Humanized monoclonal antibody targeting VE-PTP (HPTP-β)

Abstract

The present disclosure provides compositions and methods for treating ocular conditions associated with angiogenesis comprising administering in a subject an antibody that targets a tyrosine phosphatase inhibitor.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 364,381, filed on July 20, 2016, and U.S. Provisional Application No. 62 / 377,072, filed on August 19, 2016, each of which is incorporated herein by reference in its entirety. Background Art

[0003] Monoclonal antibodies produced in non-human organisms can elicit 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 potential for immunogenicity of the drug while maintaining target specificity.

[0004] Incorporation by reference

[0005] Each patent, publication, and non-patent document cited in this application is hereby incorporated by reference in its entirety, as if each were individually incorporated by reference. Summary of the invention

[0006] In some embodiments, the invention provides 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.

[0007] In some embodiments, the invention provides 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.

[0008] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:34.

[0009] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:29; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:35.

[0010] In some embodiments, the invention provides a compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:37. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 shows the rat V H Sequence (truncated V H0 ; SEQ ID NO: 47) and 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 ) sequence alignment.

[0012] Figure 2 shows the rat V L Sequence (truncated V L0 ; SEQ ID NO: 48) and four humanized V L Variant 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 ) sequence alignment.

[0013] Figure 3 Hybridoma technology for the generation of monoclonal antibodies against the extracellular domain of HPTP-β (VE-PTP / HPTP-β ECD) is outlined.

[0014] Figure 4 Shown are immunoprecipitation western blot (left panel) and flow cytometry results (right panel) of R15E6 binding to endogenous human VE-PTP (HPTP-β) in human endothelial cells.

[0015] Figure 5 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 blotting is shown.

[0016] Figure 6 Graphs show western blot (Panel A) and quantification (Panel B) of concentration-dependent phosphorylation of Tie2 by R15E6 and enhanced endothelial cell viability (Panel C).

[0017] Figure 7 Graphs show the effects 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 blotting.

[0018] Figure 8Graph depicts Tie2 activation by a polyclonal antibody against mouse VE-PTP ECD in mouse endothelial cells after 1 hour (Panel A) and 3 minutes (Panel B) incubation.

[0019] Fig. 9 Shown is the in vitro inhibition of thrombin-induced and VEGF-induced endothelial cell permeability by mouse VE-PTP ECD polyclonal antibodies.

[0020] Fig.10 Shown are 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), and immunoprecipitation of Tie2 from lung lysates of control IgG (-) or anti-VE-PTP antibody-injected mice (+) (Panel C).

[0021] Fig.11 Depicted are the effects of intraocular administration of anti-mouse VE-PTP monoclonal antibody 109.1 on retinal neovascularization (Panels A and B) and choroidal neovascularization (Panel C).

[0022] Fig.12 Shown are quantifications of the mean areas of retinal neovascularization (Panels A and B) and choroidal neovascularization (Panel C) following intraocular administration of anti-mouse VE-PTP monoclonal antibody 109.1.

[0023] Fig.13 Depicted are reducing (Panel A) and non-reducing (Panel B) SDS-PAGE of the anti-human VE-PTP (anti-HPTP-β) humanized variant of R15E6.

[0024] Fig.14 The binding of the HC1 variant of R15E6 and HCOLC0 to HPTP-β is shown.

[0025] Fig.15 Binding of the HC2 variant of R15E6 and HCOLC0 to HPTP-β is shown.

[0026] Fig.16 Binding of the HC3 variant of R15E6 and HCOLC0 to HPTP-β is shown.

[0027] Fig.17 Binding of the HC4 variant of R15E6 and HCOLC0 to HPTP-β is shown.

[0028] Fig.18 Shown are Tie2 phosphorylation (left panel) and Akt activation (right panel) by various humanized antibodies in the absence of Ang1 as determined by western blotting.

[0029] Fig.19 Graphs show concentration-dependent Tie2 activation (upper panel) and Akt activation (lower panel) of HC2LC4 and HC2LC1 in the absence of ligand as determined by western blot.

[0030] Fig. 20 Graphs show Tie2 phosphorylation (left panel) and Akt activation (right panel) of HC2LC4 and HC2LC1 in the presence of Ang1 and Ang2 as determined by western blotting.

[0031] Fig.21 Graphs show Tie2 phosphorylation (Panel A), human VE-PTP (HPTP-β) expression (Panel B), and Akt activation (Panel C) of HC2LC4 and HC2LC1 in the presence of Ang1 and / or Ang2 as determined by western blotting.

[0032] Fig. 22 Graphs show Akt activation by HC2LC1 alone and HC2LC1 with excess recombinant human VE-PTP ectodomain proteins (βECD 6His or βECD Fc) as determined by western blotting.

[0033] Fig.23 Graph showing enhanced endothelial cell viability by HC2LC1 and HC0LC0 compared to a parenteral antibody (R15E6) and mouse IgG1 control.

[0034] Fig.24 Figure 3 shows the enhanced efficacy of anti-mouse VE-PTP antibody / aflibercept combination therapy for the treatment of retinal detachment in a mouse model.

[0035] Fig.25 The graph shows that subcutaneous administration of monoclonal antibody 109.1 reduces ischemic neovascularization in mice compared to vehicle administration. DETAILED DESCRIPTION

[0036] The present disclosure provides compositions and methods for targeting vascular endothelial protein tyrosine phosphatase (VE-PTP or VEPTP) or human protein tyrosine phosphatase-β (HPTP-β) for treating ocular disorders characterized by, for example, vascular instability, angiogenesis, neovascularization, vascular leakage, and edema. The compositions disclosed herein can activate Tie2 signaling by promoting protein phosphorylation, such as phosphorylation of Tie2 protein.

[0037] 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 exhibits structural and functional similarities to cell adhesion molecules. VE-PTP is present in a variety of species, including, for example, zebrafish, chicken, dog, mouse, marmoset, monkey and human. The human ortholog of VE-PTP is HPTP-β.

[0038] Tie2 (tyrosine kinase with immunoglobulin and epidermal growth factor homology domain 2) is a membrane receptor tyrosine kinase expressed primarily in vascular endothelial cells throughout development. The main regulators of Tie2 phosphorylation are angiopoietin 1 (Ang1) and angiopoietin 2 (Ang2). Ang1 is an agonist of Tie2, and the binding of Ang1 to Tie2 promotes receptor phosphorylation. Ang2 acts as an environmentally dependent antagonist or agonist of Tie2. The binding of Ang1 to Tie2 increases the level of endogenous Tie2 receptor phosphorylation and initiates downstream signaling to induce vascular stability through highly organized angiogenesis, tightening of endothelial 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.

[0039] Under physiological conditions, the duration of Tie2 phosphorylation is regulated by VE-PTP (HPTP-β), which removes phosphate from the Tie2 receptor. By inhibiting VE-PTP (HPTP-β), the level of Tie2 phosphorylation is greatly increased, thereby 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 an inhibitor can provide vascular stability in subjects with ocular disorders described herein.

[0040] The VE-PTP (HPTP-β) inhibitor of the present disclosure may include the murine monoclonal antibody R15E6 produced by the hybridoma cell line ATCC No. PTA-7580, which is immunoreactive to the extracellular domain of human VE-PTP (HPTP-β) (SEQ ID NO.45) and immunoreactive to the first FN3 repeat sequence of human VE-PTP (HPTP-β) (SEQ ID NO.46). The VE-PTP (HPTP-β) inhibitor may include an antibody having the same or substantially the same biological characteristics as R15E6, an antibody fragment of R15E6 (wherein the fragment comprises one or both of the heavy chain and light chain variable regions), a dimer or trimer of F(ab')2, Fab, Fv, scFv of R15E6, and a diabody, triabody or tetrabody derived from R15E6. In some embodiments, fragments of the compounds described herein may be analyzed for any biological activity described herein. In some embodiments, fragments of the compounds described herein can activate Tie2, or provide the biological activity of the corresponding intact antibody at equivalent, increased, or reduced levels.

[0041] Extracellular domain of human VE-PTP (HPTP-β) (SEQ ID NO: 45):

[0042]

[0043] The first FN3 repeat sequence of human VE-PTP (HPTP-β) (SEQ ID NO: 46):

[0044] LAEPERCNFTLAESKASSHSVSIQWRILGSPCNFSLIYSSDTLGAALCPTFRIDNTTYGCNLQDLQAGTIYNFRIISLDEERTVVLQTD

[0045] The VE-PTP (HPTP-β) inhibitors of the present disclosure may include antibodies or antibody fragments, variants or derivatives thereof, alone or in combination with other amino acid sequences. The inhibitors may be modified, such as enzymatically cleaved and post-translationally modified.

[0046] The VE-PTP (HPTP-β) inhibitors of the present disclosure may bind to a dominant negative isoform of VE-PTP (HPTP-β). In some embodiments, the dominant negative isoform may correspond to a VE-PTP (HPTP-β) form that is defective in phosphatase activity and is able to compete with endogenous VE-PTP (HPTP-β). Functional assessment of dominant negative VE-PTP (HPTP-β) may be performed by delivering a transgene and measuring the effect on Tie2 phosphorylation.

[0047] The VE-PTP (HPTP-β) inhibitor of the present disclosure may have multiple VE-PTP (HPTP-β) binding sites. In some embodiments, the VE-PTP (HPTP-β) inhibitor may bind to two VE-PTP (HPTP-β) molecules at the same time, thereby bringing the two VE-PTP (HPTP-β) molecules into close proximity. The VE-PTP (HPTP-β) inhibitor may bind to three VE-PTP (HPTP-β) molecules at the same time, thereby bringing the three VE-PTP (HPTP-β) molecules into close proximity.

[0048] The VE-PTP (HPTP-β) inhibitors of the present disclosure may be covalently or non-covalently conjugated to another moiety or vehicle. For example, for the inhibitor, the moiety or vehicle may inhibit degradation, increase half-life, enhance absorption, reduce toxicity, reduce immunogenicity and / or increase biological activity. Non-limiting examples of such moieties include the Fc domain of an immunoglobulin, polymers such as polyethylene glycol (PEG), polylysine and dextran, lipids, cholesterols such as steroids, carbohydrates, dendrimers, oligosaccharides and peptides.

[0049] 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. Tie2 activation can lead to vascular stabilization, which can be beneficial for treating diabetes-related conditions and eye 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 hemorrhage. The compound can also effectively treat peripheral arterial disease, including, for example, wound healing, blood flow instability, cardiac fibrosis, erectile dysfunction, cardiomyopathy, ischemic injury, cardiac hypertrophy, interstitial fibrosis, nephropathy, proteinuria, glomerular sclerosis, renal fibrosis, neuropathy, and neuronal inflammation.

[0050] Antibody

[0051] The antibody contains two identical heavy chain (H) polypeptide sequences and two identical light chain (L) polypeptide sequences. Each heavy chain contains an N-terminal variable (V H ) region and three C-terminal constant (C H 1. C H 2 and C H 3) region. Each light chain contains an N-terminal variable (V L ) region and a C-terminal constant (C L ) region. The light chain variable region is aligned with the heavy chain variable region, and the light chain constant region is aligned with the heavy chain constant region C H1 Aligned. The pairing of the heavy chain variable region and the light chain variable region together forms a single antigen binding site. Each light chain is linked to the heavy chain by one covalent disulfide bond. Depending on the heavy chain isotype, the two heavy chains are interconnected by one or more disulfide bonds. Each heavy chain and light chain also contains regularly spaced intrachain disulfide bridges.

[0052] The light chain from any vertebrate species can be designated as κ or λ according to the amino acid sequence of the constant region. According to the amino acid sequence of the heavy chain constant region, immunoglobulins can be divided into five classes of immunoglobulins (IgA, IgD, IgE, IgG and IgM), each with a heavy chain called α, δ, ε, γ and μ, respectively. Based on the differences in the sequence and function of the heavy chain constant region, the α and γ classes are further divided into subclasses. The subclasses of IgA and IgG expressed by humans include IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.

[0053] The variable (V) region is included in the segment whose sequence can be widely different between antibodies. The variable region mediates antigen binding and limits the specificity of a particular antibody for its antigen. However, variability is not evenly distributed over the span of the variable region. On the contrary, the variable region is composed of a relatively constant fragment called a framework region (FR) of 15-30 amino acids, and the framework region is separated by a shorter region of great variability called a hypervariable region of 9-12 amino acids in length. The variable region of natural heavy chain and light chain each comprises four framework regions, mainly adopting an F3 folding configuration, connected by three hypervariable regions, which form a loop connecting the F3 folding configuration, and form a part of the F3 folding configuration in some cases. The hypervariable region in each chain is kept together by the close proximity of the framework region, and together with the hypervariable region from another chain, it helps to form the antigen binding site of the antibody. The constant domain does not directly participate in the combination of the antibody and the antigen, but exhibits various effector functions, such as antibody participation in antibody-dependent cellular toxicity (ADCC).

[0054] The hypervariable region may comprise amino acid residues from the complementarity determining regions (CDRs), e.g., around residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable region, and around residues 1-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable region, and / or residues from the hypervariable loops.

[0055] Monoclonal antibodies can be obtained from a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present in small amounts. In contrast to polyclonal antibody preparations comprising different antibodies directed against different epitopes, each monoclonal antibody is directed against a single epitope. In addition to specificity, the advantage of monoclonal antibodies is that the synthesis of each antibody can be free from contamination by other antibodies.

[0056] The monoclonal antibodies used herein may be, for example, chimeric antibodies, in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in antibodies derived 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 derived from another species or belonging to another antibody class or subclass, as well as antigen-binding fragments of such antibodies.

[0057] Antibody fragments may include a portion of a multimeric antibody, such as an antigen binding region or a variable region 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.

[0058] Non-limiting examples of epitopes include amino acids, sugars, lipids, phosphoryl groups, and sulfonyl groups. An epitope may have specific three-dimensional structural characteristics and / or specific charge characteristics. An epitope may be conformational or linear.

[0059] Humanized monoclonal antibodies

[0060] A variety of techniques can be used to identify suitable antibodies targeting VE-PTP. For example, candidate agents can be screened for binding to VE-PTP. Agents that bind to VE-PTP can 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.

[0061] The selection of suitable detection for identifying specific inhibitors depends on the properties of the candidate agent to be screened. For example, in the case where the candidate is an antibody or peptibody comprising an Fc portion, fluorescence activated cell sorting (FACS) analysis allows selection of candidate agents based on the ability to bind to cells expressing VE-PTP. Cells may endogenously express VE-PTP or may be genetically engineered to express VE-PTP. For other candidate agents such as aptamers, other techniques may be used. For example, systematic evolution of ligands by exponential enrichment (SELEX) may be used to select aptamers that specifically bind to VE-PTP, and SELEX selects specific aptamers by repeating multiple rounds of in vitro selection.

[0062] VE-PTP inhibitors can be screened for VE-PTP-mediated activity, for example, inhibition of Tie2 dephosphorylation. In a suitable assay based on western blotting, human umbilical vein endothelial cells (HUVEC) are cultured in serum-free medium in the presence or absence of various concentrations of candidate agents, cell lysates are prepared, immunoprecipitated with Tie2 antibodies, dissolved by polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to polyvinylidene fluoride (PVDF) membranes. The membrane-bound immunoprecipitated proteins are then subjected to continuous western blotting with anti-phosphotyrosine antibodies to quantify Tie2 phosphorylation, and then western blotting with Tie2 antibodies to quantify total Tie2. Tie2 phosphorylation is expressed as the ratio of anti-phosphotyrosine signal to total Tie2 signal. Higher levels of anti-phosphotyrosine signal indicate stronger inhibition of VE-PTP by the candidate agent.

[0063] Hybridoma technology is a method for producing a large number of monoclonal antibodies targeting specific antigens. Hybridoma development is initiated by injecting a mammalian host such as a mouse (murine) or a rabbit with an antigen that causes an immune response. B lymphocytes (B cells) in the host produce antibodies that bind to the antigen in response to the antigen. Then B cells are harvested from the host and fused with infinite B cell cancer cells (myeloma cells) to produce a hybrid cell line known as a hybridoma. Hybridomas retain the antibody production ability of B cells and the extraordinary life span and fecundity of myeloma. Hybridomas can be grown in culture medium from a living hybridoma cell to produce a new culture composed of genetically identical hybridomas, which in turn produces a mixture of one antibody (monoclonal) or different antibodies per culture (polyclonal). The myeloma cell line used in the process is selected for its ability to grow in tissue culture and the absence of the ability to synthesize antibodies. After cultivation, a preliminary screening process is performed to identify and select hybridomas that produce antibodies with the highest specificity. Non-limiting examples of antibody screening techniques include ELISA and immunocytochemical screening. Lead hybridomas identified from the screen can then be characterized for reactivity, binding affinity, specificity, and cross-reactivity.

[0064] In some embodiments, the binding affinity (K) of the compounds of the present disclosure for HPTP-β D ) is about 70 pM to about 70 nM, 1 nM to about 70 nM, or at least as strong as about 1 nM. In some embodiments, the binding affinity (K) of the compounds of the present disclosure for HPTP-β is D ) is from about 4 nM to about 70 nM.

[0065] Other recombinant antibody engineering techniques involve the use of viruses or yeast rather than mammals. These techniques rely on the rapid cloning of immunoglobulin gene segments to produce 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 under various environmental conditions, therapeutic efficacy, and detectability in diagnostic applications.

[0066] For human administration, monoclonal antibodies produced by non-human species can be further optimized through the humanization process to maintain target specificity while reducing the possibility of immunogenicity. The humanization process involves incorporating human DNA into the gene sequence of the gene that produces the individual antibody. The recombinant DNA is then cloned and expressed in cells to produce new humanized antibodies on a large scale.

[0067] The example of humanized antibody is modified chimeric antibody. Chimeric antibody is produced as described above. Chimeric antibody is further mutated outside CDR to replace the non-human sequence in variable region with homologous human sequence. Another example of humanized antibody is CDR transplanted antibody, wherein non-human CDR sequence is introduced into the human heavy chain and light chain variable sequence of human antibody scaffold to replace the corresponding human CDR sequence.

[0068] 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 genome that produces human antibodies inserted into the animal's genome.

[0069] Fully human monoclonal antibodies correspond to antibodies whose antigen binding residues are completely derived from human immunoglobulin sequences or fragments thereof that have undergone selection. In some embodiments, the selection can be carried out using phage display technology, in which a series of variable antibody domains are expressed and enriched on filamentous phage coat proteins to bind to target antigens. In some embodiments, the selection can be carried out using transgenic animals such as mice, rats or rabbits, in which a full set of endogenous immunoglobulin genes is replaced by a full set of human immunoglobulin genes. In some embodiments, a full set of human immunoglobulin genes can be introduced into the genome of the animal, and endogenous antibody production is made insufficient during the production of the antibody.

[0070] Humanized monoclonal antibody targeting VE-PTP (HPTP-β)

[0071] The humanization process of the mouse monoclonal antibody targeting VE-PTP (HPTP-β) described herein utilizes a combination of CDR grafting technology combined with antibody structure and a database of mature human IgG sequences. Human framework sequences are used as acceptor frameworks for CDR sequences. These acceptor sequences are all derived from mature human IgG from human sources, rather than from phage display.

[0072] Four humanized variants were designed for the heavy and light chain variable domains of a murine monoclonal antibody generated against the recombinant extracellular domain of human VE-PTP (HPTP-β). The design was based on factors including homology, T cell epitopes, key residues, and predicted structure.

[0073] The antibody numbering systems from IMGT and Kabat are used to identify the sequences disclosed herein. These two numbering systems identify different residues of murine antibodies as belonging to the CDRs, and the combined IMGT / Kabat sequence is used for optimal conservation of CDR loop conformation.

[0074] Heavy chain sequence

[0075] SEQ ID NO: 1 is a mouse monoclonal antibody R15E6 (VH0 ) H domain, which includes the mouse signal peptide sequence (underlined). SEQ ID NO: 47 is the V domain of R15E6 without the mouse signal peptide sequence H SEQ ID NO: 2 is a truncated sequence of the mouse 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 additional germline V regions similar to SEQ ID NO: 2. The peptide sequences of SEQ ID NO: 1-4 are shown in Table 1.

[0076] Table 1

[0077]

[0078]

[0079] The BLAST search algorithm was used to search online databases of human IgG sequences for comparison with mouse V H The domains were compared and candidate variable domains were selected from the top 200 BLAST results. Based on a combination of framework homology, maintained framework residues and canonical loop structure, the juxtaposed human variable domains were reduced to four candidates. 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.

[0080] Table 2

[0081]

[0082]

[0083] V H The CDRs of SEQ ID NO: 5-8 were grafted into the acceptor framework to convert these sequences into humanized variants, which are shown in Table 3. SEQ ID NO: 9 is V H1 , SEQ ID NO: 10 is V H2 SEQ ID NO: 11 is V H3 , and SEQ ID NO: 12 is V H4 .

[0084] Table 3

[0085]

[0086] Figure 1 shows the rat V H Sequence (truncated V H0 ; SEQ ID NO: 47) and 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 ) were compared. H / V K Residues important for interfaces and canonical loop structure.

[0087] Humanized variants and mouse V H The homology percentages are shown in Table 4. The order of homology of the humanized variants is V H2 >V H1 >V H3 >V H4 .

[0088] Table 4

[0089] Humanized variants Same amino acid Common amino acids <![CDATA[V H1 (SEQ ID NO:9)]]> 89.3％ 94.3％ <![CDATA[V H2 (SEQ ID NO:10)]]> 90.2％ 96.7％ <![CDATA[V H3 (SEQ ID NO:11)]]> 87.7％ 95.1％ <![CDATA[V H4 (SEQ ID NO:12)]]> 86.9％ 93.4％

[0090] Light chain sequence

[0091] In Table 5 below, SEQ ID NO: 13 is the mouse V L Domain (V L0 ), which includes the mouse signal peptide sequence (underlined). SEQ ID NO: 48 is the V sequence of R15E6 without the mouse signal peptide sequence L SEQ ID NO: 14 is a truncated sequence of the mouse V domain of R15E6. L The closest human germline gene V region domain is Homo sapiens IGKV1-16. SEQ ID NO: 15 is IGKV4-1 and is very similar to IGKV1-16.

[0092] Table 5

[0093]

[0094] The BLAST search algorithm was used to search an online database of human IgK sequences for comparison with the mouse VL domain, and candidate variable domains were selected from the top 200 BLAST results. These juxtaposed human variable domains were reduced to four candidates based on a combination of framework homology, maintenance framework residues, and canonical loop structure. 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.

[0095] Table 6

[0096]

[0097] V L The CDRs of V were grafted into these acceptor frameworks to convert these sequences into humanized variants, which are shown in Table 7. SEQ ID NO: 20 is V L1 , SEQ ID NO: 21 is V L2 , SEQ ID NO:22 is V L3 , and SEQ ID NO: 23 is V L4 .

[0098] Table 7

[0099]

[0100]

[0101] Figure 2 shows the rat V L Sequence (truncated V L0 ; SEQ ID NO: 48) and four humanized variants 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 ) were compared. L / V K Residues important for interfaces and canonical loop structure.

[0102] Humanized variants and mouse V L The homology percentages are shown in Table 8. The order of homology of the humanized variants is V L3 >V L1 >V L2 >V L4 .

[0103] Table 8

[0104] Humanized variants The same amino acid Common amino acids <![CDATA[V L1 (SEQ ID NO:20)]]> 81.3％ 88.8％ <![CDATA[V L2 (SEQ ID NO:21)]]> 79.4％ 88.8％ <![CDATA[V L3 (SEQ ID NO:22)]]> 82.2％ 86.9％ <![CDATA[V L4 (SEQ ID NO:23)]]> 78.5％ 89.7％

[0105] Design of humanized antibodies: the four Vs H and 4 V L Chain combination

[0106] As mentioned above, humanization mouse variable chain is carried out by transplanting mouse CDR sequence on suitable human antibody donor sequence under the background of IgG4 heavy chain, to express complete antibody protein.Then one group of full-length humanized antibodies is codon optimized to express in Chinese hamster ovary (CHO) cell line.Use different people's donor sequence to produce the matrix of 16 kinds of human antibodies for expressing for four kinds of variants designed for each variable chain, and its heavy chain and light chain pairing are shown in Table 9.Produce the complete mouse variable domain (V) with transplanting to human IgG4 constant domain. H0- V L0 ) served as positive controls in binding and functional assays.

[0107] Table 9

[0108] <![CDATA[V H1 -V L1 ]]> SEQ ID NO:9-SEQ ID NO:20 <![CDATA[V H3 -V L1 ]]> SEQ ID NO:11-SEQ ID NO:20 <![CDATA[V H1 -V L2 ]]> SEQ ID NO:9-SEQ ID NO:21 <![CDATA[V H3 -V L2 ]]> SEQ ID NO:11-SEQ ID NO:21 <![CDATA[V H1 -V L3 ]]> SEQ ID NO:9-SEQ ID NO:22 <![CDATA[V H3 -V L3 ]]> SEQ ID NO:11-SEQ ID NO:22 <![CDATA[V H1 -V L4 ]]> SEQ ID NO:9-SEQ ID NO:23 <![CDATA[V H3 -V L4 ]]> SEQ ID NO:11-SEQ ID NO:23 <![CDATA[V H2 -V L1 ]]> SEQ ID NO:10-SEQ ID NO:20 <![CDATA[V H4 -V L1 ]]> SEQ ID NO:12-SEQ ID NO:20 <![CDATA[V H2 -V L2 ]]> SEQ ID NO:10-SEQ ID NO:21 <![CDATA[V H4 -V L2 ]]> SEQ ID NO:12-SEQ ID NO:21 <![CDATA[V H2 -V L3 ]]> SEQ ID NO:10-SEQ ID NO:22 <![CDATA[V H4 -V L3 ]]> SEQ ID NO:12-SEQ ID NO:22 <![CDATA[V H2 -V L4 ]]> SEQ ID NO:10-SEQ ID NO:23 <![CDATA[V H4 -V L4 ]]> SEQ ID NO:12-SEQ ID NO:23

[0109] T cell epitope screening

[0110] Heavy chain

[0111] Presentation of peptide sequences in the groove of MHC class II molecules causes activation of CD8+ T cells and immunogenic responses. In order to reduce this response, therapeutic proteins can be designed to avoid incorporating T cell epitopes that can activate T cells by reducing the affinity of binding to MHC class II molecules.

[0112] Screening of murine antibodies against MHC II binding peptides V H and V L As well as humanized variant sequences, to use in silico algorithms to determine that the humanization process has removed peptide sequences with high affinity. Table 10 shows the screening results, with high affinity T cell epitope cores indicated 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 indicated in italics. CDRs are underlined.

[0113] Table 10

[0114]

[0115] Light chain sequence

[0116] Table 11 shows the screening results, with the high affinity T cell epitope core indicated in bold (IC 50 <50 nM). 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 indicated in italics. CDRs are underlined.

[0117] Table 11

[0118]

[0119] Rat V L0 A T cell epitope is contained within CDR3 / framework region 4 of the light chain that is not present in the three humanized variants.

[0120] Post-translational modification

[0121] Fv glycosylation

[0122] The N-linked glycosylation motif is NXS / T, where X is any amino acid except proline. This motif is not present in R15E6 V H or V L in mouse or humanized variants.

[0123] Deamidation

[0124] The amino acid motifs SNG, ENN, LNG and LNN are susceptible to deamidation of asparagine to provide aspartic acid. None of these four motifs are present in R15E6 V H or V L in mouse or humanized variants.

[0125] Signal peptide

[0126] Murine antibody signal peptides can lead to higher levels of expression in CHO cells. The following signal peptides can be incorporated into the humanized variants.

[0127] Heavy chain signal peptide (SEQ ID NO: 24): MGWTLVFLFLLSVTAGVHS

[0128] Light chain signal peptide (SEQ ID NO: 25): MVSSAQFLGLLLLCFQGTRC

[0129] Each V HThe domains can all be synthesized in-frame with the human IgG4 isotype constant domain sequence with the stabilizing S228P mutation. The entire heavy chain sequence can be codon optimized and the DNA sequence verified. Amino acid sequence of IgG4 constant domain with S228P mutation (SEQ ID NO:26) is: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK.

[0130] Each V L The domain can be synthesized in frame with the human IgK isotype constant domain sequence. The entire light chain sequence can be codon optimized and the DNA sequence can be verified. The amino acid sequence of the IgK constant domain (SEQ ID NO: 27) is: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

[0131] The humanized antibody of the present invention can be used for the expression of the humanized antibody of the present invention.Then, can continue with transient transfection and the expression of every kind of humanized antibody.A kind of chimeric antibody can be expressed as a positive control, and can have mouse variable domain, people Ig constant domain.16 kinds of humanized variants only have humanized variable domain and people Ig constant domain.Table 12 shows heavy chain and light chain pairing, and it provides 16 kinds of humanized variants with humanized variable domain and people Ig constant domain.

[0132] Table 12

[0133]

[0134] Tables 13 and 14 list the complete amino acid sequence of each humanized heavy and light chain.

[0135] Table 13

[0136]

[0137]

[0138]

[0139] Table 14

[0140]

[0141]

[0142] The target sequence can 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 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, or at least about 91% homology. In some embodiments, the present invention relates to a polypeptide having a homology to a gene or a sequence of the present invention, wherein the homology is 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.

[0143] Various methods and software programs can be used to determine homology between two or more peptides or nucleic acids, such as NCBIBLAST, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, or another suitable method or algorithm.

[0144] Pharmaceutical composition

[0145] The pharmaceutical composition of the present invention can 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 helps to administer the compound to an organism. The pharmaceutical composition can be administered as a pharmaceutical composition in a therapeutically effective amount in various forms and routes, including, for example, intravenous, intramuscular, oral, parenteral, ophthalmic and topical administration.

[0146] The pharmaceutical composition can be applied to the eye by any suitable form or approach, including, for example, topical, oral, systemic, intravitreal, intracameral, subconjunctival, subtenon fascia, behind the eyeball, intraocular, near sclera, periocular, subretinal and suprachoroidal administration. The composition can be applied by injecting the preparation into any part of the eye including the anterior chamber, posterior chamber, vitreous chamber (intravitreal), retina itself and / or subretinal space. The composition can be delivered by non-invasive methods. The non-invasive mode of applying the preparation can include the use of a needle-free injection device. A variety of routes of administration can be used to effectively deliver the pharmaceutical composition.

[0147] The pharmaceutical composition can be targeted to any suitable ocular cell, including, for example, endothelial cells such as vascular endothelial cells, retinal cells such as retinal pigment epithelial cells (RPE), corneal cells, fibroblasts, astrocytes, glial cells, pericytes, iris epithelial cells, neurogenic cells, ciliary epithelial cells, Müller cells, surrounding and attached muscle cells of the eye such as lateral rectus muscle cells, orbital fat cells, scleral and episcleral cells, trabecular meshwork cells, and connective tissue cells.

[0148] The pharmaceutical composition can be administered in a local manner, for example, via direct injection of the compound into an organ, optionally in the form of a reservoir or sustained release formulation or implant. The pharmaceutical composition can be provided in the form of a quick release formulation, in the form of an extended release formulation, or in the form of a medium-speed release formulation. The quick release form can provide immediate release. Extended release formulations can provide controlled release or sustained delayed release.

[0149] The pharmaceutical preparation for administration may include an aqueous solution of the active compound in a water-soluble form. The suspension of the active compound may be prepared as an oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. The suspension may also contain a suitable stabilizer or an agent that increases the solubility of the compound to allow the preparation of a highly concentrated solution. Alternatively, the active ingredient may be in powder form, reconstituted with a suitable vehicle such as sterile pyrogen-free water before use.

[0150] In implementing the treatment or use methods provided herein, a therapeutically effective amount of a compound described herein is administered to a subject suffering from a disease or condition to be treated in the form of a pharmaceutical composition. In some embodiments, the subject is a mammal, such as a human. The therapeutically effective amount can vary greatly depending on the severity of the subject's disease, age, and relative health, the effectiveness of the compound used, and other factors.

[0151] In some embodiments, the compounds described herein can be used alone or as a component of a mixture in combination with one or more therapeutic agents. For example, the VE-PTP (HPTP-β) inhibitors of the present disclosure can be co-formulated or co-administered with antibodies such as anti-VEGF agents. 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 Ranibizumab Aflibercept In some embodiments, the compounds described herein can be used before, during, or after treatment with an anti-VEGF agent.

[0152] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers comprising excipients and adjuvants that facilitate processing of the active compound into a pharmaceutically acceptable preparation. The formulation can vary depending on the chosen route of administration. Pharmaceutical compositions comprising the compounds described herein can be prepared, for example, by mixing, dissolving, emulsifying, encapsulating, embedding or compressing processes.

[0153] The pharmaceutical composition may comprise at least one pharmaceutically acceptable carrier, diluent or excipient and a compound described herein as a free base or a pharmaceutically acceptable salt form. The pharmaceutical composition may contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

[0154] Methods for preparing compositions comprising the compounds described herein include formulating the compounds with one or more inert, pharmaceutically acceptable excipients or carriers to form solid, semisolid or liquid compositions. 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 as disclosed herein. Semisolid compositions include, for example, gels, suspensions and creams. The compositions may be liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquids prior to use, or as emulsions. These compositions may also contain small amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, and other pharmaceutically acceptable additives.

[0155] Non-limiting examples of dosage forms suitable for use with the present invention include liquids, powders, gels, nanosuspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0156] Non-limiting examples of pharmaceutically 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, antimicrobial agents, spheronizers, and any combination thereof.

[0157] Compositions of the present invention can be, for example, quick-release forms or controlled-release preparations. Quick-release preparations can be formulated to allow compounds to work quickly. Non-limiting examples of quick-release preparations include easily soluble preparations. Controlled-release preparations can be such pharmaceutical preparations, which have been adjusted so that the release rate and release profile of the active agent can match physiological requirements and long-term treatment requirements, or have been formulated to realize the release of the active agent at a programmed rate. Non-limiting examples of controlled-release preparations include particles, delayed-release particles, hydrogels (e.g., synthetic or natural origin), other gelling agents (e.g., dietary fibers forming gels), preparations based on matrix (e.g., preparations comprising a polymer material having at least one active ingredient dispersed therein), particles in the matrix, polymer mixtures, and particulate matter.

[0158] In some embodiments, the controlled release formulation is a delayed release form. The delayed release form can be formulated to delay the effect of the compound for an extended period of time. The delayed release form can be formulated to delay the release of one or more compounds of an effective dose, for example, for about 4, about 8, about 12, about 16, or about 24 hours.

[0159] The controlled release formulation can be a sustained release form. The sustained release form can be formulated to maintain the effect of, for example, a compound over an extended period of time. The sustained release form can be formulated to provide an effective dose of any compound described herein (e.g., to provide a physiologically effective blood profile) within about 4, about 8, about 12, about 16, or about 24 hours.

[0160] The disclosed compositions may optionally contain a pharmaceutically acceptable preservative.

[0161] Non-limiting examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th ed. (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated herein by reference in its entirety.

[0162] The compounds described herein can be conveniently formulated into pharmaceutical compositions consisting of one or more pharmaceutically acceptable carriers. See, for example, Remington's Pharmaceutical Sciences, latest edition, EW Martin Mack Pub. Co., Easton, PA, incorporated by reference in its entirety, which discloses typical carriers and conventional methods for preparing pharmaceutical compositions. Such carriers can be carriers for administering the composition to humans and non-humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH. The pharmaceutical composition may also contain one or more additional active ingredients, such as antimicrobial agents, anti-inflammatory agents, and anesthetics.

[0163] The limiting examples of pharmaceutically acceptable carriers include saline, Ringer's solution and dextrose solution. In some embodiments, the pH value of the solution can be from about 5 to about 8, and can be from about 7 to about 7.5. Other carriers include sustained release formulations, such as semipermeable matrices of solid hydrophobic polymers comprising the compound. The matrix can be a shaped article, such as a film, a liposome, a microparticle or a microcapsule.

[0164] The disclosed method involves administering an antibody targeting VE-PTP (HPTP-β) as part of a pharmaceutical composition. Compositions suitable for topical administration can be used. In some embodiments, the composition of the present invention may include a liquid containing an active agent in a solution, a suspension, or both. The liquid composition may include a gel. The liquid composition may be, for example, aqueous. The composition is an aqueous composition that can be gelled in situ. In iterations, the composition is an aqueous solution that can be gelled in situ. Such a composition may include a gelling agent at a concentration that effectively promotes gelation when in contact with the eye or with tears outside the eye. The aqueous composition may have an eye-compatible pH and weight molar osmotic pressure concentration (osmolality). The composition may include an ophthalmic reservoir preparation containing an active agent for subconjunctival administration. Microparticles containing an active agent may be embedded in a biocompatible pharmaceutically acceptable polymer or in a lipid encapsulating agent. The reservoir preparation may be suitable for releasing all or substantially all of the active substance over an extended period of time. The polymer or lipid matrix (if present) may be suitable for fully degrading to transfer from the application site after releasing all or substantially all of the active agent. The reservoir formulation may be a liquid formulation comprising a pharmaceutically acceptable polymer and a dissolved or dispersed active agent. During injection, the polymer forms a reservoir at the injection site by, for example, gelling or precipitation. The composition may include a solid product that may be inserted into a suitable position in the eye, such as between the eye and the eyelid or in the conjunctival sac, where the product releases the active agent. The solid product suitable for being implanted in the eye in this manner may include a polymer and may be bioerodible or non-bioerodible.

[0165] In addition to the agents disclosed herein, the pharmaceutical formulations may contain additional carriers, as well as thickeners, diluents, buffers, preservatives and surfactants.

[0166] The pH of the disclosed compositions is in the range of about 3 to about 12. The pH of the composition can be, for example, about 3 to about 4, about 4 to about 5, about 5 to about 6, about 6 to about 7, about 7 to about 8, about 8 to about 9, about 9 to about 10, about 10 to about 11, or about 11 to about 12 pH units. The pH of the composition can be, for example, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, or 12 pH units. The pH of the composition can be, for example, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or at least 12 pH units. The pH of the composition can be, for example, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, at most 10, at most 11, or at most 12 pH units. If the pH is outside the range desired by the formulator, the pH can be adjusted by using sufficient pharmaceutically acceptable acids and bases.

[0167] Depending on the intended mode of administration, the pharmaceutical composition may be in the form of a solid, semisolid or liquid dosage form, e.g., tablet, suppository, pill, capsule, powder, liquid, suspension, lotion, cream or gel, e.g., in a unit dosage form suitable for single administration of a precise dose.

[0168] For solid compositions, nontoxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0169] Non-limiting examples of pharmaceutically active agents suitable for combination with the compositions of the present disclosure include anti-infectives (i.e., aminoglycosides), antiviral agents, antimicrobial agents, anticholinergics / antispasmodics, antidiabetic agents, antihypertensive agents, antineoplastic agents, cardiovascular agents, central nervous system agents, coagulation regulators, hormones, immunological agents, immunosuppressants, and ophthalmic preparations.

[0170] In some embodiments, the pharmaceutical compositions provided herein comprise a therapeutically effective amount of a compound in admixture with a pharmaceutically acceptable carrier and / or excipient, such as saline, phosphate buffered saline, phosphate and amino acids, polymers, polyols, sugars, buffers, preservatives and other proteins. Exemplary agents include octylphenoxypolyethoxyethanol compounds, polyethylene glycol monostearate compounds, polyoxyethylene sorbitan fatty acid esters, sucrose, fructose, dextrose, maltose, glucose, mannitol, dextran, sorbitol, inositol, galactitol, xylitol, lactose, trehalose, bovine or human serum albumin, citrate, acetate, Ringer's solution and Hank's solution, cysteine, arginine, carnitine, alanine, glycine, lysine, valine, leucine, polyvinyl pyrrolidone, polyethylene and ethylene glycol.

[0171] Methods of administration and treatment

[0172] The pharmaceutical compositions described herein can be used for preventive and / or therapeutic treatment. In therapeutic applications, the composition can be applied to a subject who has suffered from the disease or condition in an amount sufficient to cure or at least partially prevent the symptoms of the disease or condition, or in an amount that cures, recovers, improves or alleviates the condition. Compositions can also be used to reduce the possibility of the condition occurring, infecting or worsening. The effective amount for this purpose can be changed according to the severity and process of the disease or condition, previous therapy, the health status of the subject, body weight and the response to the drug and the judgment of the treating physician.

[0173] The multiple therapeutic agents can be administered in any order or simultaneously. If administered simultaneously, the multiple therapeutic agents can be provided in a single unified form or in multiple forms (e.g., 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 administered in multiple doses. If not administered simultaneously, the timing between multiple administrations can vary by up to about one month.

[0174] The therapeutic agent described herein can be applied before, during or after the disease or condition occurs, and the time selection of applying the composition containing the therapeutic agent can be varied. For example, the composition can be used as a prophylactic, and can be continuously applied to a subject susceptible to a condition or disease to reduce the possibility of the disease or condition occurring. The composition can be applied to the subject as early as possible during or after the onset of symptoms. The administration of the therapeutic agent can be started within the first 48 hours of symptom onset, within the first 24 hours of symptom onset, within the first 6 hours of symptom onset, or within 3 hours of symptom onset. Initial administration can be carried out by any practical approach, such as by any approach described herein, using any preparation described herein. After the onset of a disease or condition is detected or suspected, once the therapeutic agent is available, it can be applied immediately, and the time length required for the disease is continuously treated, for example, from about 1 month to about 3 months. For each subject, the length of treatment can be different.

[0175] The pharmaceutical compositions described herein can be unit dosage forms suitable for single administration of precise doses. In unit dosage forms, the preparation is divided into unit doses containing an appropriate amount of one or more compounds. The unit dose can be in the form of a package containing a discrete amount of the preparation. Non-limiting examples are packaged injections, vials or ampoules. The aqueous suspension composition can be packaged in a non-resealable single dose container. Resealable multiple dose containers can be used, for example, with or without preservatives. Preparations for injection can be presented in unit dosage form, for example in ampoules, or in multiple dose containers containing preservatives.

[0176] The pharmaceutical compositions provided herein can be administered in combination with other treatments, such as chemotherapy, radiation, surgery, anti-inflammatory agents and selected vitamins. Other agents can be administered before, after or simultaneously with the pharmaceutical composition.

[0177] dose

[0178] The VE-PTP (HPTP-β) antibody can be administered at a dose 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 by weight of the subject.

[0179] The VE-PTP (HPTP-β) antibody can be administered at any desired interval. The administration of the compound may have an irregular dosing schedule to suit 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, six times a week, seven times a week, eight times a week, nine times a week, or ten times a week. The daily dosing interval may be any interval in hours, for example, every hour, every 2 hours, every 3 hours, every 4 hours, every 5 hours, every 6 hours, every 7 hours, every 8 hours, every 9 hours, every 10 hours, every 11 hours, or every 12 hours. The dosing regimen for administering VE-PTP (HPTP-β) antibodies includes, but is not limited to, once a day, three times a week, twice a week, once a week, three times a month, twice a month, once a month, once every two months, once a year, twice a year, three times a year, or four times a year. In some embodiments, the VE-PTP (HPTP-β) antibody may be administered every other week.

[0180] In addition, the amount of VE-PTP (HPTP-β) antibody administered can be the same amount in each dose, or the dose can vary between doses. For example, a first amount is administered in the morning and a second amount is administered in the evening. The administered dose can vary according to the schedule of anti-VEGF administration.

[0181] The anti-VE-PTP antibody can be co-administered with any anti-VEGF agent in any combination, for example, co-administered with an anti-VEGF agent at the beginning of treatment, at any time during treatment, or at any time after treatment. In addition, the dose of the VE-PTP (HPTP-β) inhibitor can be adjusted during treatment. Moreover, the amount of the anti-VEGF agent can be adjusted during treatment.

[0182] Further non-limiting examples of VEGF modulators include non-inflammatory agents such as dexamethasone, fluocinolone and triamcinolone. In addition, 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 before, during, or after providing an implant to a subject with a disease or condition described herein.

[0183] Anti-VEGF treatment can be administered, for example, monthly, every 3 months, every 6 months, or yearly, with the VE-PTP (HPTP-β) inhibitor being administered at any frequency in between treatments.

[0184] Also disclosed herein are methods for treating a disease or condition as disclosed herein. The method comprises administering to a subject:

[0185] a) a therapeutically effective amount of a VE-PTP (HPTP-β) inhibitor; and

[0186] b) a therapeutically effective amount of an anti-VEGF agent;

[0187] Administration of the VE-PTP (HPTP-β) inhibitor and the anti-VEGF agent can be carried out in any manner desired by the administrator, for example, as further described herein.

[0188] VE-PTP (HPTP-β) binding agents or anti-VEGF agents can be administered in any necessary or convenient amount. For example, the compounds described herein can be administered 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 0.17 mg, about 0.18 mg, about 0.19 mg, about 0.20 mg, about 0.21 mg, about 0.22 mg, about 0.23 mg, about 0.24 mg, about 0.25 mg, about 0.26 mg, about 0.27 mg, about 0.28 mg, about 0.29 mg, about 0.30 mg, about 0.31 mg, about 0.32 mg, about 0.33 mg, about 0.34 mg, about 0.35 mg, about 0.36 mg, about 0.37 mg, about 0.38 mg, about 0.39 mg, about 0.40 mg, about 0.41 mg, about 0.42 mg, about 0.43 mg, about 0.44 mg, about 0.45 mg, about 0.46 mg, about 0.47 mg, about 0.48 mg, about 0.49 mg, about 0.50 mg, .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 0.47mg, about 0.48mg, about 0.49mg, about 0.5mg , about 0.51mg, about 0.52mg, about 0.53mg, about 0.54mg, about 0.55mg, about 0.56mg, about 0.57mg, about 0.58mg, about 0.59mg, about 0.6mg, about 0.61mg, about 0.62mg, about 0.63mg, about 0.64mg, about 0.65mg, about 0.66mg, about 0.67mg, about 0.68mg, about 0.69mg, about 0.7mg, about 0.71mg, about 0.72mg, about 0.73mg, about 0.74mg, about 0.75mg, about 0.76mg, 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.About 150 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 is administered to a subject.

[0189] Intraocular delivery

[0190] Disclosed herein is a method for intraocular delivery of a composition of the present invention to a subject suffering from a disease or condition as disclosed herein. The delivery method may include an invasive method for delivering the composition directly to the eye cells. In some embodiments, a liquid pharmaceutical composition comprising an antibody is delivered via subretinal injection. In some embodiments, a liquid pharmaceutical composition comprising an antibody is delivered via intravitreal or subcutaneous injection. In some embodiments, a liquid pharmaceutical composition comprising an antibody is delivered via intracameral injection. In some embodiments, the composition is delivered via a variety of routes of administration, such as subretinal and / or intravitreal, to improve the efficiency of antibody delivery. In some embodiments, subretinal and / or intravitreal injections are performed before vitrectomy.

[0191] Intraocular injection can be carried out at any time interval to improve delivery efficiency and / or minimize or avoid damage to surrounding tissue. The time interval of intraocular injection 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.

[0192] Intraocular injection can be performed at any rate. The rate 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. μL / min, 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.

[0193] Reagent test kit

[0194] The present disclosure further relates to a kit containing the composition of the present disclosure. The kit may include:

[0195] A) a composition comprising an antibody targeting VE-PTP (HPTP-β); and

[0196] B) A vehicle for delivering the composition to a subject.

[0197] The kit can be modified to accommodate the dosing regimen prescribed for the subject being treated. The following is a non-limiting example of a kit for use by a subject receiving a composition of the disclosure by intraocular injection:

[0198] A) an aqueous composition comprising:

[0199] a) antibodies targeting the extracellular domain of VE-PTP (HPTP-β); and

[0200] b) a vector system comprising:

[0201] i) a tonicity agent; and

[0202] ii) water,

[0203] wherein the tonicity agent is present in an amount such that the reconstituted formulation comprises from about 0.5% to about 10% by weight of the tonicity agent; and

[0204] B) Components for delivering aqueous compositions.

[0205] In some embodiments, the kit of the invention comprises:

[0206] A) a composition for delivering an anti-VE-PTP antibody; and

[0207] B) Compositions for delivery of anti-VEGF agents.

[0208] The kit can be modified to accommodate the dosing regimen prescribed for the subject being treated. The following is a non-limiting example of a kit for use by a patient receiving an intravenous delivery composition comprising a disclosed compound and an anti-VEGF agent administered intravitreally. This example provides for dosing of a disclosed compound twice daily for 3 months and provides an injection of ranibizumab at week 12.

[0209] A. 3 packs, each pack contains 4 vials. Each vial contains sufficient VE-PTP (HPTP-β) inhibitor to provide 5 mg of the disclosed compound twice daily for 7 days; and

[0210] B. Ranibizumab vial for injection at the end of week 12, providing 0.5 mg of Ranibizumab.

[0211] The kit may contain any combination of elements. In addition, when the anti-VE-PTP antibodies disclosed herein are administered orally, a single container with a sufficient dose of the disclosed compound may be supplemented with the kit.

[0212] The kit may also provide written instructions for use and disposal of the composition to be delivered. The instructions may be modified between kits to reflect the prescribed dosing regimen. The instructions may describe any therapy, compound, excipient, or method of administration described herein.

[0213] The disclosed compositions may contain, for example, about 1.5% to about 1.5% by mass volume of the carrier system. Non-limiting examples of tonicity agents include dextrose, mannitol, and glycerol. The formulator may use more than one tonicity agent.

[0214] The kit may further include a device for administration, such as a syringe, a filter needle, an extension tube, an intubation tube, and a subretinal syringe. The non-limiting examples of route of administration include intraocular, parenteral, and topical. The administration of intraocular route may include, for example, intravitreal, intracameral, subconjunctival, subtenon fascia, behind the eyeball, intraocular, near the sclera, periocular, subretinal, and on the choroid. It can be delivered by, for example, a syringe, a needle, an infusion pump, or an injector. Syringes and injectors may be, for example, single doses, multiple doses, fixed doses, or variable doses. The non-limiting examples of injectors include, but are not limited to, pen injectors, automatic injectors, and electronic patch injector systems.

[0215] The kit may include suitable components for administering the composition of the present invention to a subject. In some embodiments, the composition of the present invention is present in a kit in a unit dosage form. Thus, the formulator can provide a delivery device with a higher compound concentration and adjust the delivery amount to provide a certain amount of compound that is less than the amount in the entire solution.

[0216] A set of instructions may be included in any kit described herein. The instructions may relate to the amount to be administered, the time of administration, reconstitution of the composition when the kit contains a dry composition, and methods of discarding the delivery vehicle and unused composition. The instructions may describe any treatment, compound, excipient, or method of administration described herein.

[0217] method

[0218] The present invention provides compositions and methods for treating or preventing ocular diseases or conditions such as ocular edema, diabetic macular edema, vascular leakage, age-related macular degeneration (wet form), age-related macular degeneration (dry form), choroidal neovascularization, diabetic retinopathy, ocular ischemia, uveitis, retinal vein occlusion (central or branch), ocular trauma, surgically induced edema, surgically induced neovascularization, cystoid macular edema, ocular ischemia, and uveitis. These diseases or conditions can be characterized by changes in the ocular vasculature, whether progressive or non-progressive, whether the result of an acute disease or condition or a chronic disease or condition. These diseases can be characterized by increased plasma levels of vascular endothelial growth factor (VEGF).

[0219] The present disclosure provides methods of treating ocular neovascularization in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an antibody targeting VE-PTP (HPTP-β).

[0220] In some embodiments, the VE-PTP (HPTP-β) inhibitor stabilizes the vasculature to prevent leakage and neovascularization.

[0221] Improvement of clinical symptoms can be monitored by, for example, indirect ophthalmoscopy, fundus photography, fluorescein angiopathy, electroretinography, external eye examination, slit lamp biomicroscopy, applanation tonometry, pachymetry, optical coherence tomography, or auto-optometry. In some embodiments, the disclosed methods involve administration of a VE-PTP (HPTP-β) antibody, including administration of a composition comprising an anti-VEGF agent.

[0222] 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 to prevent leakage.

[0223] In some embodiments, the methods of the present disclosure contemplate co-administration of an anti-VE-PTP (HPTP-β) antibody with one or more anti-VEGF agents, which can stabilize the vasculature to prevent neovascularization.

[0224] In some embodiments, the anti-VE-PTP (HPTP-β) antibodies can stabilize the vasculature to prevent leakage and neovascularization.

[0225] 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 a VE-PTP (HPTP-β) antibody. Improvement of clinical symptoms can be monitored by, for example, indirect ophthalmoscopy, fundus photography, fluorescein vasculopathy, electroretinography, external eye examination, slit lamp biomicroscopy, applanation tonometry, thickness measurement, optical coherence tomography, and automated optometry. As described herein, administration may occur at any frequency determined by the administrator. After anti-VEGF agent treatment is stopped, subsequent doses may be administered, for example, once a week or once a month, for example, at a frequency of about 2-8 weeks or about 1-12 months, depending on the response.

[0226] Diseases that are direct or indirect results of diabetes include diabetic macular edema and diabetic retinopathy, among others. The ocular vascular system of a diabetic patient becomes unstable over time and leads to conditions such as non-proliferative retinopathy, macular edema, and proliferative retinopathy. As fluid leaks into the center of the macula (the part of the eye where clear, forward vision occurs), aggregates of fluid and associated proteins begin to deposit on or under the macula. This deposition leads to swelling that gradually distorts the subject's central vision. This condition is macular edema. Another condition that can occur is non-proliferative retinopathy, in which vascular changes outside the macular region of the eye, such as microaneurysms, can be observed.

[0227] These conditions may be associated with diabetic proliferative retinopathy, which is characterized by increased neovascularization. The new blood vessels are fragile and bleed easily. The result is scarring of the retina and blockage or complete blockage of the light pathway through the eye due to excessive formation of new blood vessels. Subjects with diabetic macular edema typically experience a non-proliferative phase of diabetic retinopathy; however, subjects typically begin to experience macular edema only at the onset of the proliferative phase.

[0228] Diabetic retinopathy is the most common cause of vision loss in working-age Americans. Severe vision loss occurs due to tractional retinal detachment, which complicates retinal neovascularization, but the most common cause of moderate vision loss is diabetic macular edema (DME).

[0229] VEGF is a hypoxia-regulated gene, and VEGF levels are increased in hypoxic or ischemic retina. In most cases, Ang2 binds to Tie2 but does not stimulate phosphorylation and acts as a Tie2 antagonist. In the eye, Ang2 is upregulated at the site of neovascularization and acts as a permissive factor for VEGF. Increased expression of VEGF in the retina does not stimulate the initiation of new blood vessel formation from the superficial or intermediate capillary beds of the retinal or choroidal vascular layer, but stimulates the initiation of deep capillary beds where Ang2 is constitutively expressed. Co-expression of VEGF and Ang2 on the retinal surface leads to the initiation of new blood vessel formation from superficial retinal capillaries.

[0230] Angiogenesis is the process of generating new blood vessels from pre-existing blood vessels and 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 for growth and proliferation. VEGF may be a major factor in angiogenesis by increasing vascular permeability and capillary number.

[0231] VEGF is a protein found primarily in endothelial cells and functions in angiogenesis, vasculogenesis, and vascular permeability. VEGF expression is induced by hypoxia, activated oncogenes, and cytokines. VEGF activation can lead to angiogenesis not only in normal human cells and tissues, but also in tumors, and allows tumor progression and growth. Inhibition of VEGF can inhibit tumor growth, leading to tumor regression. Various retinal diseases are associated with increased VEGF levels; ischemia of the eye leads to the induction of VEGF production due to lack of oxygen. This increase in VEGF can lead to excessive proliferation of blood vessels in the retina and can lead to blindness. The disclosed anti-VE-PTP (HPTP-β) can be used to stabilize the ocular vasculature and can counteract stimulation by VEGF and other inflammatory agents present in the diseased retina. In some embodiments, after the withdrawal of anti-VEGF drug administration to the subject, the administration of anti-VE-PTP (HPTP-β) to the subject can maintain the level of disease reversal.

[0232] Macular degeneration is characterized by the gradual loss or impairment of vision caused by cell and tissue degeneration in the macula of the center of the retina. Macular degeneration is usually characterized by one of two types: non-exudative (dry type) or exudative (wet type). Although these two types are bidirectional and progressive, each type can reflect different pathological processes. Wet age-related macular degeneration (AMD) is the most common form of choroidal neovascularization and the leading cause of blindness in the elderly. AMD has affected millions of Americans over the age of 60 and is the main cause of new blindness in the elderly.

[0233] Choroidal neovascular membrane (CNVM) is a problem associated with a variety of retinal diseases, but is most commonly associated with age-related macular degeneration. With CNVM, abnormal blood vessels originating from the choroid (a blood vessel-rich tissue layer just below the retina) grow through the retinal layers. These new blood vessels are very fragile and easily rupture, allowing blood and fluid to pool within the retinal layers.

[0234] Diabetes 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 (commonly referred to as juvenile-onset diabetes) and type 2 diabetes (commonly referred to as adult-onset diabetes). Type 1 diabetes results from the body's inability to produce insulin due to the absence of insulin-producing cells, and currently requires patients to inject insulin. Type 2 diabetes usually results from insulin resistance, a condition in which cells cannot use insulin normally. Diabetes can be associated with a large number of other conditions, including conditions or diseases of the eye, including diabetic retinopathy and diabetic macular edema (DME).

[0235] Diabetic retinopathy is a complication of diabetes caused by damage to the blood vessels of the light-sensitive tissue at the back of the eye (retina). At first, diabetic retinopathy may cause no symptoms or only mild vision problems. Eventually, diabetic retinopathy can lead to blindness. Diabetic retinopathy can occur in anyone with type 1 diabetes or type 2 diabetes.

[0236] In the early stages of non-proliferative retinopathy, microaneurysms occur in the tiny blood vessels of the retina. As the disease progresses, more of these blood vessels become damaged or blocked, and these areas of the retina send signals to the regional tissue to grow new blood vessels for nutrition. This stage is called proliferative retinopathy. New blood vessel growth grows along the retina and along the surface growth of the clear glassy gel that fills the inside of the eye. These blood vessels have thin, brittle walls, and in the absence of timely treatment, these new blood vessels may leak blood (e.g., whole blood or some of its components), and may cause severe vision loss or even blindness. In addition, fluid can leak into the center of the macula, i.e. the part in the eye where clear straight-ahead vision occurs. Fluid and related proteins begin to deposit on or under the macula swelling, and the central vision of the subject becomes distorted. This situation is called macular edema, and can occur at any stage of diabetic retinopathy, but it is more likely to occur as the disease progresses.

[0237] Uveitis is a condition in which the uvea becomes inflamed. The eye is hollow on the inside, with three different tissue layers surrounding the central cavity. The outermost is the sclera (the white outer layer 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 new blood vessel formation (neovascularization).

[0238] Eye trauma is any form of physical or chemical damage to the eye. Eye trauma can affect anyone, and the main symptoms include redness, swelling, or pain in the affected eye. Symptoms are less likely to occur if the trauma is caused by a tiny projectile.

[0239] Surgery-induced edema is the swelling of the eye tissue after surgery on the retina or other parts of the eye. Cystoid macular edema (CME) is an example of this phenomenon. CME can occur not only in people who have undergone cataract surgery, but also in those who suffer from diabetes, retinitis pigmentosa, AMD, or conditions that cause chronic inflammation in the eye. The main symptom of CME is blurred or decreased central vision.

[0240] Ocular ischemic syndrome (OIS) includes signs and symptoms caused by chronic vascular insufficiency. This condition results from insufficient ocular perfusion caused by occlusion or stenosis of the common or internal carotid arteries. OIS typically affects subjects aged 50-80 years who typically display systemic diseases such as hypertension or diabetes. The main symptoms of OIS are orbital pain, visual loss, visual field changes, asymmetric cataracts, and unresponsiveness to light.

[0241] Retinal vein occlusion (RVO) is the most common retinal vascular disease after diabetic retinopathy. According to the area where the retinal venous drainage is effectively blocked, the condition is generally classified as central retinal vein occlusion (CRVO), hemispheric retinal vein occlusion (HRVO) or branch retinal vein occlusion (BRVO). The manifestation of RVO is any combination of fundus manifestations with variable painless visual loss accompanied by: retinal vascular distortion, retinal hemorrhage (spot-shaped and flame-shaped), cotton wool-like exudate points, optic disc swelling and macular edema. In CRVO, retinal hemorrhage can be seen in all four quadrants of the fundus, while in HRVO, this hemorrhage is confined to the upper or lower fundus hemisphere. In BRVO, hemorrhage is mainly confined to the area drained by the occluded branch retinal vein. Vision loss occurs secondary to macular edema or ischemia.

[0242] The compositions of the present disclosure are used 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 diseased retina. In some embodiments, administration of an antibody of the present disclosure to a subject can be used to maintain a level of disease reversal after withdrawal of anti-VEGF drug administration to the subject.

[0243] Treatment of subjects

[0244] In some embodiments, a single administration of a composition of the disclosure in a subject having a disease or condition as disclosed herein results in sustained intraocular expression of a VE-PTP (HPTP-β) inhibitor at levels sufficient to provide long-term inhibition of ocular neovascularization.

[0245] For example, the level of VE-PTP (HPTP-β) inhibitor produced in the host ocular cells can 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 700 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 can 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.

[0246] The protein level may 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 present 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.

[0247] Foveal thickness

[0248] Also disclosed herein is a method for reducing the foveal thickness (CFT) of a subject suffering from a disease or condition as disclosed herein. The method comprises administering to the eye an antibody targeting VE-PTP (HPTP-β), wherein administration of the antibody can be performed in any manner.

[0249] The reduction level of the foveal thickness may be, for example, about 50 μm to about 1000 μm. The reduction level of 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.

[0250] Visual acuity

[0251] Further disclosed herein are methods for improving visual acuity in a subject suffering from a disease or condition as disclosed herein.

[0252] Visual acuity is the sharpness or clarity of vision, which depends on the sharpness of the retina focus in the eye and the sensitivity of the brain's interpretive ability. Visual acuity is a measure of the spatial resolution of the visual processing system. Visual acuity is tested by asking the subject to identify characters (usually numbers or letters) on a chart at a set distance. The graphic characters are represented as black symbols on a white background. The distance between the subject's eyes and the test chart is set at a sufficient distance to approximate infinity in a way that the lens attempts to focus. Twenty feet or 6 meters is essentially infinity from an optical point of view. In the present invention, the improvement in visual acuity is evaluated by the increase in the number of letters that can be read from the chart.

[0253] A non-limiting test for measuring visual acuity is to use the ESV-3000 ETDRS test set and self-calibrating test lighting. The ESV-3000 set incorporates LED light source technology. The self-calibrating circuitry continuously monitors the LED light source and calibrates to 85cd / m 2 or 3cd / m 2 Test brightness.

[0254] Although clinical trials were designed in which large-format ETDRS testing (up to 20 / 200) was 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 ETDRS, testing should be performed under standard lighting conditions, e.g., 85 cd / m 2 The visual acuity test is performed at the photopic level of . The scoring of visual acuity can be completed in any manner selected by the monitor. After providing a baseline assessment, the increase or decrease in the number of letters that can be identified by the test subject during treatment provides a measure of improvement or decrease in visual acuity.

[0255] Disclosed herein is a method for improving the visual acuity of a subject suffering from an ocular disease or condition disclosed herein. The method includes administering an antibody targeting VE-PTP (HPTP-β) to a subject suffering from an ocular disease or condition, wherein the administration of the antibody can be performed 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.

[0256] Example

[0257] Example 1: Production of monoclonal antibodies against the extracellular domain of VE-PTP (HPTP-β) using hybridoma technology.

[0258] 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, such as Figure 3 Schematic diagram shown. Mice are challenged with human VE-PTP (HPTP-β) ECD protein to produce antibodies against the target protein. B cells are then harvested from the spleen and lymph nodes and fused with myeloma cells to produce hybridoma cell lines. The fused cells are cultured in a culture medium in which only myeloma cells with genes derived from antigen-exposed mice are able to grow. Colonies derived from a single myeloma cell are produced by limiting dilution. The antibodies produced by the hybridoma colonies are then screened by ELISA for binding to human VE-PTP (HPTP-β) ECD. One hybridoma, R15E6, was selected for further analysis. The R15E6 antibody binds to endogenous human VE-PTP (HPTP-β) from human umbilical vein endothelial cells (HUVEC), as Figure 4 Immunoprecipitation (IP) western blot (left panel) and flow cytometry (right panel) were performed in Figure 1. No binding of R15E6 to the bead control (protein A / G beads without antibody; BC) or VEGFR2 or Tie2 (R2 / T2) was observed.

[0259] Example 2: Tie2 activation in HUVEC using R15E6

[0260] R15E6 activated Tie2 in a concentration-dependent manner, such as Figure 6 As demonstrated by the IP experiments shown in panels A and B. As shown in panel C, the antibody also enhanced the viability of serum-starved endothelial cells in HUVECs in a concentration-dependent manner.

[0261] Example 3: Tie2 activation and cell permeability experiments were performed using mouse monoclonal and polyclonal anti-mouse VE-PTP ECD antibodies.

[0262] To characterize the in vitro therapeutic effects of VE-PTP antibodies, 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 a VE-PTP-Fc fusion protein with eight FN3 repeats at the N-terminus 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 (pAb) against the extracellular fibronectin type III (FN3)-like domains 1-8 of mouse VE-PTP were selected for further analysis.

[0263] Figure 7The effect of mAb 109.1 on Tie2 activation (upper left), mouse VE-PTP expression (lower left), and Akt (right) in mouse endothelial cells (bEnd) as determined by western blot is shown. Treatment with mAb 109.1 resulted in activation of Tie2 as shown by Tie2 phosphorylation (pTyr; upper left). Akt was constitutively activated in the tested mouse endothelial cells as demonstrated by the lack of increase in phosphorylation and the increase in antibody concentration (right).

[0264] In vitro experiments with mouse pAb 1-8 also enhanced Tie2 (Tie-2) activation and downstream signaling in endothelial cells. Cultured mouse endothelial cells (bEnd.5) were treated with pAb 1-8 or pre-immune (control) antibodies for 1 hour, followed by immunoprecipitation for Tie2 and immunoblotting with anti-phosphotyrosine antibody (pTyr) and anti-Tie2 antibodies. Aliquots of cell lysates with equal protein content were directly immunoblotted for VE-PTP and Tie2 (bottom). Figure 8 Schematic representation of immunoprecipitation experiments in which pAb rapidly induced Tie2 activation in bEnd.5 cells treated with anti-mouse VE-PTP polyclonal antibody or preimmune antibody for 1 h (panel A) or 3 min (panel B).

[0265] Consistent with the vascular stabilization role of Tie2 activation, VE-PTP ECD antibodies reduced thrombin- and VEGF-induced permeability of endothelial monolayers in vitro. Fig. 9 As shown. Paracellular permeability of cultured HUVEC monolayers grown in transwell filters to 250 kD FITC-dextran was determined. Permeability was induced with thrombin (Figure A) or VEGF (Figure B). The 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 pre-immune antibody (control Ab). Data represent two independent experiments.

[0266] Consistent with the vascular stabilization role of Tie2 activation, VE-PTP ECD antibody also blocked VEGF-induced skin vascular permeability in vivo, e.g. Fig.10As shown. As demonstrated by the Miles assay, both monoclonal and polyclonal VE-PTP ECD antibodies mAb109.1 (Figure A) and pAb PTP 1-8 (Figure B) inhibited VEGF-induced skin vascular permeability. The Miles assay can be used as an in vivo model of vascular permeability, which has the characteristics of vascular leakage and new blood vessel formation. Mice were injected intravenously with 100 μg control IgG or anti-mouse VE-PTP antibody 30 minutes before starting the Miles assay. Evans blue was then injected intravenously into the mice, and VEGF (striped bars) or PBS (solid black bars) were injected intradermally 10 minutes later. After 30 minutes, the mice were killed and the dye was extracted from the skin samples and quantified. The data sets were checked for normality and equal variance. Values ​​are expressed as mean ± standard error of the mean (SEM). As shown in Figure C, as demonstrated in the lung tissue of mice treated with pAb PTP 1-8, the blockade of permeability was associated with Tie2 phosphorylation.

[0267] Example 4: Effects of mouse VE-PTP ECD monoclonal antibody on mouse retinal and choroidal neovascularization.

[0268] In an ischemic retinopathy model that mimics aspects of proliferative diabetic retinopathy, VE-PTP ECD monoclonal antibody (mAb 109.1) significantly reduced retinal neovascularization after administration of the antibody, such as Fig.11 (Figure A) and as Fig.12 (Figure A) quantified. At P12, mice with ischemic retinopathy were administered intravitreal injections of 0.1, 0.5 or 2 μg of mAb 109.1 or 2 μg of IgG isotype control (n≥12 per group). At P17, extensive GSA-stained retinal neovascularization was observed in control IgG-injected eyes, while significantly reduced retinal neovascularization was observed in eyes injected with 2 μg of anti-VE-PTP. *P<0.001.

[0269] Similarly, in two models that mimic different aspects of wet age-related macular edema (retinal angiomatous proliferation and choroidal neovascularization), a single 2 μg intravitreal administration of a VE-PTP ECD monoclonal antibody (mAb109.1) significantly reduced retinal neovascularization in both models ( Fig.11 Figure B, and in Fig.12(quantified in Figure B). At P15, six Rho-VEGF transgenic mice were given intravitreal injections of 0.5 or 2 μg of mAb 109.1 in one eye and the corresponding dose of control IgG in the contralateral eye. At P21, significantly reduced GSA-stained subretinal neovascularization was observed in eyes treated with 0.5 or 2 μg of mAb 109.1 compared with control IgG-treated eyes. Unpaired t-test compared with IgG control contralateral eyes yielded *P=0.01. Scale bar: 100 μm.

[0270] Compared with control IgG, intravitreal injection of 2 μg of mAb 109.1 significantly reduced the area of ​​choroidal neovascularization at the site of Bruch's membrane rupture ( Fig.11 Figure C, and in Fig.12 quantified in Figure C of Figure 1). *P<0.001 by one-way ANOVA with Bonferroni correction. Scale bar: 100 μm.

[0271] Subcutaneous administration of mAb 109.1 also reduced ischemic neovascularization in a mouse model of ischemic retinopathy. The mean area of ​​retinal neovascularization was reduced by approximately 30% compared with vehicle, as shown in Figure 2. Fig.25 Mice with ischemic retinopathy were administered 3 subcutaneous injections of 2 mg / kg qod (every other day).

[0272] Example 5: Effect of mouse VE-PTP ECD monoclonal antibody in combination with anti-VEGF agent on retinal detachment in mice.

[0273] To evaluate the effects of combination therapy using the anti-VE-PTP antibodies disclosed herein, the Tet / opsin / VEGF mouse model was used. Tet / opsin / VEGF transgenic mice suffer from severe subretinal neovascularization and vascular leakage, and develop exudative retinal detachment mediated by inducible retinal overexpression of human VEGF. Fig.24 The enhanced effect of the combination of anti-mouse VE-PTP antibody (mAb 109.1) and aflibercept on preventing retinal detachment in mice compared to either treatment alone is shown as indicated by double asterisks. Rat IgG was used as a control.

[0274] Example 6: Production and purification of humanized antibody against human VE-PTP (HPTP-β) from R15E6.

[0275] R15E6 is a 150 kDa antibody containing a heavy chain and a light chain. Sixteen different humanized antibody variants were synthesized using different combinations of heavy chain sequences and light chain sequences as shown in Table 12.

[0276] The R15E6 humanized variants described in Example 1 were cloned into the pVitro DHFR3 mammalian expression vector. A mid-scale transient transfection expression assay was performed to determine the expression yield from CHO cells. In a 500 mL vented flask, the adapted CHO cells were suspended at 2.0-3.0 x 10 5 Cells / mL were cultured in Pro CHO4 serum-free medium supplemented with 8 mM L-glutamine and 10 mL / L hypoxanthine / thymidine at 85% CO2, 37°C, 150 rpm. Maximal preparations of each construct were prepared using the Nucleobond pc500 kit according to the manufacturer's instructions. TM The vector DNA was quantified spectrophotometrically.

[0277] In a 500 mL vented flask, cells were transiently transfected with 1.25 μg / mL of plasmid DNA at a final concentration of 1.0 x 10 6 The transfected cultures were incubated at 85% CO2, 37°C, 150 rpm for 8 to 11 days and then harvested by centrifugation at 4,000 rpm, 4°C for 40 min.

[0278] After centrifugation, the culture 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. All humanized antibody purifications were performed using a 1-mL HiTrap Protein A column. All purifications were performed using Fusion Antibodies internal wash and elution buffers. After loading the protein into the column, any bound antibodies were eluted using glycine / Tris buffer (pH 3). All eluted 1 mL portions were neutralized with 100 μL Tris buffer (pH 8.5). The eluted portion corresponding to the elution peak was selected for dialyzation in PBS overnight. After dialysis, the eluted portion was dialyzed overnight in PBS using a NanoDrop Lite TM Measure the antibody concentration using a spectrophotometer.

[0279] Example 7: Evaluation of humanized antibodies against human VE-PTP (HPTP-β).

[0280] By reducing and non-reducing SDS-PAGE ( Fig.13) Analyze samples of humanized antibodies to determine the presence of heavy and light chains and the purity of the antibodies. The mouse / human chimeric antibody HCOLC0 is derived from R15E6 by fusing the mouse heavy and light chain variable regions to the human Fc region. In both SDS-PAGE gels, lane 1: protein standard prestained with SeeBlue Plus2; lane 2: HCOLC0#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 1 Lane 3: 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 that all analyzed samples were of high purity. In reducing SDS-PAGE, the heavy and light chains were visible, and the heavy chain was at the expected molecular weight of 50 kDa and the light chain was at the expected molecular weight of 25 kDa. In non-reducing SDS-PAGE, no free heavy or light chains were observed at approximately 50 kDa or 25 kDa, respectively. The gel showed small amounts of protein for all HC4 variants as well as the HC1LC3, HC2LC3 and HC3LC3 variants.

[0281] An extinction coefficient of 13.7 was used as the IgG standard reference and the NanoDrop Lite TM Purified R15E6 was quantified by spectrophotometer at 280 nm. The results of SDS-PAGE analysis are summarized in Table 16. Consistent with the SDS-PAGE analysis, all heavy chain 4 variants expressed little to no antibody.

[0282] Table 16

[0283]

[0284] Example 8: Selectivity of R15E6 and humanized variants for human VE-PTP (HPTP-β), cynomolgus VE-PTP (cynomolgus PTP-β) and human HPTP-η.

[0285] Tissue culture supernatants containing antibodies from hybridomas were tested against the targets human VE-PTP (human β1 / 2 ECD), cynomolgus VE-PTP (cynomolgus β), and human HVTP-η (human Eta) to determine binding affinity by ELISA.

[0286] HEK293 cells were used to produce and purify human HPTP-β, cynomolgus monkey PTPβ and human HVTP-η respectively. HEK293 cells were inoculated in a shake flask for 24 hours and then transfected with a plasmid expressing species-specific VE-PTP or HPTP-η. HEK293 cells were cultured using a serum-free chemically defined culture medium. The DNA of species-specific VE-PTP or HPTP-η constructs was transiently transfected into 30 mL HEK293 suspensions. After 24 hours, cells were counted to assess the viability of the cells and determine the viable cell count. Additional readings were obtained throughout the transient transfection process. Cell culture was harvested on the 5th day of transient transfection. The conditioned medium supernatant harvested from transient transfection was clarified by centrifugation. The supernatant was filtered using a 0.2 μm membrane filter. The protein was then purified by anti-His tag affinity chromatography. After purification, the protein was exchanged with PBS (pH 7.4) buffer. Then, SDS-gel capillary electrophoresis (CE-SDS) was performed to analyze and purify the protein. The amino acid sequences of the DNA plasmid inserts, the DNA plasmid inserts and the corresponding wild-type sequences are shown below in Table 15. The signal peptide of the protein sequence is underlined.

[0287] Table 15

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298] HPTP-β, cynomolgus monkey PTP-β and HPTP-η proteins were coated on ELISA plates in coating buffer (pH 9.4). Each protein was prepared as 1 μg / mL protein stock in 50mM carbonate-bicarbonate. The coated protein was aliquoted into sterile transparent 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 3 times with 200 μL / well of 1x PBS-T (PBS containing 0.05% Tween 20). The immune protein was human VE-PTP (HPTP-β). The plates were then sealed with 5% blotting grade blocking agent skim milk powder in 200 μL / well of PBS-T and incubated for 1 hour at room temperature.

[0299] The primary antibody sample anti-hCD20-hIgG4S228P for humanized variants and negative controls was prepared using 1 μg / well in 5% blotting grade blocking agent non-fat dry milk in PBS. Samples were tested in triplicate. Fab antibody fragments were treated the same as IgG antibodies. The plate was then washed 3 times with PBS-T and 100 μL of the primary antibody preparation was added to each well. The plate was then incubated at room temperature for 1 hour.

[0300] The secondary antibody samples were prepared by diluting a peroxidase-conjugated affinity-purified donkey anti-human IgG specific secondary antibody at a 1:2,000 dilution in a blotting grade blocking agent, non-fat dry milk in PBS. The plates were then washed three times with PBS-T and 100 μL of the secondary antibody preparation was added to each well. The plates were then incubated at room temperature for 1 hour. After incubation, the plates were washed three times with PBS-T.

[0301] Then, 75 μg / 1-Step was added to each well. TM Ultra TMB-ELISA, and allow the plate to develop for 5 minutes at room temperature. After 5 minutes, add 50 μL of 2M sulfuric acid to each well to stop the reaction. The plate is then analyzed on a plate reader at an absorbance wavelength of 450 nm.

[0302] Figure 5 Western blots of R15E6 screened for species-specific VE-PTP and HPTP-η are shown. R15E6 has high selectivity for human VE-PTP (HPTP-β; lanes 1 and 4). No significant binding to human PTP-η or cynomolgus VE-PTP (cynomolgus PTP-β) was observed. The recombinant hexa-histidine-tagged proteins (6-His) shown were probed sequentially with R15E6 and commercially available 6-His antibodies. The commercially available 6-His antibody showed binding to all protein targets and showed poor selectivity for human VE-PTP (HPTP-β).

[0303] The affinity of each humanized variant to the antigen human VE-PTP (HPTP-β) was measured using ELISA. 100 mg / well of VE-PTP (HPTP-β) antigen was fixed overnight at 4 ° C on a 96-well Maxisorp plate in coating buffer (0.5 mM NaHCO3, pH was reached to 9.6 by adding 0.5 mM Na2CO3). The coating buffer was then removed and 200 μL / well of blocking solution (3% w / v semi-skimmed milk powder, PBS) was added and shaken at room temperature for 2 hours. The plate was washed 4 times with PBS-T (1% v / v Tween 20). 100 μL / well of purified R15E6 variant was added, which was serially diluted from 10,000 ng / mL to 9.765 ng / mL in PBS-TB (0.05% v / v Tween 20, 0.5% w / v BSA). After shaking at room temperature for 2 hours, the plate was washed 4 times with PBS-T. Then 100 μL / well of goat anti-human HRP (Fc specific) diluted 1:60,000 in PBS was added and the plate was incubated for another hour at room temperature with shaking. The plate was then washed 6 times with PBS-T and once in PBS. Next, 100 μL / well of TMB substrate solution was added and incubated at 37°C for 10 minutes. 50 μL of 1M HCl was added to each well and the plate was immediately read at 450nm on a Tecan Sunrise plate reader. ELISA results were obtained at Figure 14-17 . Compared to the chimeric HCOLC0 antibody, the HC1, HC2 and HC3 variants generally showed similar binding properties. Compared to HCOLC0, the binding of the HC4 variant was much lower than that of the other heavy chain variants ( Fig.17 ).

[0304] Samples of each variant were further evaluated by Biacore analysis to determine binding affinity. TM Antibody binding experiments were performed on . 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 for capturing the ligand was 5 μL / min. The flow rate for kinetic analysis was 50 μL / min. Flow cells 1 and 2 of the CM5 chip were coated with the maximum amount of goat anti-human IgG. As shown, approximately 150 RU of the test antibody was captured on flow cells 2, 3, and 4. The antigen flowed through the chip. The binding of the antigen to the antibody was monitored in real time. According to the observed k on and k off Determine the value of K D .

[0305] Probing assays were performed using single analyte concentrations as indicated. At these concentrations, binding was observed even though the ligand binding was weak. The flow cell 1 response was used for reference subtraction.

[0306] Complete kinetic analysis was performed for control antibodies with 2-fold serial dilutions over the concentration range of analyte as indicated: 50, 25, 12.5, 6.25, 3.125 and 0 nM.

[0307] Table 17 shows the binding kinetic parameters for each humanized antibody binding to VE-PTP (HPTP-β) ECD, including the association rate (k) determined by Biacore binding assay. a ), dissociation rate (k d ), maximum binding capacity (R max ), binding constant (K A ) and dissociation constant (K D ). max Measured as relative response. All humanized candidates showed similar binding affinity to the mouse / human chimeric antibody HCOLC0.

[0308] Table 17

[0309]

[0310] Table 18 shows the binding selectivity of the humanized antibodies compared to the parental antibody (R15E6) and the inactive control (IgG4) determined by ELISA. Values ​​less than or equal to 0.2 indicate no binding. The lack of significant binding to human PTP-η and cynomolgus monkey PTP-β indicates that the humanized antibody candidate, parental R15E6, and mouse / human chimeric antibody (HCO:LCO) all exhibit high selectivity for human VE-PTP (HPTP-β).

[0311] Table 18

[0312]

[0313] Humanized antibodies used to target human VE-PTP (HPTP-β) can restore Tie2 activity through autophosphorylation and activate downstream effectors, including, for example, proteins in the PI3K–Akt pathway. In addition to evaluating the binding of antibodies to human VE-PTP (HPTP-β), the ability of the antibodies to activate these effector proteins was also measured. Fig.18The activation of Tie2 (left figure) and Akt (right figure) by humanized variants determined by western blotting in the absence of Tie2 ligands (e.g., Ang1 or Ang2). The activity of the humanized variants is similar to that of the parental murine antibody R15E6 and the chimeric antibody HCOLC0. Human IgG4 was used as a control. All antibodies were tested at 50 nM. Each blot in the above figure shows the phosphorylation of Tie2 or Akt, indicating activation of Tie2 or Akt. Tie2 and Akt showed stronger phosphorylation in samples treated with humanized antibodies than in those treated with control antibodies, and indicated that humanized antibodies were effective in activating Tie2 and Akt even in the absence of ligands.

[0314] Fig.19 The concentration-dependent activation of Tie2 (upper figure) and Akt (lower figure) by humanized antibodies HC2LC4 and HC1LC1 in the absence of Tie2 ligands (e.g., Ang1 or Ang2) determined by western blotting is shown. Tie2 and Akt signaling activity were observed at low nanomolar concentrations of humanized variants. Each blot in the upper figure shows phosphorylation of Tie2 or Akt, indicating activation of Tie2 or Akt. Tie2 and Akt showed stronger phosphorylation in samples treated with humanized antibodies than in samples treated with control antibodies. The results indicate that humanized antibodies are effective in activating Tie2 and Akt even in the absence of Ang1.

[0315] Fig. 20 Graphs show activation of Tie2 (left) and Akt (right) by humanized antibody candidates HC2LC4 and HC1LC1 in the presence of Ang1 and Ang2 as determined by western blot. Increased phosphorylation of Tie2 and Akt indicates that HC2LC4 and HC1LC1 activate both Tie2 and Akt in the presence of Ang2, respectively, and enhance Ang1-mediated activation of Tie2 and Akt. All antibodies were tested at 10 nM.

[0316] Fig.21 Graphs show activation of Tie2 (Figure A), human VE-PTP (HPTP-β) expression (Figure B), and Akt (Figure C) by humanized antibody candidates HC2LC1 and HC2LC4 in the presence of Ang1 or Ang2 as determined by western blotting. HC2LC1 and HC2LC4 activated Tie2 in the presence of Ang1 alone, Ang2 alone, and both Ang1 and Ang2. Antibodies were pre-incubated for 15 minutes and then stimulated by Ang1 or Ang2 at a concentration of 600 ng / mL for 10 min. All antibodies were tested at 10 nM.

[0317] Fig. 22 Activation of Akt in HUVEC using the humanized antibody candidate HC2LC1 is illustrated. The first three blots in the figure above show phosphorylation of Akt at increasing concentrations of HC2LC1, indicating Akt activation. Consistent with the binding affinity studies, HC2LC1 activated Akt at nanomolar concentrations of the antibody. Activation of Akt by HC2LC1 was blocked by pre-incubation of the antibody with the extracellular domain of human VE-PTP (HPTP-β) (βECD6His and βECD Fc). This result suggests that HC2LC1-mediated activation of Akt occurs through binding of HC2LC1 to the extracellular domain of human VE-PTP (HPTP-β).

[0318] Fig.23 Graph showing enhanced viability of serum starved HUVECs using humanized antibody candidates HC2LC1 and HC0LC0 compared to parental antibody R15E6 and mIgG1 control.

[0319] Implementation

[0320] 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] Embodiment 5. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:9.

[0325] Embodiment 6. The compound of any one of embodiments 1-5, wherein the sequence is SEQ ID NO:9.

[0326] Embodiment 7. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:10.

[0327] Embodiment 8. The compound of any one of embodiments 1-4 and 7, wherein the sequence is SEQ ID NO:10.

[0328] Embodiment 9. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO: 11.

[0329] Embodiment 10. The compound of any one of embodiments 1-4 and 9, wherein the sequence is SEQ ID NO:11.

[0330] Embodiment 11. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:12.

[0331] Embodiment 12. The compound of any one of embodiments 1-4 and 11, wherein the sequence is SEQ ID NO:12.

[0332] Embodiment 13. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:29.

[0333] Embodiment 14. The compound of embodiment 1, wherein the sequence is SEQ ID NO:29.

[0334] Embodiment 15. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:30.

[0335] Embodiment 16. The compound of embodiment 1, wherein the sequence is SEQ ID NO:30.

[0336] Embodiment 17. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:31.

[0337] Embodiment 18. The compound of embodiment 1, wherein the sequence is SEQ ID NO:31.

[0338] Embodiment 19. The compound of embodiment 1, wherein the sequence is at least 80% identical to SEQ ID NO:32.

[0339] Embodiment 20. The compound of embodiment 1, wherein the sequence is SEQ ID NO:32.

[0340] Embodiment 21. A compound as described in any of embodiments 1-20, wherein the compound inhibits tyrosine phosphatase.

[0341] Embodiment 22. The compound of any one of embodiments 1-21, wherein the compound inhibits HPTP-β.

[0342] Embodiment 23. A compound as described in any of embodiments 1-22, wherein the compound inhibits VE-PTP.

[0343] Embodiment 24. The compound of any one of embodiments 1-23, wherein the compound activates Tie2.

[0344] Embodiment 25. The compound of any one of embodiments 1-24, wherein the compound activates Akt.

[0345] Embodiment 26. The compound of any one of embodiments 1-25, wherein the compound binds to the extracellular domain of HPTP-β.

[0346] Embodiment 27. The compound of any one of embodiments 1-26, wherein the compound binds to the first FN3 repeat sequence of the extracellular domain of HPTP-β.

[0347] Embodiment 28. A compound as described in any one of embodiments 1-27, wherein the binding affinity (K) of the compound to the extracellular domain of HPTP-β is D ) is about 70 pM to about 70 nM.

[0348] Embodiment 29. A compound comprising a sequence at least 80% identical to SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 or SEQ ID NO:23.

[0349] 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.

[0350] Embodiment 31. A compound as described in any 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.

[0351] Embodiment 32. The compound of any one of embodiments 29-31, 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.

[0352] Embodiment 33. The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:20.

[0353] Embodiment 34. A compound as described in any of embodiments 29-33, wherein the sequence is SEQ ID NO:20.

[0354] Embodiment 35. The compound of embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:21.

[0355] Embodiment 36. The compound of any one of embodiments 29-32 and 35, wherein the sequence is SEQ ID NO:21.

[0356] Embodiment 37. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:22.

[0357] Embodiment 38. The compound of any one of embodiments 29-32 and 37, wherein the sequence is SEQ ID NO:22.

[0358] Embodiment 39. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:23.

[0359] Embodiment 40. The compound of any one of embodiments 29-32 and 39, wherein the sequence is SEQ ID NO:23.

[0360] Embodiment 41. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:34.

[0361] Embodiment 42. The compound of Embodiment 29, wherein the sequence is SEQ ID NO:34.

[0362] Embodiment 43. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:35.

[0363] Embodiment 44. The compound of Embodiment 29, wherein the sequence is SEQ ID NO:35.

[0364] Embodiment 45. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:36.

[0365] Embodiment 46. The compound of embodiment 29, wherein the sequence is SEQ ID NO:36.

[0366] Embodiment 47. The compound of Embodiment 29, wherein the sequence is at least 80% identical to SEQ ID NO:37.

[0367] Embodiment 48. The compound of embodiment 29, wherein the sequence is SEQ ID NO:37.

[0368] Embodiment 49. The compound of any one of embodiments 29-48, wherein the compound inhibits tyrosine phosphatase.

[0369] Embodiment 50. The compound of any one of embodiments 29-49, wherein the compound inhibits HPTP-β.

[0370] Embodiment 51. A compound as described in any of Embodiments 29-50, wherein the compound inhibits VE-PTP.

[0371] Embodiment 52. A compound as described in any of embodiments 29-51, wherein the compound activates Tie2.

[0372] Embodiment 53. A compound as described in any of embodiments 29-52, wherein the compound activates Akt.

[0373] Embodiment 54. A compound as described in any of embodiments 29-53, wherein the compound binds to the extracellular domain of HPTP-β.

[0374] Embodiment 55. The compound of any one of embodiments 29-54, wherein the compound binds to the first FN3 repeat sequence of the extracellular domain of HPTP-β.

[0375] Embodiment 56. A compound as described in any of embodiments 29-55, wherein the binding affinity (K) of the compound to the extracellular domain of HPTP-β is D ) is about 70 pM to about 70 nM.

[0376] Embodiment 57. A compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:34.

[0377] 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.

[0378] Embodiment 59. A compound as described in any 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.

[0379] Embodiment 60. A compound as described in any 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.

[0380] Embodiment 61. A compound as described in any of Embodiments 57-60, wherein the heavy chain is SEQ ID NO:30 and the light chain is SEQ ID NO:34.

[0381] Embodiment 62. A compound as described in any of embodiments 57-61, wherein the compound inhibits tyrosine phosphatase.

[0382] Embodiment 63. A compound as described in any of embodiments 57-62, wherein the compound inhibits HPTP-β.

[0383] Embodiment 64. A compound as described in any of embodiments 57-63, wherein the compound inhibits VE-PTP.

[0384] Embodiment 65. A compound as described in any of embodiments 57-64, wherein the compound activates Tie2.

[0385] Embodiment 66. The compound of any one of embodiments 57-65, wherein the compound activates Akt.

[0386] Embodiment 67. A compound as described in any of embodiments 57-66, wherein the compound binds to the extracellular domain of HPTP-β.

[0387] Embodiment 68. The compound of any one of embodiments 57-67, wherein the compound binds to the first FN3 repeat sequence of the extracellular domain of HPTP-β.

[0388] Embodiment 69. A compound as described in any of embodiments 57-68, wherein the binding affinity (K) of the compound to the extracellular domain of HPTP-β is D ) is about 70 pM to about 70 nM.

[0389] Embodiment 70. A compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:29; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:35.

[0390] Embodiment 71. A compound as described in 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.

[0391] Embodiment 72. A compound as described in any 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.

[0392] Embodiment 73. A compound as described in any 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.

[0393] Embodiment 74. A compound as described in any of embodiments 70-73, wherein the heavy chain is SEQ ID NO:29 and the light chain is SEQ ID NO:35.

[0394] Embodiment 75. The compound of any one of embodiments 70-74, wherein the compound inhibits tyrosine phosphatase.

[0395] Embodiment 76. A compound as described in any of embodiments 70-75, wherein the compound inhibits HPTP-β.

[0396] Embodiment 77. A compound as described in any of Embodiments 70-76, wherein the compound inhibits VE-PTP.

[0397] Embodiment 78. A compound as described in any of embodiments 70-77, wherein the compound activates Tie2.

[0398] Embodiment 79. The compound of any one of embodiments 70-78, wherein the compound activates Akt.

[0399] Embodiment 80. A compound as described in any of embodiments 70-79, wherein the compound binds to the extracellular domain of HPTP-β.

[0400] Embodiment 81. The compound of any one of embodiments 70-80, wherein the compound binds to the first FN3 repeat sequence of the extracellular domain of HPTP-β.

[0401] Embodiment 82. A compound as described in any of embodiments 70-81, wherein the binding affinity (K) of the compound to the extracellular domain of HPTP-β is D ) is about 70 pM to about 70 nM.

[0402] Embodiment 83. A compound comprising: a) a heavy chain comprising a sequence at least 80% identical to SEQ ID NO:30; and b) a light chain comprising a sequence at least 80% identical to SEQ ID NO:37.

[0403] 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.

[0404] Embodiment 85. A compound as described in any 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.

[0405] Embodiment 86. A compound as described in any of Embodiments 83-85, 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.

[0406] Embodiment 87. A compound as described in any of embodiments 83-86, wherein the heavy chain is SEQ ID NO:30 and the light chain is SEQ ID NO:37.

[0407] Embodiment 88. A compound as described in any of embodiments 83-87, wherein the compound inhibits tyrosine phosphatase.

[0408] Embodiment 89. A compound as described in any of embodiments 83-88, wherein the compound inhibits HPTP-β.

[0409] Embodiment 90. A compound as described in any of embodiments 83-89, wherein the compound inhibits VE-PTP.

[0410] Embodiment 91. A compound as described in any of embodiments 83-90, wherein the compound activates Tie2.

[0411] Embodiment 92. The compound of any one of embodiments 83-91, wherein the compound activates Akt.

[0412] Embodiment 93. A compound as described in any of embodiments 83-92, wherein the compound binds to the extracellular domain of HPTP-β.

[0413] Embodiment 94. The compound of any one of embodiments 83-93, wherein the compound binds to the first FN3 repeat sequence of the extracellular domain of HPTP-β.

[0414] Embodiment 95. A compound as described in any of embodiments 83-94, wherein the binding affinity (K) of the compound to the extracellular domain of HPTP-β is D ) is about 70 pM to about 70 nM.

[0415] Embodiment 96. A method of treating a condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of Embodiments 1-95.

[0416] Embodiment 97. A method as described in Embodiment 96, wherein the condition is an ocular condition.

[0417] Embodiment 98. The method of embodiment 96 or 97, wherein the condition is diabetic retinopathy.

[0418] Embodiment 99. The method of any one of Embodiments 96-97, wherein the condition is neovascularization.

[0419] Embodiment 100. The method of any one of Embodiments 96-97, wherein the condition is vascular leakage.

[0420] Embodiment 101. The method of any one of Embodiments 96-97, wherein the condition is increased intraocular pressure.

[0421] Embodiment 102. The method of any one of Embodiments 96-97, wherein the condition is ocular edema.

[0422] Embodiment 103. A method as described in Embodiment 96 or 97, wherein the condition is diabetic macular edema.

[0423] Embodiment 104. The method of any one of Embodiments 96 or 97, wherein the condition is ocular hypertension.

[0424] Embodiment 105. The method of embodiment 96 or 97, wherein the condition is ocular inflammation.

[0425] Embodiment 106. The method of any one of Embodiments 96-105, wherein the administration is to an eye of the subject.

[0426] Embodiment 107. The method of any one of Embodiments 96-106, wherein the administration is intravitreal administration.

[0427] Embodiment 108. The method of any one of embodiments 96-105, wherein the administration is subcutaneous administration.

[0428] Embodiment 109. The method of any one of embodiments 96-106, wherein the administration is topical administration.

[0429] Embodiment 110. The method of any one of embodiments 96-109, wherein the subject is a human.

[0430] 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.

[0431] 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.

[0432] 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.

[0433] Embodiment 114. The method of any one of embodiments 96-110, wherein the therapeutically effective amount of the compound is about 50 mg to about 200 mg. Sequence Listing <110> Elpio Therapeutics Ltd. <120> Humanized monoclonal antibody targeting VE-PTP (HPTP-β) <130> 45725-724.201 <140> 15 / 654,289 <141> 2017-07-19 <150> 62 / 377,072 <151> 2016-08-19 <150> 62 / 364,381 <151> 2016-07-20 <160> 56 <170> PatentIn Version 3.5 <210> 1 <211> 141 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 1 Met Asp Phe Gly Leu Ser Trp Val Phe Phe Val Val Phe Tyr Gln Gly 1 5 10 15 Val His Cys Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Val Gln 20 25 30 Pro Lys Gly Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ala Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ala 85 90 95 Gln Asn Met Leu Tyr Leu Gln Met Asn Asp Leu Lys Thr Glu Asp Thr 100 105 110 Ala Met Tyr Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 130 135 140 <210> 2 <211> 100 <212> PRT <213> Homo sapiens <400> 2 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Ser 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Ser Lys Ala Asn Ser Tyr Ala Thr Ala Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg 100 <210> 3 <211> 100 <212> PRT <213> Homo sapiens <400> 3 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp His 20 25 30 Tyr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Thr Arg Asn Lys Ala Asn Ser Tyr Thr Thr Glu Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg 100 <210> 4 <211> 98 <212> PRT <213> Homo sapiens <400> 4 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg <210> 5 <211> 122 <212> PRT <213> Homo sapiens <400> 5 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Ser 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Ser Lys Ala Asn Asn Tyr Ala Thr Ala Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Ala Tyr 85 90 95 Tyr Cys Ile Arg Asp Tyr Tyr Gly Ala Thr Arg Gly Phe Gln His Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 6 <211> 120 <212> PRT <213> Homo sapiens <400> 6 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp His 20 25 30 Tyr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Thr Arg Asn Lys Ala Asn Ser Tyr Thr Thr Glu Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Tyr Val Val Gly Ala Thr Leu Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 7 <211> 129 <212> PRT <213> Homo sapiens <400> 7 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Ser Phe Gly Asp Tyr 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Phe Ile Arg Ser Lys Thr Tyr Gly Gly Thr Thr Glu Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ile 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Arg Asp Pro Ala Asp Phe Tyr Tyr Tyr Ser Ser Gly Gln 100 105 110 Thr Gly Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser 115 120 125 Ser <210> 8 <211> 120 <212> PRT <213> Homo sapiens <400> 8 Leu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val His Tyr Cys 85 90 95 Ala Arg Asp Gly Tyr Ser Ser Ser Trp Tyr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 9 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 9 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Ala Tyr 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 10 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 10 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 11 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 11 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ile 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 12 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 12 Leu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val His 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 13 <211> 127 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 13 Met Glu Ser Gln Thr Gln Val Phe Val Tyr Met Leu Leu Trp Leu Ser 1 5 10 15 Gly Val Glu Gly Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser 20 25 30 Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro 50 55 60 Lys Gln Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Asn Val Gln Ser Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 115 120 125 <210> 14 <211> 95 <212> PRT <213> Homo sapiens <400> 14 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro 85 90 95 <210> 15 <211> 101 <212> PRT <213> Homo sapiens <400> 15 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30 Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Thr Pro 100 <210> 16 <211> 107 <212> PRT <213> Homo sapiens <400> 16 Asp Val Val Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Leu Gly Gly Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 17 <211> 113 <212> PRT <213> Homo sapiens <400> 17 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30 Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 18 <211> 107 <212> PRT <213> Homo sapiens <400> 18 Asp Ile Gln Met Thr Gln Ser Pro Phe Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Gly Ser Ser 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Leu Gln His His Asp Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 19 <211> 113 <212> PRT <213> Homo sapiens <400> 19 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Phe Tyr Ser 20 25 30 Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Glu Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Ser Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105 110 Lys <210> 20 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 20 Asp Val Val Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> twenty one <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> twenty one Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> twenty two <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> twenty two Asp Ile Gln Met Thr Gln Ser Pro Phe Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> twenty three <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> twenty three Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Glu Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> twenty four <211> 19 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: synthetic Peptides <400> twenty four Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser <210> 25 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: synthetic Peptides <400> 25 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys 20 <210> 26 <211> 327 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: synthetic Peptides <400> 26 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105 110 Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 27 <211> 106 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 27 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 1 5 10 15 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 20 25 30 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 35 40 45 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 50 55 60 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 65 70 75 80 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 85 90 95 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 28 <211> 468 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 28 Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Val Gln 20 25 30 Pro Lys Gly Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ala Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ala 85 90 95 Gln Asn Met Leu Tyr Leu Gln Met Asn Asp Leu Lys Thr Glu Asp Thr 100 105 110 Ala Met Tyr Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Leu Gly Lys 465 <210> 29 <211> 468 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: synthetic Peptides <400> 29 Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu 50 55 60 Glu Trp Val Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr 100 105 110 Ala Ala Tyr Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Leu Gly Lys 465 <210> 30 <211> 468 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 30 Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Leu Gly Lys 465 <210> 31 <211> 468 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 31 Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Arg Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Gly Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Asn Ile Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Leu Gly Lys 465 <210> 32 <211> 468 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 32 Met Gly Trp Thr Leu Val Phe Leu Phe Leu Leu Ser Val Thr Ala Gly 1 5 10 15 Val His Ser Leu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asn Ala Asn Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr 65 70 75 80 Tyr Ala Gly Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val His Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile 115 120 125 Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Leu Gly Lys 465 <210> 33 <211> 234 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 33 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser 20 25 30 Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro 50 55 60 Lys Gln Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Asn Val Gln Ser Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 34 <211> 234 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 34 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys Asp Val Val Met Thr Gln Ser Pro Ser Phe Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro 50 55 60 Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 35 <211> 234 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 35 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala 20 25 30 Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 50 55 60 Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 36 <211> 234 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 36 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Phe Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60 Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 37 <211> 234 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 37 Met Val Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln 1 5 10 15 Gly Thr Arg Cys Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala 20 25 30 Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln His 35 40 45 Val Gly Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Glu Gln Pro Pro 50 55 60 Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Ser 100 105 110 Ser Tyr Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 38 <211> 760 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 38 Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly 1 5 10 15 Val His Ser Glu Pro Glu Arg Cys Asn Phe Thr Leu Ala Glu Ser Lys 20 25 30 Ala Ser Ser His Ser Val Ser Ile Gln Trp Arg Ile Leu Gly Ser Pro 35 40 45 Cys Asn Phe Ser Leu Ile Tyr Ser Ser Asp Thr Leu Gly Ala Ala Leu 50 55 60 Cys Pro Thr Phe Arg Ile Asp Asn Thr Thr Tyr Gly Cys Asn Leu Gln 65 70 75 80 Asp Leu Gln Ala Gly Thr Ile Tyr Asn Phe Lys Ile Ile Ser Leu Asp 85 90 95 Glu Glu Arg Thr Val Val Leu Gln Thr Asp Pro Leu Pro Pro Ala Arg 100 105 110 Phe Gly Val Ser Lys Glu Lys Thr Thr Ser Thr Gly Leu His Val Trp 115 120 125 Trp Thr Pro Ser Ser Gly Lys Val Thr Ser Tyr Glu Val Gln Leu Phe 130 135 140 Asp Glu Asn Asn Gln Lys Ile Gln Gly Val Gln Ile Gln Glu Ser Thr 145 150 155 160 Ser Trp Asn Glu Tyr Thr Phe Phe Asn Leu Thr Ala Gly Ser Lys Tyr 165 170 175 Asn Ile Ala Ile Thr Ala Val Ser Gly Gly Lys Arg Ser Phe Ser Val 180 185 190 Tyr Thr Asn Gly Ser Thr Val Pro Ser Pro Val Lys Asp Ile Gly Ile 195 200 205 Ser Thr Lys Ala Asn Ser Leu Leu Ile Ser Trp Ser His Gly Ser Gly 210 215 220 Asn Val Glu Arg Tyr Arg Leu Met Leu Met Asp Lys Gly Ile Leu Val 225 230 235 240 His Gly Gly Val Val Asp Lys His Ala Thr Ser Tyr Ala Phe His Gly 245 250 255 Leu Thr Pro Gly Tyr Leu Tyr Asn Leu Thr Val Met Thr Glu Ala Ala 260 265 270 Gly Leu Gln Asn Tyr Arg Trp Lys Leu Val Arg Thr Ala Pro Met Glu 275 280 285 Val Ser Asn Leu Lys Val Thr Asn Asp Gly Ser Leu Thr Ser Leu Lys 290 295 300 Val Lys Trp Gln Arg Pro Pro Gly Asn Val Asp Ser Tyr Asn Ile Thr 305 310 315 320 Leu Ser His Lys Gly Thr Ile Lys Glu Ser Arg Val Leu Ala Pro Trp 325 330 335 Ile Thr Glu Thr His Phe Lys Glu Leu Val Pro Gly Arg Leu Tyr Gln 340 345 350 Val Thr Val Ser Cys Val Ser Gly Glu Leu Ser Ala Gln Lys Met Ala 355 360 365 Val Gly Arg Thr Phe Pro Leu Ala Val Leu Gln Leu Arg Val Lys His 370 375 380 Ala Asn Glu Thr Ser Leu Ser Ile Met Trp Gln Thr Pro Val Ala Glu 385 390 395 400 Trp Glu Lys Tyr Ile Ile Ser Leu Ala Asp Arg Asp Leu Leu Leu Ile 405 410 415 His Lys Ser Leu Ser Lys Asp Ala Lys Glu Phe Thr Phe Thr Asp Leu 420 425 430 Val Pro Gly Arg Lys Tyr Met Ala Thr Val Thr Ser Ile Ser Gly Asp 435 440 445 Leu Lys Asn Ser Ser Ser Val Lys Gly Arg Thr Val Pro Ala Gln Val 450 455 460 Thr Asp Leu His Val Ala Asn Gln Gly Met Thr Ser Ser Leu Phe Thr 465 470 475 480 Asn Trp Thr Gln Ala Gln Gly Asp Val Glu Phe Tyr Gln Val Leu Leu 485 490 495 Ile His Glu Asn Val Val Ile Lys Asn Glu Ser Ile Ser Ser Glu Thr 500 505 510 Ser Arg Tyr Ser Phe His Ser Leu Lys Ser Gly Ser Leu Tyr Ser Val 515 520 525 Val Val Thr Thr Val Ser Gly Gly Ile Ser Ser Arg Gln Val Val Val 530 535 540 Glu Gly Arg Thr Val Pro Ser Ser Val Ser Gly Val Thr Val Asn Asn 545 550 555 560 Ser Gly Arg Asn Asp Tyr Leu Ser Val Ser Trp Leu Leu Ala Pro Gly 565 570 575 Asp Val Asp Asn Tyr Glu Val Thr Leu Ser His Asp Gly Lys Val Val 580 585 590 Gln Ser Leu Val Ile Ala Lys Ser Val Arg Glu Cys Ser Phe Ser Ser 595 600 605 Leu Thr Pro Gly Arg Leu Tyr Thr Val Thr Ile Thr Thr Arg Ser Gly 610 615 620 Lys Tyr Glu Asn His Ser Phe Ser Gln Glu Arg Thr Val Pro Asp Lys 625 630 635 640 Val Gln Gly Val Ser Val Ser Asn Ser Ala Arg Ser Asp Tyr Leu Arg 645 650 655 Val Ser Trp Val His Ala Thr Gly Asp Phe Asp His Tyr Glu Val Thr 660 665 670 Ile Lys Asn Lys Asn Asn Phe Ile Gln Thr Lys Ser Ile Pro Lys Ser 675 680 685 Glu Asn Glu Cys Val Phe Val Gln Leu Val Pro Gly Arg Leu Tyr Ser 690 695 700 Val Thr Val Thr Thr Lys Ser Gly Gln Tyr Glu Ala Asn Glu Gln Gly 705 710 715 720 Asn Gly Arg Thr Ile Pro Glu Lys Gly Asn Ser Ala Asp Ile Gln His 725 730 735 Ser Gly Gly Arg Ser Ser Leu Glu Gly Pro Arg Phe Glu Arg Thr Gly 740 745 750 Gly Gly His His His His His His 755 760 <210> 39 <211> 1997 <212> PRT <213> Homo sapiens <400> 39 Met Leu Ser His Gly Ala Gly Leu Ala Leu Trp Ile Thr Leu Ser Leu 1 5 10 15 Leu Gln Thr Gly Leu Ala Glu Pro Glu Arg Cys Asn Phe Thr Leu Ala 20 25 30 Glu Ser Lys Ala Ser Ser His Ser Val Ser Ile Gln Trp Arg Ile Leu 35 40 45 Gly Ser Pro Cys Asn Phe Ser Leu Ile Tyr Ser Ser Asp Thr Leu Gly 50 55 60 Ala Ala Leu Cys Pro Thr Phe Arg Ile Asp Asn Thr Thr Tyr Gly Cys 65 70 75 80 Asn Leu Gln Asp Leu Gln Ala Gly Thr Ile Tyr Asn Phe Arg Ile Ile 85 90 95 Ser Leu Asp Glu Glu Arg Thr Val Val Leu Gln Thr Asp Pro Leu Pro 100 105 110 Pro Ala Arg Phe Gly Val Ser Lys Glu Lys Thr Thr Ser Thr Ser Leu 115 120 125 His Val Trp Trp Thr Pro Ser Ser Gly Lys Val Thr Ser Tyr Glu Val 130 135 140 Gln Leu Phe Asp Glu Asn Asn Gln Lys Ile Gln Gly Val Gln Ile Gln 145 150 155 160 Glu Ser Thr Ser Trp Asn Glu Tyr Thr Phe Phe Asn Leu Thr Ala Gly 165 170 175 Ser Lys Tyr Asn Ile Ala Ile Thr Ala Val Ser Gly Gly Lys Arg Ser 180 185 190 Phe Ser Val Tyr Thr Asn Gly Ser Thr Val Pro Ser Pro Val Lys Asp 195 200 205 Ile Gly Ile Ser Thr Lys Ala Asn Ser Leu Leu Ile Ser Trp Ser His 210 215 220 Gly Ser Gly Asn Val Glu Arg Tyr Arg Leu Met Leu Met Asp Lys Gly 225 230 235 240 Ile Leu Val His Gly Gly Val Val Asp Lys His Ala Thr Ser Tyr Ala 245 250 255 Phe His Gly Leu Thr Pro Gly Tyr Leu Tyr Asn Leu Thr Val Met Thr 260 265 270 Glu Ala Ala Gly Leu Gln Asn Tyr Arg Trp Lys Leu Val Arg Thr Ala 275 280 285 Pro Met Glu Val Ser Asn Leu Lys Val Thr Asn Asp Gly Ser Leu Thr 290 295 300 Ser Leu Lys Val Lys Trp Gln Arg Pro Pro Gly Asn Val Asp Ser Tyr 305 310 315 320 Asn Ile Thr Leu Ser His Lys Gly Thr Ile Lys Glu Ser Arg Val Leu 325 330 335 Ala Pro Trp Ile Thr Glu Thr His Phe Lys Glu Leu Val Pro Gly Arg 340 345 350 Leu Tyr Gln Val Thr Val Ser Cys Val Ser Gly Glu Leu Ser Ala Gln 355 360 365 Lys Met Ala Val Gly Arg Thr Phe Pro Asp Lys Val Ala Asn Leu Glu 370 375 380 Ala Asn Asn Asn Gly Arg Met Arg Ser Leu Val Val Ser Trp Ser Pro 385 390 395 400 Pro Ala Gly Asp Trp Glu Gln Tyr Arg Ile Leu Leu Phe Asn Asp Ser 405 410 415 Val Val Leu Leu Asn Ile Thr Val Gly Lys Glu Glu Thr Gln Tyr Val 420 425 430 Met Asp Asp Thr Gly Leu Val Pro Gly Arg Gln Tyr Glu Val Glu Val 435 440 445 Ile Val Glu Ser Gly Asn Leu Lys Asn Ser Glu Arg Cys Gln Gly Arg 450 455 460 Thr Val Pro Leu Ala Val Leu Gln Leu Arg Val Lys His Ala Asn Glu 465 470 475 480 Thr Ser Leu Ser Ile Met Trp Gln Thr Pro Val Ala Glu Trp Glu Lys 485 490 495 Tyr Ile Ile Ser Leu Ala Asp Arg Asp Leu Leu Leu Ile His Lys Ser 500 505 510 Leu Ser Lys Asp Ala Lys Glu Phe Thr Phe Thr Asp Leu Val Pro Gly 515 520 525 Arg Lys Tyr Met Ala Thr Val Thr Ser Ile Ser Gly Asp Leu Lys Asn 530 535 540 Ser Ser Ser Val Lys Gly Arg Thr Val Pro Ala Gln Val Thr Asp Leu 545 550 555 560 His Val Ala Asn Gln Gly Met Thr Ser Ser Leu Phe Thr Asn Trp Thr 565 570 575 Gln Ala Gln Gly Asp Val Glu Phe Tyr Gln Val Leu Leu Ile His Glu 580 585 590 Asn Val Val Ile Lys Asn Glu Ser Ile Ser Ser Glu Thr Ser Arg Tyr 595 600 605 Ser Phe His Ser Leu Lys Ser Gly Ser Leu Tyr Ser Val Val Val Thr 610 615 620 Thr Val Ser Gly Gly Ile Ser Ser Arg Gln Val Val Val Glu Gly Arg 625 630 635 640 Thr Val Pro Ser Ser Val Ser Gly Val Thr Val Asn Asn Ser Gly Arg 645 650 655 Asn Asp Tyr Leu Ser Val Ser Trp Leu Leu Ala Pro Gly Asp Val Asp 660 665 670 Asn Tyr Glu Val Thr Leu Ser His Asp Gly Lys Val Val Gln Ser Leu 675 680 685 Val Ile Ala Lys Ser Val Arg Glu Cys Ser Phe Ser Ser Leu Thr Pro 690 695 700 Gly Arg Leu Tyr Thr Val Thr Ile Thr Thr Arg Ser Gly Lys Tyr Glu 705 710 715 720 Asn His Ser Phe Ser Gln Glu Arg Thr Val Pro Asp Lys Val Gln Gly 725 730 735 Val Ser Val Ser Asn Ser Ala Arg Ser Asp Tyr Leu Arg Val Ser Trp 740 745 750 Val His Ala Thr Gly Asp Phe Asp His Tyr Glu Val Thr Ile Lys Asn 755 760 765 Lys Asn Asn Phe Ile Gln Thr Lys Ser Ile Pro Lys Ser Glu Asn Glu 770 775 780 Cys Val Phe Val Gln Leu Val Pro Gly Arg Leu Tyr Ser Val Thr Val 785 790 795 800 Thr Thr Lys Ser Gly Gln Tyr Glu Ala Asn Glu Gln Gly Asn Gly Arg 805 810 815 Thr Ile Pro Glu Pro Val Lys Asp Leu Thr Leu Arg Asn Arg Ser Thr 820 825 830 Glu Asp Leu His Val Thr Trp Ser Gly Ala Asn Gly Asp Val Asp Gln 835 840 845 Tyr Glu Ile Gln Leu Leu Phe Asn Asp Met Lys Val Phe Pro Pro Phe 850 855 860 His Leu Val Asn Thr Ala Thr Glu Tyr Arg Phe Thr Ser Leu Thr Pro 865 870 875 880 Gly Arg Gln Tyr Lys Ile Leu Val Leu Thr Ile Ser Gly Asp Val Gln 885 890 895 Gln Ser Ala Phe Ile Glu Gly Phe Thr Val Pro Ser Ala Val Lys Asn 900 905 910 Ile His Ile Ser Pro Asn Gly Ala Thr Asp Ser Leu Thr Val Asn Trp 915 920 925 Thr Pro Gly Gly Gly Asp Val Asp Ser Tyr Thr Val Ser Ala Phe Arg 930 935 940 His Ser Gln Lys Val Asp Ser Gln Thr Ile Pro Lys His Val Phe Glu 945 950 955 960 His Thr Phe His Arg Leu Glu Ala Gly Glu Gln Tyr Gln Ile Met Ile 965 970 975 Ala Ser Val Ser Gly Ser Leu Lys Asn Gln Ile Asn Val Val Gly Arg 980 985 990 Thr Val Pro Ala Ser Val Gln Gly Val Ile Ala Asp Asn Ala Tyr Ser 995 1000 1005 Ser Tyr Ser Leu Ile Val Ser Trp Gln Lys Ala Ala Gly Val Ala 1010 1015 1020 Glu Arg Tyr Asp Ile Leu Leu Leu Thr Glu Asn Gly Ile Leu Leu 1025 1030 1035 Arg Asn Thr Ser Glu Pro Ala Thr Thr Lys Gln His Lys Phe Glu 1040 1045 1050 Asp Leu Thr Pro Gly Lys Lys Tyr Lys Ile Gln Ile Leu Thr Val 1055 1060 1065 Ser Gly Gly Leu Phe Ser Lys Glu Ala Gln Thr Glu Gly Arg Thr 1070 1075 1080 Val Pro Ala Ala Val Thr Asp Leu Arg Ile Thr Glu Asn Ser Thr 1085 1090 1095 Arg His Leu Ser Phe Arg Trp Thr Ala Ser Glu Gly Glu Leu Ser 1100 1105 1110 Trp Tyr Asn Ile Phe Leu Tyr Asn Pro Asp Gly Asn Leu Gln Glu 1115 1120 1125 Arg Ala Gln Val Asp Pro Leu Val Gln Ser Phe Ser Phe Gln Asn 1130 1135 1140 Leu Leu Gln Gly Arg Met Tyr Lys Met Val Ile Val Thr His Ser 1145 1150 1155 Gly Glu Leu Ser Asn Glu Ser Phe Ile Phe Gly Arg Thr Val Pro 1160 1165 1170 Ala Ser Val Ser His Leu Arg Gly Ser Asn Arg Asn Thr Thr Asp 1175 1180 1185 Ser Leu Trp Phe Asn Trp Ser Pro Ala Ser Gly Asp Phe Asp Phe 1190 1195 1200 Tyr Glu Leu Ile Leu Tyr Asn Pro Asn Gly Thr Lys Lys Glu Asn 1205 1210 1215 Trp Lys Asp Lys Asp Leu Thr Glu Trp Arg Phe Gln Gly Leu Val 1220 1225 1230 Pro Gly Arg Lys Tyr Val Leu Trp Val Val Thr His Ser Gly Asp 1235 1240 1245 Leu Ser Asn Lys Val Thr Ala Glu Ser Arg Thr Ala Pro Ser Pro 1250 1255 1260 Pro Ser Leu Met Ser Phe Ala Asp Ile Ala Asn Thr Ser Leu Ala 1265 1270 1275 Ile Thr Trp Lys Gly Pro Pro Asp Trp Thr Asp Tyr Asn Asp Phe 1280 1285 1290 Glu Leu Gln Trp Leu Pro Arg Asp Ala Leu Thr Val Phe Asn Pro 1295 1300 1305 Tyr Asn Asn Arg Lys Ser Glu Gly Arg Ile Val Tyr Gly Leu Arg 1310 1315 1320 Pro Gly Arg Ser Tyr Gln Phe Asn Val Lys Thr Val Ser Gly Asp 1325 1330 1335 Ser Trp Lys Thr Tyr Ser Lys Pro Ile Phe Gly Ser Val Arg Thr 1340 1345 1350 Lys Pro Asp Lys Ile Gln Asn Leu His Cys Arg Pro Gln Asn Ser 1355 1360 1365 Thr Ala Ile Ala Cys Ser Trp Ile Pro Pro Asp Ser Asp Phe Asp 1370 1375 1380 Gly Tyr Ser Ile Glu Cys Arg Lys Met Asp Thr Gln Glu Val Glu 1385 1390 1395 Phe Ser Arg Lys Leu Glu Lys Glu Lys Ser Leu Leu Asn Ile Met 1400 1405 1410 Met Leu Val Pro His Lys Arg Tyr Leu Val Ser Ile Lys Val Gln 1415 1420 1425 Ser Ala Gly Met Thr Ser Glu Val Val Glu Asp Ser Thr Ile Thr 1430 1435 1440 Met Ile Asp Arg Pro Pro Pro Pro Pro Pro His Ile Arg Val Asn 1445 1450 1455 Glu Lys Asp Val Leu Ile Ser Lys Ser Ser Ile Asn Phe Thr Val 1460 1465 1470 Asn Cys Ser Trp Phe Ser Asp Thr Asn Gly Ala Val Lys Tyr Phe 1475 1480 1485 Thr Val Val Val Arg Glu Ala Asp Gly Ser Asp Glu Leu Lys Pro 1490 1495 1500 Glu Gln Gln His Pro Leu Pro Ser Tyr Leu Glu Tyr Arg His Asn 1505 1510 1515 Ala Ser Ile Arg Val Tyr Gln Thr Asn Tyr Phe Ala Ser Lys Cys 1520 1525 1530 Ala Glu Asn Pro Asn Ser Asn Ser Lys Ser Phe Asn Ile Lys Leu 1535 1540 1545 Gly Ala Glu Met Glu Ser Leu Gly Gly Lys Cys Asp Pro Thr Gln 1550 1555 1560 Gln Lys Phe Cys Asp Gly Pro Leu Lys Pro His Thr Ala Tyr Arg 1565 1570 1575 Ile Ser Ile Arg Ala Phe Thr Gln Leu Phe Asp Glu Asp Leu Lys 1580 1585 1590 Glu Phe Thr Lys Pro Leu Tyr Ser Asp Thr Phe Phe Ser Leu Pro 1595 1600 1605 Ile Thr Thr Glu Ser Glu Pro Leu Phe Gly Ala Ile Glu Gly Val 1610 1615 1620 Ser Ala Gly Leu Phe Leu Ile Gly Met Leu Val Ala Val Val Ala 1625 1630 1635 Leu Leu Ile Cys Arg Gln Lys Val Ser His Gly Arg Glu Arg Pro 1640 1645 1650 Ser Ala Arg Leu Ser Ile Arg Arg Asp Arg Pro Leu Ser Val His 1655 1660 1665 Leu Asn Leu Gly Gln Lys Gly Asn Arg Lys Thr Ser Cys Pro Ile 1670 1675 1680 Lys Ile Asn Gln Phe Glu Gly His Phe Met Lys Leu Gln Ala Asp 1685 1690 1695 Ser Asn Tyr Leu Leu Ser Lys Glu Tyr Glu Glu Leu Lys Asp Val 1700 1705 1710 Gly Arg Asn Gln Ser Cys Asp Ile Ala Leu Leu Pro Glu Asn Arg 1715 1720 1725 Gly Lys Asn Arg Tyr Asn Asn Ile Leu Pro Tyr Asp Ala Thr Arg 1730 1735 1740 Val Lys Leu Ser Asn Val Asp Asp Asp Pro Cys Ser Asp Tyr Ile 1745 1750 1755 Asn Ala Ser Tyr Ile Pro Gly Asn Asn Phe Arg Arg Glu Tyr Ile 1760 1765 1770 Val Thr Gln Gly Pro Leu Pro Gly Thr Lys Asp Asp Phe Trp Lys 1775 1780 1785 Met Val Trp Glu Gln Asn Val His Asn Ile Val Met Val Thr Gln 1790 1795 1800 Cys Val Glu Lys Gly Arg Val Lys Cys Asp His Tyr Trp Pro Ala 1805 1810 1815 Asp Gln Asp Ser Leu Tyr Tyr Gly Asp Leu Ile Leu Gln Met Leu 1820 1825 1830 Ser Glu Ser Val Leu Pro Glu Trp Thr Ile Arg Glu Phe Lys Ile 1835 1840 1845 Cys Gly Glu Glu Gln Leu Asp Ala His Arg Leu Ile Arg His Phe 1850 1855 1860 His Tyr Thr Val Trp Pro Asp His Gly Val Pro Glu Thr Thr Gln 1865 1870 1875 Ser Leu Ile Gln Phe Val Arg Thr Val Arg Asp Tyr Ile Asn Arg 1880 1885 1890 Ser Pro Gly Ala Gly Pro Thr Val Val His Cys Ser Ala Gly Val 1895 1900 1905 Gly Arg Thr Gly Thr Phe Ile Ala Leu Asp Arg Ile Leu Gln Gln 1910 1915 1920 Leu Asp Ser Lys Asp Ser Val Asp Ile Tyr Gly Ala Val His Asp 1925 1930 1935 Leu Arg Leu His Arg Val His Met Val Gln Thr Glu Cys Gln Tyr 1940 1945 1950 Val Tyr Leu His Gln Cys Val Arg Asp Val Leu Arg Ala Arg Lys 1955 1960 1965 Leu Arg Ser Glu Gln Glu Asn Pro Leu Phe Pro Ile Tyr Glu Asn 1970 1975 1980 Val Asn Pro Glu Tyr His Arg Asp Pro Val Tyr Ser Arg His 1985 1990 1995 <210> 40 <211> 2283 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic Polynucleotide <400> 40 atggaatgga gctgggtctt tctcttcttc ctgtcagtaa cgactggtgt ccactccgag 60 cccgagagat gcaacttcac cctggccgag tccaaggcct cctcccactc cgtgtctatc 120 cagtggcgga tcctgggctc cccctgcaac ttctctctga tctactcctc cgacaccctg 180 ggcgctgccc tgtgccctac cttcagaatc gacaacacca cctacggctg caacctgcag 240 gatctgcagg ccggcaccat ctacaacttc aagatcatct ccctggacga ggaacggacc 300 gtggtgctgc agaccgatcc tctgcctcct gccagattcg gcgtgtccaa agaaaagacc 360 acctccaccg gactgcacgt gtggtggacc ccttccagcg gcaaagtgac ctcctacgag 420 gtgcagctgt tcgacgagaa caaccagaaa atccagggcg tgcagatcca ggaatccacc 480 tcctggaacg agtacacctt cttcaacctg accgccggct ccaagtacaa tatcgccatc 540 accgccgtgt ccggcggcaa gagatccttc tccgtgtaca ccaacggctc caccgtgccc 600 agccccgtga aggacatcgg catctccacc aaggccaact ccctgctgat ctcctggtcc 660 cacggctccg gcaacgtgga acggtacaga ctgatgctga tggacaaggg catcctggtg 720 cacggcggcg tggtggataa gcacgccacc tcttacgcct tccacggcct gacccctggc 780 tacctgtaca atctgaccgt gatgaccgag gccgctggac tgcagaacta ccggtggaag 840 ctcgtgcgga ccgcccccat ggaagtgtcc aacctgaaag tgaccaacga cggctccctg 900 acctctctga aagtgaagtg gcagaggccc cctggcaatg tggactccta caacatcacc 960 ctgtcccaca agggcaccat caaagaatcc cgggtgctgg ccccttggat caccgagaca 1020 cacttcaaag aactggtgcc tggccggctg taccaagtga ccgtgtcctg tgtgtctggc 1080 gagctgtccg cccagaaaat ggccgtgggc agaaccttcc ctctggccgt gctgcagctg 1140 agagtgaagc acgctaacga gacatccctg tccatcatgt ggcagacccc cgtggccgag 1200 tgggagaagt acatcatcag cctggccgac cgggacctgc tgctgatcca caagtccctg 1260 agcaaggacg ccaaagagtt caccttcacc gacctggtgc ccggcagaaa gtacatggcc 1320 accgtgacct ccatctccgg cgacctgaag aactcctcca gcgtgaaggg caggaccgtg 1380 cctgcccaag tgacagacct gcacgtggcc aaccagggca tgacctcctc cctgttcacc 1440 aactggaccc aggctcaggg cgacgtggaa ttctaccagg tgctgctgat tcatgagaac 1500 gtcgtgatca agaacgagtc catctcctcc gagacaagcc ggtactcctt ccactccctg 1560 aagtccggca gcctgtactc cgtggtcgtg accacagtgt ccgggggcat ctcctctaga 1620 caggtggtgg tggaaggccg caccgtgcct agttcagtgt caggcgtgac cgtgaacaac 1680 agcggccgga acgactacct gtccgtgtct tggctgctgg ctcctgggga cgtggacaac 1740 tacgaagtga ccctgagcca cgacggcaag gtggtgcagt ctctcgtgat cgccaagtcc 1800 gtgcgcgagt gctccttcag ctctctgaca cctggcagac tgtataccgt gaccatcacc 1860 accagatccg ggaagtacga gaaccacagc ttctcccagg aacgcacagt gcccgacaag 1920 gtgcagggcg tgtcagtgtc taactccgcc agatctgact acctgcgggt gtcctgggtg 1980 cacgctaccg gcgacttcga ccattatgaa gtgacaatca agaacaagaa caacttcatc 2040 cagaccaagt ccatccccaa gtccgagaac gagtgcgtgt tcgtgcagct ggtgccaggc 2100 agactgtact ctgtgacagt gaccaccaag tccggccagt acgaggccaa cgagcagggc 2160 aacggcagga ccatccctga gaagggcaac tccgccgaca tccagcactc tggcggcaga 2220 tcctctgg aaggccccag attcgagaga accggcggag gccaccacca tcatcaccat 2280 is 2283 <210> 41 <211> 736 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 41 Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly 1 5 10 15 Val His Ser Glu Arg Cys Asn Phe Thr Leu Ala Glu Ser Lys Ala Ser 20 25 30 Ser His Ser Val Ser Ile Arg Trp Arg Ile Trp Gly Ser Pro Cys Asn 35 40 45 Phe Asn Leu Thr Tyr Ser Ser Asp Thr Leu Gly Ala Ala Ser Cys Pro 50 55 60 Pro Phe Arg Leu Asp Asn Thr Thr Tyr Tyr Gly Cys Asn Leu Gln Asp Leu 65 70 75 80 Gln Ala Gly Thr Ile Tyr Asn Phe Arg Ile Val Ser Leu Asp Gly Glu 85 90 95 Glu Arg Thr Val Val Leu Gln Thr Asp Pro Leu Pro Pro Ala Arg Phe 100 105 110 Gly Val Ser Lys Glu Lys Thr Thr Ser Thr Ser Leu His Val Trp Trp 115 120 125 Thr Pro Ser Pro Gly Lys Val Thr Ser Tyr Glu Val Gln Leu Phe Asp 130 135 140 Glu Asn Asn Gln Lys Ile Gln Gly Val Gln Ile Gln Glu Ser Thr Ser 145 150 155 160 Trp Asn Lys Tyr Thr Phe Phe Asn Leu Thr Ala Gly Ser Lys Tyr Asn 165 170 175 Ile Thr Ile Thr Ala Val Ser Gly Gly Lys Arg Ser Ser Ser Val Tyr 180 185 190 Thr Asn Gly Ser Thr Val Pro Ser Pro Val Lys Asp Ile Gly Ile Ser 195 200 205 Thr Lys Ala Asn Ser Leu Leu Val Ser Trp Ser His Gly Ser Gly Asn 210 215 220 Val Glu Arg Tyr Arg Leu Met Leu Met Asp Lys Gly Ile Leu Val His 225 230 235 240 Gly Ser Val Val Asp Arg Gln Ala Thr Ser Tyr Thr Phe Asn Gly Leu 245 250 255 Thr Pro Gly Tyr Leu Tyr Asn Leu Thr Val Val Thr Glu Ala Ala Gly 260 265 270 Leu Gln Asn Tyr Lys Trp Lys Leu Val Arg Thr Ala Pro Met Glu Val 275 280 285 Ser Asn Leu Lys Val Thr Asn Asp Gly Ser Leu Thr Ser Leu Lys Val 290 295 300 Lys Trp Gln Arg Pro Pro Gly Asn Val Asp Ser Tyr Asn Ile Thr Leu 305 310 315 320 Ser His Lys Gly Thr Ile Lys Glu Ser Arg Val Leu Ala Pro Arg Val 325 330 335 Thr Glu Thr His Phe Lys Glu Leu Thr Pro Gly Arg Leu Tyr Gln Val 340 345 350 Thr Val Ser Cys Val Ser Gly Glu Leu Ser Ala Gln Arg Met Ala Val 355 360 365 Gly Arg Thr Phe Pro Leu Pro Val Leu Gln Leu Arg Val Lys His Ala 370 375 380 Asn Glu Thr Ser Leu Ser Ile Ile Trp Gln Pro Pro Val Ala Glu Trp 385 390 395 400 Glu Glu Tyr Ile Ile Ser Leu Ala Asp Arg Asp Leu Arg Leu Ile His 405 410 415 Lys Ser Leu Ser Lys Asp Ala Lys Glu Phe Thr Phe Thr Asp Leu Val 420 425 430 Pro Gly Arg Lys Tyr Met Ala Thr Val Thr Ser Ile Ser Gly Asp Leu 435 440 445 Lys Asn Ser Ser Ser Val Lys Gly Arg Thr Val Pro Ala Gln Val Thr 450 455 460 Asp Leu His Val Ala Asn Gln Gly Met Thr Ser Ser Leu Phe Thr Asn 465 470 475 480 Trp Thr Gln Ala Gln Gly Asp Val Glu Phe Tyr Gln Val Leu Leu Ile 485 490 495 His Glu Asn Val Val Ile Lys Asn Glu Ser Ile Pro Ser Glu Thr Ser 500 505 510 Gly Tyr Asn Phe His Phe Leu Lys Ser Gly Ser Leu Tyr Ser Val Val 515 520 525 Val Thr Thr Val Ser Gly Gly Ile Ser Ser Arg Gln Val Val Val Glu 530 535 540 Gly Arg Thr Val Pro Ser Ser Val Ser Gly Val Thr Val Asn Asn Ser 545 550 555 560 Gly Arg Asn Asp Tyr Leu Ser Val Ser Trp Leu Pro Ala Pro Gly Asp 565 570 575 Val Asp Asn Tyr Glu Val Thr Leu Ser His Asp Gly Arg Val Val Gln 580 585 590 Ser Leu Val Ile Ala Lys Ser Val Arg Glu Cys Ser Phe Ser Ser Leu 595 600 605 Thr Pro Gly Arg Leu Tyr Thr Val Thr Ile Thr Thr Arg Ser Gly Lys 610 615 620 Tyr Glu Asn His Ser Phe Ser Gln Glu Arg Thr Val Pro Asp Lys Val 625 630 635 640 Gln Gly Val Ser Val Ser Asn Ser Ala Arg Ser Asp Tyr Leu Arg Val 645 650 655 Ser Trp Val His Ala Thr Gly Asp Phe Asp His Tyr Glu Val Thr Ile 660 665 670 Lys Asn Lys Asn Asn Phe Ile Glu Thr Lys Ser Ile Pro Lys Ser Glu 675 680 685 Asn Glu Cys Val Phe Val Gln Leu Val Pro Gly Arg Leu Tyr Ser Val 690 695 700 Thr Val Thr Thr Lys Ser Gly Gln Tyr Glu Ala Ser Glu Gln Gly Asn 705 710 715 720 Gly Arg Thr Gly Gly Gly His His His His His His His 725 730 735 <210> 42 <211> 2210 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Polynucleotide <400> 42 atggaatgga gctgggtctt tctcttcttc ctgtcagtaa cgactggtgt ccactccgag 60 cggtgcaact ttaccctggc cgagtccaag gcctcctccc actccgtgtc tatccggtgg 120 cggatctggg gctccccctg caacttcaac ctgacctact cctccgatac cctgggcgct 180 gcctcctgtc ctcctttccg gctggagaac accacctacg gctgcaacct gcaggatctg 240 caggccggca ccatctacaa cttccggatc gtgtccctgg acggcgagga acggacagtg 300 gtgctccaga ccgatcctct gccccctgcc agattcggcg tgtccaaaga aaagaccacc 360 tccacctccc tgcacgtgtg gtggacccct agccctggca aagtgacctc ctacgaggtg 420 cagctgttcg acgagaacaa ccagaaaatc cagggcgtgc agatccagga atccacctcc 480 tggaacaagt acaccttctt caatctgacc gccggctcca agtacaacat caccatcacc 540 gccgtgtccg gcggcaagag atcctcctcc gtgtacacca acggctccac cgtgcccagc 600 cccgtgaagg acatcggcat ctccaccaag gccaactccc tgctggtgtc ctggtcccac 660 ggctccggca acgtggaacg gtacagactg atgctgatgg acaagggcat cctggtgcac 720 ggcagcgtgg tggatagaca ggccacctcc tacaccttca acggcctgac ccccggctac 780 ctgtataacc tgaccgtcgt gaccgaggcc gctggactgc agaactacaa gtggaagctc 840 gtgcggaccg cccccatgga agtgtccaac ctgaaagtga ccaacgacgg ctccctgacc 900 tctctgaaag tgaagtggca gaggccccct ggcaatgtgg acagctacaa tatcaccctg 960 tcccacaagg gcaccatcaa agaatcccgg gtgctggccc ccagagtgac cgagacacac 1020 ttcaaagagc tgacccctgg ccggctgtac caagtgaccg tgtcctgtgt gtctggcgag 1080 ctgtctgccc agagaatggc cgtgggcaga accttccctc tgcccgtgct gcagctgaga 1140 gtgaagcacg ccaacgagac atccctgtcc atcatctggc agccccctgt ggccgagtgg 1200 gagagtaca tcatcagcct ggccgaccgg gacctgcggc tgatccacaa gtccctgagc 1260 1320 gtgacctcca tctccggcga cctgaagaac tcctccagcg tgaagggcag gaccgtgcct 1380 gcccaagtga cagacctgca tgtggccaac cagggcatga cctccagcct gttcaccaac 1440 tggacccagg ctcagggcga cgtggaattc taccaggtgc tgctgatcca tgagaacgtc 1500 gtgatcaaga acgagtccat cccctccgag acaagcggct acaactttca cttcctgaag 1560 tccggcagcc tgtactccgt ggtcgtgacc acagtgtccg ggggcatctc ctctagaagg 1620 tggtggtgga aggccgcacc gtgcctagtt cagtgtcagg cgtgaccgtg aacaacagcg 1680 gccggaacga ctacctgtcc gtgtcttggc tgcctgcccc tggggacgtg gacaactacg 1740 aagtgaccct gtctcacgac ggccgggtgg tgcagtctct cgtgatcgct aagtccgtgc 1800 gcgagtgctc cttcagcagc ctgacacctg gcagactgta taccgtgacc atcaccacca 1860 gatccgggaa gtacgagaac cacagcttct cccaggaacg aaccgtgccc gacaaggtgc 1920 agggcgtgtc agtgtctaac tccgccagat ctgactacct gagagtgtcc tgggtgcacg 1980 ccaccggcga cttcgaccat tatgaagtga caatcaagaa caagaacaac ttcatcgaga 2040 caaagagcat ccccaagtcc gagaacgagt gcgtgttcgt gcagctggtg ccaggcaggc 2100 tgtattctgt gacagtgacc accaagtccg gccagtacga ggcctctgag cagggcaatg 2160 gcagaaccgg cggtggacac caccatcatc accatcacca ccatcactag 2210 <210> 43 <211> 971 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 43 Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly 1 5 10 15 Val His Ser Ala Gly Gly Thr Pro Ser Pro Ile Pro Asp Pro Ser Val 20 25 30 Ala Thr Val Ala Thr Gly Glu Asn Gly Ile Thr Gln Ile Ser Ser Thr 35 40 45 Ala Glu Ser Phe His Lys Gln Asn Gly Thr Gly Thr Pro Gln Val Glu 50 55 60 Thr Asn Thr Ser Glu Asp Gly Glu Ser Ser Gly Ala Asn Asp Ser Leu 65 70 75 80 Arg Thr Pro Glu Gln Gly Ser Asn Gly Thr Asp Gly Ala Ser Gln Lys 85 90 95 Thr Pro Ser Ser Thr Gly Pro Ser Pro Val Phe Asp Ile Lys Ala Val 100 105 110 Ser Ile Ser Pro Thr Asn Val Ile Leu Thr Trp Lys Ser Asn Asp Thr 115 120 125 Ala Ala Ser Glu Tyr Lys Tyr Val Val Lys His Lys Met Glu Asn Glu 130 135 140 Lys Thr Ile Thr Val Val His Gln Pro Trp Cys Asn Ile Thr Gly Leu 145 150 155 160 Arg Pro Ala Thr Ser Tyr Val Phe Ser Ile Thr Pro Gly Ile Gly Asn 165 170 175 Glu Thr Trp Gly Asp Pro Arg Val Ile Lys Val Ile Thr Glu Pro Ile 180 185 190 Pro Val Ser Asp Leu Arg Val Ala Leu Thr Gly Val Arg Lys Ala Ala 195 200 205 Leu Ser Trp Ser Asn Gly Asn Gly Thr Ala Ser Cys Arg Val Leu Leu 210 215 220 Glu Ser Ile Gly Ser His Glu Glu Leu Thr Gln Asp Ser Arg Leu Gln 225 230 235 240 Val Asn Ile Ser Gly Leu Lys Pro Gly Val Gln Tyr Asn Ile Asn Pro 245 250 255 Tyr Leu Leu Gln Ser Asn Lys Thr Lys Gly Asp Pro Leu Gly Thr Glu 260 265 270 Gly Gly Leu Asp Ala Ser Asn Thr Glu Arg Ser Arg Ala Gly Ser Pro 275 280 285 Thr Ala Pro Val His Asp Glu Ser Leu Val Gly Pro Val Asp Pro Ser 290 295 300 Ser Gly Gln Gln Ser Arg Asp Thr Glu Val Leu Leu Val Gly Leu Glu 305 310 315 320 Pro Gly Thr Arg Tyr Asn Ala Thr Val Tyr Ser Gln Ala Ala Asn Gly 325 330 335 Thr Glu Gly Gln Pro Gln Ala Ile Glu Phe Arg Thr Asn Ala Ile Gln 340 345 350 Val Phe Asp Val Thr Ala Val Asn Ile Ser Ala Thr Ser Leu Thr Leu 355 360 365 Ile Trp Lys Val Ser Asp Asn Glu Ser Ser Ser Asn Tyr Thr Tyr Lys 370 375 380 Ile His Val Ala Gly Glu Thr Asp Ser Ser Asn Leu Asn Val Ser Glu 385 390 395 400 Pro Arg Ala Val Ile Pro Gly Leu Arg Ser Ser Thr Phe Tyr Asn Ile 405 410 415 Thr Val Cys Pro Val Leu Gly Asp Ile Glu Gly Thr Pro Gly Phe Leu 420 425 430 Gln Val His Thr Pro Pro Val Pro Val Ser Asp Phe Arg Val Thr Val 435 440 445 Val Ser Thr Thr Glu Ile Gly Leu Ala Trp Ser Ser His Asp Ala Glu 450 455 460 Ser Phe Gln Met His Ile Thr Gln Glu Gly Ala Gly Asn Ser Arg Val 465 470 475 480 Glu Ile Thr Thr Asn Gln Ser Ile Ile Ile Gly Gly Leu Phe Pro Gly 485 490 495 Thr Lys Tyr Cys Phe Glu Ile Val Pro Lys Gly Pro Asn Gly Thr Glu 500 505 510 Gly Ala Ser Arg Thr Val Cys Asn Arg Thr Val Pro Ser Ala Val Phe 515 520 525 Asp Ile His Val Val Tyr Val Thr Thr Thr Glu Met Trp Leu Asp Trp 530 535 540 Lys Ser Pro Asp Gly Ala Ser Glu Tyr Val Tyr His Leu Val Ile Glu 545 550 555 560 Ser Lys His Gly Ser Asn His Thr Ser Thr Tyr Asp Lys Ala Ile Thr 565 570 575 Leu Gln Gly Leu Ile Pro Gly Thr Leu Tyr Asn Ile Thr Ile Ser Pro 580 585 590 Glu Val Asp His Val Trp Gly Asp Pro Asn Ser Thr Ala Gln Tyr Thr 595 600 605 Arg Pro Ser Asn Val Ser Asn Ile Asp Val Ser Thr Asn Thr Thr Ala 610 615 620 Ala Thr Leu Ser Trp Gln Asn Phe Asp Asp Ala Ser Pro Thr Tyr Ser 625 630 635 640 Tyr Cys Leu Leu Ile Glu Lys Ala Gly Asn Ser Ser Asn Ala Thr Gln 645 650 655 Val Val Thr Asp Ile Gly Ile Thr Asp Ala Thr Val Thr Glu Leu Ile 660 665 670 Pro Gly Ser Ser Tyr Thr Val Glu Ile Phe Ala Gln Val Gly Asp Gly 675 680 685 Ile Lys Ser Leu Glu Pro Gly Arg Lys Ser Phe Cys Thr Asp Pro Ala 690 695 700 Ser Met Ala Ser Phe Asp Cys Glu Val Val Pro Lys Glu Pro Ala Leu 705 710 715 720 Val Leu Lys Trp Thr Cys Pro Pro Gly Ala Asn Ala Gly Phe Glu Leu 725 730 735 Glu Val Ser Ser Gly Ala Trp Asn Asn Ala Thr His Leu Glu Ser Cys 740 745 750 Ser Ser Glu Asn Gly Thr Glu Tyr Arg Thr Glu Val Thr Tyr Leu Asn 755 760 765 Phe Ser Thr Ser Tyr Asn Ile Ser Ile Thr Thr Val Ser Cys Gly Lys 770 775 780 Met Ala Ala Pro Thr Arg Asn Thr Cys Thr Thr Gly Ile Thr Asp Pro 785 790 795 800 Pro Pro Pro Asp Gly Ser Pro Asn Ile Thr Ser Val Ser His Asn Ser 805 810 815 Val Lys Val Lys Phe Ser Gly Phe Glu Ala Ser His Gly Pro Ile Lys 820 825 830 Ala Tyr Ala Val Ile Leu Thr Thr Gly Glu Ala Gly His Pro Ser Ala 835 840 845 Asp Val Leu Lys Tyr Thr Tyr Glu Asp Phe Lys Lys Gly Ala Ser Asp 850 855 860 Thr Tyr Val Thr Tyr Leu Ile Arg Thr Glu Glu Lys Gly Arg Ser Gln 865 870 875 880 Ser Leu Ser Glu Val Leu Lys Tyr Glu Ile Asp Val Gly Asn Glu Ser 885 890 895 Thr Thr Leu Gly Tyr Tyr Asn Gly Lys Leu Glu Pro Leu Gly Ser Tyr 900 905 910 Arg Ala Cys Val Ala Gly Phe Thr Asn Ile Thr Phe His Pro Gln Asn 915 920 925 Lys Gly Leu Ile Asp Gly Ala Glu Ser Tyr Val Ser Phe Ser Arg Tyr 930 935 940 Ser Asp Ala Val Ser Leu Pro Gln Asp Pro Gly Val Ile Cys Gly Gly 945 950 955 960 Gly His His His His His His His His His His 965 970 <210> 44 <211> 2916 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic Polynucleotide <400> 44 atggaatgga gctgggtctt tctcttcttc ctgtcagtaa cgactggtgt ccactccgca 60 ggtggcaccc ctagtccaat tcctgaccct tcagtagcaa ctgttgccac aggggaaaat 120 ggcataacgc agatcagcag tacagcagaa tcctttcata aacagaatgg aactggaaca 180 cctcaggtgg aaacaaacac cagtgaggat ggtgaaagct ctggagccaa cgatagttta 240 agaacacctg aacaaggatc taatgggact gatggggcat ctcaaaaaac tcccagtagc 300 actgggccca gtcctgtgtt tgacattaaa gctgtttcca tcagtccaac caatgtgatc 360 ttaacttgga aaagtaatga cacagctgct tctgagtaca agtatgtagt aaagcataag 420 atggaaaatg agaagacaat tactgttgtg catcaaccat ggtgtaacat cacaggctta 480 cgtccagcga cttcatatgt attctccatc actccaggaa taggcaatga gacttgggga 540 gatcccagag tcataaaagt catcacagag ccgatcccag tttctgatct ccgtgttgcc 600 ctcacgggtg tgaggaaggc tgctctctcc tggagcaatg gcaatggcac tgcctcctgc 660 cgggttcttc ttgaaagcat tggaagccat gaggagttga ctcaagactc aagacttcag 720 gtcaatatct cgggcctgaa gccaggggtt caatacaaca tcaacccgta tcttctacaa 780 tcaaataaga caaagggaga ccccttgggc acagaaggtg gcttggatgc cagcaataca 840 gagagaagcc gggcagggag ccccaccgcc cctgtgcatg atgagtccct cgtgggacct 900 gtggacccat cctccggcca gcagtcccga gacacggaag tcctgcttgt cgggttagag 960 cctggcaccc gatacaatgc caccgtttat tcccaagcag cgaatggcac agaaggacag 1020 ccccaggcca tagagttcag gacaaatgct attcaggttt ttgacgtcac cgctgtgaac 1080 atcagtgcca caagcctgac cctgatctgg aaagtcagcg ataacgagtc gtcatctaac 1140 tatacctaca agatacatgt ggcgggggag acagattctt ccaatctcaa cgtcagtgag 1200 cctcgcgctg tcatccccgg actccgctcc agcaccttct acaacatcac agtgtgtcct 1260 gtcctaggtg acatcgaggg cacgccgggc ttcctccaag tgcacacccc ccctgttcca 1320 gtttctgact tccgagtgac agtggtcagc acgacggaga tcggcttagc atggagcagc 1380 catgatgcag aatcatttca gatgcatatc acacaggagg gagctggcaa ttctcgggta 1440 gaaataacca ccaaccaaag tattatcatt ggtggcttgt tccctggaac caagtattgc 1500 tttgaaatag ttccaaaagg accaaatggg actgaagggg catctcggac agtttgcaat 1560 agaactgttc ccagtgcagt gtttgacatc cacgtggtct acgtcaccac cacggagatg 1620 tggctggact ggaagagccc tgacggtgct tccgagtatg tctaccattt agtcatagag 1680 tccaagcatg gctctaacca cacaagcacg tatgacaaag cgattactct ccagggcctg 1740 attccgggca ccttatataa catcaccatc tctccagaag tggaccacgt ctggggggac 1800 cccaactcca ctgcacagta cacacggccc agcaatgtgt ccacattga tgtaagtacc 1860 aacaccacag cagcaactt aagttggcag aactttgatg acgccctcc cacgtactcc 1920 tactgccttc ttattgagaa ggctggaat tccacaacg tactgccttc aggcaccacg 1980 attggaatta ctgacgctac agtcactgaa ttaatactg gctcatcata cacagtggag 2040 atctttgcac aagtaggga tgggatcaag tcactggac ctggccgga gtcattctct 2100 acgatcctg cgtccatggc ctccttcgac tgcgaagtgg tcccaaaga gccagccctg 2160 gttctcaaat ggacctgccc tcctggcgcc aatgcaggct tgagctgga ggtcagcagt 2220 ggagcctgga acatgcgac ccacctggag agctgctcct ctgagaatgg cactgagtat 2280 agaacggaag tcacgtattt gatttttct acctcgtaca acatcagcat caccactgtg 2340 tcctgtggaa agatggcagc cccacccgg aacacctgca ctactggcat cacagatccc 2400 cctcctccag atggatcccc taatattaca tctgtcagtc acaatcagt aaggtcag 2460 ttcagtggat ttgaagccag ccacggaccc atcaaagcct atgctgtcat tctcaccacc 2520 ggggaagctg gtcacccttc tgcagatgtc ctgaaataca cgtatgagga tttcaaaaag 2580 ggagcctcag atacttatgt gacatacctc ataagaacag aagaaaaggg acgttctcag 2640 agcttgtctg aagttttgaa atatgaaatt gacgttggga atgagtcaac cacacttggt 2700 tattacaatg ggaagctgga acctctgggc tcctaccggg cttgtgtggc tggcttcacc 2760 aacattacct tccaccctca aaacaagggg ctcattgatg gggctgagag ctatgtgtcc 2820 ttcagtcgct actcagatgc tgtttccttg ccccaggatc caggtgtcat ctgtggcggt 2880 ggacaccacc atcatcacca tcaccaccat cactag 2916 <210> 45 <211> 1621 <212> PRT <213> Homo sapiens <400> 45 Met Leu Ser His Gly Ala Gly Leu Ala Leu Trp Ile Thr Leu Ser Leu 1 5 10 15 Leu Gln Thr Gly Leu Ala Glu Pro Glu Arg Cys Asn Phe Thr Leu Ala 20 25 30 Glu Ser Lys Ala Ser Ser His Ser Val Ser Ile Gln Trp Arg Ile Leu 35 40 45 Gly Ser Pro Cys Asn Phe Ser Leu Ile Tyr Ser Ser Asp Thr Leu Gly 50 55 60 Ala Ala Leu Cys Pro Thr Phe Arg Ile Asp Asn Thr Thr Tyr Gly Cys 65 70 75 80 Asn Leu Gln Asp Leu Gln Ala Gly Thr Ile Tyr Asn Phe Arg Ile Ile 85 90 95 Ser Leu Asp Glu Glu Arg Thr Val Val Leu Gln Thr Asp Pro Leu Pro 100 105 110 Pro Ala Arg Phe Gly Val Ser Lys Glu Lys Thr Thr Ser Thr Ser Leu 115 120 125 His Val Trp Trp Thr Pro Ser Ser Gly Lys Val Thr Ser Tyr Glu Val 130 135 140 Gln Leu Phe Asp Glu Asn Asn Gln Lys Ile Gln Gly Val Gln Ile Gln 145 150 155 160 Glu Ser Thr Ser Trp Asn Glu Tyr Thr Phe Phe Asn Leu Thr Ala Gly 165 170 175 Ser Lys Tyr Asn Ile Ala Ile Thr Ala Val Ser Gly Gly Lys Arg Ser 180 185 190 Phe Ser Val Tyr Thr Asn Gly Ser Thr Val Pro Ser Pro Val Lys Asp 195 200 205 Ile Gly Ile Ser Thr Lys Ala Asn Ser Leu Leu Ile Ser Trp Ser His 210 215 220 Gly Ser Gly Asn Val Glu Arg Tyr Arg Leu Met Leu Met Asp Lys Gly 225 230 235 240 Ile Leu Val His Gly Gly Val Val Asp Lys His Ala Thr Ser Tyr Ala 245 250 255 Phe His Gly Leu Thr Pro Gly Tyr Leu Tyr Asn Leu Thr Val Met Thr 260 265 270 Glu Ala Ala Gly Leu Gln Asn Tyr Arg Trp Lys Leu Val Arg Thr Ala 275 280 285 Pro Met Glu Val Ser Asn Leu Lys Val Thr Asn Asp Gly Ser Leu Thr 290 295 300 Ser Leu Lys Val Lys Trp Gln Arg Pro Pro Gly Asn Val Asp Ser Tyr 305 310 315 320 Asn Ile Thr Leu Ser His Lys Gly Thr Ile Lys Glu Ser Arg Val Leu 325 330 335 Ala Pro Trp Ile Thr Glu Thr His Phe Lys Glu Leu Val Pro Gly Arg 340 345 350 Leu Tyr Gln Val Thr Val Ser Cys Val Ser Gly Glu Leu Ser Ala Gln 355 360 365 Lys Met Ala Val Gly Arg Thr Phe Pro Asp Lys Val Ala Asn Leu Glu 370 375 380 Ala Asn Asn Asn Gly Arg Met Arg Ser Leu Val Val Ser Trp Ser Pro 385 390 395 400 Pro Ala Gly Asp Trp Glu Gln Tyr Arg Ile Leu Leu Phe Asn Asp Ser 405 410 415 Val Val Leu Leu Asn Ile Thr Val Gly Lys Glu Glu Thr Gln Tyr Val 420 425 430 Met Asp Asp Thr Gly Leu Val Pro Gly Arg Gln Tyr Glu Val Glu Val 435 440 445 Ile Val Glu Ser Gly Asn Leu Lys Asn Ser Glu Arg Cys Gln Gly Arg 450 455 460 Thr Val Pro Leu Ala Val Leu Gln Leu Arg Val Lys His Ala Asn Glu 465 470 475 480 Thr Ser Leu Ser Ile Met Trp Gln Thr Pro Val Ala Glu Trp Glu Lys 485 490 495 Tyr Ile Ile Ser Leu Ala Asp Arg Asp Leu Leu Leu Ile His Lys Ser 500 505 510 Leu Ser Lys Asp Ala Lys Glu Phe Thr Phe Thr Asp Leu Val Pro Gly 515 520 525 Arg Lys Tyr Met Ala Thr Val Thr Ser Ile Ser Gly Asp Leu Lys Asn 530 535 540 Ser Ser Ser Val Lys Gly Arg Thr Val Pro Ala Gln Val Thr Asp Leu 545 550 555 560 His Val Ala Asn Gln Gly Met Thr Ser Ser Leu Phe Thr Asn Trp Thr 565 570 575 Gln Ala Gln Gly Asp Val Glu Phe Tyr Gln Val Leu Leu Ile His Glu 580 585 590 Asn Val Val Ile Lys Asn Glu Ser Ile Ser Ser Glu Thr Ser Arg Tyr 595 600 605 Ser Phe His Ser Leu Lys Ser Gly Ser Leu Tyr Ser Val Val Val Thr 610 615 620 Thr Val Ser Gly Gly Ile Ser Ser Arg Gln Val Val Val Glu Gly Arg 625 630 635 640 Thr Val Pro Ser Ser Val Ser Gly Val Thr Val Asn Asn Ser Gly Arg 645 650 655 Asn Asp Tyr Leu Ser Val Ser Trp Leu Leu Ala Pro Gly Asp Val Asp 660 665 670 Asn Tyr Glu Val Thr Leu Ser His Asp Gly Lys Val Val Gln Ser Leu 675 680 685 Val Ile Ala Lys Ser Val Arg Glu Cys Ser Phe Ser Ser Leu Thr Pro 690 695 700 Gly Arg Leu Tyr Thr Val Thr Ile Thr Thr Arg Ser Gly Lys Tyr Glu 705 710 715 720 Asn His Ser Phe Ser Gln Glu Arg Thr Val Pro Asp Lys Val Gln Gly 725 730 735 Val Ser Val Ser Asn Ser Ala Arg Ser Asp Tyr Leu Arg Val Ser Trp 740 745 750 Val His Ala Thr Gly Asp Phe Asp His Tyr Glu Val Thr Ile Lys Asn 755 760 765 Lys Asn Asn Phe Ile Gln Thr Lys Ser Ile Pro Lys Ser Glu Asn Glu 770 775 780 Cys Val Phe Val Gln Leu Val Pro Gly Arg Leu Tyr Ser Val Thr Val 785 790 795 800 Thr Thr Lys Ser Gly Gln Tyr Glu Ala Asn Glu Gln Gly Asn Gly Arg 805 810 815 Thr Ile Pro Glu Pro Val Lys Asp Leu Thr Leu Arg Asn Arg Ser Thr 820 825 830 Glu Asp Leu His Val Thr Trp Ser Gly Ala Asn Gly Asp Val Asp Gln 835 840 845 Tyr Glu Ile Gln Leu Leu Phe Asn Asp Met Lys Val Phe Pro Pro Phe 850 855 860 His Leu Val Asn Thr Ala Thr Glu Tyr Arg Phe Thr Ser Leu Thr Pro 865 870 875 880 Gly Arg Gln Tyr Lys Ile Leu Val Leu Thr Ile Ser Gly Asp Val Gln 885 890 895 Gln Ser Ala Phe Ile Glu Gly Phe Thr Val Pro Ser Ala Val Lys Asn 900 905 910 Ile His Ile Ser Pro Asn Gly Ala Thr Asp Ser Leu Thr Val Asn Trp 915 920 925 Thr Pro Gly Gly Gly Asp Val Asp Ser Tyr Thr Val Ser Ala Phe Arg 930 935 940 His Ser Gln Lys Val Asp Ser Gln Thr Ile Pro Lys His Val Phe Glu 945 950 955 960 His Thr Phe His Arg Leu Glu Ala Gly Glu Gln Tyr Gln Ile Met Ile 965 970 975 Ala Ser Val Ser Gly Ser Leu Lys Asn Gln Ile Asn Val Val Gly Arg 980 985 990 Thr Val Pro Ala Ser Val Gln Gly Val Ile Ala Asp Asn Ala Tyr Ser 995 1000 1005 Ser Tyr Ser Leu Ile Val Ser Trp Gln Lys Ala Ala Gly Val Ala 1010 1015 1020 Glu Arg Tyr Asp Ile Leu Leu Leu Thr Glu Asn Gly Ile Leu Leu 1025 1030 1035 Arg Asn Thr Ser Glu Pro Ala Thr Thr Lys Gln His Lys Phe Glu 1040 1045 1050 Asp Leu Thr Pro Gly Lys Lys Tyr Lys Ile Gln Ile Leu Thr Val 1055 1060 1065 Ser Gly Gly Leu Phe Ser Lys Glu Ala Gln Thr Glu Gly Arg Thr 1070 1075 1080 Val Pro Ala Ala Val Thr Asp Leu Arg Ile Thr Glu Asn Ser Thr 1085 1090 1095 Arg His Leu Ser Phe Arg Trp Thr Ala Ser Glu Gly Glu Leu Ser 1100 1105 1110 Trp Tyr Asn Ile Phe Leu Tyr Asn Pro Asp Gly Asn Leu Gln Glu 1115 1120 1125 Arg Ala Gln Val Asp Pro Leu Val Gln Ser Phe Ser Phe Gln Asn 1130 1135 1140 Leu Leu Gln Gly Arg Met Tyr Lys Met Val Ile Val Thr His Ser 1145 1150 1155 Gly Glu Leu Ser Asn Glu Ser Phe Ile Phe Gly Arg Thr Val Pro 1160 1165 1170 Ala Ser Val Ser His Leu Arg Gly Ser Asn Arg Asn Thr Thr Asp 1175 1180 1185 Ser Leu Trp Phe Asn Trp Ser Pro Ala Ser Gly Asp Phe Asp Phe 1190 1195 1200 Tyr Glu Leu Ile Leu Tyr Asn Pro Asn Gly Thr Lys Lys Glu Asn 1205 1210 1215 Trp Lys Asp Lys Asp Leu Thr Glu Trp Arg Phe Gln Gly Leu Val 1220 1225 1230 Pro Gly Arg Lys Tyr Val Leu Trp Val Val Thr His Ser Gly Asp 1235 1240 1245 Leu Ser Asn Lys Val Thr Ala Glu Ser Arg Thr Ala Pro Ser Pro 1250 1255 1260 Pro Ser Leu Met Ser Phe Ala Asp Ile Ala Asn Thr Ser Leu Ala 1265 1270 1275 Ile Thr Trp Lys Gly Pro Pro Asp Trp Thr Asp Tyr Asn Asp Phe 1280 1285 1290 Glu Leu Gln Trp Leu Pro Arg Asp Ala Leu Thr Val Phe Asn Pro 1295 1300 1305 Tyr Asn Asn Arg Lys Ser Glu Gly Arg Ile Val Tyr Gly Leu Arg 1310 1315 1320 Pro Gly Arg Ser Tyr Gln Phe Asn Val Lys Thr Val Ser Gly Asp 1325 1330 1335 Ser Trp Lys Thr Tyr Ser Lys Pro Ile Phe Gly Ser Val Arg Thr 1340 1345 1350 Lys Pro Asp Lys Ile Gln Asn Leu His Cys Arg Pro Gln Asn Ser 1355 1360 1365 Thr Ala Ile Ala Cys Ser Trp Ile Pro Pro Asp Ser Asp Phe Asp 1370 1375 1380 Gly Tyr Ser Ile Glu Cys Arg Lys Met Asp Thr Gln Glu Val Glu 1385 1390 1395 Phe Ser Arg Lys Leu Glu Lys Glu Lys Ser Leu Leu Asn Ile Met 1400 1405 1410 Met Leu Val Pro His Lys Arg Tyr Leu Val Ser Ile Lys Val Gln 1415 1420 1425 Ser Ala Gly Met Thr Ser Glu Val Val Glu Asp Ser Thr Ile Thr 1430 1435 1440 Met Ile Asp Arg Pro Pro Pro Pro Pro Pro His Ile Arg Val Asn 1445 1450 1455 Glu Lys Asp Val Leu Ile Ser Lys Ser Ser Ile Asn Phe Thr Val 1460 1465 1470 Asn Cys Ser Trp Phe Ser Asp Thr Asn Gly Ala Val Lys Tyr Phe 1475 1480 1485 Thr Val Val Val Arg Glu Ala Asp Gly Ser Asp Glu Leu Lys Pro 1490 1495 1500 Glu Gln Gln His Pro Leu Pro Ser Tyr Leu Glu Tyr Arg His Asn 1505 1510 1515 Ala Ser Ile Arg Val Tyr Gln Thr Asn Tyr Phe Ala Ser Lys Cys 1520 1525 1530 Ala Glu Asn Pro Asn Ser Asn Ser Lys Ser Phe Asn Ile Lys Leu 1535 1540 1545 Gly Ala Glu Met Glu Ser Leu Gly Gly Lys Cys Asp Pro Thr Gln 1550 1555 1560 Gln Lys Phe Cys Asp Gly Pro Leu Lys Pro His Thr Ala Tyr Arg 1565 1570 1575 Ile Ser Ile Arg Ala Phe Thr Gln Leu Phe Asp Glu Asp Leu Lys 1580 1585 1590 Glu Phe Thr Lys Pro Leu Tyr Ser Asp Thr Phe Phe Ser Leu Pro 1595 1600 1605 Ile Thr Thr Glu Ser Glu Pro Leu Phe Gly Ala Ile Glu 1610 1615 1620 <210> 46 <211> 89 <212> PRT <213> Homo sapiens <400> 46 Leu Ala Glu Pro Glu Arg Cys Asn Phe Thr Leu Ala Glu Ser Lys Ala 1 5 10 15 Ser Ser His Ser Val Ser Ile Gln Trp Arg Ile Leu Gly Ser Pro Cys 20 25 30 Asn Phe Ser Leu Ile Tyr Ser Ser Asp Thr Leu Gly Ala Ala Leu Cys 35 40 45 Pro Thr Phe Arg Ile Asp Asn Thr Thr Tyr Gly Cys Asn Leu Gln Asp 50 55 60 Leu Gln Ala Gly Thr Ile Tyr Asn Phe Arg Ile Ile Ser Leu Asp Glu 65 70 75 80 Glu Arg Thr Val Val Leu Gln Thr Asp 85 <210> 47 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 47 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Val Gln Pro Lys Gly 1 5 10 15 Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ala Gln Asn Met 65 70 75 80 Leu Tyr Leu Gln Met Asn Asp Leu Lys Thr Glu Asp Thr Ala Met Tyr 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 48 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 48 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Gln Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 49 <400> 49 000 <210> 50 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 50 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Val Gln Pro Lys Gly 1 5 10 15 Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Ala Asn 20 25 30 Ala Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Arg Thr Lys Ser Asn Asn Tyr Ala Thr Tyr Tyr Ala Gly 50 55 60 Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ala Gln Asn Met 65 70 75 80 Leu Tyr Leu Gln Met Asn Asp Leu Lys Thr Glu Asp Thr Ala Met Tyr 85 90 95 Tyr Cys Val Arg Asp Tyr Tyr Gly Ser Ser Ala Trp Ile Thr Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 51 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 51 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln His Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Gln Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 52 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic 6xHis tag <400> 52 His His His His His 1 5 <210> 53 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic 10xHis Tag <400> 53 His His His His His His His His 1 5 10 <210> 54 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 54 Lys Ala Ser Gln His Val Gly Thr Ala Val Ala 1 5 10 <210> 55 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 55 Trp Ala Ser Thr Arg His Thr 1 5 <210> 56 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic Peptides <400> 56 Gln Gln Tyr Ser Ser Tyr Pro Phe Thr 1 5

Claims

1. An antibody comprising: (a) a heavy chain comprising the sequence of SEQ ID NO: 10 or 12; and (b) a light chain comprising the sequence of any one of SEQ ID NOs: 20, 22 and 23.

2. The antibody of claim 1, wherein the light chain comprises the sequence of SEQ ID NO:

20.

3. The antibody of claim 1, wherein the light chain comprises the sequence of SEQ ID NO:

22.

4. The antibody of claim 1, wherein the light chain comprises the sequence of SEQ ID NO:

23.

5. The antibody of claim 1, comprising: (a) a heavy chain comprising the sequence of SEQ ID NO: 10; and (b) a light chain comprising the sequence of SEQ ID NO:

23.

6. A pharmaceutical composition comprising a therapeutically effective amount of the antibody according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

7. Use of the antibody of any one of claims 1-5 in the preparation of a medicament for treating a condition in a subject in need thereof, wherein the condition is diabetic retinopathy, retinal neovascularization, ischemic retinopathy, retinal angiomatous proliferation, choroidal neovascularization, or wet age-related macular degeneration.

8. The use of claim 7, wherein the medicament is formulated for administration to the eye of a subject.

9. The use according to claim 7, wherein the medicament is for intravitreal administration.

10. The use according to claim 7, wherein the drug is for subcutaneous administration.

11. The use according to claim 7, wherein the medicament is for topical administration.

12. The use according to claim 7, wherein the subject is a human.

13. The use according to claim 7, wherein the medicament comprises the antibody in an amount of 0.25 mg to 200 mg.

14. The use according to claim 7, wherein the medicament comprises the antibody in an amount of 1 mg / kg to 10 mg / kg.

15. The use according to claim 7, wherein the medicament comprises the antibody in an amount of 1 mg to 50 mg.

16. The use according to claim 7, wherein the medicament comprises the antibody in an amount of 50 mg to 200 mg.

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