The kit can include components suitable for administering the composition of the present invention to a subject.In some embodiments, the composition of the present invention is present in the kit as a unit dosage form.Therefore, the prescriber can provide a delivery device with a higher concentration of compound, and adjust the delivery volume to provide a smaller amount of compound than that in the entire solution.

[0137] Any kit described herein may include a set of instructions.The instructions may relate to dosage, timing of administration, reconstitution of composition when kit contains dry composition, and delivery vehicle and method of disposal of unused composition.The instructions may describe any therapy, compound, excipient, or administration method described herein.

[0138] method The present invention provides compositions and methods for treating or preventing eye diseases or conditions, such as eye edema, diabetic macular edema, vascular leakage, age-related macular degeneration (wet type), age-related macular degeneration (dry type), choroidal neovascularization, diabetic retinopathy, ocular ischemia, uveitis, retinal vein occlusion (central or branch), ocular trauma, surgery-induced edema, surgery-induced neovascularization, cystoid macular edema, ocular ischemia, and uveitis.These diseases or conditions can be characterized by changes in ocular vasculature, whether they are progressive or non-progressive, whether they are the result of acute disease or condition, or chronic disease or condition.These diseases can be characterized by an increase in the level of vascular endothelial growth factor (VEGF) in plasma.

[0139] The present disclosure provides a method of treating ocular neovascularization in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody targeting VE-PTP (HPTP-β).

[0140] In some embodiments, VE-PTP (HPTP-β) inhibitors stabilize the vasculature against leakage and neovascularization.

[0141] Improvement in clinical symptoms may be monitored, for example, by indirect ophthalmoscopy, fundus photography, fluorescein vasculopathy, electroretinography, external eye examination, slit-lamp biomicroscopy, applanation tonometry, pachymetry, optical coherence tomography, or autorefraction. In some embodiments, the disclosed methods involve administration of a VE-PTP (HPTP-β) antibody, including a composition comprising an anti-VEGF agent.

[0142] In some embodiments, the methods of the disclosure include co-administration of a VE-PTP (HPTP-β) antibody with one or more anti-VEGF agents, which can stabilize the vasculature against leakage.

[0143] In some embodiments, the methods of the disclosure are directed to co-administration of anti-VE-PTP (HPTP-β) antibodies with one or more anti-VEGF agents, which can stabilize the vasculature against neovascularization.

[0144] In some embodiments, anti-VE-PTP (HPTP-β) antibodies can stabilize the vasculature against leakage and neovascularization.

[0145] In some embodiments, a human subject having at least one visually impaired eye is treated with about 0.1 mg to about 100 mg of VE-PTP (HPTP-β) antibody. Clinical improvement can be monitored, for example, by indirect ophthalmoscopy, fundus photography, fluorescein vasculopathy, electroretinography, exophthalmoscopy, slit-lamp biomicroscopy, applanation tonometry, corneal thickness measurement, optical coherence tomography, and autorefraction. As described herein, dosing can be done at any frequency determined by the administering person. After discontinuation of anti-VEGF agent treatment, subsequent doses can be administered, for example, weekly or monthly, for example, at frequencies spaced apart by about 2-8 weeks or about 1-12 months depending on the response.

[0146] Diseases that are direct or indirect results of diabetes include, among others, diabetic macular edema and diabetic retinopathy. The vascular system of the eye of a diabetic patient becomes unstable over time, resulting in conditions such as non-proliferative retinopathy, macular edema, and proliferative retinopathy. When fluid leaks into the center of the macula, the part of the eye where clear and straight vision occurs, an accumulation of fluid and associated proteins begins to deposit on or under the macula. This deposition creates swelling that gradually distorts the subject's central vision. This condition is macular edema. Another condition that can occur is non-proliferative retinopathy, where vascular changes such as microaneurysms can be observed outside the macula of the eye.

[0147] These conditions may be associated with diabetic proliferative retinopathy, which is characterized by increased neovascularization. The new blood vessels are fragile and prone to bleeding. The result is obstruction or complete blockage of the light pathway through the eye due to retinal scarring and excessive formation of new blood vessels. Subjects with diabetic macular edema often suffer from the non-proliferative phase of diabetic retinopathy, but subjects often only begin to develop macular edema at the onset of the proliferative phase.

[0148] Diabetic retinopathy is the most common cause of vision loss in working-age Americans. Severe vision loss occurs due to tractional retinal detachment exacerbated by retinal neovascularization, while the most common cause of moderate vision loss is diabetic macular edema (DME).

[0149] VEGF is a hypoxia-regulated gene, and VEGF levels are increased in hypoxic or ischemic retina. Under most circumstances, Ang2 binds to Tie2 but does not stimulate phosphorylation, acting as a Tie2 antagonist. In the eye, Ang2 is upregulated at sites of neovascularization and acts as a permissive factor for VEGF. Increased expression of VEGF in the retina does not stimulate neovascular sprouting from the superficial or intermediate capillary beds of the retina or choriocapillaris, but does stimulate sprouting from the deep capillary beds where constitutive expression of Ang2 occurs. Coexpression of VEGF and Ang2 at the surface of the retina causes neovascular sprouting from superficial retinal capillaries.

[0150] Angiogenesis, the process of generating new blood vessels from pre-existing vessels, is essential for a wide range of physiological and pathological events, including embryonic development, menstruation, wound healing, and tumor growth. Most, if not all, tumors require angiogenesis to grow and proliferate. VEGF may be a major factor in angiogenesis by increasing vascular permeability and the number of capillaries.

[0151] VEGF is a protein found primarily in endothelial cells and has functions in vasculogenesis, angiogenesis, and vascular permeabilization. VEGF expression is induced by hypoxia, activated oncogenes, and cytokines. VEGF activation can lead to angiogenesis in normal human cells and tissues, but also in tumors, allowing tumor progression and growth. Inhibition of VEGF can inhibit tumor growth and lead to tumor regression. Various retinopathies are associated with increased levels of VEGF, and ischemia in the eye leads to induction of VEGF production due to lack of oxygen. This increase in VEGF can cause hyperproliferation of blood vessels in the retina, leading to blindness. The disclosed anti-VE-PTP (HPTP-β) antibodies can act to stabilize the ocular vasculature and counter the irritation caused by VEGF and other inflammatory agents present in the diseased retina. In some embodiments, administration of an anti-VE-PTP (HPTP-β) antibody to a subject can maintain a level of disease reversal after administration of the anti-VEGF drug to the subject is discontinued.

[0152] Macular degeneration is characterized by the gradual loss or impairment of vision due to cellular and tissue degeneration of the macula in the center of the retina. Macular degeneration is often characterized as one of two types: nonexudative (dry) or exudative (wet). Both types are bilateral and progressive, but each type may reflect a different pathological process. The wet form of age-related macular degeneration (AMD) is the most common type of choroidal neovascularization and the leading cause of blindness in the elderly. AMD affects millions of Americans over the age of 60 and is the leading cause of new blindness among the elderly.

[0153] Choroidal neovascular membranes (CNVM) are a problem associated with a wide variety of retinal diseases, but are most commonly associated with age-related macular degeneration. In CNVM, abnormal blood vessels originating from the choroid (a blood vessel-rich layer of tissue just below the retina) grow through the retinal layers. These new blood vessels are very fragile and break easily, allowing blood and fluid to pool within the layers of the retina.

[0154] Diabetes mellitus (DM) is a metabolic disease caused by the inability of the pancreas to produce or use insulin. The most common types of diabetes are type 1 diabetes (often referred to as juvenile-onset diabetes) and type 2 diabetes (often referred to as adult-onset diabetes). Type 1 diabetes results from the body's inability to produce insulin due to the loss of insulin-producing cells, and currently requires humans to inject insulin. Type 2 diabetes generally results from insulin resistance, a condition in which cells are unable to properly use insulin. Diabetes can be correlated with a number of other conditions, including eye conditions or diseases, including diabetic retinopathy and diabetic macular edema (DME).

[0155] Diabetic retinopathy is a complication of diabetes that results from damage to the blood vessels of the light-sensitive tissue at the back of the eye (retina). Initially, diabetic retinopathy may not cause any symptoms or only mild vision problems. Eventually, diabetic retinopathy can lead to blindness. Diabetic retinopathy can occur in any person with type 1 or type 2 diabetes.

[0156] In the earliest stage of non-proliferative retinopathy, microaneurysms develop in the very small blood vessels of the retina. As the disease progresses, many of these blood vessels become damaged or blocked, and these areas of the retina send signals to local tissues to grow new blood vessels for nutrients. This stage is called proliferative retinopathy. New blood vessels grow along the retina and along the surface of the clear vitreous gel that fills the inside of the eye. The blood vessels have thin, fragile walls, and without timely treatment, the new blood vessels can leak blood, e.g., whole blood or some of its components, resulting in severe vision loss and even blindness. Fluid can also leak into the center of the macula, the part of the eye where clear and straight visual vision occurs. When fluid and associated proteins begin to deposit on or under the swelling of the macula, the subject's central vision becomes distorted. This condition is called macular edema, and can occur at any stage of diabetic retinopathy, but is more likely to occur as the disease progresses.

[0157] Uveitis is a condition in which the uvea becomes inflamed. The eye is hollow inside and has three distinct layers of tissue that surround a central cavity. The outermost is the sclera (the white covering of the eye) and the innermost is the retina. The middle layer between the sclera and the retina is called the uvea. The uvea contains many blood vessels that nourish the eye. Complications of uveitis include glaucoma, cataracts, and the formation of new blood vessels (neovascularization).

[0158] Eye trauma is any kind of physical or chemical damage to the eye. Eye trauma can affect anyone, and the main symptoms include redness or pain in the affected eye. This symptom tends not to occur when a very small projectile causes the trauma.

[0159] Surgery-induced edema is the occurrence of swelling in ocular tissues following surgery to the retina or other parts of the eye. Cystoid macular edema (CME) is an example of this phenomenon. CME can occur in people who have had cataract surgery, as well as those with diabetes, retinitis pigmentosa, AMD, or conditions that cause chronic inflammation of the eye. The main symptom of CME is blurred or reduced central vision.

[0160] Ocular ischemic syndrome (OIS) encompasses signs and symptoms resulting from chronic vascular insufficiency. The condition is caused by ocular hypoperfusion due to occlusion or stenosis of the common or internal carotid arteries. OIS generally affects subjects aged 50-80 years who often exhibit systemic diseases such as hypertension or diabetes. The main symptoms of OIS are orbital pain, vision loss, changes in visual field, asymmetric cataracts, and sluggish response to light.

[0161] Retinal vein occlusion (RVO) is the most common retinal vascular disease following diabetic retinopathy. Depending on the area of ​​effectively occluded retinal venous drainage, the condition is broadly classified as central retinal vein occlusion (CRVO), hemiretinal vein occlusion (HRVO), or branch retinal vein occlusion (BRVO). The manifestation of RVO involves variable painless vision loss accompanied by any combination of fundus findings consisting of retinal vascular tortuosity, retinal hemorrhage (patchy and flake-like), cotton-wool spots, optic disc swelling, and macular edema. In CRVO, retinal hemorrhage can be found in all four quadrants of the fundus, whereas hemorrhage is restricted to either the superior or inferior fundus hemisphere in HRVO. In BRVO, hemorrhage is primarily localized in the area drained by the occluded branch retinal vein. Vision loss occurs secondary to macular edema or ischemia.

[0162] The compositions of the present disclosure act to stabilize the ocular vasculature, and in some embodiments, the agents of the present disclosure can counteract the irritation caused by VEGF and other inflammatory agents that may be present in the affected retina. In some embodiments, administration of the antibodies of the present disclosure to a subject can be used to maintain a level of disease reversal after administration of an anti-VEGF drug to the subject has been discontinued.

[0163] Treatment of Subjects In some embodiments, a single administration of a composition of the present disclosure in a subject having a disease or condition disclosed herein results in sustained intraocular expression of a VE-PTP (HPTP-β) inhibitor at levels sufficient for long-term inhibition of ocular neovascularization.

[0164] For example, the level of VE-PTP (HPTP-β) inhibitor produced in the host ocular cells may be at least 100 pg / mL, at least 200 pg / mL, at least 300 pg / mL, at least 400 pg / mL, at least 500 pg / mL, at least 600 pg / mL, at least 100 pg / mL, at least 800 pg / mL, at least 900 pg / mL, at least 1000 pg / mL, at least 2000 pg / mL, at least 3000 pg / mL, at least 4000 pg / mL, at least 5000 pg / mL, at least 6000 pg / mL, at least 7000 pg / mL, at least 8000 pg / mL, at least 9000 pg / mL or at least 10,000 pg / mL. The level of VE-PTP (HPTP-β) inhibitor produced in the host ocular cells may be at most 100 pg / mL, at most 200 pg / mL, at most 300 pg / mL, at most 400 pg / mL, at most 500 pg / mL, at most 600 pg / mL, at most 700 pg / mL, at most 800 pg / mL, at most 900 pg / mL, at most 1000 pg / mL, at most 2000 pg / mL, at most 3000 pg / mL, at most 4000 pg / mL, at most 5000 pg / mL, at most 6000 pg / mL, at most 7000 pg / mL, at most 8000 pg / mL, at most 9000 pg / mL, or at most 10,000 pg / mL.

[0165] Protein levels can be measured at least about 0.1, at least about 0.2, at least about 0.3, at least about 0.4, at least about 0.5, at least about 0.6, at least about 0.7, at least about 0.8, at least about 0.9, at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 14, at least about 21, at least about 30, at least about 50, at least about 75, at least about 100, at least about 125, at least about 150, at least about 175, at least about 200, at least about 225, at least about 250, at least about 275, at least about 300, at least about 325, at least about 350, or at least about 365 days after administration of a pharmaceutical composition of the disclosure. Protein levels may be measured at most about 0.1, at most about 0.2, at most about 0.3, at most about 0.4, at most about 0.5, at most about 0.6, at most about 0.7, at most about 0.8, at most about 0.9, at most about 1, at most about 2, at most about 3, at most about 4, at most about 5, at most about 6, at most about 7, at most about 14, at most about 21, at most about 30, at most about 50, at most about 75, at most about 100, at most about 125, at most about 150, at most about 175, at most about 200, at most about 225, at most about 250, at most about 275, at most about 300, at most about 325, at most about 350, or at most about 365 days after administration of a pharmaceutical composition of the present disclosure.

[0166] Central Foveal Thickness Also disclosed herein is a method of reducing central foveal thickness (CFT) in a subject having a disease or condition disclosed herein. The method includes administering a VE-PTP (HPTP-β) targeting antibody to the eye, and administration of the antibody may be performed in any manner.

[0167] The level of reduction in the foveal thickness may be, for example, about 50 μm to about 1000 μm. The level of reduction in the foveal thickness may be, for example, about 50 μm to about 500 μm, about 50 μm to about 750 μm, about 150 μm to about 500 μm, about 200 μm to about 500 μm, about 200 μm to about 1000 μm, about 250 μm to about 650 μm, or about 400 μm to about 700 μm.

[0168] eyesight Further disclosed herein are methods of increasing vision in a subject having a disease or condition disclosed herein.

[0169] Visual acuity is the acuteness or clearness of vision and depends on the sharpness of the retinal focus in the eye and the sensitivity of the interpretative faculty of the brain. Visual acuity is a measure of the spatial resolution of the visual processing system. Visual acuity is tested by requiring the subject to identify features, typically numbers or letters, on a chart from a set distance. The chart features are represented as black symbols against a white background. The distance between the subject's eye and the test chart is set far enough to approach infinity in the way the lens tries to focus. 20 feet, or 6 meters, is essentially infinity from visual perspective. In this disclosure, improvement in visual acuity was assessed by an increase in the number of letters that could be read from the chart.

[0170] One non-limiting test for measuring visual acuity is the use of the ESV-3000 ETDRS testing device and self-calibrating test illumination. The ESV-3000 device incorporates LED light source technology. Auto-calibration circuitry constantly monitors the LED light source and adjusts the test luminance to 85 cd / m 2 or 3 cd / m 2 Calibrate to.

[0171] Although designed for clinical trials where the large ETDRS test (up to 20 / 200) is performed at 4 meters, the device can be used in non-research settings, i.e., hospitals or clinics where ocular disease monitoring is performed. To properly evaluate the ETDRS, the test should be performed under standardized lighting conditions, e.g., 85 cd / m 2 The visual acuity should be scored at a photopic test level of 100x100x100mm. Scoring of visual acuity can be accomplished in any manner selected by the monitor. After providing a baseline assessment, an increase or decrease in the number of letters the test subject can distinguish is a measure of the increase or decrease in visual acuity during treatment.

[0172] Disclosed herein is a method of increasing visual acuity in a subject having an ocular disease or condition disclosed herein. The method includes administering a VE-PTP (HPTP-β) targeting antibody to a subject having an ocular disease or condition, where administration of the antibody can be in any manner described herein. The increase in the number of letters recognized by the treated eye can be, for example, about 1 to about 30 letters, about 5 to about 25 letters, about 5 to about 20 letters, about 5 to about 15 letters, about 5 to about 10 letters, about 10 to about 25 letters, about 15 to about 25 letters, or about 20 to about 25 letters. The increase in visual acuity can be about 1 letter, about 5 letters, about 10 letters, about 15 letters, about 20 letters, or about 25 letters. EXAMPLES

[0173] Example 1: Generation of monoclonal antibodies against the VE-PTP (HPTP-β) extracellular domain using hybridoma technology.

[0174] Hybridoma technology was used to generate monoclonal antibodies against the N-terminal extracellular domain (ECD) of human VE-PTP (HPTP-β) or mouse VE-PTP, as shown in the schematic diagram in Figure 3. Mice were challenged with human VE-PTP (HPTP-β) ECD protein to generate antibodies against the target protein. B cells were then harvested from the spleen and lymph nodes and fused with myeloma cells to generate hybridoma cell lines. The fused cells were cultured in a medium in which only myeloma cells carrying genes derived from the antigen-exposed mouse could grow. Colonies derived from single myeloma cells were generated by limiting dilution. Antibodies produced by the hybridoma colonies were then screened for binding to human VE-PTP (HPTP-β) ECD by ELISA. One hybridoma, R15E6, was selected for further analysis. The R15E6 antibody bound to endogenous human VE-PTP (HPTP-β) from human umbilical vein endothelial cells (HUVEC) as shown by immunoprecipitation (IP) Western blot (left panel) and flow cytometry (right panel) in Figure 4. No binding of R15E6 to bead controls (protein A / G beads without antibody; BC) or to VEGFR2 or Tie2 (R2 / T2) was observed.

[0175] Example 2: Tie2 activation in HUVECs by R15E6.

[0176] R15E6 activated Tie2 in a concentration-dependent manner, as demonstrated by IP experiments shown in Figure 6, panels A and B. The antibody also increased the viability of serum-starved endothelial cells in a concentration-dependent manner in HUVEC, as shown in panel C.

[0177] Example 3: Tie2 activation and cell permeability experiments using murine monoclonal and polyclonal anti-mouse VE-PTP ECD antibodies.

[0178] To characterize the therapeutic effects of VE-PTP antibodies in vitro, Tie2 activation and cell permeability experiments were performed using anti-mouse VE-PTP antibodies. Monoclonal and polyclonal antibodies against mouse VE-PTP ECD were generated by immunizing rats with VE-PTP-Fc fusion protein of the N-terminal 8 FN3 repeats of the extracellular domain. Immunization, hybridoma fusion, and screening were performed as described above. In Example 3, monoclonal antibody 109.1 (mAb 109.1) and polyclonal antibodies (pAbs) against the extracellular fibronectin type III (FN3)-like domains 1-8 of mouse VE-PTP were selected for further analysis.

[0179] Figure 7 shows the effect of mAb 109.1 on Tie2 activation (upper left panel), mouse VE-PTP protein expression (lower left panel), and Akt (right panel) in mouse endothelial cells (bEnd) as determined by Western blot. Treatment with mAb 109.1 resulted in activation of Tie2, as indicated by Tie2 phosphorylation (pTyr; upper left panel). Akt was constitutively activated in the mouse endothelial cells tested, as confirmed by the absence of increased phosphorylation with increasing concentrations of antibody (right panel).

[0180] In vitro experiments with mouse pAb 1-8 also enhanced Tie2 (Tie-2) activation and downstream signaling in endothelial cells. Cultured mouse endothelial (bEnd.5) cells were treated with pAb1-8 or preimmune (control) antibodies for 1 hour, followed by immunoprecipitation for Tie2 and then immunoblotting with anti-phosphotyrosine (pTyr) and antibodies against Tie2. Aliquots of cell lysates with identical protein content were directly immunoblotted for VE-PTP and Tie2 (lower panel). Figure 8 shows immunoprecipitation experiments in which pAbs rapidly induced Tie2 activation in bEnd.5 cells treated with polyclonal or preimmune antibodies against mouse VE-PTP for 1 hour (panel A) or 3 minutes (panel B).

[0181] Consistent with the vascular stabilizing effect of Tie2 activation, VE-PTP ECD antibodies reduced thrombin- and VEGF-induced permeability of endothelial monolayers in vitro, as shown in FIG. 9. Paracellular permeability for 250 kD FITC-dextran was determined for cultured HUVEC monolayers grown in transwell filters. Permeability was induced by either thrombin (panel A) or VEGF (panel B). Permeability of PBS-treated cells was set to 100%. For VE-PTP targeting, cells were treated with anti-VE-PTP pAb. As a control, cells were treated with preimmune antibody (control Ab). Data represent two independent experiments.

[0182] Consistent with the vascular stabilizing effect of Tie2 activation, VE-PTP ECD antibodies also blocked VEGF-induced skin vascular permeability in vivo, as shown in FIG. 10. Both monoclonal and polyclonal VE-PTP ECD antibodies, mAb 109.1 (panel A) and pAb PTP 1-8 (panel B), inhibited VEGF-induced skin vascular permeability as demonstrated by the Miles assay. The Miles assay can be used as an in vivo model of vascular permeability, a characteristic of vascular leakage and neovascularization. Mice were intravenously injected with 100 μg of control IgG or anti-mouse VE-PTP antibody 30 minutes prior to initiating the Miles assay. Mice were then intravenously injected with Evans Blue, followed 10 minutes later by intradermal injection of VEGF (striped bars) or PBS (solid black bars). After 30 minutes, mice were sacrificed and the dye was extracted from the skin samples and quantified. Data sets were examined for normality and equal variance. Values ​​are presented as the mean ± standard error of the mean (SEM). As shown in panel C, permeability block correlates with Tie2 phosphorylation, as demonstrated in lung tissue of mice treated with pAb PTP1-8.

[0183] Example 4: Mouse VE-PTP on retinal and choroidal neovascularization in mice Effect of ECD monoclonal antibody.

[0184] In an ischemic retinopathy model that mimics aspects of proliferative diabetic retinopathy, VE-PTP ECD monoclonal antibody (mAb 109.1) significantly reduced retinal neovascularization following antibody administration, as shown in Figure 11 (Panel A) and quantified in Figure 12 (Panel A). At P12, mice with ischemic retinopathy were administered 0.1, 0.5, or 2 μg of mAb 109.1 or 2 μg of IgG isotype control (n≧12 for each) via intravitreal injection. At P17, extensive GSA-stained retinal neovascularization was observed in eyes injected with control IgG, and significantly less retinal neovascularization was observed in eyes injected with 2 μg of anti-VE-PTP. * P<0.001.

[0185] Similarly, in two models mimicking different aspects of wet age-related macular edema (retinal hemangioma proliferation and choroidal neovascularization), VE-PTP ECD monoclonal antibody (mAb A single intravitreal dose of 2 μg of mAb 109.1 significantly reduced both retinal neovascularizations (quantified in FIG. 11 panel B and FIG. 12 panel B). At P15, six Rho-VEGF transgenic mice were intravitreally injected with 0.5 or 2 μg of mAb 109.1 in one eye and the other eye received the corresponding dose of control IgG. At P21, significantly less GSA-stained subretinal neovascularization was observed in eyes treated with 0.5 or 2 μg of mAb 109.1 than in eyes treated with control IgG. * P=0.01 by unpaired t-test for comparison with IgG control fellow eye. Scale bar: 100 μm.

[0186] Intravitreal injection of 2 μg of mAb 109.1 significantly reduced the area of ​​choroidal neovascularization at the site of Bruch's membrane rupture compared to control IgG (quantified in FIG. 11, panel C and FIG. 12, panel C). * P<0.001 by one-way ANOVA with Bonferroni correction. Scale bar: 100 μm.

[0187] Subcutaneous administration of mAb 109.1 also reduced ischemic neovascularization in a mouse model of ischemic retinopathy. The average area of ​​retinal neovascularization was reduced by about 30% compared to vehicle, as shown in Figure 25. Mice with ischemic retinopathy were administered subcutaneous injections of 2 mg / kg qod (every other day) x 3.

[0188] Example 5: Effect of mouse VE-PTP ECD monoclonal antibody in combination with an anti-VEGF agent on retinal detachment in mice.

[0189] To evaluate the efficacy of combination therapy using anti-VE-PTP antibodies disclosed herein, a Tet / opsin / VEGF mouse model was used. Tet / opsin / VEGF transgenic mice develop severe subretinal neovascularization and vascular leakage, leading to exudative retinal detachment mediated by inducible retinal overexpression of human VEGF. Figure 24 shows the enhanced efficacy of the combination of anti-mouse VE-PTP antibody (mAb 109.1) and aflibercept compared to either therapy alone to prevent retinal detachment in mice, as indicated by double asterisks. Rat IgG was used as a control.

[0190] Example 6: Generation and purification of humanized antibodies derived from R15E6 against human VE-PTP (HPTP-β).

[0191] R15E6 is a 150 kDa antibody containing heavy and light chains. Sixteen different humanized variants of the antibody were synthesized using different combinations of heavy and light chain sequences, as shown in Table 12.

[0192] The R15E6 humanized variants described in Example 1 were cloned into the pVitro DHFR3 mammalian expression vector. A medium-scale transient transfection expression assay was performed to determine expression yields from CHO cells. Suspension-adapted CHO cells were cultured at 2.0-3.0 × 10 in Pro CHO4 serum-free medium supplemented with 8 mM L-glutamine and 10 mL / L hypoxanthine / thymidine in 500 mL vented flasks at 37 °C, 150 rpm and 85% CO2. 5 The vectors were cultured at 1000 cells / mL. Maxi-preps of each construct were prepared using the Nucleobond pc500 kit according to the manufacturer's instructions. Vector DNA was quantified using a NanoDrop Lite™ spectrophotometer.

[0193] 1.0×10 6 Cells at a final density of 100 mL / mL were transiently transfected with 1.25 μg / mL plasmid DNA in Pro CHO5 serum-free medium supplemented with 8 mM L-glutamine and 10 mL / L hypoxanthine / thymidine in 500 mL vented flasks. Transfected cultures were incubated at 37°C, 150 rpm, 85% CO2 for 8-11 days and then harvested by centrifugation at 4,000 rpm, 4°C for 40 min.

[0194] After centrifugation, the medium was filtered through a 0.8 μm cellulose acetate filter. Each batch was purified using an Amersham Biosciences AKTA chromatography system. The expressed humanized antibodies were purified by Protein A affinity chromatography. A 1 mL HiTrap Protein A column was used for all humanized antibody purifications. All purifications were performed using Fusion Antibodies in-house washing and elution buffers. After the protein was loaded onto the column, any bound antibodies were eluted using glycine / Tris buffer (pH 3). All eluted 1 mL fractions were neutralized with 100 μL Tris buffer (pH 8.5). The elution fractions corresponding to the elution peak were selected for overnight dialysis in PBS. After dialysis, the concentration of the antibody was measured by a NanoDrop Lite™ spectrophotometer.

[0195] Example 7: Evaluation of humanized antibodies against human VE-PTP (HPTP-β).

[0196] Samples of the humanized antibodies were analyzed by reducing and non-reducing SDS-PAGE (Figure 13) to determine the presence of heavy and light chains and the purity of the antibodies. The mouse / human chimeric antibody, HC0LC0, was derived from R15E6 by fusion of the murine heavy and light chain variable regions with a human Fc region. In both SDS-PAGE gels, lane 1: SeeBlue Plus2 prestained protein standard; lane 2: HC0LC0 #1; lane 3: HC1LC1 #2; lane 4: HC1LC2 #1; lane 5: HC1LC3 #1; lane 6: HC1LC3 #2; lane 7: HC1LC4 #1; lane 8: HC2LC1 #1; lane 9: HC2LC2 #1; lane 10: HC2LC3 #1; lane 11: HC2LC3 #2; lane 12: HC2LC4 #1; lane 13: HC3LC1 #1; lane 14: HC3LC2 #1; lane 15: HC3LC3 #1; lane 16: HC3LC4 #1; lane 17: HC4LC1 #1; lane 18: HC4LC1 #2; lane 19: HC4LC2 #1; lane 20: HC4LC3 #1; lane 21: HC4LC3 #2; lane 22: HC4LC4 #1; lane 23: HC4LC4 #2; and lane 24: SeeBlue Plus2 prestained protein standard. SDS-PAGE gel analysis showed high purity of all analyzed samples. In reducing SDS-PAGE, heavy and light chains were visible with estimated molecular weights of 50 kDa for the heavy chain and 25 kDa for the light chain. In non-reducing SDS-PAGE, no free heavy or light chains were observed at approximately 50 kDa or 25 kDa, respectively. The gel shows low amounts of protein for all HC4 variants as well as HC1LC3, HC2LC3 and HC3LC3 variants.

[0197] Purified R15E6 was quantified using a NanoDrop Lite™ spectrophotometer, using an extinction coefficient of 13.7 as the standard for IgG at 280 nm. Table 16 summarizes the results of the SDS-PAGE analysis. Consistent with the SDS-PAGE analysis, all four variants of the heavy chain expressed little or no antibody. [Table 15]

[0198] Example 8: Selectivity of R15E6 and humanized variants for human VE-PTP (HPTP-β), cynomolgus monkey VE-PTP (cyno PTP-β), and human HPTP-η.

[0199] Tissue culture supernatants containing antibodies from the hybridomas were tested against targets human VE-PTP (human beta1 / 2ECD), cyno VE-PTP (Cyno beta), and human HPTP-η (human eta) to determine binding affinity by ELISA.

[0200] Human HPTP-β, cyno PTPβ, and human HPTP-η were independently produced and purified using HEK293 cells. HEK293 cells were seeded in shake flasks for 24 hours and then transfected with plasmids expressing species-specific VE-PTP or HPTP-η. HEK293 cells were grown using serum-free, chemically defined medium. DNA for species-specific VE-PTP or HPTP-η constructs was transiently transfected into 30 mL suspensions of HEK293. After 24 hours, cells were counted to assess cell viability and determine viable cell numbers. Further readings were taken throughout the transient transfection process. Cell cultures were harvested on the fifth day of transient transfection. Conditioned medium supernatants harvested from transient transfections were clarified by centrifugation. The supernatants were filtered using a 0.2 μm membrane filter. Proteins were then purified by anti-His tag affinity chromatography. After purification, the protein was buffer exchanged into PBS (pH 7.4). SDS-gel capillary electrophoresis (CE-SDS) analysis was then performed to purify the protein. The amino acid sequence of the DNA plasmid insert, the DNA plasmid insert, and the corresponding wild-type sequence are shown in Table 15 below. The signal peptide of the protein sequence is underlined. [Table 16-1] [Table 16-2] [Table 16-3] [Table 16-4] [Table 16-5] [Table 16-6] [Table 16-7] [Table 16-8] [Table 16-9] [Table 16-10]

[0201] HPTP-β, cyno PTP-β, and HPTP-η proteins were coated onto ELISA plates in coating buffer (pH 9.4). Each protein was prepared as a 1 μg / mL protein stock in 50 mM carbonate-bicarbonate. The coated proteins were aliquoted into sterile clear polystyrene flat-bottom 96-well plates at a volume of 100 μL / well. The plates were incubated overnight at 4° C. After overnight incubation, the plates were washed three times with 200 μL / well of 1×PBS-T (PBS containing 0.05% Tween 20). The immunizing protein was human VE-PTP (HPTP-β). The plates were then blocked with 200 μL / well of 5% blotting grade blocker nonfat dry milk in PBS-T and incubated at room temperature for 1 hour.

[0202] Primary antibody samples were prepared for the humanized variants and negative control, anti-hCD20-hIgG4S228P, using 1 μg / well of 5% blotting grade blocker nonfat dry milk in PBS. Samples were tested in triplicate. Fab antibody fragments were treated as IgG antibodies. Plates were then washed three times with PBS-T and 100 μL of primary antibody preparation was added to each well. Plates were then incubated at room temperature for 1 hour.

[0203] Secondary antibody samples were prepared by diluting peroxidase-conjugated, affinity-purified donkey anti-human IgG specific secondary antibody at a 1:2,000 dilution in blotting grade blocker nonfat dry milk in PBS. The plate was then washed 3 times with PBS-T and 100 μL of the secondary antibody preparation was added to each well. The plate was then incubated for 1 hour at room temperature. After incubation, the plate was washed 3 times with PBS-T.

[0204] 75 μg of 1-Step™ Ultra TMB-ELISA was then added to each well and the plate was allowed to develop for 5 minutes at room temperature. After 5 minutes, 50 μL of 2M sulfuric acid was added to each well to stop the reaction. The plate was then analyzed by a plate reader at an absorbance wavelength of 450 nm.

[0205] Figure 5 shows a Western blot of R15E6 screened against species-specific VE-PTP and HPTP-η. R15E6 was highly selective for human VE-PTP (HPTP-β; lanes 1 and 4). No significant binding to human PTP-η or cyno VE-PTP (cyno PTP-β) was observed. The indicated recombinant hexahistidine-tagged proteins (6-His) were sequentially probed with R15E6 and a commercially available 6-His antibody. The commercially available 6-His antibody showed binding to all protein targets and showed poor selectivity for human VE-PTP (HPTP-β).

[0206] Each humanized variant was measured for affinity to the antigen, human VE-PTP (HPTP-β), using ELISA. 100 mg / well of VE-PTP (HPTP-β) antigen was immobilized on a 96-well Maxisorp plate in coating buffer (0.5 mM NaHCO3, pH 9.6 by addition of 0.5 mM Na2CO3) overnight at 4°C. Coating buffer was then removed and 200 μL / well of blocking solution (3% w / v semi-skimmed milk powder, PBS) was added and agitated for 2 hours at room temperature. Plates were washed 4 times with PBS-T (1% v / v Tween20). 100 μL / well was added to purified R15E6 variants serially diluted from 10,000 ng / mL to 9.765 ng / mL in PBS-TB (0.05% v / v Tween20, 0.5% w / v BSA). After 2 hours of agitation at room temperature, the plate was washed 4 times with PBS-T. 100 μL / well of goat anti-human HRP (Fc specific) was then added at a ratio of 1:60,000 PBS and the plate was incubated for an additional hour with agitation at room temperature. The plate was then washed 6 times with PBS-T and once in PBS. 100 μL / well of TMB substrate solution was then added and incubated for 10 minutes at 37°C. 50 μL of 1M HCl was added per well and the plate was immediately read at 450 nm on a Tecan Sunrise plate reader. The ELISA results are shown in Figures 14-17. The HC1, HC2 and HC3 variants generally showed similar binding properties compared to the chimeric HC0LC0 antibody. The binding of the HC4 variant was much lower than the binding of the other heavy chain variants compared to HC0LC0 (Figure 17).

[0207] Samples of each variant were further evaluated by Biacore analysis to determine binding affinity. Antibody binding experiments were performed on a Biacore 3000™ at 25° C. Assay buffer: 10 mM HEPES buffer (pH 7.4), 150 mM NaCl, 3 mM EDTA, 0.05% P20 (polyoxyethylene sorbitan). Regeneration buffer: 10 mM glycine HCl (pH 1.75). Conjugation buffer: 10 mM sodium acetate buffer (pH 5). The flow rate used to capture the ligand was 5 μL / min. The flow rate for kinetic analysis was 50 μL / min. Flow cells 1 and 2 of a CM5 chip were coated with a maximum amount of goat anti-human IgG. Approximately 150 RU of test antibody was captured on flow cells 2, 3, and 4 as indicated. Antigen was flowed over the chip. Binding of antigen to antibody was monitored in real time. K D I observed on and k off was determined from the value.

[0208] Scouting analysis was performed using single analyte concentrations as indicated. At these concentrations, binding can be observed even if the ligand binding is weak. The response of flow cell 1 was used for baseline subtraction.

[0209] A complete kinetic analysis was performed on a control antibody in two-fold serial dilutions across the following analyte concentration range: 50, 25, 12.5, 6.25, 3.125, and 0 nM.

[0210] Table 17 shows the binding rates (k a ), dissociation rate (k d ), maximum binding capacity (R max ), coupling constant (K A ), and the dissociation constant (K D The binding kinetic parameters of each humanized antibody binding to VE-PTP (HPTP-β) ECD are shown. max Responses were measured as relative responses (RU). All of the humanized candidates showed similar binding affinities to the mouse / human chimeric antibody, HC0LC0. [Table 17]

[0211] Table 18 shows the binding selectivity of the humanized antibodies compared to the parent antibody (R15E6) and the inactive control (IgG4) as determined by ELISA. Values ​​less than or equal to 0.2 indicate no binding. The humanized antibody candidates, the parent R15E6, and the mouse / human chimeric antibody (HC0:LC0) all showed high selectivity for human VE-PTP (HPTP-β) as shown by the lack of significant binding to human PTP-η and cyno PTP-β. [Table 18]

[0212] The humanized antibodies used to target human VE-PTP (HPTP-β) can restore Tie2 activity by autophosphorylation and initiate downstream effectors, including, for example, proteins in the PI3K-Akt pathway. In addition to assessing the binding of the antibodies to human VE-PTP (HPTP-β), the ability of the antibodies to activate these effector proteins was measured. Figure 18 shows the activation of Tie2 (left panel) and Akt (right panel) by humanized variants in the absence of Tie2 ligand (e.g., Ang1 or Ang2) as determined by Western blot. The activity of the humanized variants was similar to that of the parent murine antibody R15E6 and the chimeric antibody HC0LC0. Human IgG4 was used as a control. All antibodies were tested at 50 nM. The top panel of each blot shows phosphorylation of either Tie2 or Akt, indicating Tie2 or Akt activation. Tie2 and Akt showed higher phosphorylation in samples treated with the humanized antibodies than in samples treated with the control antibody, indicating that the humanized antibodies were effective in activating Tie2 and Akt even in the absence of ligand.

[0213] Figure 19 shows the concentration-dependent activation of Tie2 (upper panel) and Akt (lower panel) by humanized antibodies, HC2LC4 and HC1LC1, in the absence of Tie2 ligand (e.g., Ang1 or Ang2), as determined by Western blot. Tie2 and Akt signaling activity was observed at low nanomolar concentrations of humanized variants. The upper panel of each blot shows phosphorylation of either Tie2 or Akt, indicating Tie2 or Akt activation. Tie2 and Akt showed higher phosphorylation in samples treated with humanized antibodies than that shown in samples treated with control antibodies. This result indicates that the humanized antibodies were effective in activating Tie2 and Akt even in the absence of Ang1.

[0214] Figure 20 shows activation of Tie2 (left panel) and Akt (right panel) by humanized antibody candidates, HC2LC4 and HC1LC1, in the presence of Ang1 and Ang2, as determined by Western blot. HC2LC4 and HC1LC1 activated both Tie2 and Akt in the presence of Ang2, enhancing Ang1-mediated Tie2 and Akt activation, as indicated by increased phosphorylation of Tie2 and Akt, respectively. All antibodies were tested at 10 nM.

[0215] Figure 21 shows activation of Tie2 (panel A), human VE-PTP (HPTP-β) expression (panel B), and Akt (panel C) in the presence of Ang1 or Ang2 by humanized antibody candidates, HC2LC1 and HC2LC4, as determined by Western blot. HC2LC1 and HC2LC4 activated Tie2 in the presence of Ang1 alone, Ang2 alone, and both Ang1 and Ang2. Antibodies were preincubated for 15 minutes and then stimulated with Ang1 or Ang2 at a concentration of 600 ng / mL for 10 minutes. All antibodies were tested at 10 nM.

[0216] FIG. 22 shows activation of Akt in HUVEC using the humanized antibody candidate, HC2LC1. The top panel of the first three blots shows phosphorylation of Akt, indicating Akt activation, at increasing concentrations of HC2LC1. Consistent with the binding affinity studies, HC2LC1 activated Akt at nanomolar concentrations of antibody. Akt activation by HC2LC1 was blocked by preincubation of the antibody with human VE-PTP (HPTP-β) extracellular domain (βECD 6His and βECD Fc). This result indicates that HC2LC1-mediated Akt activation occurs via HC2LC1 binding to the human VE-PTP (HPTP-β) extracellular domain.

[0217] FIG. 23 shows improved viability in serum-starved HUVEC using humanized antibody candidates HC2LC1 and HC0LC0 compared to the parental antibody, R15E6, and the mIgG1 control.

[0218] Embodiment Embodiment 1. A compound comprising a sequence that is at least 80% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12.

[0219] Embodiment 2. The compound of embodiment 1, wherein the sequence is at least 85% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12.

[0220] Embodiment 3. The compound of any one of embodiments 1-2, wherein the sequence is at least 90% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12.

[0221] Embodiment 4. The compound of any one of embodiments 1-3, wherein the sequence is at least 95% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12.

[0222] Embodiment 5. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:9.

[0223] Embodiment 6. The compound of any one of embodiments 1 to 5, wherein the sequence is SEQ ID NO:9.

[0224] Embodiment 7. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:10.

[0225] Embodiment 8. The compound of any one of embodiments 1-4 and 7, wherein the sequence is SEQ ID NO:10.

[0226] Embodiment 9. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:11.

[0227] Embodiment 10. The compound of any one of embodiments 1-4 and 9, wherein the sequence is SEQ ID NO:11.

[0228] Embodiment 11. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:12.

[0229] Embodiment 12. The compound of any one of embodiments 1-4 and 11, wherein the sequence is SEQ ID NO:12.

[0230] Embodiment 13. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:29.

[0231] Embodiment 14. The compound of embodiment 1, wherein the sequence is SEQ ID NO:29.

[0232] Embodiment 15. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:30.

[0233] Embodiment 16 The compound of embodiment 1, wherein the sequence is SEQ ID NO:30.

[0234] Embodiment 17. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:31.

[0235] Embodiment 18 The compound of embodiment 1, wherein the sequence is SEQ ID NO:31.

[0236] Embodiment 19. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:32.

[0237] Embodiment 20 The compound of embodiment 1, wherein the sequence is SEQ ID NO:32.

[0238] Embodiment 21. A compound of any one of embodiments 1 to 20, which inhibits a tyrosine phosphatase.

[0239] Embodiment 22. A compound of any one of embodiments 1 to 21, which inhibits HPTP-β.

[0240] Embodiment 23. A compound of any one of embodiments 1 to 22, which inhibits VE-PTP.

[0241] Embodiment 24. A compound of any one of embodiments 1 to 23, which activates Tie2.

[0242] Embodiment 25. A compound of any one of embodiments 1 to 24 which activates Akt.

[0243] Embodiment 26. A compound of any one of embodiments 1 to 25, which binds to the extracellular domain of HPTP-β.

[0244] Embodiment 27. A compound of any one of embodiments 1 to 26, which binds to the first FN3 repeat of the extracellular domain of HPTP-β.

[0245] 28. Binding affinity of a compound to the extracellular domain of HPTP-β (K D 28. The compound of any one of embodiments 1-27, wherein the .DELTA.I.I.

[0246] Embodiment 29. A compound comprising a sequence that is at least 80% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23.

[0247] Embodiment 30. The compound of embodiment 29, wherein the sequence is at least 85% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23.

[0248] Embodiment 31. The compound of any one of embodiments 29-30, wherein the sequence is at least 90% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23.

[0249] Embodiment 32. The compound of any one of embodiments 29 to 31, wherein the sequence is at least 95% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23.

[0250] Embodiment 33 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:20.

[0251] Embodiment 34. The compound of any one of embodiments 29 to 33, wherein the sequence is SEQ ID NO: 20.

[0252] Embodiment 35 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:21.

[0253] Embodiment 36 The compound of any one of embodiments 29 to 32 and 35, wherein the sequence is SEQ ID NO: 21.

[0254] Embodiment 37 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:22.

[0255] Embodiment 38. The compound of any one of embodiments 29 to 32 and 37, wherein the sequence is SEQ ID NO: 22.

[0256] Embodiment 39. The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:23.

[0257] Embodiment 40. The compound of any one of embodiments 29 to 32 and 39, wherein the sequence is SEQ ID NO: 23.

[0258] Embodiment 41 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:34.

[0259] Embodiment 42. The compound of embodiment 29, wherein the sequence is SEQ ID NO:34.

[0260] Embodiment 43 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:35.

[0261] Embodiment 44 The compound of embodiment 29, wherein the sequence is SEQ ID NO:35.

[0262] Embodiment 45 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:36.

[0263] Embodiment 46 The compound of embodiment 29, wherein the sequence is SEQ ID NO:36.

[0264] Embodiment 47 The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:37.

[0265] Embodiment 48 The compound of embodiment 29, wherein the sequence is SEQ ID NO:37.

[0266] Embodiment 49. A compound of any one of embodiments 29 to 48, which inhibits a tyrosine phosphatase.

[0267] Embodiment 50. A compound of any one of embodiments 29-49, which inhibits HPTP-β.

[0268] Embodiment 51. A compound of any one of embodiments 29-50, which inhibits VE-PTP.

[0269] Embodiment 52. A compound of any one of embodiments 29 to 51, which activates Tie2.

[0270] Embodiment 53. A compound of any one of embodiments 29 to 52, which activates Akt.

[0271] Embodiment 54. A compound of any one of embodiments 29 to 53, which binds to the extracellular domain of HPTP-β.

[0272] Embodiment 55. The compound of any one of embodiments 29 to 54, which binds to the first FN3 repeat of the extracellular domain of HPTP-β.

[0273] Binding affinity of a compound to the extracellular domain of HPTP-β (K D 56. The compound of any one of embodiments 29-55, wherein the .times. ...

[0274] Embodiment 57. A compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:34.

[0275] Embodiment 58. The compound of embodiment 57, wherein the heavy chain is at least 85% identical to SEQ ID NO:30 and the light chain is at least 85% identical to SEQ ID NO:34.

[0276] Embodiment 59. The compound of any one of embodiments 57-58, wherein the heavy chain is at least 90% identical to SEQ ID NO:30 and the light chain is at least 90% identical to SEQ ID NO:34.

[0277] Embodiment 60. The compound of any one of embodiments 57-59, wherein the heavy chain is at least 95% identical to SEQ ID NO:30 and the light chain is at least 95% identical to SEQ ID NO:34.

[0278] Embodiment 61. The compound of any one of embodiments 57-60, wherein the heavy chain is SEQ ID NO: 30 and the light chain is SEQ ID NO: 34.

[0279] Embodiment 62. A compound of any one of embodiments 57-61, which inhibits a tyrosine phosphatase.

[0280] Embodiment 63. A compound of any one of embodiments 57-62, which inhibits HPTP-β.

[0281] Embodiment 64. A compound of any one of embodiments 57-63, which inhibits VE-PTP.

[0282] Embodiment 65. A compound of any one of embodiments 57-64, which activates Tie2.

[0283] Embodiment 66. A compound of any one of embodiments 57-65, which activates Akt.

[0284] Embodiment 67. A compound of any one of embodiments 57-66, which binds to the extracellular domain of HPTP-β.

[0285] Embodiment 68. The compound of any one of embodiments 57 to 67, which binds to the first FN3 repeat of the extracellular domain of HPTP-β.

[0286] 69. The binding affinity (K D 69. The compound of any one of embodiments 57-68, wherein the .times. ...

[0287] Embodiment 70. A compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:29; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:35.

[0288] Embodiment 71. The compound of embodiment 70, wherein the heavy chain is at least 85% identical to SEQ ID NO:29 and the light chain is at least 85% identical to SEQ ID NO:35.

[0289] Embodiment 72. The compound of any one of embodiments 70-71, wherein the heavy chain is at least 90% identical to SEQ ID NO:29 and the light chain is at least 90% identical to SEQ ID NO:35.

[0290] Embodiment 73. The compound of any one of embodiments 70-72, wherein the heavy chain is at least 95% identical to SEQ ID NO:29 and the light chain is at least 95% identical to SEQ ID NO:35.

[0291] Embodiment 74. The compound of any one of embodiments 70 to 73, wherein the heavy chain is SEQ ID NO: 29 and the light chain is SEQ ID NO: 35.

[0292] Embodiment 75. A compound of any one of embodiments 70-74, which inhibits a tyrosine phosphatase.

[0293] Embodiment 76. A compound of any one of embodiments 70-75, which inhibits HPTP-β.

[0294] Embodiment 77. A compound of any one of embodiments 70-76, which inhibits VE-PTP.

[0295] Embodiment 78. A compound of any one of embodiments 70 to 77 which activates Tie2.

[0296] Embodiment 79. A compound of any one of embodiments 70 to 78, which activates Akt.

[0297] Embodiment 80. A compound of any one of embodiments 70-79, which binds to the extracellular domain of HPTP-β.

[0298] Embodiment 81. The compound of any one of embodiments 70 to 80, which binds to the first FN3 repeat of the extracellular domain of HPTP-β.

[0299] Embodiment 82. Binding affinity of a compound to the extracellular domain of HPTP-β (K D 82. The compound of any one of embodiments 70-81, wherein the .times. ...

[0300] Embodiment 83. A compound comprising: a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence that is at least 80% identical to SEQ ID NO:37.

[0301] Embodiment 84. The compound of embodiment 83, wherein the heavy chain is at least 85% identical to SEQ ID NO:30 and the light chain is at least 85% identical to SEQ ID NO:37.

[0302] Embodiment 85. The compound of any one of embodiments 83-84, wherein the heavy chain is at least 90% identical to SEQ ID NO:30 and the light chain is at least 90% identical to SEQ ID NO:37.

[0303] Embodiment 86 The compound of any one of embodiments 83 to 85, wherein the heavy chain is at least 95% identical to SEQ ID NO: 30 and the light chain is at least 95% identical to SEQ ID NO: 37.

[0304] Embodiment 87. The compound of any one of embodiments 83 to 86, wherein the heavy chain is SEQ ID NO: 30 and the light chain is SEQ ID NO: 37.

[0305] Embodiment 88. A compound of any one of embodiments 83 to 87, which inhibits a tyrosine phosphatase.

[0306] Embodiment 89. A compound of any one of embodiments 83-88, which inhibits HPTP-β.

[0307] Embodiment 90. A compound of any one of embodiments 83-89, which inhibits VE-PTP.

[0308] Embodiment 91. A compound of any one of embodiments 83 to 90 which activates Tie2.

[0309] Embodiment 92. A compound of any one of embodiments 83 to 91, which activates Akt.

[0310] Embodiment 93. A compound of any one of embodiments 83 to 92, which binds to the extracellular domain of HPTP-β.

[0311] Embodiment 94. The compound of any one of embodiments 83 to 93, which binds to the first FN3 repeat of the extracellular domain of HPTP-β.

[0312] Embodiment 95. The binding affinity (K D ) is from about 70 pM to about 70 nM.

[0313] Embodiment 96. A method of treating a condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of any one of embodiments 1 to 95.

[0314] Embodiment 97. The method of embodiment 96, wherein the condition is an ophthalmic condition.

[0315] Embodiment 98. The method of embodiment 96 or 97, wherein the condition is diabetic retinopathy.

[0316] Embodiment 99. The method of any one of embodiments 96-97, wherein the condition is neovascularization.

[0317] Embodiment 100. The method of any one of embodiments 96-97, wherein the condition is vascular leakage.

[0318] Embodiment 101. The method of any one of embodiments 96-97, wherein the condition is elevated intraocular pressure.

[0319] Embodiment 102. The method of any one of embodiments 96-97, wherein the condition is ocular edema.

[0320] Embodiment 103. The method of embodiment 96 or 97, wherein the condition is diabetic macular edema.

[0321] Embodiment 104. The method of any one of embodiments 96 or 97, wherein the condition is ocular hypertension.

[0322] Embodiment 105. The method of embodiment 96 or 97, wherein the condition is ocular inflammation.

[0323] Embodiment 106. The method of any one of embodiments 96 to 105, wherein administration is to an eye of the subject.

[0324] Embodiment 107. The method of any one of embodiments 96 to 106, wherein administration is intravitreal.

[0325] Embodiment 108. The method of any one of embodiments 96 to 105, wherein administration is subcutaneous.

[0326] Embodiment 109. The method of any one of embodiments 96 to 106, wherein administration is local.

[0327] Embodiment 110. The method of any one of embodiments 96-109, wherein the subject is a human.

[0328] Embodiment 111. The method of any one of embodiments 96-110, wherein the therapeutically effective amount of the compound is from about 0.25 mg to about 200 mg.

[0329] Embodiment 112. The method of any one of embodiments 96-110, wherein the therapeutically effective amount of the compound is from about 1 mg / kg to about 10 mg / kg.

[0330] Embodiment 113. The method of any one of embodiments 96-110, wherein the therapeutically effective amount of the compound is from about 1 mg to about 50 mg.

[0331] Embodiment 114. The method of any one of embodiments 96-110, wherein the therapeutically effective amount of the compound is from about 50 mg to about 200 mg. The present invention provides, for example, the following items. (Item 1) A compound comprising a sequence that is at least 80% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12. (Item 2) 2. The compound according to item 1, wherein the sequence is at least 85% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12. (Item 3) 2. The compound according to item 1, wherein the sequence is at least 90% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12. (Item 4) 2. The compound according to item 1, wherein the sequence is at least 95% identical to SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 or SEQ ID NO:12. (Item 5) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:9. (Item 6) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO:9. (Item 7) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:10. (Item 8) 2. The compound according to item 1, wherein the sequence is SEQ ID NO: 10. (Item 9) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:11. (Item 10) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO:11. (Item 11) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:12. (Item 12) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO: 12. (Item 13) 2. The compound according to claim 1, wherein the sequence is at least 80% identical to SEQ ID NO:29. (Item 14) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO: 29. (Item 15) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:30. (Item 16) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO: 30. (Item 17) 2. The compound according to item 1, wherein the sequence is at least 80% identical to SEQ ID NO:31. (Item 18) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO: 31. (Item 19) 2. The compound according to claim 1, wherein the sequence is at least 80% identical to SEQ ID NO:32. (Item 20) 2. The compound according to claim 1, wherein the sequence is SEQ ID NO: 32. (Item 21) 2. The compound according to item 1, which inhibits tyrosine phosphatase. (Item 22) The compound according to item 1, which inhibits HPTP-β. (Item 23) Item 1. A compound according to item 1 which inhibits VE-PTP. (Item 24) 2. The compound according to item 1, which activates Tie2. (Item 25) The compound according to item 1, which activates Akt. (Item 26) Item 2. The compound according to item 1, which binds to the extracellular domain of HPTP-β. (Item 27) 2. The compound according to item 1, which binds to the first FN3 repeat of the extracellular domain of HPTP-β. (Item 28) The binding affinity of the compound to the extracellular domain of HPTP-β (K D 2. The compound according to item 1, wherein the concentration of 1000 mM is about 70 pM to about 70 nM. (Item 29) A compound comprising a sequence that is at least 80% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23. (Item 30) 30. The compound according to item 29, wherein the sequence is at least 85% identical to SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or SEQ ID NO: 23. (Item 31) 30. The compound according to item 29, wherein the sequence is at least 90% identical to SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or SEQ ID NO: 23. (Item 32) 30. The compound according to item 29, wherein the sequence is at least 95% identical to SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or SEQ ID NO: 23. (Item 33) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 20. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 20. (Item 35) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 21. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 21. (Item 37) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 22. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 22. (Item 39) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 23. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 23. (Item 41) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 34. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 34. (Item 43) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 35. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 35. (Item 45) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 36. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 36. (Item 47) The compound according to claim 29, wherein the sequence is at least 80% identical to SEQ ID NO: 37. 30. The compound according to item 29, wherein the sequence is SEQ ID NO: 37. (Item 49) 30. The compound according to item 29, which inhibits tyrosine phosphatase. (Item 50) 30. The compound according to item 29, which inhibits HPTP-β. (Item 51) 30. The compound according to item 29, which inhibits VE-PTP. (Item 52) 30. The compound according to item 29, which activates Tie2. (Item 53) 30. The compound according to item 29, which activates Akt. (Item 54) 30. The compound according to item 29, which binds to the extracellular domain of HPTP-β. (Item 55) 30. The compound according to item 29, which binds to the first FN3 repeat of the extracellular domain of HPTP-β. (Item 56) The binding affinity of the compound to the extracellular domain of HPTP-β (K D 30. The compound according to item 29, wherein the concentration of 1000 mM is about 70 pM to about 70 nM. (Item 57) a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO: 30; b) a light chain comprising a sequence at least 80% identical to SEQ ID NO: 34; A compound comprising: (Item 58) 58. The compound according to item 57, wherein the heavy chain is at least 85% identical to SEQ ID NO: 30 and the light chain is at least 85% identical to SEQ ID NO: 34. (Item 59) 58. The compound according to item 57, wherein the heavy chain is at least 90% identical to SEQ ID NO: 30 and the light chain is at least 90% identical to SEQ ID NO: 34. (Item 60) 58. The compound according to item 57, wherein the heavy chain is at least 95% identical to SEQ ID NO: 30 and the light chain is at least 95% identical to SEQ ID NO: 34. (Item 61) 58. The compound according to item 57, wherein the heavy chain is SEQ ID NO: 30 and the light chain is SEQ ID NO: 34. (Item 62) 58. The compound according to item 57, which inhibits tyrosine phosphatase. (Item 63) 58. The compound according to item 57, which inhibits HPTP-β. (Item 64) 58. The compound according to item 57, which inhibits VE-PTP. (Item 65) 58. The compound according to item 57, which activates Tie2. (Item 66) 58. The compound according to item 57, which activates Akt. (Item 67) 58. The compound according to item 57, which binds to the extracellular domain of HPTP-β. (Item 68) 58. The compound according to item 57, which binds to the first FN3 repeat of the extracellular domain of HPTP-β. (Item 69) The binding affinity of the compound to the extracellular domain of HPTP-β (K D 58. The compound according to item 57, wherein the concentration of 1000 mM is about 70 pM to about 70 nM. (Item 70) a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO: 29; b) a light chain comprising a sequence at least 80% identical to SEQ ID NO: 35; A compound comprising: (Item 71) 71. The compound according to item 70, wherein the heavy chain is at least 85% identical to SEQ ID NO: 29 and the light chain is at least 85% identical to SEQ ID NO: 35. (Item 72) 71. The compound according to item 70, wherein the heavy chain is at least 90% identical to SEQ ID NO: 29 and the light chain is at least 90% identical to SEQ ID NO: 35. (Item 73) 71. The compound according to item 70, wherein the heavy chain is at least 95% identical to SEQ ID NO: 29 and the light chain is at least 95% identical to SEQ ID NO: 35. (Item 74) 71. The compound according to item 70, wherein the heavy chain is SEQ ID NO: 29 and the light chain is SEQ ID NO: 35. (Item 75) 71. The compound according to item 70, which inhibits tyrosine phosphatase. (Item 76) 71. The compound according to item 70, which inhibits HPTP-β. (Item 77) 71. The compound according to item 70, which inhibits VE-PTP. (Item 78) 71. The compound according to item 70, which activates Tie2. (Item 79) 71. The compound according to item 70, which activates Akt. (Item 80) 71. The compound according to item 70, which binds to the extracellular domain of HPTP-β. (Item 81) 71. The compound according to item 70, which binds to the first FN3 repeat of the extracellular domain of HPTP-β. (Item 82) The binding affinity of the compound to the extracellular domain of HPTP-β (K D 71. The compound according to item 70, wherein the concentration of 1000 mM is about 70 pM to about 70 nM. (Item 83) a) a heavy chain comprising a sequence that is at least 80% identical to SEQ ID NO: 30; b) a light chain comprising a sequence at least 80% identical to SEQ ID NO: 37; A compound comprising: (Item 84) 84. The compound according to item 83, wherein the heavy chain is at least 85% identical to SEQ ID NO: 30 and the light chain is at least 85% identical to SEQ ID NO: 37. (Item 85) 84. The compound according to item 83, wherein the heavy chain is at least 90% identical to SEQ ID NO: 30 and the light chain is at least 90% identical to SEQ ID NO: 37. (Item 86) 84. The compound according to item 83, wherein the heavy chain is at least 95% identical to SEQ ID NO: 30 and the light chain is at least 95% identical to SEQ ID NO: 37. (Item 87) 84. The compound according to item 83, wherein the heavy chain is SEQ ID NO: 30 and the light chain is SEQ ID NO: 37. (Item 88) 84. The compound according to item 83, which inhibits tyrosine phosphatase. (Item 89) 84. The compound according to item 83, which inhibits HPTP-β. (Item 90) 84. The compound according to item 83, which inhibits VE-PTP. (Item 91) 84. The compound according to item 83, which activates Tie2. (Item 92) 84. The compound according to item 83, which activates Akt. (Item 93) 84. The compound according to item 83, which binds to the extracellular domain of HPTP-β. (Item 94) 84. The compound according to item 83, which binds to the first FN3 repeat of the extracellular domain of HPTP-β. (Item 95) The binding affinity of the compound to the extracellular domain of HPTP-β (K D 84. The compound according to item 83, wherein the concentration of 1000 mM is about 70 pM to about 70 nM. (Item 96) 11. A method of treating a condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of items 1 to 95. (Item 97) Item 97. The method of item 96, wherein the condition is an ophthalmic condition. (Item 98) 97. The method of claim 96, wherein the condition is diabetic retinopathy. (Item 99) 97. The method of item 96, wherein the condition is neovascularization. (Item 100) 97. The method of claim 96, wherein the condition is vascular leakage. (Item 101) 97. The method of item 96, wherein the condition is elevated intraocular pressure. (Item 102) 97. The method of claim 96, wherein the condition is ocular edema. (Item 103) 97. The method of claim 96, wherein the condition is diabetic macular edema. (Item 104) 97. The method of item 96, wherein the condition is ocular hypertension. (Item 105) 97. The method of item 96, wherein the condition is ocular inflammation. (Item 106) 97. The method of claim 96, wherein the administration is to an eye of the subject. (Item 107) 97. The method of item 96, wherein said administration is intravitreal. (Item 108) 97. The method of item 96, wherein the administration is subcutaneous. (Item 109) 97. The method of item 96, wherein the administration is topical. (Item 110) 97. The method of claim 96, wherein the subject is a human. (Item 111) 97. The method of claim 96, wherein the therapeutically effective amount of the compound is from about 0.25 mg to about 200 mg. (Item 112) Item 97. The method of item 96, wherein the therapeutically effective amount of the compound is about 1 mg / kg to about 10 mg / kg. (Item 113) 97. The method of item 96, wherein the therapeutically effective amount of the compound is from about 1 mg to about 50 mg. (Item 114) 97. The method of item 96, wherein the therapeutically effective amount of the compound is from about 50 mg to about 200 mg.

Claims

[Claim 1] The invention described in this specification.

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