Novel Tissue Protective Erythropoietin Receptor (nepor) And Methods Of Use

Abstract

Methods of determining whether a patient is suitable for erythropoietin (EPO) therapy, including (A) isolating a tissue sample from said patient; (B) determining the level of expression of EPH-B4 in said sample; and (C) correlating a presence of EPH-B4 expression to a negative physiological response to EPO therapy). In addition, methods of enhancing the effectiveness of EPO therapy in a patient by administering to said patient, in conjunction with EPO therapy, an siRNA specific for EPH-B4.

Description

INFORMATION ON RELATED APPLICATIONS

[0001] This application is a continuation-in-part of International Application No. PCT/EP2008/066480 filed Nov. 28, 2008, which claims the benefit of U.S. Provisional Application 60/991,042, filed Nov. 29, 2007, both of which are herein incorporated by reference.

BACKGROUND

[0002] Approved by the FDA in 1993 for treatment of anemia, Erythropoietin (EPO) is a 193 amino acid glycoprotein hormone, produced by the kidneys to regulate red blood cell (RBC) production; a process commonly termed erythropoiesis. EPO was originally identified as a cytokine that promotes erythrocyte progenitor survival and differentiation, but has also been shown to possess neuroprotective functions, particularly in response to ischemic injury in the central nervous system, (CNS). Clinical use of EPO has been prevalent in the treatment of anemic cancer patients, while ongoing studies are exploring EPO's potential in the treatment of neurological diseases (e.g. stroke). Notwithstanding, recent clinical studies in cancer patients have begun to uncover highly worrying adverse events, suggesting that administration of recombinant human EPO (rHuEPO) can adversely effect overall patient survival. An urgent need thus exists in medical oncology to better understand and predict the prevalence or susceptibility to this effect, so that administration of rHuEPO can be contra-indicated, continued or stopped.

SUMMARY

[0003] The present invention discloses members of the ephrin family (ephrinA1 and EPH-B4) as mediators of cytoprotective EPO signalling, either as homodimers and/or as heterodimeric partners of EPOR and/or each other. Our data emphasize the importance of EPH-B4 and EphrinA1 in mediating this function. As such, NEPOR represents a novel EPO receptor derived from a unique combination (i.e. via homo- and/or hetero-dimerization) of components derived from ephrin biology and possibly the EPO receptor. See FIG. 3 for summary.

[0004] The present disclosure is based upon the data that EPH-B4 and EphrinA1 are the components of a novel EPO receptor (NEPOR). We are able to show that EPO stimulates enhanced tumor growth in a mouse tumor model system. EPO stimulates the Akt signalling pathway in cell lines lacking EPO receptor expression. These cells express EPH-B4 which is a receptor that stimulates signalling via the Akt pathway. Furthermore in a mouse tumor model it can be shown that EPO is capable of stimulating significant tumor growth. Such activity is inhibited via knock-down of the EPH-B4 receptor highlighting the EPH-B4 dependant nature of a EPO mediated tumor genesis. As such, NEPOR is primarily composed of EPH-B4 as a homodimer and/or in heterodimeric association with EPOR or an Ephrin. Furthermore, in silico analyses points to structural complementarity between EPO and Ephrin molecules, particularly Ephrin A1. Thus, NEPOR may also be composed of EphrinA1 as a homodimer and/or in heterodimeric association EPH-B4. A summary of these putative NEPOR species is provided in FIG. 3 and Table 5.

[0005] The present disclosure provides a method for assessing a tissue for expression of the tissue protective NEPOR receptor complex and/or EPH-B4 and/or Ephrin A1. In so doing, the present disclosure provides a prognostic method to stratify patients having a tumour as suitable (NEPOR not present on the tumour; NEPOR-) or non-suitable (NEPOR present on the tumour; NEPOR+) for EPO treatment. Specifically, the method for assessing tumour tissue NEPOR and/or gene expression components comprises: [0006] (a) isolating a tissue sample from an individual who is receiving or shall receive erythropoietin, [0007] (b) determining the level of expression of the NEPOR gene transcript(s) (i.e. EPH-B4, and/or Ephrin A1 mRNA) and/or the presence of the NEPOR gene products (i.e. EPH-B4, and/or Ephrin A1 proteins) from the isolated tissue, and [0008] (c) correlating the presence of these NEPOR component gene expression products to a negative physiological response to the treatment with erythropoietin.

[0009] In one aspect, methods are provided for determining whether a patient is suitable for erythropoietin (EPO) therapy, comprising (A) isolating a tissue sample from said patient; (B) determining the level of expression of EPH-B4 in said sample; and (C) correlating a presence of EPH-B4 expression to a negative physiological response to EPO therapy. In one embodiment, the level of expression is determined by measuring the amount of EPH-B4 protein (SEQ ID NO: 2) in said sample. In another embodiment, the level of expression is determined by measuring the amount of EPH-B4 mRNA (SEQ ID NO: 6) in said sample.

[0010] In another embodiment, the methods further comprise determining the level of expression in the sample of at least one of Ephrin A1 protein (SEQ ID NO: 3) or EPOR protein (SEQ ID NO: 1). Similarly, the methods can comprise determining the level of expression in the sample of at least one of Ephrin A1 mRNA (SEQ ID NO: 7) or EPOR mRNA (SEQ ID NO: 5).

[0011] Methods of determining the level of expression of EPH-B4 are further explained below.

[0012] In one embodiment, the presence of EPH-B4 expression is defined by the percentage of cells in said sample showing detectable levels of EPH-B4 protein and the concentration of EPH-B4 protein in said cells. In one example, the presence of EPH-B4 expression is defined by the formula P.times.C wherein P is the percentage of cells in said sample showing detectable levels of EPH-B4 protein and C is the relative concentration of EPH-B4 protein in said cells, wherein a score of 0, 1, 2, 3 or 4 is assigned to a sample comprising a percentage of cells showing detectable levels of EPH-B4 protein of, respectively, 0%, <25%, 25-50%, 50-75% and 75-100%, wherein a score of 1, 2 or 3 is assigned to relative concentrations of EPH-B4 protein of, respectively, weak, moderate and heavy, and wherein a resulting product of >3 denotes EPH-B4 expression in the sample.

[0013] In one embodiment of the present invention, the level of expression of EPH-B4, but not of EPH-A1 is determined. Alternatively, the level of expression of EPH-B4, but not of other components of NEPOR is determined. This includes the possibility that the level of other proteins not being part of NEPOR is determined.

[0014] In a further embodiment, only the level of expression of EPH-B4 is determined.

[0015] In one embodiment, the level of expression of EPH-B4 is determined by immunohistochemistry. In another embodiment, the level of expression of EPH-B4 is determined by ELISA. In another embodiment, the level of expression of EPH-B4 is determined by RT-PCR.

[0016] Preferably, the expression of the NEPOR component genes (i.e. EPH-B4, and/or Ephrin A1 mRNA) is determined by a molecular biological technique selected from the group consisting of PCR, QPCR, R-PCR, gene expression microarray analysis, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), capillary electrophoresis with laser induced fluorescence (CE-LIF) and combinations thereof.

[0017] Preferably, the determination of the presence of the NEPOR gene products is done by detecting the respective proteins with an immunoassay procedure, where the immunoassay procedure is selected from the group of immunoprecipitation, enzyme immunoassay (EIA), radioimmunoassay (RIA) or fluorescent immunoassay, a chemiluminescent assay, an agglutination assay, nephelometric assay, turbidimetric assay, a Western blot, a competitive immunoassay, a noncompetitive immunoassay, a homogeneous immunoassay a heterogeneous immunoassay, a bioassay and a reporter-assay such as a luciferase-assay. The immunoassay procedure is most preferably based on ELISA.

[0018] Preferably, the method for detection of NEPOR and/or EPH-B4, and/or Ephrin A1 on tumour tissue can also be an in situ imaging method, comprising administering an anti-NEPOR antibody or NEPOR binding peptide linked to a radio-ligand or other imaging agent, and measuring for tissue distribution and location of the radio-ligand or other imaging agent. Preferably, the tissue sample is selected from the cancerous tissue or circulating cells derived from same, or from a group of biological tissues and fluids such as blood, lymph, urine, cerebral fluid. Specifically, the individual is a cancer patient who is to be treated with erythropoietin or is being treated with erythropoietin. Preferably, the negative physiological effect is increased tumor progression and/or poorer patient survival. Preferably, the presence of NEPOR gene products and/or EPH-B4, and/or Ephrin A1 is indicative of increased tumor progression and/or poorer patient survival upon treatment with erythropoietin. Preferably the cancer is one of head and neck cancer, breast cancer, liver cancer, colorectal cancer, small intestine cancer, leukemia, prostate cancer, lung cancer, ovarian cancer, pancreatic cancer, endometrial cancer, stomach cancer, non-Hodgkin lymphoma, kidney cancer, Renal cell carcinoma (RCC), malignant melanoma, gallbladder cancer, bladder cancer, vulvar cancer, Penile cancer, testicular cancer, thymus cancer, Kaposi's sarcoma, eye cancer, adrenal gland cancer, brain cancer, cervical cancer, appendix cancer, adenoid cancer, bile duct cancer, urethral cancer, spinal cancer, Ewing's family of tumors, extragonal germ cell cancer, extra hepatic bile duct cancer, fallopian tube cancer, soft tissue cancers, bone cancer, Hodgkin's lymphoma, anal cancer, malignant mesothelioma, vaginal cancer skin cancer, central nervous system cancer (craniopharyngioma), pleuropulmonary blastoma, nasal cavity and paranasal sinus cancer transitional cell cancer of renal pelvis and ureter, pituitary gland cancer, squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, brain cancer, bladder cancer, and salivary gland cancer. It is particularly preferred that the cancer is selected from the group of squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, kidney cancer, brain cancer, bladder cancer and breast cancer.

[0019] The present disclosure further provides a method for designing a therapy which modulates the activity of NEPOR and/or EPH-B4, and/or Ephrin A1, comprising:

1) performing an in vitro screening assay for NEPOR and/or EPH-B4, and/or Ephrin A1 specific therapies; by measuring the binding of test compounds to a tissue protective NEPOR receptor complex and/or EPH-B4, and/or Ephrin A1 (also in comparison to EPOR homodimer complexes), wherein the test compound is labelled (binding of the labelled test compound to the receptor complexes detailed in FIG. 10) and is measured by detecting the label attached to the test compound; 2) performing a label-free screening approach such as surface plasmon resonance. In this case the test compound is not labelled and its binding to NEPOR receptor complexes (as detailed in FIG. 10) is measured by a label independent (optical) method. 3) testing NEPOR and/or EPH-B4, and/or Ephrin A1 activity by (a) contacting a test compound with a tissue protective NEPOR receptor complex (N) or tissue protective NEPOR receptor complex-expressing cell; measuring the level of the activity of (N) in the cell; identifying a test compound that increases or decreases the level of activity of (N) as compared to the level of activity of (N) measured in the absence of the test compound; and assaying the identified test compound for tissue protective activity; 4) testing the modulation of NEPOR/ligand binding and/or EPH-B4, and/or Ephrin A1 ligand binding by (a) contacting (N) with a tissue protective NEPOR receptor complex ligand and/or EPH-B4, and/or Ephrin A1 ligand attached to a first label, and an equivalent amount of a test compound attached to a second label under conditions conducive to binding, removing unbound material from (N), and detecting the level of the first and second labels, where if the second label is present the compound binds (N) and if the level of the first label decreases relative to the level of the first label when the labelled ligand is contacted with (N) under conditions conducive to binding in the absence of a test compound after removal of unbound material, then a compound that binds to (N) is identified. 5) identifying a compound that modulates a tissue protective activity in a mammal, comprising: (a) administering the compound to a first animal immediately following infliction of an injury, wherein the first animal endogenously expresses a tissue protective NEPOR receptor complex; and (b) administering the compound to a second animal immediately following infliction of the same injury as in step (a), wherein the second animal is deficient in expression of a tissue protective NEPOR receptor complex and/or EPH-B4, and/or Ephrin A1 or components thereof; such that if recovery from the injury differs in the animal of step (a) as compared to the animal of step (b), a compound that modulates a tissue protective activity is identified.

[0020] The present disclosure further provides methods for treating or preventing a disease or disorder in a human comprising administering a therapeutically effective amount of a compound that modulates the activity of a tissue protective NEPOR receptor complex to a human in need of such treatment or prevention, with the proviso that the compound is not EPO. The compound is selected from the group consisting of an antibody specific for the tissue protective NEPOR receptor complex, an antibody is specific for a tissue protective NEPOR receptor complex ligand, a small molecule, a peptide, an EPO mutant, an EPO:Ephrin_ligand_binding domain chimera, a member of a library, and a combination thereof. Preferably, such compounds negatively modulate the tissue protective function of the NEPOR receptor complex in the aforementioned mentioned cancers. Preferably such compounds positively modulate the tissue protective function of the NEPOR receptor complex wherein the disease or disorder is caused by hypoxia, seizure disorders, neurodegenerative diseases, neurotoxin poisoning, multiple sclerosis, hypotension, cardiac arrest, radiation, or hypoglycemia.

[0021] The present disclosure further provides a method for identifying compounds that modulate NEPOR's tissue protective signalling activity, comprising (a) contacting a test compound with the NEPOR receptor complex expressing cell; (b) measuring the level of tissue protective activity initiated by NEPOR activation in the cell; (c) identifying a test compound which increases or decreases the level of tissue protective NEPOR complex activity in a cell; (d) assaying the identified compounds for tissue protective activity mediated via NEPOR; and (e) assaying the identified therapeutics for NEPOR inhibitory activity. Preferably, the assay in step (d) is a tissue protective NEPOR receptor complex activity is measured by a cell proliferation/differentiation assay. More preferably, the cells in the cell proliferentiation/differentiation assay are recombinantly engineered to express EPH-B4, and/or EPOR, and/or Ephrin A1. More preferably, the cells endogenously express an EPO receptor and are transformed with a nucleic acid comprising a nucleotide sequence that (i) is operably linked to a promoter, and (ii) encodes either EPH-B4 and/or Ephrin A1. Most preferably, the cells endogenously express EPH-B4 and/or Ephrin A1 and are transformed with a nucleic acid comprising a nucleotide sequence that (i) is operably linked to a promoter, and (ii) encodes an EPO receptor polypeptide.

[0022] The present disclosure further provides a method for identifying a compound that modulates the interaction between a tissue protective NEPOR receptor complex and a tissue protective NEPOR receptor complex ligand, comprising: (a) contacting a tissue protective NEPOR receptor complex with one or more test compounds; and (b) measuring the tissue protective NEPOR receptor complex activity, whereby if the activity measured in (b) differs from the tissue protective NEPOR receptor complex activity in the absence of the one or more test compounds, then a compound that modulates the interaction between the tissue protective NEPOR receptor complex and the tissue protective NEPOR receptor complex ligand is identified. Preferably, the tissue protective NEPOR receptor complex activity is measured by cell proliferation or cell differentiation. Preferably, the tissue protective NEPOR receptor complex activity measured is the ability of the tissue protective NEPOR receptor complex to interact with a tissue protective NEPOR receptor complex ligand. Preferably, the step of assaying the identified compound for tissue protective activity comprises detecting the presence of nucleolin in the cell. Preferably, the step of assaying the identified compound for tissue protective activity comprises detecting or measuring an increased level of activity of neuroglobin or cytoglobin in a cell. Preferably, the tissue protective NEPOR receptor complex is in solution. Preferably, the tissue protective NEPOR receptor complex is in a cell. Preferably, the compound inhibits the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex. Preferably, the compound enhances the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex. Preferably, the tissue protective NEPOR receptor complex contacted in step (a) is on a cell surface. Preferably, the tissue protective NEPOR receptor complex is on an isolated cell membrane. Preferably, the tissue protective NEPOR receptor complex activity is compared to EPOR receptor activation to identify NEPOR specific compounds. Preferably, the tissue protective NEPOR receptor complex is immobilized to a solid surface and more preferably, the solid surface is a microtiter dish or a chip.

[0023] The present disclosure further provides a method for identifying a compound that binds a tissue protective NEPOR receptor complex, comprising: (a) contacting a test compound with a ligand-binding tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; and (b) contacting a test compound with a ligand-binding EPOR receptor complex fragment comprising at least two EPO receptor extracellular domains fused to an Fc fragment attached to a solid support (c) removing unbound test compounds from the solid supports; (d) identifying the compound attached to the tissue protective NEPOR receptor complex fragment, but not the EPOR receptor complex (and vice versa), whereby a compound bound to the solid support is identified as a compound that binds specifically to a tissue protective NEPOR receptor complex or a compound that binds specifically to an EPOR receptor complex.

[0024] The present disclosure further provides a method for identifying a compound that binds a tissue protective NEPOR receptor complex, comprising: (a) contacting a test compound with a ligand-binding tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; (b) removing unbound test compounds from the solid supports; (c) identifying the compound attached to the tissue protective NEPOR receptor complex fragment, whereby a compound bound to the solid support is identified as a compound that binds specifically to a tissue protective NEPOR receptor complex.

[0025] The present disclosure further provides a method for identifying a compound that binds to a tissue protective NEPOR receptor complex, comprising: (a) contacting a tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support with (i) a tissue protective NEPOR receptor complex ligand attached to a first label and (ii) an equivalent amount of a test compound attached to a second label under conditions conducive to binding; (b) removing unbound material from the tissue protective NEPOR receptor complex; and (c) detecting the level of the first and second labels wherein if the second label is present the compound binds the complex and if the level of the first label decreases relative to the level of the first label where the labelled ligand is contacted with a tissue protective NEPOR receptor complex under conditions conducive to binding in the absence of a test compound after removal of unbound material, then a compound that binds to a tissue protective NEPOR receptor complex is identified.

[0026] The present disclosure further provides a method for identifying a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex, comprising: (a) contacting a tissue protective NEPOR receptor complex ligand with a tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; in the presence of one or more test compounds under conditions conducive to binding; and (b) measuring the amount of tissue protective NEPOR receptor complex ligated bound to the tissue protective NEPOR receptor complex; whereby if the amount of bound tissue protective NEPOR receptor complex ligand measured in (b) differs from the amount of bound tissue protective NEPOR receptor complex ligand measured in the absence of the one or more test compounds, then a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to the tissue protective NEPOR receptor complex is identified.

[0027] Preferably, the amount of bound tissue protective NEPOR receptor complex ligand is measured using a tissue protective NEPOR receptor complex ligand-specific antibody. Preferably, the tissue protective NEPOR receptor complex ligand is labelled and binding of the tissue protective NEPOR receptor complex ligand to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the tissue protective NEPOR receptor complex ligand. Preferably, the tissue protective NEPOR receptor complex ligand is labelled and binding of the labelled ligand to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the tissue protective NEPOR receptor complex ligand. Preferably, the label is fluorescent. Preferably, the test compound is an antibody specific for the tissue protective NEPOR receptor complex. Preferably, the test compound is a small molecule. Preferably, the test compound is a peptide or a member of a library. Preferably, the tissue protective NEPOR receptor complex ligand is EPO, or derivatives thereof. Preferably, the compound binds the tissue protective NEPOR receptor complex or ligand thereof. Preferably, the tissue protective NEPOR receptor complex activity is compared to EPOR receptor activation to identify NEPOR specific compounds.

[0028] The present disclosure further provides a method for identifying a compound that modulates a tissue protective activity in a mammal, comprising: (a) administering the compound to a first animal immediately following infliction of an injury, wherein the first animal endogenously expresses a tissue protective NEPOR receptor complex; and (b) administering the compound to a second animal immediately following infliction of the same injury as in step (a), wherein the second animal is deficient in expression of a tissue protective NEPOR receptor complex or components thereof; such that if recovery from the injury differs in the animal of step (a) as compared to the animal of step (b), a compound that modulates a tissue protective activity is identified.

[0029] The present disclosure further provides a method for designing a compound which interferes with NEPOR's survival promoting activity, comprising: [0030] (a) providing the molecular makeup of the NEPOR species and providing amino acid sequences of a component NEPOR polypeptides; [0031] (b) using software comprised by the digital computer to design a chemical compound/protein construct which is predicted to bind to NEPOR; and [0032] (c) optionally designing protein constructs which mimic NEPOR in its dimerised/multimerised state (e.g. Fc constructs).

[0033] The present disclosure further provides a method for identifying compounds that modulate NEPOR's tissue protective signalling activity, comprising (a) contacting a test compound with the NEPOR receptor complex; (b) measuring the level of tissue protective activity initiated by NEPOR activation; (c) identifying a test compound which increases or decreases the level of tissue protective NEPOR complex activity; (d) assaying the identified therapeutics for tissue protective activity mediated via NEPOR; and (e) assaying the identified therapeutics for NEPOR inhibitory activity. Preferably, the tissue protective NEPOR receptor complex activity is measured by measuring the binding of the test compound to the NEPOR receptor complex. More preferably, the test compound is labelled and binding of the labelled test compound to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the test compound. Most preferably, the tissue protective NEPOR receptor complex activity is measured by measuring the binding of the test compound to the tissue protective NEPOR receptor complex.

[0034] The present disclosure further provides a method for imaging tumour tissue that is susceptible to enhanced survival in response to EPO treatment, comprising administering an anti-NEPOR antibody or NEPOR binding peptide linked to a radio-ligand or other imaging agent, and measuring for tissue distribution and location of the radio-ligand or other imaging agent. Preferably, the anti-NEPOR antibody is a monoclonal or polyclonal antibody selected from the group of antibodies listed in Table 6.

[0035] The present disclosure further provides a method for modulating cell survival in NEPOR positive tissue comprising administering an EPO mutants and peptides selected from the group consisting of peptides from SEQ ID NO. 17 through SEQ ID NO. 212.

[0036] The present disclosure further provides a method for modulating cell survival in NEPOR positive tissue comprising administering an effective amount of an EPO chimera, comprising an ephrin receptor ligand binding domain selected from the group consisting of SEQ ID NO. 215, and SEQ ID NO. 216.

[0037] In another aspect, methods are provided for enhancing the effectiveness of EPO therapy in a patient, comprising administering to the patient, in conjunction with EPO therapy, an siRNA specific for EPH-B4. In one embodiment, the siRNA is selected from the group of nucleic acid duplexes consisting of SEQ ID NO: 242 and SEQ ID NO: 243; SEQ ID NO: 244 and SEQ ID NO: 245; SEQ ID NO: 246 and SEQ ID NO: 247; SEQ ID NO: 248 and SEQ ID NO: 249; SEQ ID NO: 250 and SEQ ID NO: 251; SEQ ID NO: 252 and SEQ ID NO: 253; SEQ ID NO: 254 and SEQ ID NO: 255; SEQ ID NO: 256 and SEQ ID NO: 257; SEQ ID NO: 258 and SEQ ID NO: 259; and SEQ ID NO: 260 and SEQ ID NO: 261.

[0038] In another embodiment, the siRNA is a duplex of SEQ ID NO: 266 and SEQ ID NO: 267. In another, the siRNA is a duplex of ID NO: 219 and SEQ ID NO: 220.

BRIEF DESCRIPTION OF THE FIGURES

[0039] FIG. 1 shows the genomic localization of human Eph receptor (EPH) and ephrin (EFN) genes on human chromosomes.

[0040] FIG. 2 shows the domain architecture of Eph receptors and Ephrins (A and B subclasses).

[0041] FIG. 3 shows the theoretical combinations of receptors that might have EPO binding capacity.

[0042] FIG. 4 shows a process for identifying putative EPO binding transmembrane receptors. All proteins containing two membrane proximal FN3 domains were extracted (84 in all) and assessed for evidence of response to hypoxia. EPH-B4 was amongst one of four possible proteins extracted. Moreover, it is the only member of the Ephrin receptor family which is embryonic lethal, with death in embryo's preceding that of EPOR knock-outs.

[0043] FIG. 5 shows the human EPO locus showing the neighbouring EPH-B4 gene.

[0044] FIG. 6 shows a schematic of the results from analysis of the 5' and 3' UTR's (and an additional 500 bp on either side) of the EPO, EPOR and EPH-B4 genes for the presence of hypoxia inducible transcription factor binding sites. This study was performed employing the "Match" algorithm from TRANSFAC (Nucleic Acids Res. 2003 January 1, 31(1):374-8) to analyse the composition of HIF1 binding sites. Strikingly, only the EPO and EPH-B4 genes were found to contain such sites, supporting the hypothesis that EPH-B4 is indeed hypoxia inducible. The figure discloses the nucleotide sequences as SEQ ID NOS 225-233, respectively, in order of appearance and the protein sequences as SEQ ID NOS 268-273, respectively, in order of appearance.

[0045] FIG. 7A shows a structural analysis of EphrinA5:EphB2 association in comparison with that of EPO:EPOR. This structural analysis reveals several commonalities consistent with a propensity for Ephrin A1 to bind EPO. The top panel shows homology of the EPO binding region of EPOR to the human Ephrin A molecules. FIG. 7B compares the structural aspects of Ephrin A with EPOR. Figure discloses SEQ ID NOS 234-240 in the first box, residues 73-107 of SEQ ID NO: 234, 75-109 of SEQ ID NO: 235, 81-119 of SEQ ID NO: 236, 85-123 of SEQ ID NO: 237, 76-114 of SEQ ID NO: 238, 79-117 of SEQ ID NO: 239 and 76-105 of SEQ ID NO: 240 in the second box, residues 31-34 of SEQ ID NO: 234, 31-34 of SEQ ID NO: 235, 33-36 of SEQ ID NO: 236, 36-39 of SEQ ID NO: 237, 30-33 of SEQ ID NO: 238, 30-33 of SEQ ID NO: 239 and 26-32 of SEQ ID NO: 240 in the third box, and residues 45-53 of SEQ ID NO: 234, 47-55 of SEQ ID NO: 235, 50-58 of SEQ ID NO: 236, 54-62 of SEQ ID NO: 237, 47-55 of SEQ ID NO: 238, 48-56 of SEQ ID NO: 239 and 47-55 of SEQ ID NO: 240 in the fourth box.

[0046] FIG. 8 shows staining of hippocampus with anti-EPH-B4 and anti-EpoR antibodies. It should be noted that there is a striking co-expression of both proteins restricted to certain cells only. These data suggest functional coupling of EPH-B4 and EPOR activity.

[0047] FIG. 9 shows co-immunoprecipitation of EPH-B4 using flag-tagged EpoR. This finding is consistent with the notion that EPH-B4 and EPOR might heterodimerize.

[0048] FIG. 10 shows the possible various species of NEPOR (without being bound by theory). In this representation, NEPOR homo/heterodimer species are shown as Fc constructs. This mimics the dimerization of separate receptor monomers. Any method which allows the production of such NEPOR dimers can be employed in screening for NEPOR specific agonists and antagonists, including small molecules, peptides, proteins and EPO variants.

[0049] FIG. 11 shows an alignment of EPO protein mutants which are predicted to bind NEPOR more favourably than EPOR. Such mutants are predicted to be primarily tissue protective as opposed to haematopoietic, particularly those versions combining the described mutations.

[0050] FIG. 12 shows mRNA levels of ovarian cancer cell lines. RNA was isolated from a panel ovarian cancer cell lines and was reverse transcribed into cDNA. PCR was done using primers specific for EPO receptor, EPH-B4, Ephrin A1 and actin.

[0051] FIG. 13 shows protein expression in ovarian cancer cell lines. Protein extracts were isolated from a panel ovarian cancer cell lines. Samples were separated using SDS-Page gel electrophoresis. Immunoblots using antibodies for EPO Receptor (R&D biosystems), EPH-B4 (a gift from Prakash Gil), Ephrin A1 and acting (Sigma Aldrich) were used to compare protein expression.

[0052] FIG. 14 shows ESA protection from chemotherapy induced apoptosis. Ovarian cancer cell lines Hey A8, SkoV3 ip1, and HeyA8-MDR (chemoresistant) were treated with 50 U erythropoietin (EPO), 50 nM docetaxel, or a combination of EPO and docetaxes for 48 hours. Cells were then fixed and DNA stained with propidium iodide. Percentage of sub 01 cells were then quantified using flow cytometer (BD).

[0053] FIG. 15 shows signalling pathways activated in response to EPO in ovarian cancer cell lines. Cell lines previously characterized for expression levels of EPOR, EPH-B4, and Ephrin A1 were washed and grown in serum free media for two hours. Cells were then treated with 50 U EPO and collected and designated time points (0, 5 and 30 minutes). Protein extracts were isolated and analyzed by immunoblots using antibodies for phosphor-STAT5 (Invitrogen), phosphor-AKT, phosphor-ERK (Cell Signaling) and acting (Sigma Aldrich).

[0054] FIG. 16 shows erythropoietin induced tumor growth in nude mice. Mice were injected i.p. with 1.times.10.sup.6 Hey MDR ovarian cancer cells. Day eight following injections mice were injected with designated amounts of EPO (10, 50, 100 U, three mice per group) every second day. A) Mice were sacrificed at day 26 and tumor weight was measured. B) Protein extracts were isolated from tumors and analyzed by immunoblot using antibodies specific for phosphor AKT ser 473, phosphor ERK (Cell Signaling) and pSTAT5b (Invitrogen).

[0055] FIG. 17 shows EPH-B4 expression effects tumor promoting effect of EPO. Female nude mice were injected i.p. with 1.times.10.sup.6 HeyA8-MDR cells. Day eight following injection the cells were treated with control siRNA-DOPC, EPH-B4 siRNA-DOPC, EPO, or in EPO+ control or EPH-B4 siRNA-DOPC (10 per group). (50 U EPO given 3.times. week, 5 .mu.g siRNA 2.times. week). Mice were sacrificed on day 25 and tumor weights were measured. Statistics were done using students T-test. B) Distribution of tumor weight per group.

[0056] FIG. 18 shows tumor weight distributions.

[0057] FIG. 19 shows an immunoblot analysis of HeyA8 MDR and A2780cp20 cells exposed to Epo (50 U/ml) for 15 and 30 minutes or MG132 (10 .mu.M) for 30 minutes followed by co-immunoprecipitation with an anti-EPHB4 antibody.

[0058] FIG. 20 graphically shows the effect of EphB4 and EpoR inhibition on binding of iodine-125-labelled EPO in cell lines HeyA8, HeyA8 MDR and A2780cp20.

[0059] FIG. 21 graphically depicts immunohistochemical analysis of EpoR conducted on 4 .mu.m-thick formalin-fixed paraffin-embedded epithelial ovarian cancer specimens. ".largecircle." designates patients negative for EpoR, while "1" designates patients positive for EpoR. The ".dagger." symbol designates censored points, i.e. last medical follow-up for patients who have not died.

[0060] FIG. 22 graphically depicts immunohistochemical analysis of EphB4 conducted on 4 .mu.m-thick formalin-fixed paraffin-embedded epithelial ovarian cancer specimens. ".largecircle." designates patients negative for EphB4, while "1" designates patients positive for EphB4. The ".dagger." designates censored points, i.e. last medical follow-up for patients who have not died.

[0061] FIG. 23 graphically depicts immunohistochemical analysis of EphB4 and Epo-R conducted on 4 .mu.m-thick formalin-fixed paraffin-embedded epithelial ovarian cancer specimens. ".largecircle." designates patients that are both EphB4 and EpoR negative; "1" designates patients that are EphB4 positive and EpoR negative; "2" designates patients that are EphB4 negative and EpoR positive; "3" designates patients that are EphB4 positive and EpoR positive. The ".dagger." designates censored points, i.e. last medical follow-up for patients who have not died.

DETAILED DESCRIPTION

[0062] The present disclosure results from the identification of a novel EPO receptor, henceforth referred to as NEPOR. NEPOR was identified using a bioinformatics workflow encompassing both a functional and sequence based analysis of the human genome/proteome. Homology analysis involving an extracellular protein database (termed XtraCellDB) was used in conjunction text-mining and genome context analysis. These in silico predictions were subsequently verified in lab-based experiments. Thus, the present disclosure provides genomic, proteomic and experiment evidence that the protein EPH-B4 (Erythropoietin Producing Hepatoma protein B4) and/or Ephrin A1 act as EPO receptors.

EPO: Biological Function

[0063] Erythropoietin (EPO) is a 193 amino acid type I cytokine, produced by cells of the renal cortex to regulate red blood cell (RBC) production in a process termed erythropoiesis. Erythropoiesis is multistage in nature, involving the differentiation of pluripotent hematopoietic stem cells through the lineage-committed burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) progenitor cells, which give rise to a series of early and late erythroblasts, eventually leading to the formation of reticulocytes and mature erythrocytes. During this process, the sequential formation of pro-erythroblasts, basophilic, polychromatophilic, and orthochromatic erythroblasts is positively regulated by EPO. EPO induces multiple positive effects on early erythroblasts, including increased proliferation, progression through maturation, and protection from programmed cell death.

[0064] In terms of molecular mechanism, EPO binds to two identical receptors (EpoR), an event which activates several intracellular signaling pathways. These include Janus kinase 2-signal transducer and activator of transcription 5 (JAK2-STAT5), phosphatidylinositol 3-kinase (PI3K), protein kinase C (PKC), and Ras-Raf-MEK (mitogen-activated or extracellular signal-regulated protein kinase kinase)--ERK (extracellular signal-regulated protein kinase). The JAK2-STAT5 and RAS-RAF-MEK-ERK pathways are thought to be associated with Epo's mitogenic action, while the PI3K pathway, acting through Akt (PI3K-Akt), is viewed as a mediator of EPO's anti-apoptotic activities.

EPO: Clinical Use

[0065] Anemia (AmE) or an.ae butted.mia/anaemia (BrE), from the Greek ('v.alpha.i.mu.ia)(an-ha{hacek over (i)}ma) meaning "without blood", is a deficiency of red blood cells (RBCs) and/or hemoglobin. The condition is commonly observed in patients with chronic diseases, and is particularly common in cancer where about 50% of patients are anaemic at presentation and some 70-90% developing the condition during the course of treatment (typically termed chemotherapy induced anemia (CIA)). In a recent review of the European Cancer Anemia Survey (ECAS), Ludwig et al. cited a 50% baseline anemia rate (hemoglobin [Hb]<12 g/dL) among 3010 patients with hematological malignancies and a 41% baseline anemia rate among 11,453 patients with solid tumours (Blood, 2002; 100:234a-235a. Abstract 884). Further longitudinal analysis revealed that 72% of 2780 patients with haematologic malignancies and 66% of 10,067 patients with solid tumours succumbed to CIA. Other published studies have reported varying high rates in patients at different phases and with different types of treatment (Table 1). Notwithstanding, all studies demonstrate the extremely high prevalence of anemia amongst cancer patients.

TABLE-US-00001 TABLE 1 Prevalence of Anemia in Cancer Patients Undergoing Treatment Type of Cancer Prevalence of Anemia (Hb < 12 g/dL) Cervical cancer.sup.[3] 82% Solid tumors.sup.[1] 66% Colorectal cancer.sup.[3] 67% Lung cancer.sup.[3] 63% Haematological malignancies 72%

[0066] A number of factors contribute to the high incidence of anemia among cancer patients, including not only chemotherapy and radiation-induced myelosuppression, but also cytokine-mediated anemia of chronic disease, bleeding, marrow infiltration by tumour, hemolysis, and nutritional deficiencies. Whatever the source, anemia results in a reduced ability of blood to transfer oxygen to the tissues, leading to tissue hypoxia and an associated range of clinical consequences, affecting all realms of patient health: physiologic status, psychosocial well-being and quality of life. Not surprising, anemia can negatively affect a patient's response to cancer therapy, a fact which highlights the important supportive role of rHuEPO in restoring normal RBC counts.

EPO: Clinical Safety

[0067] ESA's were for many years considered to be extremely safe in their labelled indications of chronic kidney disease and chemotherapy-induced anemia. The first hints of safety issues came in 2003 when results from a pair of studies examining EPO's potentiation of radiation and chemotherapy prompted an FDA meeting in May 2004. This first study (the ENHANCE study: Lancet 2003; 362:1255-1260) suggested the relative risk of progression-free survival was worse for patients who received radiotherapy plus NeoRecormon epoetin beta from Roche than for patients receiving placebo plus radiotherapy. A randomized, double-blind, multi-institutional trial that included a study population of 351 patients who were receiving radiotherapy was performed. The patients were treated 3 times per week with either placebo or EPO in the form of epoetin beta starting 10 to 14 days before and continuing through radiation therapy. Although haemoglobin levels increased in 82% of patients receiving EPO, compared with 15% in patients receiving placebo, the rate of loco-regional progression-free survival was significantly lower. In addition, the EPO group had a higher relative risk for loco-regional progression and death.

[0068] In the second trial involving 939 breast cancer patients receiving chemotherapy (the BEST study: J. Clin. Oncol. 2005; 23:5960-5972; see table 2), those given Eprex epoetin alfa from Johnson & Johnson had a higher 4-month mortality rate and a lower 12-month survival rate than those on placebo. Both studies attempted to push the limits of hemoglobin levels beyond that permitted for marketing by the FDA--the recommended haemoglobin target for Aranesp was at the time up to 12 g/dL, while the labels for Epogen and Procrit recommended 10-12 g/dL. Henke treated men to target levels of at least 15 g/dL, while women were treated to at least 14 g/dL. The target level in the BEST study was 12-14 g/dL.

TABLE-US-00002 TABLE 2 Summary of the results from Leyland-Jones et al. (J. Clin. Oncol. 2005; 23: 5960-5972) showing that 8.7% of patients from the EPO treatment arm died within 4 months of treatment, compared to 3.4% in the non-treated arm. Table 2 Causes of Death Among Patients Who Died Within 4 Months of Random Assignment (ITT population, N = 939) Epoetin Alfa Placebo (n = 469) (n = 470) No. of No. of Outcome Patients % Patients % Alive at 4 months 428 91.3 454 96.6 Died within 4 months 41 8.7 16 3.4 ITT = Intention to treat.

[0069] Johnson & Johnson (JNJ, New Brunswick, N.J.) have since reported data from the Phase IV CHOIR trial (N. Engl. J. Med. 2006 November 16; 355(20):2085-98.) that tested whether using Procrit epoetin alfa to get hemoglobin levels to 13.5 g/dL would improve outcomes vs. treating to 11.3 g/dL (within the 10-12 g/dL range on the drug's label). Patients in the higher haemoglobin is group had a significantly increased incidence of mortality and cardiovascular events. While this study was carried out in the renal disease space, the safety implications were further emphasized in a more recent study--DAHANCA10. In February 2007, Amgen disclosed that this independent study had been halted three months earlier after interim data showed that Aranesp plus radiation missed the primary endpoint of 3-year loco-regional control vs. radiation alone. The study also showed a non-significant increase in death in the Aranesp arm. DAHANCA10 explored whether the use of Aranesp to maintain a hemoglobin level of 14-15.5 g/dL during radiotherapy could improve loco-regional disease control in patients with primary head and neck squamous cell carcinoma (HNSCC).

[0070] Safety signals also emerged from the use of Aranesp in the AoC space (study 103). In January 2007, Amgen reported that the risk/benefit profile of Aranesp was "at best neutral" in a Phase III trial in patients who had AoC and who were not receiving chemo- or radio-therapy. Here the data revealed significantly more deaths in Aranesp patients than in placebo patients. The trial, which treated patients to a haemoglobin level of 12-13 g/dL, also missed its primary endpoint of a significant reduction in transfusion frequency at 16 weeks. Study 103 enrolled patients with various cancers, including non-small cell lung cancer (NSCLC), breast cancer and prostate cancer. Canadian researchers have published similar findings (J. Clin. Oncol. 2007 March 20; 25(9):1027-32). Here the authors showed that of the 70 advanced NSCLC patients with AoC, those receiving Procrit, had a significantly higher mortality rate than those receiving placebo. A synopsis of each of these studies is provided in Table 3 below:

TABLE-US-00003 TABLE 3 Summary of results from EPO safety studies highlighting survival issues. STUDY EPO type POPULATION DESIGN STATUS DAHANCA Aranesp HNSCC; Baseline Multicenter, Terminated early by DMC (SE20029001) Hb <= 14.5 open-label trial (after 522 of 600 planned of radiotherapy +/- patients enrolled) based on Aranesp lower LRC rates and increased deaths in the ESA arm at planned interim analysis; 522 of 600 planned pts; summary results December 2006; CSR anticipated September 2008 EPO-CAN-20 Eprex/Procrit NSCLC not Double-blind, Terminated early by DSMB receiving chemo; placebo for increased deaths in ESA baseline Hb <= 12 controlled, arm; 70 of 300 patients randomized enrolled; results published in (1:1) +/- Eprex abstract in 2004 and in the journal of clinical oncology March 2007 BEST (EPO- Eprex/Procrit Metastatic breast Randomised, Terminated in April 2002, INT-76) cancer double blind, after review of data in the first placebo 938 pts by the DMC, due to controlled evidence of excess mortality in the Eprex arm RTOG 9903 Eprex/Procrit HNSCC; baseline Open-label, Terminated early by DSMB Hb 9-12.5 randomized (1:1), for trend to poorer LRC and (female), 9-13.5 chemo/radiation +/- OS in EPO arm. 148 of 372 (male) procrit patients enrolled. Results published in abstract 2004 Study 103 Aranesp NSCLC, prostate, (Amgen) breast cancer

[0071] These clinical findings have led many investigators to suggest a possible role for ESA's in promoting tumour growth through stimulation of EPO receptor survival signalling in tumour to cells, and via the stimulation of angiogenesis. Implicit in these proposed activities is the notion that the EPO receptor can somehow confer survival advantage to cancer cells, a negative side effect. This, in turn, suggests that EPO receptor is both present and activated by EPO binding in such cells. Using real-time, quantitative RT-PCR, the EPOR gene has not only been shown to be strongly expressed in bone marrow (containing the EPO-responsive erythroid progenitors), but also at significant levels in normal tissues (e.g. kidney, heart, brain, endothelium, and smooth muscle). Moreover, EPOR transcript levels in breast, colon, ovary, prostate, lung, lymphoma, ileum, stomach, and kidney tumour tissues and tumour cell lines were no higher than those levels observed in normal tissue counterparts. These findings are in concordance other reports which demonstrated that EPOR transcript levels are basically equivalent in matched tumour and non-tumour samples from patients with lung, colon and prostate cancer. From the perspective of these data, it is questionable whether the EPOR gene might somehow provide selective advantage to tumour cells, at least via abnormal expression levels.

[0072] Therefore, there is a possible role for EPOR in mediating tumour cell survival in response to EPO. From a molecular perspective, the ability of cancer cells to subvert the EPO/EPOR system would not be surprising. A number of preclinical studies have demonstrated EPO-mediated activation of the mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI.sub.3K)-Akt, JAK-STAT (Janus kinase-Signal Transducer and Activator of Transcription), and nuclear factor-kappa B (NF.kappa.B) signalling pathways in a variety of human cancers. Each of these signalling cascades has been associated with cellular functions that promote tumour progression. EPO stimulated not only chemotaxis of endothelial cells, together with migration and invasion of breast cancer and HNSCC cells, but also appears to induce cancer cell proliferation and inhibit apoptosis. Moreover, pretreatment with rHuEPO protects some cancer cell lines from the cytotoxic effects of the chemotherapeutic agent, cisplatin. Thus, EPO/EPOR signalling appears to contribute to a wide variety of tumour-promoting functions in different cancer types.

[0073] Despite this evidence, the possible contribution of EPO/EPOR signalling to cancer progression is anything but straightforward. The influence of EPO/EPOR on different cancer types appears to be quite variable and remains incompletely understood. Studies have shown that EPO does not influence the proliferation of cancer cell lines. Rosti et al. (Haematologica 1993 July-August; 78(4):208-12.), for example, investigated the proliferative potential of rHuEPO by testing the effects of this factor on clonogenic growth and DNA synthesis in 10 different cell lines derived from haematologic malignancies and solid tumours. The cell lines K-562 and HEL were included in this study, both of which express EPO receptors. Results showed that rHuEPO had no effect on either colony growth or DNA synthesis (see Table 4).

TABLE-US-00004 TABLE 4 Showing the lack of effect of rHuEPO on the percentage of cells in S phase in human cell lines. Cell line EPO (IU/ml) K-562 37.0 .+-. 2.0 37.1 .+-. 2.1 36.8 .+-. 1.7 HEL 27.3 .+-. 1.9 26.2 .+-. 1.3 25.8 .+-. 1.4 HL-60 26.4 .+-. 1.8 24.8 .+-. 2.1 25.6 .+-. 2.0 PLB 985 30.0 .+-. 1.7 27.8 .+-. 2.3 28.2 .+-. 2.5 KG-1 14.2 .+-. 1.3 14.0 .+-. 1.7 15.5 .+-. 1.8 H69 15.3 .+-. 1.5 15.8 .+-. 1.3 14.9 .+-. 1.6 N417 16.6 .+-. 1.8 17.0 .+-. 1.4 16.3 .+-. 2.2 MCF-7 20.0 .+-. 0.9 21.1 .+-. 1.2 19.7 .+-. 1.0 OCUM-1 16.1 .+-. 2.1 17.3 .+-. 2.4 15.3 .+-. 2.3 GBL-HU12 19.2 .+-. 1.5 20.9 .+-. 1.6 19.1 .+-. 2.0

[0074] In a similar study, Westphal et al. (Tumori 2002 March-April; 88(2):150-9.) investigated the effects of EPO on more than 25 different benign and malignant human cell lines. Expression of EPO receptor mRNA and protein was analyzed with RT-PCR, Western blot, and immunocytochemistry. Cellular responses to various concentrations of EPO were evaluated using tritiated thymidine uptake, Northern blot analysis of c-fos expression, and tyrosine-kinase activity assay. EPO receptor mRNA and protein were identified in the majority of the tumour cell lines evaluated. Despite these findings, treatment with rHuEPO did not significantly influence the proliferation rate of EPO-receptor-positive tumour cell lines. Moreover, treatment with EPO neither affected the gene c-fos mRNA of those cell lines nor stimulated tyrosine-kinase activation. Based on their findings, the authors concluded that expression of the EPO receptor in tumour cells does not appear to be essential for growth and therefore should not have a deleterious effect in cancer patients.

[0075] Results by Lu et al. (J. Biol. Chem., Vol. 281, Issue 11, 7002-7011, 2006) establish that receptor activation is not simply accomplished by bringing two receptors into close proximity through disulfide linkages in the transmembrane or extracellular domains. Instead, the relative orientation of the two transmembrane domains of an EpoR dimer, rather than their proximity, determines the extent of receptor activation. More specifically, these authors propose that Epo binding to the inactive, symmetric EpoR dimer causes the repositioning of the two fibronectinIII domains to an asymmetric 120.degree. relative orientation, which in turn changes the orientation of the transmembrane domains and intracellular domains, and juxtaposes the appended JAK2s to initiate the phosphorylation cascade. EPO mutants would not necessarily be expected to be capable of initiating EPOR-signalling, due to their inability to induce the correct relative conformation of the fibronectinIII domains. Interestingly, it appears that certain aspects of EPO function can be decoupled from EPOR activity. Leist et al. (Science 305, 239-242.) have shown that the haematopoietic and tissue-protective activities of Epo are distinct and separate, demonstrating for example that carbamylated Epo (CEpo) does not stimulate erythropoiesis, yet prevents tissue injury in a wide variety of in vivo and in vitro models.

[0076] EPO's efficacy in treating nervous system disease has been demonstrated in several experimental models of brain and spinal cord injury. As such, EPO has become a candidate for therapeutic neuro-protection. Notwithstanding, the use of EPO as a neuro-protectant raises several safety issues. Although recombinant EPO seems to be potentially safe at neuroprotective proven doses, cardiovascular or cerebrovascular events can occur as a result of its bone marrow stimulating activities. Interestingly, as highlighted above, EPO's neuronal protective function appears molecularly separable from the haematopoietic activity, as carbamylated EPO and certain EPO mutants are neuroprotective but fail to induce haematopoiesis. Such mutants fail to bind EPOR (Leist et al. Science 305, 239-242).

[0077] EPO was for a long time considered to act solely on haematopoietic cells, a fact which led to its emergence as a leading treatment for chemotherapy-induced anemia. However, emerging evidence has shown that EPO is expressed in a variety of tissue and cell types, including cancer, vascular endothelial, and neuronal cells. Expression of EPO is induced in response to hypoxia, an event mediated by the HIF-1 transcription factor. EPO is prototypically thought to exert its biological effects via binding to its cell surface receptor EPOR, resulting in tyrosine phosphorylation of the receptor and other intracellular proteins, including JAK2 and STAT5. The JAK/STAT pathway is utilized both in haematopoietic and non-haematopoietic cells (including brain cells) following binding of EPO to the EPO receptor. The recent findings of EPO-receptor expression in human breast and renal cancer cells, as well as in several tumour cell lines, have raised important questions in the oncology setting about a possible tumour-growth-promoting effect of rHuEPO on EPO-receptor-bearing tumours. This possibility has been borne out in several clinical trials. Interestingly, other studies have shown that certain EPO mutants which are cytoprotective but not longer able to induce haematopoiesis, function independently of EPOR. This suggests that another EPO receptor may exist which lacks EPOR's strict binding conformation requirements.

Ephrin and Ephrin Receptor Biology

[0078] Erythropoietin-producing hepatocellular carcinoma (Eph) receptors form the largest family of receptor tyrosine kinases. Eph receptors are divided into two groups (Eph-A's and Eph-B's) based on the similarity of their extracellular domain sequences and the distinct structural properties of the ephrin ligands (Eph Nomenclature Committee, 1997). About 16 ephrin receptor genes (EphA1-10, EphB1-6) have been identified in the vertebrate genome (Pasquale, Nat. Rev., Mol. Cell Biol. 6 (2005), pp. 462-475.), 14 of which are present in humans (FIG. 1) and other mammals (EphA1-8, EphA10, EphB1-4, EphB6).

[0079] Eph receptors are single-pass transmembrane proteins with highly conserved extracellular and intracellular domains. The former domains consists of an N-terminal ligand binding domain, a cysteine-rich EGF-like region and two fibronectin type III repeats (Yamaguchi and Pasquale, Curr. Opin. Neurobiol. 14 (2004), pp. 288-296.). Intracellularly, the juxtamembrane region is followed by a tyrosine kinase domain, followed by a sterile-.alpha.-motif (SAM), and a type-II PSD-95/Disc large/ZO-1 (PDZ) binding motif at the carboxyl terminus (Kullander and Klein, Nat. Rev., Mol. Cell Biol. 3 (2002), pp. 475-486.). The tyrosine kinase domain of one receptor from each class (EphA10 and EphB6) lacks residues that are essential for catalytic activity. Eph receptor variants are generated by alternative splicing and their structures differ from the prototypical domain structure. The domain architecture of Eph receptors and Ephrins (A and B subclasses) are shown in FIG. 2.

[0080] Eph receptors can undergo cis-oriented homo- as well as heterodimerization (Freywald et al., J. Biol. Chem. 277 (2002), pp. 3823-3828.), which is mediated directly by the extracellular cysteine-rich region, the fibronectin type III repeats (Lackmann et al., J. Biol. Chem. 273 (1998), pp. 20228-20237.) and the SAM motif (Stapleton et al., Nat. Struct. Biol. 6 (1999), pp. 44-49. and Thanos et al., Science 283 (1999), pp. 833-836.) or indirectly through PDZ protein interactions (Fanning and Anderson, J. Clin. Invest. 103 (1999), pp. 767-772). Trans-oriented interactions typically occur with select ephrin molecules on opposing cells. In common with their receptors, the ephrins (named derived from Eph family receptor interacting proteins or ephoros) are divided into two distinct subclasses A and B. Ephrin-A ligands are GPI-anchored peripheral membrane molecules. In contrast, ephrin-B ligands are transmembrane molecules whose short cytoplasmic domain is capable of participating in various signalling events. The ephrin-A and ephrin-B molecules were initially described as selectively interacting with EphA and EphB receptors, respectively. However, there may be crosstalk between A and B family members. For example, ephrin-A5 is capable of binding EphB2, while EphA4 binds to ephrin-A and ephrin-B family members. Although interactions across classes are limited, within a class they are promiscuous, with multiple EphA receptors binding to a given ephrinA and vice versa.

[0081] While neither class of ephrins possesses a catalytic activity, both can activate signal transduction pathways after interaction with Eph receptors (reverse signalling). Reverse signalling activated by transmembrane ephrins includes tyrosine phosphorylation of their cytoplasmic tail and interaction with various signalling molecules. The mechanism by which GPI-linked ephrins stimulate downstream signalling is still unclear.

[0082] Signalling sometimes involves formation of signalling assemblies, a process that begins with a monovalent interaction (nanomolar affinity) between an Eph receptor and an ephrin on a juxtaposed cell. Crystallographic work has shown that the globular ephrin-binding domain of EphB2 contains a cavity that accommodates a hydrophobic protrusion from the ephrins. Structural changes occur upon binding. For example, EphB2 undergoes different structural rearrangements upon binding to ephrin-B2 or ephrin-A5.

[0083] A lower affinity binding interface is also present on the opposite side of the EphB2 ligand binding domain (Eph.sub.--1b), with complementary interfaces also present in the Eph-receptor-binding domain of ephrin-B2. While only of micromolar binding affinity, the second interface can mediate the dimerization of two Eph-ephrin dimers into a tetramer that comprises two receptor and two ephrin molecules extending from adjacent cell surfaces. The lower-affinity interface contains important determinants of subclass specificity and is not engaged in the EphB2-ephrin-A5 complex.

[0084] Signalling is initiated upon transphosphorylation via correctly orientated kinase domains. Eph receptors become extensively phosphorylated upon activation by ephrins and via src-kinase association. Phosphorylation promotes conformational order on the activation segment of the kinase domain that favours substrate binding and also disrupts intra-molecular inhibitory interactions that occur between the juxtamembrane segment and the kinase domain. Src-family mediated phosphorylation of Eph receptors has also been shown to act in a similar manner.

Discussion

[0085] Working on the theory that the adverse effects of EPO seen in many cancer patients may be mediated by a receptor complex distinct from the prototypical EPO receptor (EPOR) homodimer, we initiated an in silky discovery project to try to identify a novel EPO receptor. Should such a novel EPO receptor species exist, we hypothesized that it will be responsible for mediating EPO-induced cell survival activity, as opposed to EPO mediated haematopoietic activity. Thus, we proposed the existence of at least two species of EPO receptor; the prototypical EPOR homodimer which is primarily responsible for EPO's haematopoietic activity, and a novel EPO receptor, termed NEPOR, which is primarily responsible for EPO's cytoprotective activities. The existence of such a novel EPO receptor is compelling for three main reasons. Firstly it allows the prediction of a cancer patients response to EPO. Presence of NEPOR on a tumour cell would imply a negative response to EPO, since binding of EPO by NEPOR would induce a cascade of survival signals within tumour cells and tissues, thus contributing to cancer progression and poorer patient survival. Thus, detection of NEPOR expression in a tumour provides a novel biomarker for stratify cancer patients as suitable (i.e. NEPOR not present) or unsuitable (i.e. NEPOR present) for EPO treatment. A corollary of this model is a second interesting perspective. If NEPOR is capable of initiating survival signals on cancer cells, then it represents an excellent therapeutic target for treatment of cancers expressing this receptor. Thus, therapeutic molecules targeting and antagonizing the tissue protective function of this receptor should be efficacious anti-cancer agents. Finally, under conditions where induction of cell survival is favourable, such as in response to ischemic stroke, therapeutic molecules capable of activating NEPOR-mediated survival signals provide an efficacious path to treating a variety of neurological diseases. Definition of NEPOR's molecular composition therefore provides the molecular basis for designing such therapies.

[0086] It had previously been proposed that rHuEPO can promote tumour growth through stimulation of Epo receptor (EPOR) signalling in tumour cells, and via the stimulation of angiogenesis. Binding of EPO to EPOR homodimers was assumed to somehow confer survival advantage to cancer cells, leading to increased loco-regional progression and poorer survival rates in patients having a form of cancer. However, aware of the binding promiscuity of exogenously administered therapeutics, we were anxious to address the possibility as to whether another receptor might be responsible for the observed negative outcomes, either alone or in functional interaction with EPOR.

[0087] In an effort to identify such a novel cytoprotective EPO receptor, we developed an in silico based analysis approach specifically designed to mine the human proteome for candidate molecules. Combining the power of text-mining and in-depth bioinformatics analysis, this multi-evidence based approach successfully identified a putative novel EPO receptor. Subsequent lab-based validation supports these findings. Given its established physiological role, we propose that by impinging on this receptors activity, EPO can confer survival advantage to certain cells, including cancer cells and neurons. As a consequence, the expression of this protein on cancer cells can be used to stratify the suitability of cancer patients for EPO treatment. Patients with cancer associated NEPOR expression should be contraindicated for EPO treatment. However, a corollary of this finding is that these same individuals represent excellent candidates for treatment with antagonistic anti-NEPOR therapies. In addition, we also propose that by mediating EPO's cyto-protective activity, NEPOR represents an excellent therapeutic target for a variety of diseases involving tissue ischaemia (e.g. stroke).

[0088] Thus, in the first instance, the present disclosure provides a method for assessing a tumour for expression of NEPOR. The disclosure provides a method to stratify patients having a tumour as suitable (i.e. NEPOR not present) or non-suitable (i.e., NEPOR present) for EPO treatment. The method disclosed comprises: (a) isolating a tissue sample from an individual who is receiving or shall receive erythropoietin, (b) determining the level of expression of the NEPOR gene(s) (mRNA) and/or the presence of the NEPOR gene product (protein) from the isolated tissue, and (c) correlating the presence of an NEPOR gene expression product or the presence of NEPOR protein to a physiological response to the treatment with erythropoietin. In a second instance, the present disclosure provides a method for treating patients possessing NEPOR positive tumors. Furthermore, the present disclosure provides a method for treating stroke. Finally, by providing a means of comparing binding affinities of putative therapeutics to both NEPOR and EPOR, the present disclosure provides a method for screening for NEPOR specific therapeutics (both antagonistic therapeutics for cancer, and agonistic therapeutics for treatment of hypoxia associated disease such as stroke). Such therapeutics will lack the haematopoietic activity associated with EPOR binding and signaling.

NEPOR--Molecular Definition

[0089] We have identified a novel multimeric EPO receptor, which we term NEPOR. NEPOR comprises EPHB4 and/or Ephrin A1 molecules either as homodimers or heterodimers. Without being bound by theory, these components may also heterodimerize with the EPO receptor. A synopsis of the possible molecular compositions of NEPOR is provided in FIG. 3. Despite the room for molecular promiscuity involving other components from ephrin biology, EPH-B4 and/or EphrinA1 are components of a novel EPO receptor (NEPOR). As such NEPOR is primarily composed of EPH-B4 and Ephrin A1, either as a homodimers and/or in heterodimeric association with each other, or the EPO receptor. Without being bound by theory, given the strong functional association between EPH-B4 and Ephrin B2, NEPOR may also comprise Ephrin B2 disclosed herein as SEQ ID NO. 4 (amino acid sequence), SEQ ID NO. 8 (mRNA sequence), and SEQ ID NO. 12 (binding region).

[0090] Table 5 shows, without being bound by theory, the possible molecular composition of dimeric EPO receptors. The prototypical haematopoietic EPO receptor (EPOR) represents a homodimer of two EPOR (SEQ ID NO. 1) monomers (1). Our results suggest that a novel tissue protective EPO receptor dimer is comprised of Ephrin A1 (SEQ ID NO. 3) and EPH-B4 (SEQ ID NO.2). Possible scenarios are shown in Table 5.

TABLE-US-00005 TABLE 5 Description Monomer 1 Monomer 2 1 EPOR SEQ ID NO. 1 SEQ ID NO. 1 2 NEPOR SEQ ID NO. 1 SEQ ID NO. 2 3 NEPOR SEQ ID NO. 1 SEQ ID NO. 3 4 NEPOR SEQ ID NO. 2 SEQ ID NO. 2 5 NEPOR SEQ ID NO. 2 SEQ ID NO. 3 6 NEPOR SEQ ID NO. 3 SEQ ID NO, 3 7 NEPOR SEQ ID NO. 1 SEQ ID NO. 4 8 NEPOR SEQ ID NO. 2 SEQ ID NO. 4 9 NEPOR SEQ ID NO. 3 SEQ ID NO. 4 10 NEPOR SEQ ID NO. 4 SEQ ID NO. 4 SEQ ID NO. 1 >EPOR MDHLGASLWPQVGSLCLLLAGAAWAPPPNLPDPKFESKAALLAARGPEELLCFTERLEDL VCFWEEAASAGVGPGNYSFSYQLEDEPWKLCRLHQAPTARGAVRFWCSLPTADTSSFVPL ELRVTAASGAPRYHRVIHINEVVLLDAPVGLVARLADESGHVVLRWLPPPETPMTSHIRY EVDVSAGNGAGSVQRVEILEGRTECVLSNLRGRTRYTFAVRARMAEPSFGGFWSAWSEPV SLLTPSDLDPLILTLSLILVVILVLLTVLALLSHRRALKQKIWPGIPSPESEFEGLFTTH KGNFQLWLYQNDGCLWWSPCTPFTEDPPASLEVLSERCWGTMQAVEPGTDDEGPLLEPVG SEHAQDTYLVLDKWLLPRNPPSEDLPGPGGSVDIVAMDEGSEASSCSSALASKPSPEGAS AASFEYTILDPSSQLLRPWTLCPELPPTPPHLKYLYLVVSDSGISTDYSSGDSQGAQGGL SDGPYSNPYENSLIPAAEPLPPSYVACS SEQ ID NO. 2 >EPH-B4 MELRVLLCWASLAAALEETLLNTKLETADLKWVTFPQVDGQWEELSGLDEEQHSVRTYEV CDVQRAPGQAHWLRTGWVPRRGAVHVYATLRFTMLECLSLPRAGRSCKETFTVFYYESDA DTATALTPAWMENPYIKVDTVAAEHLTRKRPGAEATGKVNVKTLRLGPLSKAGFYLAFQD QGACMALLSLHLFYKKCAQLTVNLTRFPETVPRELVVPVAGSCVVDAVPAPGPSPSLYCR EDGQWAEQPVTGCSCAPGFEAAEGNTKCRACAQGTFKPLSGEGSCQPCPANSHSNTIGSA VCQCRVGYFRARTDPRGAPCTTPPSAPRSVVSRLNGSSLHLEWSAPLESGGREDLTYALR CRECRPGGSCAPCGGDLTFDPGPRDLVEPWVVVRGLREDFTTITEVTALNGVSSLATGPV PFEPVNVTTDREVPPAVSDIRVTRSSPSSLSLAWAVPRAPSGAVLDYEVKYHEKGAEGPS SVRFLKTSENRAELRGLKRGASYLVQVRARSEAGYGPFGQEHHSQTQLDESEGWREQLAL IAGTAVVGVVLVLVVIVVAVLCLRKQSNGREAEYSDKHGQYLIGHGTKVYIDPFTYEDPN EAVREFAKEIDVSYVKIEEVIGAGEFGEVCRGRLKAPGKKESCVAIKTLKGGYTERQRRE FLSEASIMGQFEHPNIIRLEGVVTNSMPVMILTEFMENGALDSFLRLNDGQFTVIQLVGM LRGIASGMRYLAEMSYVHRDLAARNILVNSNLVCKVSDFGLSRFLEENSSDPTYTSSLGG KIPIRWTAPEAIAFRKFTSASDAWSYGIVMWEVMSFGERPYWDMSNQDVINAIEQDYRLP PPPDCPTSLHQLMLDCWQKDRNARPRFPQVVSALDKMIRNPASLKIVARENGGASHPLLD QRQPHYSAFGSVGEWLRAIKMGRYEESFAAAGFGSFELVSQISAEDLLRIGVTLAGHQKK ILASVQHMKSQAKPGTPGGTGGPAPQY SEQ ID NO. 3 >EphrinA1 MEFLWAPLLGLCCSLAAADRHTVFWNSSNPKFRNEDYTIHVQLNDYVDIICPHYEDHSVA DAAMEQYILYLVEHEEYQLCQPQSKDQVRWQCNRPSAKHGPEKLSEKFQRFTPFTLGKEF KEGHSYYYISKPIHQHEDRCLRLKVTVSGKITHSPQAHDNPQEKRLAADDPEVRVLHSIG HSAAPRLFPLAWTVLLLPLLLLQTP SEQ ID NO. 4 >EphrinB2 MAVRRDSVWKYCWGVLMVLCRTAISKSIVLEPIYWNSSNSKFLPGQGLVLYPQIGDKLDI ICPKVDSKTVGQYEYYKVYMVDKDQADRCTIKKENTPLLNCAKPDQDIKFTIKFQEFSPN LWGLEFQKNKDYYIISTSNGSLEGLDNQEGGVCQTRAMKILMKVGQDASSAGSTRNKDPT RRPELEAGTNGRSSTTSPFVKPNPGSSTDGNSAGHSGNNILGSEVALFAGIASGCIIFIV IIITLVVLLLKYRRRHRKHSPQHTTTLSLSTLATPKRSGNNNGSEPSDIIIPLRTADSVF CPHYEKVSGDYGHPVYIVQEMPPQSPANIYYKV

[0091] The present disclosure includes any splice variant of the polypeptides of SEQ ID NOS 1-4 components possessing the extracellular EPO binding region (for EPH-B4 this region of proposed to encompass the two fibronectinIII domains; the oval structures adjacent to Epo in FIGS. 3B, D and F) and the intracellular signalling part, is also capable of mediating EPO's (and derivatives thereof) cyto-protective effect.

NEPOR: Prognostic Implications

[0092] The type I cytokine, Erythropoietin (EPO), possesses both haematopoietic and tissue protective activities. The present disclosure provides that the latter functionality is mediated via interactions of EPO with a novel EPO receptor, termed NEPOR. The model provides that binding of EPO to NEPOR receptor complexes, on NEPOR positive cancer cells, confers survival advantage to such cells. The implicit physiological outcome for patients possessing NEPOR positive cancers is therefore increased loco-regional cancer progression and poorer overall survival.

[0093] Thus, the present disclosure provides a diagnostic or prognostic test that can predict whether or not cancer patients administered EPO will respond negatively in terms of survival outcome. The prognostic test comprises determining NEPOR (i.e. EPH-B4, and/or Ephrin A1) in tumour tissue, or more particularly cancer cells. In another embodiment NEPOR component gene expression levels in tumour cells can be compared to baseline levels or levels in surrounding normal cells or tissue. Therefore, a comparative analysis looking at elevated or normal baseline expression levels of NEPOR component expression, using standard gene expression analysis methods (such as q-PCR and DNA microarray analyses) provides a diagnostic test that can determine whether or not administration of EPO to cancer patients will unwittingly enhance tumour cell survival (a negative outcome).

[0094] As stated, one method that can be used for comparing levels of gene expression of components of NEPOR and/or EPH-B4, and/or Ephrin A1 is Quantitative polymerase chain reaction (qPCR). This is a modification of PCR or polymerase chain reaction used to rapidly measure the quantity of DNA present in a tissue sample. Like other forms of polymerase chain reaction, the process is used to amplify nucleic acid samples, via the temperature-mediated enzyme DNA polymerase. PCR amplifies DNA exponentially, doubling the number of molecules present with each amplification cycle. The number of amplification cycles and the amount of PCR end-product should allow one to calculate the initial quantity of NEPOR-specific genetic material and/or EPH-B4 and/or Ephrin A1 genetic material in particular mRNA molecules using NEPOR-specific component sequences in particular and/or EPH-B4, and/or Ephrin A1 sequences for the two primers used for amplification.

[0095] In addition, gene expression analysis of NEPOR components and/or EPH-B4, and/or Ephrin A1 can be done with a microarray analysis containing a plurality of capture probes specific for sequences of the NEPOR complex in particular and/or EPH-B4, and/or Ephrin A1. As EPO is proposed to stimulate survival of NEPOR positive cancer cells and/or EPH-B4, and/or Ephrin A1 positive cells, it is important to test all cancer patients for NEPOR status and/or EPH-B4, and/or Ephrin A1 status prior to and during EPO administration. This is best done with a microarray analysis for expression status of NEPOR component genes in tumour tissue and with mRNA samples taken from tumour tissue. Ascertaining the levels of endogenous tumour associated NEPOR (i.e. EPH-B4, and/or EphrinA1) expression, provide correlations as to patient prognosis/survival rate.

[0096] The present disclosure thus provides a method to stratify patients having a tumour as suitable (i.e. NEPOR not present and/or EPH-B4, and/or Ephrin A1 present) or non-suitable (i.e., NEPOR present and/or EPH-B4, and/or Ephrin A1 present) for EPO treatment. The method disclosed comprises: (a) isolating a tissue sample from an individual who is receiving or shall receive erythropoietin, (b) determining the level of expression of EPH-B4 and/or Ephrin A1 from the isolated tissue, and (c) correlating the presence of these component gene expression products to a negative physiological response to the treatment with erythropoietin.

TABLE-US-00006 SEQ ID NO. 5 >erythropoietin receptor (EPOR), mRNA ACTTAGAGGCGCCTGGTCGGGAAGGGCCTGGTCAGCTGCGTCCGGCGGAGGCAGCTGCTGACCCAGCTGT GGACTGTGCCGGGGGCGGGGGACGGAGGGGCAGGAGCCCTGGGCTCCCCGTGGCGGGGGCTGTATCATGG ACCACCTCGGGGCGTCCCTCTGGCCCCAGGTCGGCTCCCTTTGTCTCCTGCTCGCTGGGGCCGCCTGGGC GCCCCCGCCTAACCTCCCGGACCCCAAGTTCGAGAGCAAAGCGGCCTTGCTGGCGGCCCGGGGGCCCGAA GAGCTTCTGTGCTTCACCGAGCGGTTGGAGGACTTGGTGTGTTTCTGGGAGGAAGCGGCGAGCGCTGGGG TGGGCCCGGGCAACTACAGCTTCTCCTACCAGCTCGAGGATGAGCCATGGAAGCTGTGTCGCCTGCACCA GGCTCCCACGGCTCGTGGTGCGGTGCGCTTCTGGTGTTCGCTGCCTACAGCCGACACGTCGAGCTTCGTG CCCCTAGAGTTGCGCGTCACAGCAGCCTCCGGCGCTCCGCGATATCACCGTGTCATCCACATCAATGAAG TAGTGCTCCTAGACGCCCCCGTGGGGCTGGTGGCGCGGTTGGCTGACGAGAGCGGCCACGTAGTGTTGCG CTGGCTCCCGCCGCCTGAGACACCCATGACGTCTCACATCCGCTACGAGGTGGACGTCTCGGCCGGCAAC GGCGCAGGGAGCGTACAGAGGGTGGAGATCCTGGAGGGCCGCACCGAGTGTGTGCTGAGCAACCTGCGGG GCCGGACGCGCTACACCTTCGCCGTCCGCGCGCGTATGGCTGAGCCGAGCTTCGGCGGCTTCTGGAGCGC CTGGTCGGAGCCTGTGTCGCTGCTGACGCCTAGCGACCTGGACCCCCTCATCCTGACGCTCTCCCTCATC CTCGTGGTCATCCTGGTGCTGCTGACCGTGCTCGCGCTGCTCTCCCACCGCCGGGCTCTGAAGCAGAAGA TCTGGCCTGGCATCCCGAGCCCAGAGAGCGAGTTTGAAGGCCTCTTCACCACCCACAAGGGTAACTTCCA GCTGTGGCTGTACCAGAATGATGGCTGCCTGTGGTGGAGCCCCTGCACCCCCTTCACGGAGGACCCACCT GCTTCCCTGGAAGTCCTCTCAGAGCGCTGCTGGGGGACGATGCAGGCAGTGGAGCCGGGGACAGATGATG AGGGCCCCCTGCTGGAGCCAGTGGGCAGTGAGCATGCCCAGGATACCTATCTGGTGCTGGACAAATGGTT GCTGCCCCGGAACCCGCCCAGTGAGGACCTCCCAGGGCCTGGTGGCAGTGTGGACATAGTGGCCATGGAT GAAGGCTCAGAAGCATCCTCCTGCTCATCTGCTTTGGCCTCGAAGCCCAGCCCAGAGGGAGCCTCTGCTG CCAGCTTTGAGTACACTATCCTGGACCCCAGCTCCCAGCTCTTGCGTCCATGGACACTGTGCCCTGAGCT GCCCCCTACCCCACCCCACCTAAAGTACCTGTACCTTGTGGTATCTGACTCTGGCATCTCAACTGACTAC AGCTCAGGGGACTCCCAGGGAGCCCAAGGGGGCTTATCCGATGGCCCCTACTCCAACCCTTATGAGAACA GCCTTATCCCAGCCGCTGAGCCTCTGCCCCCCAGCTATGTGGCTTGCTCTTAGGACACCAGGCTGCAGAT GATCAGGGATCCAATATGACTCAGAGAACCAGTGCAGACTCAAGACTTATGGAACAGGGATGGCGAGGCC TCTCTCAGGAGCAGGGGCATTGCTGATTTTGTCTGCCCAATCCATCCTGCTCAGGAAACCACAACCTTGC AGTATTTTTAAATATGTATAGTTTTTTTG SEQ ID NO. 6 >EPH receptor B4 (EPHB4), mRNA TTCCAGCGCAGCTCAGCCCCTGCCCGGCCCGGCCCGCCCGGCTCCGCGCCGCAGTCTCCCTCCCTCCCGC TCCGTCCCCGCTCGGGCTCCCACCATCCCCGCCCGCGAGGAGAGCACTCGGCCCGGCGGCGCGAGCAGAG CCACTCCAGGGAGGGGGGGAGACCGCGAGCGGCCGGCTCAGCCCCCGCCACCCGGGGCGGGACCCCGAGG CCCCGGAGGGACCCCAACTCCAGCCACGTCTTGCTGCGCGCCCGCCCGGCGCGGCCACTGCCAGCACGCT CCGGGCCCGCCGCCCGCGCGCGCGGCACAGACGCGGGGCCACACTTGGCGCCGCCGCCCGGTGCCCCGCA CGCTCGCATGGGCCCGCGCTGAGGGCCCCGACGAGGAGTCCCGCGCGGAGTATCGGCGTCCACCCGCCCA GGGAGAGTCAGACCTGGGGGGGCGAGGGCCCCCCAAACTCAGTTCGGATCCTACCCGAGTGAGGCGGCGC CATGGAGCTCCGGGTGCTGCTCTGCTGGGCTTCGTTGGCCGCAGCTTTGGAAGAGACCCTGCTGAACACA AAATTGGAAACTGCTGATCTGAAGTGGGTGACATTCCCTCAGGTGGACGGGCAGTGGGAGGAACTGAGCG GCCTGGATGAGGAACAGCACAGCGTGCGCACCTACGAAGTGTGTGACGTGCAGCGTGCCCCGGGCCAGGC CCACTGGCTTCGCACAGGTTGGGTCCCACGGCGGGGCGCCGTCCACGTGTACGCCACGCTGCGCTTCACC ATGCTCGAGTGCCTGTCCCTGCCTCGGGCTGGGCGCTCCTGCAAGGAGACCTTCACCGTCTTCTACTATG AGAGCGATGCGGACACGGCCACGGCCCTCACGCCAGCCTGGATGGAGAACCCCTACATCAAGGTGGACAC GGTGGCCGCGGAGCATCTCACCCGGAAGCGCCCTGGGGCCGAGGCCACCGGGAAGGTGAATGTCAAGACG CTGCGTCTGGGACCGCTCAGCAAGGCTGGCTTCTACCTGGCCTTCCAGGACCAGGGTGCCTGCATGGCCC TGCTATCCCTGCACCTCTTCTACAAAAAGTGCGCCCAGCTGACTGTGAACCTGACTCGATTCCCGGAGAC TGTGCCTCGGGAGCTGGTTGTGCCCGTGGCCGGTAGCTGCGTGGTGGATGCCGTCCCCGCCCCTGGCCCC AGCCCCAGCCTCTACTGCCGTGAGGATGGCCAGTGGGCCGAACAGCCGGTCACGGGCTGCAGCTGTGCTC CGGGGTTCGAGGCAGCTGAGGGGAACACCAAGTGCCGAGCCTGTGCCCAGGGCACCTTCAAGCCCCTGTC AGGAGAAGGGTCCTGCCAGCCATGCCCAGCCAATAGCCACTCTAACACCATTGGATCAGCCGTCTGCCAG TGCCGCGTCGGGTACTTCCGGGCACGCACAGACCCCCGGGGTGCACCCTGCACCACCCCTCCTTCGGCTC CGCGGAGCGTGGTTTCCCGCCTGAACGGCTCCTCCCTGCACCTGGAATGGAGTGCCCCCCTGGAGTCTGG TGGCCGAGAGGACCTCACCTACGCCCTCCGCTGCCGGGAGTGCCGACCCGGAGGCTCCTGTGCGCCCTGC GGGGGAGACCTGACTTTTGACCCCGGCCCCCGGGACCTGGTGGAGCCCTGGGTGGTGGTTCGAGGGCTAC GTCCTGACTTCACCTATACCTTTGAGGTCACTGCATTGAACGGGGTATCCTCCTTAGCCACGGGGCCCGT CCCATTTGAGCCTGTCAATGTCACCACTGACCGAGAGGTACCTCCTGCAGTGTCTGACATCCGGGTGACG CGGTCCTCACCCAGCAGCTTGAGCCTGGCCTGGGCTGTTCCCCGGGCACCCAGTGGGGCTGTGCTGGACT ACGAGGTCAAATACCATGAGAAGGGCGCCGAGGGTCCCAGCAGCGTGCGGTTCCTGAAGACGTCAGAAAA CCGGGCAGAGCTGCGGGGGCTGAAGCGGGGAGCCAGCTACCTGGTGCAGGTACGGGCGCGCTCTGAGGCC GGCTACGGGCCCTTCGGCCAGGAACATCACAGCCAGACCCAACTGGATGAGAGCGAGGGCTGGCGGGAGC AGCTGGCCCTGATTGCGGGCACGGCAGTCGTGGGTGTGGTCCTGGTCCTGGTGGTCATTGTGGTCGCAGT TCTCTGCCTCAGGAAGCAGAGCAATGGGAGAGAAGCAGAATATTCGGACAAACACGGACAGTATCTCATC GGACATGGTACTAAGGTCTACATCGACCCCTTCACTTATGAAGACCCTAATGAGGCTGTGAGGGAATTTG CAAAAGAGATCGATGTCTCCTACGTCAAGATTGAAGAGGTGATTGGTGCAGGTGAGTTTGGCGAGGTGTG CCGGGGGCGGCTCAAGGCCCCAGGGAAGAAGGAGAGCTGTGTGGCAATCAAGACCCTGAAGGGTGGCTAC ACGGAGCGGCAGCGGCGTGAGTTTCTGAGCGAGGCCTCCATCATGGGCCAGTTCGAGCACCCCAATATCA TCCGCCTGGAGGGCGTGGTCACCAACAGCATGCCCGTCATGATTCTCACAGAGTTCATGGAGAACGGCGC CCTGGACTCCTTCCTGCGGCTAAACGACGGACAGTTCACAGTCATCCAGCTCGTGGGCATGCTGCGGGGC ATCGCCTCGGGCATGCGGTACCTTGCCGAGATGAGCTACGTCCACCGAGACCTGGCTGCTCGCAACATCC TAGTCAACAGCAACCTCGTCTGCAAAGTGTCTGACTTTGGCCTTTCCCGATTCCTGGAGGAGAACTCTTC CGATCCCACCTACACGAGCTCCCTGGGAGGAAAGATTCCCATCCGATGGACTGCCCCGGAGGCCATTGCC TTCCGGAAGTTCACTTCCGCCAGTGATGCCTGGAGTTACGGGATTGTGATGTGGGAGGTGATGTCATTTG GGGAGAGGCCGTACTGGGACATGAGCAATCAGGACGTGATCAATGCCATTGAACAGGACTACCGGCTGCC CCCGCCCCCAGACTGTCCCACCTCCCTCCACCAGCTCATGCTGGACTGTTGGCAGAAAGACCGGAATGCC CGGCCCCGCTTCCCCCAGGTGGTCAGCGCCCTGGACAAGATGATCCGGAACCCCGCCAGCCTCAAAATCG TGGCCCGGGAGAATGGCGGGGCCTCACACCCTCTCCTGGACCAGCGGCAGCCTCACTACTCAGCTTTTGG CTCTGTGGGCGAGTGGCTTCGGGCCATCAAAATGGGAAGATACGAAGAAAGTTTCGCAGCCGCTGGCTTT GGCTCCTTCGAGCTGGTCAGCCAGATCTCTGCTGAGGACCTGCTCCGAATCGGAGTCACTCTGGCGGGAC ACCAGAAGAAAATCTTGGCCAGTGTCCAGCACATGAAGTCCCAGGCCAAGCCGGGAACCCCGGGTGGGAC AGGAGGACCGGCCCCGCAGTACTGACCTGCAGGAACTCCCCACCCCAGGGACACCGCCTCCCCATTTTCC GGGGCAGAGTGGGGACTCACAGAGGCCCCCAGCCCTGTGCCCCGCTGGATTGCACTTTGAGCCCGTGGGG TGAGGAGTTGGCAATTTGGAGAGACAGGATTTGGGGGTTCTGCCATAATAGGAGGGGAAAATCACCCCCC AGCCACCTCGGGGAACTCCAGACCAAGGGTGAGGGCGCCTTTCCCTCAGGACTGGGTGTGACCAGAGGAA AAGGAAGTGCCCAACATCTCCCAGCCTCCCCAGGTGCCCCCCTCACCTTGATGGGTGCGTTCCCGCAGAC CAAAGAGAGTGTGACTCCCTTGCCAGCTCCAGAGTGGGGGGGCTGTCCCAGGGGGCAAGAAGGGGTGTCA GGGCCCAGTGACAAAATCATTGGGGTTTGTAGTCCCAACTTGCTGCTGTCACCACCAAACTCAATCATTT TTTTCCCTTGTAAATGCCCCTCCCCCAGCTGCTGCCTTCATATTGAAGGTTTTTGAGTTTTGTTTTTGGT CTTAATTTTTCTCCCCGTTCCCTTTTTGTTTCTTCCTTTTGTTTTTCTACCGTCCTTGTCATAACTTTGT GTTGGAGGGAACCTGTTTCACTATGGCCTCCTTTGCCCAAGTTGAAACAGGGGCCCATCATCATGTCTGT TTCCAGAACAGTGCCTTGGTCATCCCACATCCCCGGACCCCGCCTGGGACCCCCAAGCTGTGTCCTATGA AGGGGTGTGGGGTGAGGTAGTGAAAAGGGCGGTAGTTGGTGGTGGAACCCAGAAACGGACGCCGGTGCTT GGAGGGGTTCTTAAATTATATTTAAAAAAGTAACTTTTTGTATAAATAAAAGAAAATGGGACGTGTCCCA GCTCCAGGGGTAAAAAAAAAAAAAAAAAA SEQ ID NO. 7 >Ephrin-A1 (EFNA1) mRNA GCCAGATCTGTGAGCCCAGCGCTGACTGCGCCGCGGAGAAAGCCAGTGGGAACCCAGACCCATAGGAGAC CCGCGTCCCCGCTCGGCCTGGCCAGGCCCCGCGCTATGGAGTTCCTCTGGGCCCCTCTCTTGGGTCTGTG CTGCAGTCTGGCCGCTGCTGATCGCCACACCGTCTTCTGGAACAGTTCAAATCCCAAGTTCCGGAATCAG GACTACACCATACATGTGCAGCTGAATGACTACGTGGACATCATCTGTCCGCACTATGAAGATCACTCTG TGGCAGACGCTGCCATGGAGCAGTACATACTGTACCTGGTGGAGCATGAGGAGTACCAGCTGTGCCAGCC CCAGTCCAAGGACCAAGTCCGCTGGCAGTGCAACCGGCCCAGTGCCAAGCATGGCCCGGAGAAGCTGTCT GAGAAGTTCCAGCGCTTCACACCTTTCACCCTGGGCAAGGAGTTCAAAGAAGGACACAGCTACTACTACA TCTCCAAACCCATCCACCAGCATGAAGACCGCTGCTTGAGGTTGAAGGTGACTGTCAGTGGCAAAATCAC TCACAGTCCTCAGGCCCATGACAATCCACAGGAGAAGAGACTTGCAGCAGATGACCCAGAGGTGCGGGTT CTACATAGCATCGGTCACAGTGCTGCCCCACGCCTCTTCCCACTTGCCTGGACTGTGCTGCTCCTTCCAC TTCTGCTGCTGCAAACCCCGTGAAGGTGTATGCCACACCTGGCCTTAAAGAGGGACAGGCTGAAGAGAGG GACAGGCACTCCAAACCTGTCTTGGGGCCACTTTCAGAGCCCCCAGCCCTGGGAACCACTCCCACCACAG GCATAAGCTATCACCTAGCAGCCTCAAAACGGGTCAGTATTAAGGTTTTCAACCGGAAGGAGGCCAACCA GCCCGACAGTGCCATCCCCACCTTCACCTCGGAGGGATGGAGAAAGAAGTGGAGACAGTCCTTTCCCACC ATTCCTGCCTTTAAGCCAAAGAAACAAGCTGTGCAGGCATGGTCCCTTAAGGCACAGTGGGAGCTGAGCT GGAAGGGGCCACGTGGATGGGCAAAGCTTGTCAAAGATGCCCCCTCCAGGAGAGAGCCAGGATGCCCAGA TGAACTGACTGAAGGAAAAGCAAGAAACAGTTTCTTGCTTGGAAGCCAGGTACAGGAGAGGCAGCATGCT TGGGCTGACCCAGCATCTCCCAGCAAGACCTCATCTGTGGAGCTGCCACAGAGAAGTTTGTAGCCAGGTA CTGCATTCTCTCCCATCCTGGGGCAGCACTCCCCAGAGCTGTGCCAGCAGGGGGGCTGTGCCAACCTGTT CTTAGAGTGTAGCTGTAAGGGCAGTGCCCATGTGTACATTCTGCCTAGAGTGTAGCCTAAAGGGCAGGGC CCACGTGTATAGTATCTGTATATAAGTTGCTGTGTGTCTGTCCTGATTTCTACAACTGGAGTTTTTTTAT ACAATGTTCTTTGTCTCAAAATAAAGCAATGTGTTTTTTCGGACATGCTTTTCTGCCACTCCATATTAAA ACATATGACCATTGAGTCCCTGCTAAAAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO. 8 >ephrin-B2 (EFNB2), mRNA GCGCGGAGCTGGGAGTGGCTTCGCCATGGCTGTGAGTAGGGACTCCGTGTGGAAGTACTGCTGGGGTGTT TTGATGGTTTTATGCAGAACTGCGATTTCCAAATCGATAGTTTTAGAGCCTATCTATTGGAATTCCTCGA ACTCCAAATTTCTACCTGGACAAGGACTGGTACTATACCCACAGATAGGAGACAAATTGGATATTATTTG CCCCAAAGTGGACTCTAAAACTGTTGGCCAGTATGAATATTATAAAGTTTATATGGTTGATAAAGACCAA GCAGACAGATGCACTATTAAGAAGGAAAATACCCCTCTCCTCAACTGTGCCAAACCAGACCAAGATATCA AATTCACCATCAAGTTTCAAGAATTCAGCCCTAACCTCTGGGGTCTAGAATTTCAGAAGAACAAAGATTA TTACATTATATCTACATCAAATGGGTCTTTGGAGGGCCTGGATAACCAGGAGGGAGGGGTGTGCCAGACA AGAGCCATGAAGATCCTCATGAAAGTTGGACAAGATGCAAGTTCTGCTGGATCAACCAGGAATAAAGATC

CAACAAGACGTCCAGAACTAGAAGCTGGTACAAATGGAAGAAGTTCGACAACAAGTCCCTTTGTAAAACC AAATCCAGGTTCTAGCACAGACGGCAACAGCGCCGGACATTCGGGGAACAACATCCTCGGTTCCGAAGTG GCCTTATTTGCAGGGATTGCTTCAGGATGCATCATCTTCATCGTCATCATCATCACGCTGGTGGTCCTCT TGCTGAAGTACCGGAGGAGACACAGGAAGCACTCGCCGCAGCACACGACCACGCTGTCGCTCAGCACACT GGCCACACCCAAGCGCAGCGGCAACAACAACGGCTCAGAGCCCAGTGACATTATCATCCCGCTAAGGACT GCGGACAGCGTCTTCTGCCCTCACTACGAGAAGGTCAGCGGGGACTACGGGCACCCGGTGTACATCGTCC AGGAGATGCCCCCGCAGAGCCCGGCGAACATTTACTACAAGGTCTGAGAGGGACCCTGGTGGTACCTGTG CTTTCCCAGAGGACACCTAATGTCCCGATGCCTCCCTTGAGGGTTTGAGAGCCCGCGTGCTGGAGAATTG ACTGAAGCACAGCACCGGGGGAGAGGGACACTCCTCCTCGGAAGAGCCCGTCGCGCTGGACAGCTTACCT AGTCTTGTAGCATTCGGCCTTGGTGAACACACACGCTCCCTGGAAGCTGGAAGACTGTGCAGAAGACGCC CATTCGGACTGCTGTGCCGCGTCCCACGTCTCCTCCTCGAAGCCATGTGCTGCGGTCACTCAGGCCTCTG CAGAAGCCAAGGGAAGACAGTGGTTTGTGGACGAGAGGGCTGTGAGCATCCTGGCAGGTGCCCCAGGATG CCACGCCTGGAAGGGCCGGCTTCTGCCTGGGGTGCATTTCCCCCGCAGTGCATACCGGACTTGTCACACG GACCTCGGGCTAGTTAAGGTGTGCAAAGATCTCTAGAGTTTAGTCCTTACTGTCTCACTCGTTCTGTTAC CCAGGGCTCTGCAGCACCTCACCTGAGACCTCCACTCCACATCTGCATCACTCATGGAACACTCATGTCT GGAGTCCCCTCCTCCAGCCGCTGGCAACAACAGCTTCAGTCCATGGGTAATCCGTTCATAGAAATTGTGT TTGCTAACAAGGTGCCCTTTAGCCAGATGCTAGGCTGTCTGCGAAGAAGGCTAGGAGTTCATAGAAGGGA GTGGGGCTGGGGAAAGGGCTGGCTGCAATTGCAGCTCACTGCTGCTGCCTCTGAAACAGAAAGTTGGAAA GGAAAAAAGAAAAAAGCAATTAGGTAGCACAGCACTTTGGTTTTGCTGAGATCGAAGAGGCCAGTAGGAG ACACGACAGCACACACAGTGGATTCCAGTGCATGGGGAGGCACTCGCTGTTATCAAATAGCGATGTGCAG GAAGAAAAGCCCCTCTTCATTCCGGGGAACAAAGACGGGTATTGTTGGGAAAGGAACAGGCTTGGAGGGA AGGGAGAAAGTAGGCCGCTGATGATATATTCGGGCAGGACTGTTGTGGTACTGGCAATAAGATACACAGC TCCGAGCTGTAGGAGAGTCGGTCTGCTTTGGATGATTTTTTAAGCAGACTCAGCTGCTATACTTATCACA TTTTATTAAACACAGGGAAAGCATTTAGGAGAATAGCAGAGAGCCAAATCTGACCTAAAAGTTGAAAAGC CAAAGGTCAAACAGGCTGTAATTCCATCATCATCGTTGTTATTAAAGAATCCTTATCTATAAAAGGTAGG TCAGATCCCCCTCCCCCCAGGTTCCTCCTTCCCCTCCCGATTGAGCCTTACGACACTTTGGTTTATGCGG TGCTGTCCGGGTGCCAGGGCTGCAGGGTCGGTACTGATGGAGGCTGCAGCGCCCGGTGCTCTGTGTCAAG GTGAAGCACATACGGCAGACCTCTTAGAGTCCTTAAGACGGAAGTAAATTATGATGTCCAGGGGGAGAAG GAAGATAGGACGTATTTATAATAGGTATATAGAACACAAGGGATATAAAATGAAAGATTTTTACTAATAT ATATTTTAAGGTTGCACACAGTACACACCAGAAGATGTGAAATTCATTTGTGGCAATTAAGTGGTCCCAA TGCTCAGCGCTTAAAAAAACAAATTGGACAGCTACTTCTGGGAAAAACAACATCATTCCAAAAAGAACAA TAATGAGAGCAAATGCAAAAATAACCAAGTCCTCCGAAGGCATCTCACGGAACCGTAGACTAGGAAGTAC GAGCCCCACAGAGCAGGAAGCCGATGTGACTGCATCATATATTTAACAATGACAAGATGTTCCGGCGTTT ATTTCTGCGTTGGGTTTTCCCTTGCCTTATGGGCTGAAGTGTTCTCTAGAATCCAGCAGGTCACACTGGG GGCTTCAGGTGACGATTTAGCTGTGGCTCCCTCCTCCTGTCCTCCCCCGCACCCCCTCCCTTCTGGGAAA CAAGAAGAGTAAACAGGAAACCTACTTTTTATGTGCTATGCAAAATAGACATCTTTAACATAGTCCTGTT ACTATGGTAACACTTTGCTTTCTGAATTGGAAGGGAAAAAAAATGTAGCGACAGCATTTTAAGGTTCTCA GACCTCCAGTGAGTACCTGCAAAAATGAGTTGTCACAGAAATTATGATCCTCTATTTCCTGAACCTGGAA ATGATGTTGGTCCAAAGTGCGTGTGTGTATGTGTGAGTGGGTGCGTGGTATACATGTGTACATATATGTA TAATATATATCTACAATATATATTATATATATCTATATCATATTTCTGTGGAGGGTTGCCATGGTAACCA GCCACAGTACATATGTAATTCTTTCCATCACCCCAACCTCTCCTTTCTGTGCATTCATGCAAGAGTTTCT TGTAAGCCATCAGAAGTTACTTTTAGGATGGGGGAGAGGGGCGAGAAGGGGAAAAATGGGAAATAGTCTG ATTTTAATGAAATCAAATCTATGTATCATCAGTTGGCTACGTTTTGGTTCTATGCTAAACTGTGAAAAAT CAGATGAATTGATAAAAGAGTTCCCTGCAACCAATTGAAAAGTGTTCTGTGCGTCTGTTTTGTGTCTGGT GCAGAATATGACAATCTACCAACTGTCCCTTTGTTTGAAGTTGGTTTAGCTTTGGAAAGTTACTGTAAAT GCCTTGCTTGTATGATCGTCCCTGGTCACCCGACTTTGGAATTTGCACCATCATGTTTCAGTGAAGATGC TGTAAATAGGTTCAGATTTTACTGTCTATGGATTTGGGGTGTTACAGTAGCCTTATTCACCTTTTTAATA AAAATACACATGAAAACAAGAAAGAAATGGCTTTTCTTACCCAGATTGTGTACATAGAGCAATGTTGGTT TTTTATAAAGTCTAAGCAAGATGTTTTGTATAAAATCTGAATTTTGCAATGTATTTAGCTACAGCTTGTT TAACGGCAGTGTCATTCCCCTTTGCACTGTAATGAGGAAAAAATGGTATAAAAGGTTGCCAAATTGCTGC ATATTTGTGCCGTAATTATGTACCATGAATATTTATTTAAAATTTCGTTGTCCAATTTGTAAGTAACACA GTATTATGCCTGAGTTATAAATATTTTTTTCTTTCTTTGTTTTATTTTAATAGCCTGTCATAGGTTTTAA ATCTGCTTTAGTTTCACATTGCAGTTAGCCCCAGAAAATGAAATCCGTGAAGTCACATTCCACATCTGTT TCAAACTGAATTTGTTCTTAAAAAAATAAAATATTTTTTTCCTATGGAAAAAAAAAAAAAAAAAA

[0097] Detection of NEPOR component mRNA (SEQ ID NOs 5-8) should preferentially be performed using probes complementary to the sub-region of SEQ ID NO's 5-8, encoding the EPO binding domain and is particular SEQ Id NO. 6 and/or 7 encoding EPH-B4 and Ephrin A1. This implies for EPH-B4, probes complementary to SEQ ID NO. 10.; for Ephrin A1, probes complementary to SEQ ID NO. 11.

TABLE-US-00007 SEQ ID NO. 9 >epor_epobinding_coding region AGCAAAGCGGCCTTGCTGGCGGCCCGGGGGCCCGAAGAGCTTCTGTGCTTCACCGAGCGGTTGGAGGACTTGGT- GTG TTTCTGGGAGGAAGCGGCGAGCGCTGGGGTGGGCCCGGGCAACTACAGCTTCTCCTACCAGCTCGAGGATGAGC- CAT GGAAGCTGTGTCGCCTGCACCAGGCTCCCACGGCTCGTGGTGCGGTGCGCTTCTGGTGTTCGCTGCCTACAGCC- GAC ACGTCGAGCTTCGTGCCCCTAGAGTTGCGCGTCACAGCAGCCTCCGGCGCTCCGCGATATCACCGTGTCATCCA- CAT CAATGAAGTAGTGCTCCTAGACGCCCCCGTGGGGCTGGTGGCGCGGTTGGCTGACGAGAGCGGCCACGTAGTGT- TGC GCTGGCTCCCGCCGCCTGAGACACCCATGACGTCTCACATCCGCTACGAGGTGGACGTCTCGGCCGGCAACGGC- GCA GGGAGCGTACAGAGGGTGGAGATCCTGGAGGGCCGCACCGAGTGTGTGCTGAGCAACCTGCGGGGCCGGACGCG- CTA CACCTTCGCCGTCCGCGCGCGTATGGCTGAGCCGAGCTTCGGCGGCTTCTGGAGCGCCTGGTCGGAGCCTGTGT- CGC TGCTGACGCCTAGCGACCTGGACCCC SEQ ID NO. 10 >ephb4_epobinding_coding region CCTTCGGCTCCGCGGAGCGTGGTTTCCCGCCTGAACGGCTCCTCCCTGCACCTGGAATGGAGTGCCCCCCTGGA- GTC TGGTGGCCGAGAGGACCTCACCTACGCCCTCCGCTGCCGGGAGTGCCGACCCGGAGGCTCCTGTGCGCCCTGC GGGGGAGACCTGACTTTTGACCCCGGCCCCCGGGACCTGGTGGAGCCCTGGGTGGTGGTTCGAGGGCTACGTCC- TGA CTTCACCTATACCTTTGAGGTCACTGCATTGAACGGGGTATCCTCCTTAGCCACGGGGCCCGTCCCATTTGAGC- CTG TCAATGTCACCACTGACCGAGAGGTACCTCCTGCAGTGTCTGACATCCGGGTGACGCGGTCCTCACCCAGCAGC- TTG AGCCTGGCCTGGGCTGTTCCCCGGGCACCCAGTGGGGCTGTGCTGGACTACGAGGTCAAATACCATGAGAAGGG- CGC CGAGGGTCCCAGCAGCGTGCGGTTCCTGAAGACGTCAGAAAACCGGGCAGAGCTGCGGGGGCTGAAGCGGGGAG- CCA GCTACCTGGTGCAGGTACGGGCGCGCTCTGAGGCCGGCTACGGGCCCTTCGGCCAGGAACATCACAGCCAGACC- CAA CTGGATGAGAGCGAGGGCTGGCGGGAGCAGCTGGCCCTG SEQ ID NO. 11 >ephrinA1_epobinding_coding region CTGGCCGCTGCTGATCGCCACACCGTCTTCTGGAACAGTTCAAATCCCAAGTTCCGGAATGAGGACTACACCAT- ACA TGTGCAGCTGAATGACTACGTGGACATCATCTGTCCGCACTATGAAGATCACTCTGTGGCAGACGCTGCCATGG- AGC AGTACATACTGTACCTGGTGGAGCATGAGGAGTACCAGCTGTGCCAGCCCCAGTCCAAGGACCAAGTCCGCTGG- CAG TGCAACCGGCCCAGTGCCAAGCATGGCCCGGAGAAGCTGTCTGAGAAGTTCCAGCGCTTCACACCTTTCACCCT- GGG CAAGGAGTTCAAAGAAGGACACAGCTACTACTACATCTCCAAACCCATCCACCAGCATGAAGACCGCTGCTTGA- GGT TGAAGGTGACTGTCAGTGGCAAAATCACTCAC SEQ ID NO. 12 >ephrinb2_epobinding_coding region TCCAAATCGATAGTTTTAGAGCCTATCTATTGGAATTCCTCGAACTCCAAATTTCTACCTGGACAAGGACTGGT- ACT ATACCCACAGATAGGAGACAAATTGGATATTATTTGCCCCAAAGTGGACTCTAAAACTGTTGGCCAGTATGAAT- ATT ATAAAGTTTATATGGTTGATAAAGACCAAGCAGACAGATGCACTATTAAGAAGGAAAATACCCCTCTCCTCAAC- TGT GCCAAACCAGACCAAGATATCAAATTCACCATCAAGTTTCAAGAATTCAGCCCTAACCTCTGGGGTCTAGAATT- TCA GAAGAACAAAGATTATTACATTATATCTACATCAAATGGGTCTTTGGAGGGCCTGGATAACCAGGAGGGAGGGG- TGT GCCAGACAAGAGCCATGAAGATCCTCATGAAAGTTGGACAA

[0098] The determination of the presence of the Ephrin A1 and/or the determination of the presence of the EPH-B4 gene product (mRNA) may be done by using a hybridization technique or an amplification technique. It is preferred that the technique is selected from the group of, real-time-PCR, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), capillary electrophoresis with laser induced fluorescence (CE-LIF) and combinations thereof.

[0099] Specifically, the individual is a cancer patient who is to be treated with erythropoietin or is being treated with erythropoietin. Preferably, the negative physiological effect is poorer patient survival due to enhanced tumor progression. Preferably, the presence of a higher level of NEPOR component genes (mRNA) and/or the presence of NEPOR component gene expression products (proteins) and/or EPH-B4 and/or Ephrin A1 on tumor tissues is indicative of poorer survival prognosis upon treatment with erythropoietin.

[0100] Preferably, the determination of the presence of the NEPOR dimer complex is done by detecting the respective NEPOR proteins with an immunoassay. Also peptides thereof may be detected. The immunoassay is selected from the group of immunoprecipitation, a protein array or binding to a mass microbalance instrument (for example, Q-Sense or Attana), enzyme immunoassay (EIA), radioimmunoassay (RIA) or fluorescent immunoassay, a chemiluminescent assay, an agglutination assay, nephelometric assay, turbidimetric assay, a Western blot, a competitive immunoassay, a noncompetitive immunoassay, a homogeneous immunoassay a heterogeneous immunoassay, a bioassay and a reporter-assay such as a luciferase-assay. Preferably, the immunoassay is an ELISA. Preferably, the anti-NEPOR antibody and/or EPH-B4 and/or Ephrin A1 antibody is a monoclonal or polyclonal antibody, for example selected from--or similar to--the antibodies listed in Table 6.

[0101] Preferably, detection of NEPOR component proteins should preferentially be performed using antibodies detecting the sub-regions of SEQ ID NOs 6 and 7, representing the EPO binding domain. This implies for EPH-B4, antibodies specific to SEQ ID NO. 14.; for Ephrin A1, antibodies specific to SEQ ID NO. 15.

TABLE-US-00008 SEQ ID NO. 13 >epor_epobinding_region SKAALLAARGPEELLCFTERLEDLVCFWEEAASAGVGPGNYSFSYQLEDEPWKLCRLHQAPTARGAVRFWCSLP- TAD TSSFVPLELRVTAASGAPRYHRVIHINEVVLLDAPVGLVARLADESGHVVLRWLPPPETPMTSHIRYEVDVSAG- NGA GSVQRVEILEGRTECVLSNLRGRTRYTFAVRARMAEPSFGGFWSAWSEPVSLLTPSDLDP SEQ ID NO. 14 >ephb4_epobinding_region PSAPRSVVSRLNGSSLHLEWSAPLESGGREDLTYALRCRECRPGGSCAPCGGDLTFDPGPRDLVEPWVVVRGLR- PDF TYTFEVTALNGVSSLATGPVPFEPVNVTTDREVPPAVSDIRVTRSSPSSLSLAWAVPRAPSGAVLDYEVKYHEK- GAE GPSSVRFLKTSENRAELRGLKRGASYLVQVRARSEAGYGPFGQEHHSQTQLDESEGWREQLAL SEQ ID NO. 15 >ephrinA1_epobinding_region LAAADRHTVFWNSSNPKFRNEDYTIHVQLNDYVDIICPHYEDHSVADAAMEQYILYLVEHEEYQLCQPQSKDQV- RWQ CNRPSAKHGPEKLSEKFQRFTPFTLGKEFKEGHSYYYISKPIHQHEDRCLRLKVTVSGKITH SEQ ID NO. 16 >ephrinb2_epobinding_region SKSIVLEPIYWNSSNSKFLPGQGLVLYPQIGDKLDIICPKVDSKTVGQYEYYKVYMVDKDQADRCTIKKENTPL- LNC AKPDQDIKFTIKFQEFSPNLWGLEFQKNKDYYIISTSNGSLEGLDNQEGGVCQTRAMKILMKVGQ

[0102] Preferably, the individual is a cancer patient who is to be treated with erythropoietin or is being treated with erythropoietin. The tissue sample may be selected from the group of biological tissues and fluids such as blood, lymph, urine, cerebral fluid. The tissue sample may also be a tumor biopsy sample. It is preferred that the tissue sample is from the cancer tissue or circulating cells derived from same.

[0103] It is preferred that the cancer of the cancer patient is selected from the group of, head and neck cancer, breast cancer, liver cancer, colorectal cancer, small intestine cancer, leukemia, prostate cancer, lung cancer, ovarian cancer, pancreatic cancer, endometrial cancer, stomach cancer, non-Hodgkin lymphoma, kidney cancer, Renal cell carcinoma (RCC), malignant melanoma, gallbladder cancer, bladder cancer, vulvar cancer, Penile cancer, testicular cancer, thymus cancer, Kaposi's sarcoma, eye cancer, adrenal gland cancer, brain cancer, cervical cancer, appendix cancer, adenoid cancer, bile duct cancer, urethral cancer, spinal cancer, Ewing's family of tumors, extragonal germ cell cancer, extra hepatic bile duct cancer, fallopian tube cancer, soft tissue cancers, bone cancer, Hodgkin's lymphoma, anal cancer, malignant mesothelioma, vaginal cancer skin cancer, central nervous system cancer (craniopharyngioma), pleuropulmonary blastoma, nasal cavity and paranasal sinus cancer transitional cell cancer of renal pelvis and ureter, pituitary gland cancer, squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, brain cancer, bladder cancer, and salivary gland cancer. It is particularly preferred that the cancer is selected from the group of squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, kidney cancer, brain cancer and bladder cancer.

NEPOR and Disease Intervention and Therapy Design/Screening.

[0104] Without being bound by theory, NEPOR is proposed to mediate the cyto-protective effects of EPO and its variants. Thus, EPO and variants that have been shown to possess cyto-protective (but not haematopoietic) activity can affect NEPOR function. Therefore, the present disclosure provides knowledge of NEPOR's composition that can be used to optimize the structure and efficacy of such therapeutic molecules (that is, better manage the structure-activity relationship or SAR of the EPO pharmacophore). Moreover, the present disclosure provides knowledge of NEPOR's composition that can be used to identify novel NEPOR regulating compounds. For example, in diseases associated with hypoxic conditions (e.g., stroke, heart attack), NEPOR binding compounds of enhanced efficacy can be developed to mimic the effects of EPO on NEPOR. Similarly, NEPOR specific antagonists (such as those molecules that bind the active site of NEPOR yet do not transducer signal are antagonists of EPO function. Such EPO antagonist agents, when concomitantly administered with EPO, can allow for EPO effects to improve haematopoiesis (that is, treat the anaemia) yet prevent the side effect of promoting tumour cell growth, survival and angiogenesis in NEPOR positive cancers such as HNSCC. Moreover, contrasting the relative activity of compounds to the tissue protective NEPOR receptor complex in comparison to the EPOR receptor homodimer provides for generating NEPOR specific/directed therapies.

[0105] Definition of NEPOR provides methods for identifying therapeutic molecules that modulate NEPOR's tissue protective signalling activity. This comprises: (a) contacting a test compound with the NEPOR receptor complex and/or EPH-B4 and/or Ephrin A1 and an EPOR homodimer complex; (b) measuring and comparing the level of tissue protective activity initiated by NEPOR activation with the activation of EPOR homodimer signalling; (c) identifying a test compound which increases or decreases the level of tissue protective NEPOR complex activity as compared to the level of EPOR complex activation; and (d) assaying the identified therapeutics for tissue protective activity mediated via NEPOR, but lack of EPOR activation and (e) assaying the identified therapeutics for NEPOR inhibitory activity. The method is useful for identifying therapeutics that modulates the interaction between a tissue protective NEPOR complex and/or EPH-B4 and/or Ephrin A1 and the EPO ligand. The method is furthermore useful for identifying therapies for treating diseases of the central nervous system or peripheral nervous system which have primarily neurological or psychiatric symptoms, ophthalmic diseases, cardiovascular diseases, cardiopulmonary diseases, respiratory diseases, kidney, urinary and reproductive diseases, bone diseases, skin diseases, gastrointestinal diseases and endocrine and metabolic abnormalities and cancer.

[0106] More specifically, identification of NEPOR provides a method identifying (I1) a compound that modulates the tissue protective activity of NEPOR, comprising: [0107] (a) contacting a test compound with a tissue protective NEPOR receptor complex (N) and/or EPH-B4 and/or Ephrin A1 or tissue protective cytokine receptor complex-expressing cell; measuring the level of the activity of (N) in the cell; identifying a test compound that increases or decreases the level of activity of (N) as compared to the level of activity of (N) measured in the absence of the test compound; and assaying the identified test compound for tissue protective activity; [0108] (b) contacting a test compound with a cell that is recombinantly engineered to express (N), where the cell or the recombinant cell is transformed with a nucleic acid comprising a nucleotide sequence that is functionally linked to a promoter and encodes EPH-B4 and/or Ephrin A1 polypeptides; measuring the level of activity of (N) in the cell; and [0109] (c) contacting a test compound with a tissue protective NEPOR receptor complex-expressing cell, where the cell is transformed with a nucleic acid comprising a nucleotide sequence that encodes a reporter gene functionally linked to regulatory element associated with the activity of (N); identifying a test compound that increases or decreases the level of reporter gene expression relative to the level of reporter gene expression measured in the absence of the test compound; and assaying the identified test compound for a tissue protective activity.

[0110] The present disclosure further provides a method for identifying (I2) a compound that binds to (N), comprising: [0111] (a) contacting (N) with a tissue protective NEPOR receptor complex ligand and/or EPH-B4 and/or Ephrin A1 ligand attached to a first label, and an equivalent amount of a test compound attached to a second label under conditions conducive to binding, removing unbound material from (N), and detecting the level of the first and second labels, where if the second label is present the compound binds (N) and if the level of the first label decreases relative to the level of the first label when the labelled ligand is contacted with (N) under conditions conducive to binding in the absence of a test compound after removal of unbound material, then a compound that binds to (N) is identified; or [0112] (b) contacting a test compound with a ligand-binding tissue protective receptor NEPOR complex fragment comprising at least one EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to a Fc fragment attached to a solid support, removing unbound test compounds from the solid support, and identifying the compound attached to the tissue protective NEPOR receptor complex fragment, such that a compound bound to the solid support is identified as a compound that binds to a tissue protective NEPOR receptor complex; and identifying (I3) a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to (N), or compound that modulates the interaction between (N) and tissue protective cytokine receptor complex ligand, involves (i) contacting a tissue protective NEPOR receptor complex ligand with (N) in the presence of one or more test compounds under conditions conducive to binding, and measuring the amount of tissue protective cytokine receptor complex ligand bound to (N).

[0113] The present disclosure further provides novel tissue protective NEPOR receptor complexes in particular EPH-B4 and/or Ephrin A1 containing complexes that can be used to provide an in vitro screening assay for NEPOR specific therapies; by measuring the binding of test compounds to the tissue protective NEPOR receptor complex in comparison to EPOR homodimer complexes. The test compound is labelled and binding of the labelled test compound to the receptor complexes detailed in FIG. 10 is measured by detecting the label attached to the test compound. Alternatively, a label free detection approach such as surface plasmon resonance may be employed. Such an approach can provide for novel neuroprotective therapies (i.e. NEPOR agonists) which lack haematopoietic activity. Such an approach can also provide for novel onco-therapies (i.e. NEPOR antagonists i.e. at least a and/or EPH-B4 and/or Ephrin A1 agonist) which do not inhibit haematopoiesis. The nature of such screening arrays involving recombinant receptor constructs is demonstrated in FIG. 10 (in the exemplified case as Fc constructs).

Use

[0114] (I1) is useful for identifying a compound that modulates NEPOR's tissue protective activity. (I2) is useful for identifying a compound that binds to NEPOR. (I3) is useful for identifying a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to (N), or compound that modulates the interaction between (N) and tissue protective cytokine receptor complex ligand (claimed). The compounds identified using (I1)-(I3) are useful for treating various conditions of the central and peripheral nervous systems (e.g., hypoxia, and/or ischemia, epilepsy, chronic seizure disorders, neurotoxin poisoning, septic shock, anaphylactic shock), neuropsychologic disorders (senile dementia, Alzheimer's disease, Parkinson's disease, dermentia, multiple sclerosis, Creutzfeldt-Jakob disease, Huntington's disease), inflammatory diseases (e.g., chronic bronchitis, rheumatoid arthritis, glomerulonephritis, encephalitis, meningitis, polymyositis), opthalamic diseases (e.g., angiitis, retinal ischemia), cardiovascular diseases (e.g., myocardial infraction, myocarditis), cardiopulmonary diseases (e.g., asthma, pulmonary thrombosis), respiratory diseases, kidney, urinary, and reproductive diseases (e.g., myasthenia gravis, diabetes, autoimmune diseases), bone diseases (e.g., osteopenia, Paget's disease), gastrointestinal diseases and endocrine and metabolic abnormalities.

[0115] The compounds identified using (I1)-(I3) are also useful for treating NEPOR positive cancers in particular and/or EPH-B4 and/or Ephrin A1 positive cancers including, head and neck cancer, breast cancer, liver cancer, colorectal cancer, small intestine cancer, leukemia, prostate cancer, lung cancer, ovarian cancer, pancreatic cancer, endometrial cancer, stomach cancer, non-Hodgkin lymphoma, kidney cancer, Renal cell carcinoma (RCC), malignant melanoma, gallbladder cancer, bladder cancer, vulvar cancer, Penile cancer, testicular cancer, thymus cancer, Kaposi's sarcoma, eye cancer, adrenal gland cancer, brain cancer, cervical cancer, appendix cancer, adenoid cancer, bile duct cancer, urethral cancer, spinal cancer, Ewing's family of tumors, extragonal germ cell cancer, extra hepatic bile duct cancer, fallopian tube cancer, soft tissue cancers, bone cancer, Hodgkin's lymphoma, anal cancer, malignant mesothelioma, vaginal cancer skin cancer, central nervous system cancer (craniopharyngioma), pleuropulmonary blastoma, nasal cavity and paranasal sinus cancer transitional cell cancer of renal pelvis and ureter, pituitary gland cancer, squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, brain cancer, bladder cancer, and salivary gland cancer. It is particularly preferred that the cancer is selected from the group of squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, kidney cancer, brain cancer and bladder cancer.

NEPOR in Oncology Therapy

[0116] The hypothesis of the present disclosure is that EPO results in poorer survival outcomes (at least in some cancers) because of its effects on NEPOR activity i.e. in particular EPH-B4 and/or Ephrin A1 activity. Therefore, treatment of these NEPOR positive patients with a NEPOR targeted therapy is a prudent path to disease intervention. Specific approaches to antagonising NEPOR mediated survival signals include, for example: [0117] a) NEPOR specific antagonistic antibodies. Such antibodies block and antagonise the extracellular regions of the molecule specifically associated with the mediation of NEPOR's cyto-protective activity. [0118] b) NEPOR specific small-molecules. Such small molecules block and antagonise the extracellular regions of the molecule specifically associated with the mediation of NEPOR's cytoprotective activity. [0119] c) high-affinity peptides which specifically target NEPOR to block and antagonise the is mediation of EPO's cytoprotective activity. [0120] d) Small molecules targeting EPH-B4's intracellular tyrosine kinase domain (e.g. Dasatinib), including: 1: CID: 1095868, AKI-STT-00166305; ZINC00818264; BAS 09636496 IUPAC: N-[5-[(3-chlorophenyl)methyl]-1,3-thiazol-2-yl]-2-(4,6-dimethylpyr- imidin-2-yl)sulfanylacetamide. MW: 404.93678|MF: C18H17ClN4OS2. (MW is molecular weight and MF is molecular formula) 2: CID: 1465558, IUPAC: 2-[(3-chlorobenzoyl)amino]-4-methyl-N-pyridin-3-yl-1,3-thiazole-5-carboxa- mide, MW: 372.82872|MF: C17H13ClN4O2S. 3: CID: 1468201, IUPAC: N-[5-[(2-chlorophenyl)carbamoyl]-4-methyl-1,3-thiazol-2-yl]pyridine-4-car- boxamide, MW: 372.82872|MF: C17H13ClN4O2S. 4: CID: 3062316, Dasatinib; Sprycel; BMS Dasatinib, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 5: CID: 3072360, 142287-40-9; Pyrimido(4,5-d)pyrimidin-4(1H)-one, 7-methyl-1-phenyl-2-((3-(4-(2-thiazolyl)-1-piperazinyl)propyl)thio)-IUPAC- : 2-methyl-8-phenyl-7-[3-[4-(1,3-thiazol-2-yl)piperazin-1-yl]propylsulfany- l]pyrimido[6,5-d]pyrimidin-5-one, MW: 479.6209|MF: C23H25N7OS2. 6: CID: 5041467, STK154706; ZINC04687922, IUPAC: [2-[(2-methylphenyl)amino]-1,3-thiazol-4-yl]-(4-pyrimidin-2-ylpiperazin-1- -yl)methanone, MW: 380.4667|MF: C19H2ON6OS. 7: CID: 9822929, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-imidazol-1-ylpyridazin-3-yl)amino]-1,3-- thiazole-5-carboxamide, MW: 411.869|MF: C18H14ClN7OS. 8: CID: 9927718, IUPAC: N-(2-chloro-6-methylphenyl)-2-(cyclopropanecarbonylamino)-1,3-thia- zole-5-carboxamide, MW: 335.809|MF: C15H14ClN3O2S. 9: CID: 10006113, IUPAC: N-[4-chloro-2-[(5-chloropyridin-2-yl)carbamoyl]phenyl]-5-methyl-6,- 7-dihydro-4H-[1,3]thiazolo[5,4-c]pyridine-2-carboxamide hydrochloride, MW: 498.81322|MF: C20H18C13N5O2S. 10: CID: 10006114, IUPAC: N-[4-chloro-2-[(5-chloropyridin-2-yl)carbamoyl]phenyl]-5-methyl-6,7-dihyd- ro-4H-[1,3]thiazolo[5,4-c]pyridine-2-carboxamide, MW: 462.35228|MF: C20H17Cl2N5O2S. 11: CID: 10052635, IUPAC: 2-[[2-methyl-5-[[6-[(4-methylpiperazin-1-yl)methyl]pyridin-2-yl]amino]phe- nyl]amino]-N-(2-methylphenyl)-1,3-thiazole-5-carboxamide, MW: 527.68362|MF: C29H33N7OS. 12: CID: 10195898, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[[(2S)-2-hydroxy-2-pyrimidin-2-ylethyl]-met- hylamino]methyl]-4-methyl-7-oxothieno[2,3-e]pyridine-6-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 13: CID: 10206276, IUPAC: N-[4-[(5-chloropyridin-2-yl)carbamoyl]-2-phenyl-1,3-thiazol-5-yl]-1-propa- n-2-ylpiperidine-4-carboxamide, MW: 484.01354|MF: C24H26ClN5O2S. 14: CID: 10252208, IUPAC: 2-[4-(5-amino-1,3-thiazol-2-yl)phenyl]-3-(5-chloropyridin-2-yl)quinazolin- -4-one, MW: 431.89746|MF: C22H14ClN5OS. 15: CID: 10253695, IUPAC: 2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazole-- 5-carboxamide, MW: 459.90756|MF: C23H14ClN5O2S. 16: CID: 10301604, IUPAC: N-[4-[(5-chloropyridin-2-yl)carbamoyl]-2-(3,4-difluorophenyl)-1,3-thiazol- -5-yl]-1-propan-2-ylpiperidine-4-carboxamide, MW: 519.994466|MF: C24H24ClF2N5O2S. 17: CID: 10344807, IUPAC: N-[2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazo- l-4-yl]acetamide, MW: 473.93414|MF: C24H16ClN5O2S. 18: CID: 10368624, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[(2-hydroxy-2-pyrimidin-2-ylethyl)-m- ethylamino]methyl]-7-methyl-4-oxothieno[3,2-e]pyridine-5-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 19: CID: 10370949, IUPAC: (3Z)-4-[[(2S)-2-(3-chlorophenyl)-2-hydroxyethyl]amino]-3-[6-methyl-2-[4-(- 1,3-thiazol-2-ylmethyl)piperazin-1-yl]-7,9-dihydropurin-8-ylidene]pyridin-- 2-one, MW: 578.08832|MF: C27H28ClN9O2S. 20: CID: 10412586, IUPAC: N-[2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazo- l-5-yl]acetamide, MW: 473.93414|MF: C24H16ClN5O2S. 21: CID: 10413555, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[[(2R)-2-hydroxy-2-pyrimidin-2-yleth- yl]-methylamino]methyl]-7-methyl-4-oxothieno[3,2-e]pyridine-5-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 22: CID: 10456156, IUPAC: 4-[(3-chlorothiophen-2-yl)methylamino]-2-[(4-morpholin-4-ylphenyl)amino]p- yrimidine-5-carboxamide, MW: 444.93774|MF: C20H21ClN6O2S. 23: CID: 10458706, IUPAC: N-[5-[2-[(4-chlorophenyl)amino]pyrimidin-4-yl]-4-methyl-1,3-thiazol-2-yl]- -3-(2-morpholin-4-ylethylamino)propanamide, MW: 502.03212|MF: C23H28ClN7O2S. 24: CID: 11153014, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2,6-dimethylpyrimidin-4-yl)amino]-1,3-thi- azole-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 25: CID: 11167695, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-(2-morpholin-4-ylethyla- mino)pyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 26: CID: 11168231, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-N-- [(4-methoxyphenyl)methyl]-1,3-thiazole-5-carboxamide, MW: 514.42684|MF: C24H21Cl2N5O2S. 27: CID: 11200510, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-hydroxyethylamino)pyridin-2-yl]amino- ]-1,3-thiazole-5-carboxamide, MW: 403.88582|MF: C18H18ClN5O2S. 28: CID: 11247793, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(methyl-(3-methylaminopropyl)amino)pyri- din-2-yl]amino]-1,3-thiazole-5-carboxamide, MW: 444.9808|MF: C21H25ClN6OS. 29: CID: 11260009, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(hydroxymethyl)piperidin-1-yl]-2-met- hylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 472.9909|MF: C22H25ClN6O2S. 30: CID: 11269410, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-1,- 3-thiazole-5-carboxamide, MW: 394.27832|MF: C16H13Cl2N5OS. 31: CID: 11282881, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-morpholin-4-ylethylamino)pyrimidin-4- -yl]amino]-1,3-thiazole-5-carboxamide, MW: 473.97896|MF: C21H24ClN7O2S 32: CID: 11283174, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(3-morpholin-4-ylpropylamino)pyridin-2-- yl]amino]-1,3-thiazole-5-carboxamide, MW: 487.01748|MF: C23H27ClN6O2S. 33: CID: 11328827, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(3-imidazol-1-ylpropylamino)pyridin-2-y- l]amino]-1,3-thiazole-5-carboxamide, MW: 467.97438|MF: C22H22ClN7OS. 34: CID: 11407465, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin- -4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 418.90046|MF: C18H19ClN6O2S. 35: CID: 11466196, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-(3-morpholin-4-ylpropylamino)p- yrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide. MW: 502.03212|MF: C23H28ClN7O2S. 36: CID: 11466607, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrochloride, MW: 524.46648|MF: C22H27Cl2N7O2S. 37: CID: 11487256, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-morpholin-4-ylpyrimidin-4-yl)amino]-1,3- -thiazole-5-carboxamide, MW: 430.91116|MF: C19H19ClN6O2S. 38: CID: 11505502, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]pyrimidin-4-yl]amino]-N-[2-methyl- -5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide. MW: 626.65257|MF: C29H29F3N8O3S. 39: CID: 11512538, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]ethyl 2,2-dimethylpropanoate, MW: 572.12196|MF: C27H34ClN7O3S. 40: CID: 11539665, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-[(5-fluoro-1,3-thiazol-2-yl)amino]pyridi- n-2-yl]methyl]piperazin-1-yl]methanone, MW: 449.904626|MF: C20H18ClF2N5OS. 41: CID: 11540687, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrate, MW: 506.02082 MF: C22H28ClN7O3S. 42: CID: 11569328, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[3-[4-(2-hydroxyethyl)piperazin-1-yl]-5-me- thylphenyl]amino]-1,3-thiazole-5-carboxamide, MW: 486.02942|MF: C24H28ClN5O2S. 43: CID: 11570976, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-N-- [2-methyl-5-[[3-(trifluoromethyl)phenyl]carbamoyl]phenyl]-1,3-thiazole-5-c- arboxamide, MW: 640.67915|MF: C30H31F3N8O3S. 44: CID: 11577776, IUPAC: 2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin-4-yl]amino]-N-[2-methyl-5-[- [3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 571.57407|MF: C26H24F3N7O3S. 45: CID: 11590089, IUPAC: (3-chloro-2-fluorophenyl)-[4-[5-methyl-6-(1,3-thiazol-2-ylamino)pyridin-2- -yl]piperazin-1-yl]methanone, MW: 431.914163|MF: C20H19ClFN5OS. 46: CID: 11606973, IUPAC: N-[5-[[3-[4-(2-hydroxyethyl)piperazin-1-yl]-5-(trifluoromethyl)benzoyl]am- ino]-2-methylphenyl]-2-(pyridin-2-ylamino)-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 47: CID: 11650098, IUPAC: 2-[[6-(4-methylpiperazin-1-yl)pyrimidin-4-yl]amino]-N-[2-methyl-5-[[3-(tr- ifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 596.62659|MF: C28H27F3N8O2S. 48: CID: 11650132, IUPAC: pentyl N-[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]-N-[6-[4-(2-hy- droxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]carbamate, MW: 602.14794|MF: C28H36ClN7O4S. 49: CID: 11650511, IUPAC: N-[5-[[3-(4-ethylpiperazin-1-yl)-5-(trifluoromethyl)benzoyl]amino]-2-meth- ylphenyl]-2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin-4-yl]amino]-1,3-th- iazole-5-carboxamide, MW: 683.74695|MF: C32H36F3N9O3S. 50: CID: 11664355, IUPAC: 2-[(2-methyl-6-morpholin-4-ylpyrimidin-4-yl)amino]-N-[2-methyl-5-[- [3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 597.61135|MF: C28H26F3N7O3S. 51: CID: 11664511, IUPAC: 2-[[4-[4-(2-hydroxyethyl)piperazin-1-yl]pyridin-2-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 52: CID: 11669430, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2-methyl-6-piperazin-1-ylpyrimidin-4-yl)a- mino]-1,3-thiazole-5-carboxamide, MW: 443.95298|MF: C20H22ClN7OS. 53: CID: 11676373, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-(1,3-thiazol-2-ylamino)pyridin-2-yl]meth- yl]piperazin-1-yl]methanone, MW: 431.914163|MF: C20H19ClFN5OS. 54: CID: 11684148, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-[(5-chloro-1,3-thiazol-2-yl)amino]pyridi- n-2-yl]methyl]piperazin-1-yl]methanone, MW: 466.359223|MF: C20H18Cl2FN5OS. 55: CID: 11700117, IUPAC: 2-[[6-(4-ethylpiperazin-1-yl)-2-methylpyrimidin-4-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 624.67975|MF: C30H31F3N8O2S. 56: CID: 11707091, IUPAC: 2-[[2-methyl-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl]amino]-N-[2-methyl-- 5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 610.65317|MF: C29H29F3N8O2S. 57: CID: 11714286, IUPAC: 2-[[5-[4-(2-hydroxyethyl)piperazin-1-yl]pyridin-2-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 58: CID: 11714353, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-N-- [2-methyl-5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carb- oxamide, MW: 640.67915|MF: C30H31F3N8O3S. 59: CID: 11752136, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[5-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 60: CID: 11772766, IUPAC: 4-[2-(3-chlorophenyl)ethylamino]-2-pyridin-4-yl-1,3-thiazole-5-carboxamid- e, MW: 358.8452|MF: C17H15ClN4OS. 61: CID: 11775143, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2-methyl-6-morpholin-4-ylpyrimidin-4-yl)a- mino]-1,3-thiazole-5-carboxamide, MW: 444.93774|MF: C20H21ClN6O2S. 62: CID: 11854012, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]acetic acid, MW: 501.98906|MF: C22H24ClN7O3S. 63: CID: 11854269, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]ethyl hydrogen sulfate, MW: 568.06874|MF: C22H26ClN7O5S2. 64: CID: 11854270, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[2-(2-hydroxyethylamino)ethyl amino]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 461.96826|MF: C20H24ClN7O2S 65: CID: 11854271, IUPAC: 2-[[6-(2-aminoethylamino)-2-methylpyrimidin-4-yl]amino]-N-(2-chloro-6-met- hylphenyl)-1,3-thiazole-5-carboxamide, MW: 417.9157|MF: C18H20ClN7OS. 66: CID: 11854272, IUPAC: 2-[[2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]ami- no]-2-methylpyrimidin-4-yl]piperazin-1-yl]acetyl]amino]ethanesulfonic acid, MW: 609.12066|MF: C24H29ClN8O5S2. 67: CID: 11854533, IUPAC: N-(2-chloro-4-hydroxy-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-- 1-yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 68: CID: 11854534, IUPAC: N-[2-chloro-6-(hydroxymethyl)phenyl]-2-[[6-[4-(2-hydroxyethyl)piperazin-1- -yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 69: CID: 11854535, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-4-oxidopiperazin-4-- ium-1-yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 70: CID: 11854536, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]-1-oxidopiperazin-1-ium-1-yl]acetic acid, MW: 517.98846|MF: C22H24ClN7O4S. 71: CID: 11949914, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-dimethylaminopyrimidin-4-yl]amino]-N-[2- -(2-hydroxyethylamino)ethyl]pyridine-3-carboxamide, MW: 473.931003|MF: C22H25ClFN7O2. 72: CID: 11951866, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-pyrrolidin-1-ylpyrimidin-4-yl]amino]-N-- (2-hydroxyethyl)pyridine-3-carboxamide, MW: 456.900483|MF: C22H22ClFN6O2. 73: CID: 11952045, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-pyrrolidin-1-ylpyrimidin-4-yl]amino]-N-- [(2S)-2-hydroxypropyl]pyridine-3-carboxamide, MW: 470.927063|MF: C23H24ClFN6O2. 74: CID: 15979866, IUPAC: 5-[2-[[4-(4-acetylpiperazin-1-yl)pyridin-2-yl]amino]-1,3-thiazol-5-yl]-N-- methylpyridine-3-carboxamide, MW: 437.51802|MF: C21H23N7O2S. 75: CID: 15980109, IUPAC: N-(2-aminoethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thiazol-5- -yl]pyridine-3-carboxamide, MW: 425.50732|MF: C20H23N7O2S 76: CID: 15980233, IUPAC: N-(2-hydroxyethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thiazol- -5-yl]pyridine-3-carboxamide, MW: 426.49208|MF: C20H22N6O3S. 77: CID: 15980347, IUPAC: N-(2-methylaminoethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thi- azol-5-yl]pyridine-3-carboxamide, MW: 439.5339|MF: C21H25N7O2S. 78: CID: 15980351, IUPAC: 5-[2-[[4-[4-(2-hydroxyacetyl)piperazin-1-yl]pyridin-2-yl]amino]-1,3-thiaz- ol-5-yl]-N-(2,2,2-trifluoroethyl)pyridine-3-carboxamide, MW: 521.51539|MF: C22H22F3N7O3S. 79: CID: 15982537, IUPAC: (3-chloro-2-fluorophenyl)-[4-[6-[(5-fluoro-1,3-thiazol-2-yl)amino]-5-meth- ylpyridin-2-yl]piperazin-1-yl]methanone, MW: 449.904626|MF: C20H18ClF2N5OS. 80: CID: 16034848, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 2,3-dihydroxybutanedioic acid, MW: 638.09238|MF: C26H32ClN7O8S. 81: CID: 16037977, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-5-me-

thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 82: CID: 16061431, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 4-[(4-methylpiperazin-1-yl)methyl]-N-[4-methyl-3-[(4-pyridin-3-ylpyrimidi- n-2-yl)amino]phenyl]benzamide, MW: 981.60828|MF: C51H57ClN14O3S. 83: CID: 16223227, IUPAC: but-2-enedioic acid; N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 604.0777|MF: C26H30ClN7O6S. 84: CID: 16223228, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrobromide, MW: 568.91748|MF: C22H27BrClN7O2S. 85: CID: 16223229, IUPAC: but-2-enedioic acid; N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl- ]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 604.0777|MF: C26H30ClN7O6S. 86: CID: 16223316, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; methanesulfonic acid, MW: 584.1112|MF: C23H30ClN7O5S2. 87: CID: 16223317, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; phosphoric acid, MW: 586.00072|MF: C22H29ClN7O6PS. 88: CID: 16223318, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 2-hydroxybenzoic acid, MW: 626.12628|MF: C29H32ClN7O5S. 89: CID: 16223319, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; sulfuric acid, MW: 586.08402|MF: C22H28ClN7O6S2. 90: CID: 16223320, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 4-methylbenzenesulfonic acid, MW: 660.20716|MF: C29H34ClN7O5S2. 91: CID: 16584134, AKE-PB223730486, IUPAC: N-(4-chlorophenyl)-2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methylpyrim- idine-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 92: CID: 16584137, AKE-PB223730492, IUPAC: N-(3-chlorophenyl)-2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methylpyrim- idine-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 93: CID: 16584139, AKE-PB223730496, IUPAC: 2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methyl-N-(2-methylphenyl)pyrim- idine-5-carboxamide, MW: 353.44136|MF: C18H19N5OS. 94: CID: 16655683, IUPAC: 2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-N-(2,6-dichlorophenyl)-- 1,3-thiazole-5-carboxamide, MW: 414.6968|MF: C15H10Cl3N5OS. 95: CID: 16655839, IUPAC: N-(2,6-dichlorophenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylp- yrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 508.42402|MF: C21H23Cl2N7O2S. 96: CID: 16660745, IUPAC: N-(4-fluorophenyl)-4-(2-hydroxyethyl amino)-6-methylsulfanyl-2-pyridin-4-ylpyrimidine-5-carboxamide, MW: 399.441923|MF: C19H18FN5O2S. 97: CID: 16660747, IUPAC: N-(4-ethylphenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin-4-yl- pyrimidine-5-carboxamide, MW: 409.50462|MF: C21H23N5O2S. 98: CID: 16660907, IUPAC: 4-(2-hydroxyethylamino)-N-(4-methylphenyl)-6-methylsulfanyl-2-pyridin-4-y- lpyrimidine-5-carboxamide, MW: 395.47804|MF: C20H21N5O2S. 99: CID: 16661063, IUPAC: N-(4-chlorophenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin-4-y- lpyrimidine-5-carboxamide, MW: 415.89652|MF: C19H18ClN5O2S. 100: CID: 16661212, IUPAC: N-(2,4-dimethylphenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin- -4-ylpyrimidine-5-carboxamide, MW: 409.50462|MF: C21H23N5O2S. 101: CID: 16661214, IUPAC: 4-(1-hydroxybutan-2-ylamino)-N-(4-methylphenyl)-6-methylsulfanyl-2-pyridi- n-4-ylpyrimidine-5-carboxamide, MW: 423.5312|MF: C22H25N5O2S.

[0121] Herein, CID is the compound identifier as defined in Pubchem. [0122] e) Small molecules targeting and antagonising downstream components of the NEPOR signalling pathway, particularly EPH-B4 tyrosine kinase inhibitors. [0123] f) Combination therapies involving one or more of approaches a-e. [0124] g) The present disclosure also provides a combination therapy. Co-administration of EPO with an intracellular inhibitor of NEPOR signalling (e.g. Dasatinib) is proposed to maintain EPO signalling via EPOR (and thus promote haematopoiesis) while inhibiting survival of NEPOR positive tumour cells.

NEPOR Based Therapeutics to Treat Neuronal Insults

[0125] Without being bound by theory, the present disclosure provides that EPO is neuroprotective because of its effects on NEPOR activity, i.e. in particular and/or EPH-B4 and/or Ephrin A1 activity. Therefore, the present disclosure provides a method for treating ischemic stroke, trauma, epilepsy, neurodegenerative diseases, and cognitive dysfunction with an agonistic NEPOR targeted therapy. Specific approaches to positively enhance NEPOR mediated survival signals include: [0126] a) NEPOR specific antibodies. Such antibodies bind and initiate/enhance the mediation of NEPOR's cyto-protective activity. [0127] b) NEPOR specific small-molecules. Such small molecules bind and initiate/enhance the mediation of NEPOR's cytoprotective activity. [0128] c) NEPOR-targeting EPO mutants and glycosylated versions thereof. Due to EPOR's strict conformational requirements for mediating signalling in response to EPO, the following to EPOR mutants (SEQ ID NO. 17-SEQ ID NO. 212.) favour binding to NEPOR as opposed to EPOR and thus primarily act as tissue protective.

TABLE-US-00009 [0128] SEQ ID NO. 17 APPRLICDSRVLERYLLEAKEAENITRVGQQAVEVWQGLALLSEAVLRGQALLVNSSQPWEPLQLHVDKAVSGL- RSL TTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGEACRTGDR SEQ ID NO. 18 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQALLVNSSQPWEPLQL- HVD KAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGEACRTGDR SEQ ID NO. 19 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYALLVNSSQPWEPLQLHVDKAVSGLR- SLT TLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGEACRTGDR SEQ ID NO. 20 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEPWEPLQLHVDKAVSGLRSL- TTL LRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGEACRTGDR SEQ ID NO. 21 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPPGVGQLFPAVGAPAAACG SEQ ID NO. 22 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENNHC SEQ ID NO. 23 APPRLICDSRVLEAYLLEAKEAENIT SEQ ID NO. 24 APPRLICDSRVLEAYLLEAKEAENIT SEQ ID NO. 25 APPRLICDSRVLEEYLLEAKEAENIT SEQ ID NO. 26 APPRLICDSRVLERYL SEQ ID NO. 27 APPRLI SEQ ID NO. 28 APPRLICDSRVLERYILEAKEAENVTMGCAEGPRLSENITVPDTKVNFYAWKRMEKELMSPPDTTPPAPLRTLT- VDT FCKLFRVYANFLRGKLKLYTGEVCRRGDR

Deletions of hWT 4 EPOR Interaction Sites

TABLE-US-00010 [0129] SEQ ID NO. 29 APPRLICEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAVLRGQALLVNSSQP- WEP LQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGEACRTG- DR SEQ ID NO. 30 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDFYAWKRMEVGQQAVEVWQGLALLSEAVLRGQ- ALL VNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKL- YTG EACRTGDR SEQ ID NO. 31 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKLKLYTGE- ACR TGDR SEQ ID NO. 32 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFGKLKLYTGE- ACR TGDR

C-Term Deletions Beginning at the Last Cysteine Bridge C161

TABLE-US-00011 [0130] SEQ ID NO. 33 APPRLICDSRVLERYLLEAKEAENITTGCAEHOSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEA SEQ ID NO. 34 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGE SEQ ID NO. 35 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTG SEQ ID NO. 36 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYT SEQ ID NO. 37 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLITLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LY SEQ ID NO. 38 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK L SEQ ID NO. 39 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK SEQ ID NO. 40 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KL SEQ ID NO. 41 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- K SEQ ID NO. 42 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG SEQ ID NO. 43 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLR SEQ ID NO. 44 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFL SEQ ID NO. 45 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNF SEQ ID NO. 46 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSN SEQ ID NO. 47 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYS SEQ ID NO. 48 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVY SEQ ID NO. 49 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLASLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRV SEQ ID NO. 50 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFR SEQ ID NO. 51 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLF SEQ ID NO. 52 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKL SEQ ID NO. 53 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRK SEQ ID NO. 54 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFR SEQ ID NO. 55 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTF SEQ ID NO. 56 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADT SEQ ID NO. 57 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITAD SEQ ID NO. 58 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITA SEQ ID NO. 59 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTIT SEQ ID NO. 60 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTI SEQ ID NO. 61 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRT SEQ ID NO. 62 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLR SEQ ID NO. 63 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPL SEQ ID NO. 64 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAP SEQ ID NO. 65 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAA SEQ ID NO. 66 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASA SEQ ID NO. 67 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAAS SEQ ID NO. 68 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAA SEQ ID NO. 69 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDA SEQ ID NO. 70 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPD

SEQ ID NO. 71 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPP SEQ ID NO. 72 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISP SEQ ID NO. 73 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAIS SEQ ID NO. 74 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAI SEQ ID NO. 75 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEA SEQ ID NO. 76 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKE SEQ ID NO. 77 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQK SEQ ID NO. 78 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQ SEQ ID NO. 79 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGA SEQ ID NO. 80 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALG SEQ ID NO. 81 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRAL SEQ ID NO. 82 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRA SEQ ID NO. 83 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLR SEQ ID NO. 84 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLL SEQ ID NO. 85 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTL SEQ ID NO. 86 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTT SEQ ID NO. 87 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLT SEQ ID NO. 88 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSL SEQ ID NO. 89 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRS SEQ ID NO. 90 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLR SEQ ID NO. 91 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGL SEQ ID NO. 92 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSG SEQ ID NO. 93 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVS SEQ ID NO. 94 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAV SEQ ID NO. 95 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKA SEQ ID NO. 96 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDK SEQ ID NO. 97 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVD SEQ ID NO. 98 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHV SEQ ID NO. 99 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLH SEQ ID NO. 100 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQL SEQ ID NO. 101 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQ SEQ ID NO. 102 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPL SEQ ID NO. 103 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEP SEQ ID NO. 104 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWE SEQ ID NO. 105 APPRLICDSRVLERYLLEAKEAENTTTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPW SEQ ID NO. 106 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQP SEQ ID NO. 107 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQ SEQ ID NO. 108 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSS SEQ ID NO. 109 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNS SEQ ID NO. 110 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVN SEQ ID NO. 111 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLV SEQ ID NO. 112 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALL

SEQ ID NO. 113 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QAL SEQ ID NO. 114 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QA SEQ ID NO. 115 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG Q SEQ ID NO. 116 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG SEQ ID NO. 117 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LR SEQ ID NO. 118 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- L SEQ ID NO. 119 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV SEQ ID NO. 120 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEA SEQ ID NO. 121 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSE SEQ ID NO. 122 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLS SEQ ID NO. 123 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALL SEQ ID NO. 124 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLAL SEQ ID NO. 125 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLA SEQ ID NO. 126 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGL SEQ ID NO. 127 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQG SEQ ID NO. 128 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQ SEQ ID NO. 129 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVW SEQ ID NO. 130 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEV SEQ ID NO. 131 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVE SEQ ID NO. 132 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAV SEQ ID NO. 133 APPRLICDSRVLERYLLEAREAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQA SEQ ID NO. 134 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQ SEQ ID NO. 135 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQ SEQ ID NO. 136 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVG SEQ ID NO. 137 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEV SEQ ID NO. 138 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRME SEQ ID NO. 139 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRM SEQ ID NO. 140 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKR SEQ ID NO. 141 APPRLICDSRVIERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWK SEQ ID NO. 142 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAW SEQ ID NO. 143 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYA SEQ ID NO. 144 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFY SEQ ID NO. 145 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNF SEQ ID NO. 146 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVN SEQ ID NO. 147 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKV SEQ ID NO. 148 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTK SEQ ID NO. 149 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDT SEQ ID NO. 150 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPD SEQ ID NO. 151 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVP SEQ ID NO. 152 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITV SEQ ID NO. 153 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENIT SEQ ID NO. 154 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENI SEQ ID NO. 155 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNEN SEQ ID NO. 156 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNE SEQ ID NO. 157 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLN SEQ ID NO. 158 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSL SEQ ID NO. 159 APPRLICDSRVLERYLLEAKEAENITTGCAEHCS SEQ ID NO. 160 APPRLICDSRVLERYLLEAKEAENITTGCAEHC SEQ ID NO. 161 APPRLICDSRVLERYLLEAKEAENITTGCAEH SEQ ID NO. 162 APPRLICDSRVLERYLLEAKEAENITTGCAE SEQ ID NO. 163 APPRLICDSRVLERYLLEAKEAENITTGCA SEQ ID NO. 164 APPRLICDSRVLERYLLEAKEAENITTGC SEQ ID NO. 165 APPRLICDSRVLERYLLEAKEAENITTG SEQ ID NO. 166 APPRLICDSRVLERYLLEAKEAENITT SEQ ID NO. 167 APPRLICDSRVLERYLLEAKEAENIT SEQ ID NO. 168 APPRLICDSRVLERYLLEAKEAENI SEQ ID NO. 169 APPRLICDSRVLERYLLEAKEAEN SEQ ID NO. 170 APPRLICDSRVLERYLLEAKEAE SEQ ID NO. 171 APPRLICDSRVLERYLLEAKEA SEQ ID NO. 172 APPRLICDSRVLERYLLEAKE SEQ ID NO. 173 APPRLICDSRVLERYLLEAK SEQ ID NO. 174 APPRLICDSRVLERYLLEA SEQ ID NO. 175 APPRLICDSRVLERYLLE SEQ ID NO. 176 APPRLICDSRVLERYLL SEQ ID NO. 177 APPRLICDSRVLERYL SEQ ID NO. 178 APPRLICDSRVLERY SEQ ID NO. 179 APPRLICDSRVLER SEQ ID NO. 180 APPRLICDSRVLE SEQ ID NO. 181 APPRLICDSRVL SEQ ID NO. 182 APPRLICDSRV SEQ ID NO. 183 APPRLICDSR SEQ ID NO. 184 APPRLICDS SEQ ID NO. 185 APPRLICD SEQ ID NO. 186 APPRLIC

[0131] Single Amino Acid Mutations (Ala/Conversions) and all combinations/permutations thereof and all glycosylated versions of same. All possible combinations/permutations of mutations contained in Single mutations of SEQ ID NOs. 187-208 and glycosylated versions thereof.

TABLE-US-00012 SEQ ID NO. 187 APPRLICASRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 188 APPRLICRSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 189 APPRLICDSRVLEAYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 190 APPRLICDSRVLEEYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKRAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 191 APPRLICDSRVLERYLLEAAEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 192 APPRLICDSRVLERYLLEAEEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 193 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDAKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 194 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTAVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 195 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTEVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 196 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKANFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 197 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVAFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 198 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDAAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 199 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDEAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 200 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVAGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 201 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLASLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 202 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEWGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLESLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 203 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRALTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 204 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFAVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 205 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFEVYSNFLRG- KLK LYTGEACRTGDR SEQ ID NO. 206 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLASLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSAFLRG- KLK LYTGEACRTGDR SEQ ID NO. 207 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLAG- KLK LYTGEACRTGDR SEQ ID NO. 208 APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAV- LRG QALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLEG- KLK LYTGEACRTGDR

EPO Peptides Overlapping Interaction Regions

TABLE-US-00013 [0132] SEQ ID NO. 209 APPRLICDSRVLERYLLEAKEAENITT SEQ ID NO. 210 NENITVPDTKVNFYAWKRMEV SEQ ID NO. 211 NSSQPWEPLQLHVDKAVSGLRSLTTLL SEQ ID NO. 212 FRKLFRVYSNFLRGKLKL

[0133] d) NEPOR-targeting EPO chimera's. Such mutants bind and initiate/enhance the mediation of NEPOR's cytoprotective activity. For example, in a scenario where NEPOR constitutes an Ephrin A1 molecule (either as a homodimer or in heterodimeric association with EPOR), then chimeric proteins involving fusions of part of EPH-B4's Ephrin-ligand-binding domain and part of the EPO molecule may be developed as optimised binding partners. This implies fusing an N-terminal portion of EPO (derived from SEQ IDNO. 213) to a C-terminal portion of EPH-B4's Ephrin ligand binding domain (SEQ IDNO. 214), giving a sequence similar to SEQ IDNO. 215, or fusing an N-terminal portion of EPH-B4's Ephrin ligand binding domain (derived from SEQ ID NO. 214) to a C-terminal portion of EPO (SEQ ID NO. 213), giving a sequence similar to SEQ ID NO. 216. [0134] e) high-affinity peptides which specifically target NEPOR to initiate/enhance the mediation of EPO's cytoprotective activity. [0135] f) Small molecules targeting and enhancing the activity of downstream components of NEPOR. [0136] g) Combination therapies involving one or more of approaches a-f.

TABLE-US-00014 [0136] SEQ ID NO. 213 >P01588|EPO_HUMAN Erythropoietin - Homo sapiens (Human). MGVHECPAWLWLLLSLLSLPLGLPVLGAPPRLICDSRVLERYLLEAKEAENITTGCAEHC SLNENITVPDTKVNFYAWKRMEVGQQAVEVWQGLALLSEAVLRGQALLVNSSQPWEPLQL HVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRKLFRVYSNFLRGKL KLYTGEACRTGDR SEQ ID NO. 214 >EPH-B4_ephrin_ligand_binding_domain EETLLNTKLETADLKWVTFPQVDGQWEELSGLDEEQHSVRTYEVCDVQRAPGQAHWLRTG WVPRRGAVHVYATLRFTMLECLSLPRAGRSCKETFTVFYYESDADTATALTPAWMENPYI KVDTVAAEHLTRKRPGAEATGKVNVKTLRLGPLSKAGFYLAFQDQGACMALLSLHLFYKK C SEQ ID NO. 215 >NtermEPO_CtermEPHB4LBD APPRLICDSRVLERYLLEAKEAENITTGCAEHCSLNENITVPDTKVNFYAWKRMEVGQQA VEVWQGLALLSEAVLRGQALLVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAIS PPDAASALTPAWMENPYIKVDTVAAEHLTRKRPGAEATGKVNVKTLRLGPLSKAGFYLAF QDQGACMALLSLHLFYKKC SEQ ID NO. 216 >NtermEPHB4LBD_CtermEPO EETLLNTKLETADLKWVTFPQVDGQWEELSGLDEEQHSVRTYEVCDVQRAPGQAHWLRTG WVPRRGAVHVYATLRFTMLECLSLPRAGRSCKETFTVFYYESDADTATALSEAVLRGQAL LVNSSQPWEPLQLHVDKAVSGLRSLTTLLRALGAQKEAISPPDAASAAPLRTITADTFRK LFRVYSNFLRGKLKLYTGEACRTGDR

Compounds in Combination with EPO

[0137] Such compounds, in combination with EPO, inhibit EPH-B4's tyrosine kinase activity while permitting EPOR mediated signalling/haematopoiesis. The following 101 compounds, either alone or in combination, inhibit the tyrosine kinase activity of EPH-B4 containing NEPOR dimers. Therefore, the present disclosure provides a combination therapeutic agent of a tyrosine kinase inhibitor in combination with EPO to provide the hematopoietic properties of EPO along with the prevention of NEPOR signalling so as to block the potentially fatal side effect of EPO to promote tumour survival and angiogenesis.

1: CID: 1095868, AKI-STT-00166305; ZINC00818264; BAS 09636496 IUPAC: N-[5-[(3-chlorophenyl)methyl]-1,3-thiazol-2-yl]-2-(4,6-dimethylpyrimidin-- 2-yl)sulfanylacetamide. MW: 404.93678|MF: C18H17ClN4OS2. (MW is molecular weight and MF is molecular formula). 2: CID: 1465558, IUPAC: 2-[(3-chlorobenzoyl)amino]-4-methyl-N-pyridin-3-yl-1,3-thiazole-5-carboxa- mide, MW: 372.82872|MF: C17H13ClN4O2S. 3: CID: 1468201, IUPAC: N-[5-[(2-chlorophenyl)carbamoyl]-4-methyl-1,3-thiazol-2-yl]pyridine-4-car- boxamide, MW: 372.82872|MF: C17H13ClN4O2S. 4: CID: 3062316, Dasatinib; Sprycel; BMS Dasatinib, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 5: CID: 3072360, 142287-40-9; Pyrimido(4,5-d)pyrimidin-4(1H)-one, 7-methyl-1-phenyl-2-((3-(4-(2-thiazolyl)-1-piperazinyl)propyl)thio)-IUPAC- : 2-methyl-8-phenyl-7-[3-[4-(1,3-thiazol-2-yl)piperazin-1-yl]propylsulfany- l]pyrimido[6,5-d]pyrimidin-5-one, MW: 479.6209|MF: C23H25N7OS2. 6: CID: 5041467, STK154706; ZINC04687922, IUPAC: [2-[(2-methylphenyl)amino]-1,3-thiazol-4-yl]-(4-pyrimidin-2-ylpiperazin-1- -yl)methanone, MW: 380.4667|MF: C19H20N6OS. 7: CID: 9822929, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-imidazol-1-ylpyridazin-3-yl)amino]-1,3-- thiazole-5-carboxamide, MW: 411.869|MF: C18H14ClN7OS. 8: CID: 9927718, IUPAC: N-(2-chloro-6-methylphenyl)-2-(cyclopropanecarbonylamino)-1,3-thia- zole-5-carboxamide, MW: 335.809|MF: C15H14ClN3O2S. 9: CID: 10006113, IUPAC: N-[4-chloro-2-[(5-chloropyridin-2-yl)carbamoyl]phenyl]-5-methyl-6,- 7-dihydro-4H-[1,3]thiazolo[5,4-c]pyridine-2-carboxamide hydrochloride, MW: 498.81322|MF: C20H18Cl3N5O2S. 10: CID: 10006114, IUPAC: N-[4-chloro-2-[(5-chloropyridin-2-yl)carbamoyl]phenyl]-5-methyl-6,7-dihyd- ro-4H-[1,3]thiazolo[5,4-c]pyridine-2-carboxamide, MW: 462.35228|MF: C20H17Cl2N5O2S. 11: CID: 10052635, IUPAC: 2-[[2-methyl-5-[[6-[(4-methylpiperazin-1-yl)methyl]pyridin-2-yl]amino]phe- nyl]amino]-N-(2-methylphenyl)-1,3-thiazole-5-carboxamide, MW: 527.68362 MF: C29H33N7OS. 12: CID: 10195898, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[[(2S)-2-hydroxy-2-pyrimidin-2-ylethyl]-met- hylamino]methyl]-4-methyl-7-oxothieno[2,3-e]pyridine-6-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 13: CID: 10206276, IUPAC: N-[4-[(5-chloropyridin-2-yl)carbamoyl]-2-phenyl-1,3-thiazol-5-yl]-1-propa- n-2-ylpiperidine-4-carboxamide, MW: 484.01354|MF: C24H26ClN5O2S. 14: CID: 10252208, IUPAC: 2-[4-(5-amino-1,3-thiazol-2-yl)phenyl]-3-(5-chloropyridin-2-yl)quinazolin- -4-one, MW: 431.89746|MF: C22H14ClN5OS. 15: CID: 10253695, IUPAC: 2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazole-- 5-carboxamide, MW: 459.90756|MF: C23H14ClN5O2S. 16: CID: 10301604, IUPAC: N-[4-[(5-chloropyridin-2-yl)carbamoyl]-2-(3,4-difluorophenyl)-1,3-thiazol- -5-yl]-1-propan-2-ylpiperidine-4-carboxamide, MW: 519.994466|MF: C24H24ClF2N5O2S. 17: CID: 10344807, IUPAC: N-[2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazo- l-4-yl]acetamide, MW: 473.93414|MF: C24H16ClN5O2S. 18: CID: 10368624, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[(2-hydroxy-2-pyrimidin-2-ylethyl)-m- ethylamino]methyl]-7-methyl-4-oxothieno[3,2-e]pyridine-5-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 19: CID: 10370949, IUPAC: (3Z)-4-[[(2S)-2-(3-chlorophenyl)-2-hydroxyethyl]amino]-3-[6-methyl-2-[4-(- 1,3-thiazol-2-ylmethyl)piperazin-1-yl]-7,9-dihydropurin-8-ylidene]pyridin-- 2-one, MW: 578.08832|MF: C27H28ClN9O2S. 20: CID: 10412586, IUPAC: N-[2-[4-[3-(5-chloropyridin-2-yl)-4-oxoquinazolin-2-yl]phenyl]-1,3-thiazo- l-5-yl]acetamide, MW: 473.93414|MF: C24H16ClN5O2S. 21: CID: 10413555, IUPAC: N-[(4-chlorophenyl)methyl]-2-[[[(2R)-2-hydroxy-2-pyrimidin-2-yleth- yl]-methylamino]methyl]-7-methyl-4-oxothieno[3,2-e]pyridine-5-carboxamide, MW: 497.99706|MF: C24H24ClN5O3S. 22: CID: 10456156, IUPAC: 4-[(3-chlorothiophen-2-yl)methylamino]-2-[(4-morpholin-4-ylphenyl)amino]p- yrimidine-5-carboxamide, MW: 444.93774|MF: C20H21ClN6O2S. 23: CID: 10458706, IUPAC: N-[5-[2-[(4-chlorophenyl)amino]pyrimidin-4-yl]-4-methyl-1,3-thiazol-2-yl]- -3-(2-morpholin-4-ylethylamino)propanamide, MW: 502.03212|MF: C23H28ClN7O2S. 24: CID: 11153014, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2,6-dimethylpyrimidin-4-yl)amino]-1,3-thi- azole-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 25: CID: 11167695, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-(2-morpholin-4-ylethyla- mino)pyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 26: CID: 11168231, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-N-- [(4-methoxyphenyl)methyl]-1,3-thiazole-5-carboxamide, MW: 514.42684|MF: C24H21Cl2N5O2S 27: CID: 11200510, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-hydroxyethylamino)pyridin-2-yl]amino- ]-1,3-thiazole-5-carboxamide, MW: 403.88582|MF: C18H18ClN5O2S. 28: CID: 11247793, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(methyl-(3-methylaminopropyl)amino)pyri- din-2-yl]amino]-1,3-thiazole-5-carboxamide, MW: 444.9808|MF: C21H25ClN6OS. 29: CID: 11260009, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(hydroxymethyl)piperidin-1-yl]-2-met- hylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 472.9909|MF: C22H25ClN6O2S. 30: CID: 11269410, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-1,- 3-thiazole-5-carboxamide, MW: 394.27832|MF: C16H13Cl2N5OS. 31: CID: 11282881, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-morpholin-4-ylethylamino)pyrimidin-4- -yl]amino]-1,3-thiazole-5-carboxamide, MW: 473.97896|MF: C21H24ClN7O2S. 32: CID: 11283174, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(3-morpholin-4-ylpropylamino)pyridin-2-- yl]amino]-1,3-thiazole-5-carboxamide, MW: 487.01748|MF: C23H27ClN6O2S. 33: CID: 11328827, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(3-imidazol-1-ylpropylamino)pyridin-2-y- l]amino]-1,3-thiazole-5-carboxamide, MW: 467.97438|MF: C22H22ClN7OS. 34: CID: 11407465, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin- -4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 418.90046|MF: C18H19ClN6O2S. 35: CID: 11466196, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-(3-morpholin-4-ylpropylamino)p- yrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide. MW: 502.03212|MF: C23H28ClN7O2S. 36: CID: 11466607, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrochloride, MW: 524.46648|MF: C22H27Cl2N7O2S. 37: CID: 11487256, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(6-morpholin-4-ylpyrimidin-4-yl)amino]-1,3- -thiazole-5-carboxamide, MW: 430.91116|MF: C19H19ClN6O2S 38: CID: 11505502, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]pyrimidin-4-yl]amino]-N-[2-methyl- -5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide. MW: 626.65257|MF: C29H29F3N8O3S. 39: CID: 11512538, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]ethyl 2,2-dimethylpropanoate, MW: 572.12196|MF: C27H34ClN7O3S. 40: CID: 11539665, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-[(5-fluoro-1,3-thiazol-2-yl)amino]pyridi- n-2-yl]methyl]piperazin-1-yl]methanone, MW: 449.904626|MF: C20H18ClF2N5OS. 41: CID: 11540687, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrate, MW: 506.02082 MF: C22H28ClN7O3S. 42: CID: 11569328, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[3-[4-(2-hydroxyethyl)piperazin-1-yl]-5-me- thylphenyl]amino]-1,3-thiazole-5-carboxamide, MW: 486.02942|MF: C24H28ClN5O2S. 43: CID: 11570976, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-N-- [2-methyl-5-[[3-(trifluoromethyl)phenyl]carbamoyl]phenyl]-1,3-thiazole-5-c- arboxamide, MW: 640.67915|MF: C30H31F3N8O3S. 44: CID: 11577776, IUPAC: 2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin-4-yl]amino]-N-[2-methyl-5-[- [3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 571.57407|MF: C26H24F3N7O3S. 45: CID: 11590089, IUPAC: (3-chloro-2-fluorophenyl)-[4-[5-methyl-6-(1,3-thiazol-2-ylamino)pyridin-2- -yl]piperazin-1-yl]methanone, MW: 431.914163|MF: C20H19ClFN5OS. 46: CID: 11606973, IUPAC: N-[5-[[3-[4-(2-hydroxyethyl)piperazin-1-yl]-5-(trifluoromethyl)benzoyl]am- ino]-2-methylphenyl]-2-(pyridin-2-ylamino)-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 47: CID: 11650098, IUPAC: 2-[[6-(4-methylpiperazin-1-yl)pyrimidin-4-yl]amino]-N-[2-methyl-5-[[3-(tr- ifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 596.62659|MF: C28H27F3N8O2S. 48: CID: 11650132, IUPAC: pentyl N-[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]-N-[6-[4-(2-hy- droxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]carbamate, MW: 602.14794|MF: C28H36ClN7O4S. 49: CID: 11650511, IUPAC: N-[5-[[3-(4-ethylpiperazin-1-yl)-5-(trifluoromethyl)benzoyl]amino]-2-meth- ylphenyl]-2-[[6-(2-hydroxyethylamino)-2-methylpyrimidin-4-yl]amino]-1,3-th- iazole-5-carboxamide, MW: 683.74695|MF: C32H36F3N9O3S. 50: CID: 11664355, IUPAC: 2-[(2-methyl-6-morpholin-4-ylpyrimidin-4-yl)amino]-N-[2-methyl-5-[- [3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 597.61135|MF: C28H26F3N7O3S. 51: CID: 11664511, IUPAC: 2-[[4-[4-(2-hydroxyethyl)piperazin-1-yl]pyridin-2-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 52: CID: 11669430, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2-methyl-6-piperazin-1-ylpyrimidin-4-yl)a- mino]-1,3-thiazole-5-carboxamide, MW: 443.95298|MF: C20H22ClN7OS. 53: CID: 11676373, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-(1,3-thiazol-2-ylamino)pyridin-2-yl]meth- yl]piperazin-1-yl]methanone, MW: 431.914163|MF: C20H19ClFN5OS. 54: CID: 11684148, IUPAC: (3-chloro-2-fluorophenyl)-[4-[[6-[(5-chloro-1,3-thiazol-2-yl)amino]pyridi- n-2-yl]methyl]piperazin-1-yl]methanone, MW: 466.359223|MF: C20H18Cl2FN5OS. 55: CID: 11700117, IUPAC: 2-[[6-(4-ethylpiperazin-1-yl)-2-methylpyrimidin-4-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 624.67975|MF: C30H31F3N8O2S. 56: CID: 11707091, IUPAC: 2-[[2-methyl-6-(4-methylpiperazin-1-yl]pyrimidin-4-yl]amino]-N-[2-methyl-- 5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 610.65317|MF: C29H29F3N8O2S. 57: CID: 11714286, IUPAC: 2-[[5-[4-(2-hydroxyethyl)piperazin-1-yl]pyridin-2-yl]amino]-N-[2-methyl-5- -[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carboxamide, MW: 625.66451|MF: C30H30F3N7O3S. 58: CID: 11714353, IUPAC: 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-N-- [2-methyl-5-[[3-(trifluoromethyl)benzoyl]amino]phenyl]-1,3-thiazole-5-carb- oxamide, MW: 640.67915 j MF: C30H31F3N8O3S. 59: CID: 11752136, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[5-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 60: CID: 11772766, IUPAC: 4-[2-(3-chlorophenyl)ethylamino]-2-pyridin-4-yl-1,3-thiazole-5-carboxamid- e, MW: 358.8452|MF: C17H15ClN4OS. 61: CID: 11775143, IUPAC: N-(2-chloro-6-methylphenyl)-2-[(2-methyl-6-morpholin-4-ylpyrimidin-4-yl)a- mino]-1,3-thiazole-5-carboxamide, MW: 444.93774|MF: C20H21ClN6O2S. 62: CID: 11854012, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]acetic acid, MW: 501.98906|MF: C22H24ClN7O3S. 63: CID: 11854269, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]piperazin-1-yl]ethyl hydrogen sulfate, MW: 568.06874 MF: C22H26ClN7O5S2. 64: CID: 11854270, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[2-(2-hydroxyethylamino)ethylamino]-2-m- ethylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 461.96826|MF: C20H24ClN7O2S 65: CID: 11854271, IUPAC: 2-[[6-(2-aminoethylamino)-2-methylpyrimidin-4-yl]amino]-N-(2-chloro-6-met- hylphenyl)-1,3-thiazole-5-carboxamide, MW: 417.9157|MF: C18H20ClN7OS. 66: CID: 11854272, IUPAC: 2-[[2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]ami- no]-2-methylpyrimidin-4-yl]piperazin-1-yl]acetyl]amino]ethanesulfonic acid, MW: 609.12066|MF: C24H29ClN8O5S2. 67: CID: 11854533, IUPAC: N-(2-chloro-4-hydroxy-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-- 1-yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 68: CID: 11854534, IUPAC: N-[2-chloro-6-(hydroxymethyl)phenyl]-2-[[6-[4-(2-hydroxyethyl)piperazin-1- -yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 69: CID: 11854535, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-4-oxidopiperazin-4-- ium-1-yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 504.00494|MF: C22H26ClN7O3S. 70: CID: 11854536, IUPAC: 2-[4-[6-[[5-[(2-chloro-6-methylphenyl)carbamoyl]-1,3-thiazol-2-yl]amino]-- 2-methylpyrimidin-4-yl]-1-oxidopiperazin-1-ium-1-yl]acetic acid, MW: 517.98846|MF: C22H24ClN7O4S. 71: CID: 11949914, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-dimethylaminopyrimidin-4-yl]amino]-N-[2- -(2-hydroxyethylamino)ethyl]pyridine-3-carboxamide, MW: 473.931003|MF: C22H25ClFN7O2. 72: CID: 11951866, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-pyrrolidin-1-ylpyrimidin-4-yl]amino]-N-- (2-hydroxyethyl)pyridine-3-carboxamide, MW: 456.900483|MF: C22H22ClFN6O2. 73: CID: 11952045, IUPAC: 4-[[2-(5-chloro-2-fluorophenyl)-5-pyrrolidin-1-ylpyrimidin-4-yl]amino]-N-- [(2S)-2-hydroxypropyl]pyridine-3-carboxamide, MW: 470.927063|MF: C23H24ClFN6O2. 74: CID: 15979866, IUPAC: 5-[2-[[4-(4-acetylpiperazin-1-yl)pyridin-2-yl]amino]-1,3-thiazol-5-yl]-N-- methylpyridine-3-carboxamide, MW: 437.51802|MF: C21H23N7O2S. 75: CID: 15980109, IUPAC: N-(2-aminoethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thiazol-5- -yl]pyridine-3-carboxamide, MW: 425.50732|MF: C20H23N7O2S 76: CID: 15980233, IUPAC: N-(2-hydroxyethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thiazol- -5-yl]pyridine-3-carboxamide, MW: 426.49208|MF: C20H22N6O3S. 77: CID: 15980347, IUPAC: N-(2-methylaminoethyl)-5-[2-[(4-morpholin-4-ylpyridin-2-yl)amino]-1,3-thi- azol-5-yl]pyridine-3-carboxamide, MW: 439.5339|MF: C21H25N7O2S. 78: CID: 15980351, IUPAC: 5-[2-[[4-[4-(2-hydroxyacetyl)piperazin-1-yl]pyridin-2-yl]amino]-1,3-thiaz- ol-5-yl]-N-(2,2,2-trifluoroethyl)pyridine-3-carboxamide, MW: 521.51539|MF: C22H22F3N7O3S. 79: CID: 15982537, IUPAC: (3-chloro-2-fluorophenyl)-[4-[6-[(5-fluoro-1,3-thiazol-2-yl)amino]-5-meth- ylpyridin-2-yl]piperazin-1-yl]methanone, MW: 449.904626|MF: C20H18ClF2N5OS. 80: CID: 16034848, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 2,3-dihydroxybutanedioic acid, MW: 638.09238|MF: C26H32ClN7O8S. 81: CID: 16037977, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-5-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 488.00554|MF: C22H26ClN7O2S. 82: CID: 16061431, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 4-[(4-methylpiperazin-1-yl)methyl]-N-[4-methyl-3-[(4-pyridin-3-ylpyrimidi- n-2-yl)amino]phenyl]benzamide, MW: 981.60828|MF: C51H57ClN14O3S. 83: CID: 16223227, IUPAC: but-2-enedioic acid; N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 604.0777|MF: C26H30ClN7O6S. 84: CID: 16223228, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide hydrobromide, MW: 568.91748|MF: C22H27BrClN7O2S. 85: CID: 16223229, IUPAC: but-2-enedioic acid; N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl- ]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 604.0777|MF: C26H30ClN7O6S. 86: CID: 16223316, IUPAC:

N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; methanesulfonic acid, MW: 584.1112|MF: C23H30ClN7O5S2. 87: CID: 16223317, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; phosphoric acid, MW: 586.000721|MF: C22H29ClN7O6PS. 88: CID: 16223318, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 2-hydroxybenzoic acid, MW: 626.12628|MF: C29H32ClN7O5S. 89: CID: 16223319, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; sulfuric acid, MW: 586.08402|MF: C22H28ClN7O6S2. 90: CID: 16223320, IUPAC: N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-me- thylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide; 4-methylbenzenesulfonic acid, MW: 660.20716|MF: C29H34ClN7O5S2. 91: CID: 16584134, AKE-PB223730486, IUPAC: N-(4-chlorophenyl)-2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methylpyrim- idine-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 92: CID: 16584137, AKE-PB223730492, IUPAC: N-(3-chlorophenyl)-2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methylpyrim- idine-5-carboxamide, MW: 373.85984|MF: C17H16ClN5OS. 93: CID: 16584139, AKE-PB223730496, IUPAC: 2-[(4,5-dimethyl-1,3-thiazol-2-yl)amino]-4-methyl-N-(2-methylphenyl)pyrim- idine-5-carboxamide, MW: 353.44136|MF: C18H19N5OS. 94: CID: 16655683, IUPAC: 2-[(6-chloro-2-methylpyrimidin-4-yl)amino]-N-(2,6-dichlorophenyl)-- 1,3-thiazole-5-carboxamide, MW: 414.6968|MF: C15H10Cl3N5OS. 95: CID: 16655839, IUPAC: N-(2,6-dichlorophenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylp- yrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide, MW: 508.42402|MF: C21H23Cl2N7O2S. 96: CID: 16660745, IUPAC: N-(4-fluorophenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin-4-y- lpyrimidine-5-carboxamide, MW: 399.441923|MF: C19H18FN5O2S. 97: CID: 16660747, IUPAC: N-(4-ethylphenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin-4-yl- pyrimidine-5-carboxamide, MW: 409.50462|MF: C21H23N5O2S. 98: CID: 16660907, IUPAC: 4-(2-hydroxyethyl amino)-N-(4-methylphenyl)-6-methylsulfanyl-2-pyridin-4-ylpyrimidine-5-car- boxamide, MW: 395.47804|MF: C20H21N5O2S. 99: CID: 16661063, IUPAC: N-(4-chlorophenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin-4-y- lpyrimidine-5-carboxamide, MW: 415.89652|MF: C19H18ClN5O2S 100: CID: 16661212, IUPAC: N-(2,4-dimethylphenyl)-4-(2-hydroxyethylamino)-6-methylsulfanyl-2-pyridin- -4-ylpyrimidine-5-carboxamide, MW: 409.50462|MF: C21H23N5O2S. 101: CID: 16661214, IUPAC: 4-(1-hydroxybutan-2-ylamino)-N-(4-methylphenyl)-6-methylsulfanyl-2-pyridi- n-4-ylpyrimidine-5-carboxamide, MW: 423.5312|MF: C22H25N5O2S.

NEPOR: Combined Prognostic and Therapeutic Value in Cancer Treatment.

[0138] Without being bound by theory, the observation that EPO treated patients often have poorer survival outcomes (at least in some cancers) means that treatment of these patients with a NEPOR targeted therapy provides a pharmacogenetic approach to targeted cancer treatment providing tumour tissue can be assessed for expression of NEPOR. Such a therapeutic perspective changes the balance in favour of performing biopsies under all suitable circumstances--meaning for cancers where EPOR, EPH-B4 and/or EphrinA1 are typically expressed.

[0139] The present disclosure further provides a method for imaging tumour tissue that is susceptible to enhanced survival in response to EPO treatment, comprising administering an anti-NEPOR antibody or NEPOR binding peptide linked to a radio-ligand or other imaging agent, and measuring for tissue distribution and location of the radio-ligand or other imaging agent.

[0140] If a tumour is NEPOR positive, then EPO is contraindicated and a NEPOR targeted therapy is administered. If NEPOR is not present, then it is safe to administer EPO. Both outcomes stand to benefit patient outcome, regardless of whether a patient is NEPOR positive or negative. Again, this shifts the balance in favour of performing routine biopsies.

[0141] In one embodiment the invention relates to an siRNA molecule specific to EPH-B4 and/or Ephrin A1 for use in treating a cancer patient that is or will receive EPO.

EPH-B4 siRNAs and Antisense Oligodeoxynucleotides

[0142] Various EphB4-specific anti-sense phosphorothioate-modified oligodeoxynucleotides (ODNs) and siRNA may be synthesized from (e.g. by Qiagen. The most active antisense ODN and siRNA that knocks down EphB4 expression in the transiently transfected 293T cell line is chosen. The antisense ODN that may be used is AS-10 which spans nucleotides 1980 to 1999 with a sequence 5'-ATG GAG GCC TCG CTC AGA AA-3' (SEQ ID NO. 217). To eliminate cytokine responses, the cytosine at the CpG site may be methylated (AS-10M) without any loss in EphB4 knockdown efficiency (data not shown). Scrambled ODNs containing random nucleotide sequence and a similar CpG site, 5'-TAC CTG AAG GTC AGG CGA AC-3' (SEQ ID NO. 218), may be used as control. siRNA 465 corresponding to the sequences 5'-GGU GAA UGU CAA GAC GCU GUU-3' (SEQ ID NO. 219) and 3'-UUC CAC UUA CAG UUC UGC GAC-5' (SEQ ID NO. 220) may be used for RNA interference. Control siRNA may be generated by mutating three bases in this sequence to effectively abrogate EphB4 knockdown. This mutated siRNA (siRNA.DELTA.) had the sequences 5'-AGU UAA UAU CAA GAC GCU GUU-3' (SEQ ID NO. 221) and 3'-UUU CAA UUA UAG UUC UGC GAC-5' (SEQ ID NO. 222). Additionally, siRNA directed against green fluorescent protein with sequences 5'-CGC UGA CCC UGA AGU UCA TUU-3' (SEQ ID NO. 223) and 3'-UUG CGA CUG GGA CUU CAA GUA-5' (SEQ ID NO. 224) may be used as a negative control.

[0143] In one aspect, one or more of the following EPHB4-specific siRNA, which are depicted in double-stranded form, can be administered to a patient to enhance the effectiveness of Epo therapy.

TABLE-US-00015 5'-caauagccacucuaacaccuu-3' (SEQ ID NO: 242) 3'-uuguuaucggugagauugugg-5' (SEQ ID NO: 243) 5'-ggggcccgucccauuugaguu-3' (SEQ ID NO: 244) 3'-uuccccgggcaggguaaacuc-5' (SEQ ID NO: 245) 5'-cugaucugaagugggugacuu-3' (SEQ ID NO: 246) 3'-uugacuagacuucacccacug-5' (SEQ ID NO: 247) 5'-aagacccuaaugaggcuguuu-3' (SEQ ID NO: 248) 3'-uuuucugggauuacuccgaca-5' (SEQ ID NO: 249) 5'-ucgaugucuccuacgucaauu-3' (SEQ ID NO: 250) 3'-uuagcuacagaggaugcaguu-5' (SEQ ID NO: 251) 5'-auugaagaggugauugguguu-3' (SEQ ID NO: 252) 3'-uuuaacuucuccacuaaccac-5' (SEQ ID NO: 253) 5'-ggaguuacgggauugugauuu-3' (SEQ ID NO: 254) 3'-uuccucaaugcccuaacacua-5' (SEQ ID NO: 255) 5'-gguacuaaggucuacaucguu-3' (SEQ ID NO: 256) 3'-uuccaugauuccagauguagc-5' (SEQ ID NO: 257) 5'-guccugacuucaccuauacuu-3' (SEQ ID NO: 258) 3'-uucaggacugaaguggauaug-5' (SEQ ID NO: 259) 5'-ugccgcgucggguacuuccuu-3' (SEQ ID NO: 260) 3'-uuacggcgcagcccaugaagg-5' (SEQ ID NO: 261)

[0144] In other examples, siRNA can be obtained from commercial sources, such as Sigma-Aldrich (St. Louis, Mo.) and used to enhance Epo therapy. For example, the following siRNA's are commercially available from Sigma-Aldrich:

TABLE-US-00016 siRNA_ID entrezgene_ID approx_start_nucleotide EPHRIN A1 SASI_Hs01_00211016 NM_004428 247 SASI_Hs01_00211017 NM_004428 223 SASI_Hs01_00211018 NM_004428 248 SASI_Hs01_00211019 NM_004428 1071 SASI_Hs01_00211020 NM_004428 256 SASI_Hs01_00211021 NM_004428 208 SASI_Hs01_00211022 NM_004428 829 SASI_Hs01_00211023 NM_004428 1015 SASI_Hs01_00211024 NM_004428 846 SASI_Hs01_00211025 NM_004428 225 SASI_Hs01_00071683 NM_182685 248 SASI_Hs01_00071684 NM_182685 214 SASI_Hs01_00071685 NM_182685 242 SASI_Hs01_00071686 NM_182685 1000 SASI_Hs01_00071687 NM_182685 263 SASI_Hs01_00071688 NM_182685 203 SASI_Hs01_00071689 NM_182685 769 SASI_Hs01_00071690 NM_182685 948 SASI_Hs01_00071691 NM_182685 778 SASI_Hs01_00071692 NM_182685 227 EPHB4 SASI_Hs01_00039855 NM_004444 1756 SASI_Hs01_00039856 NM_004444 577 SASI_Hs01_00039857 NM_004444 1373 SASI_Hs01_00039858 NM_004444 2290 SASI_Hs01_00039859 NM_004444 2318 SASI_Hs01_00039860 NM_004444 2353 SASI_Hs01_00039861 NM_004444 2898 SASI_Hs01_00039862 NM_004444 2245 SASI_Hs01_00039863 NM_004444 1679 SASI_Hs01_00039864 NM_004444 1416

[0145] In another aspect, methods are provided for enhancing the effectiveness of EPO therapy in a patient, comprising administering to said patient, in conjunction with EPO therapy, antisense molecules specific for EPH-B4 mRNA. In one embodiment, the antisense molecule is an oligonucleotide having the nucleic acid sequence of SEQ ID NO. 217.

Antibodies to NEPOR

[0146] The present disclosure includes several antibodies that bind to NEPOR components. The following Table 6 provides a list of such antibodies and their availability.

TABLE-US-00017 TABLE 6 Catalog Company Item Antigen Number Applications Type EPOR Abcam Goat Anti-EPO Receptor Polyclonal Antibody, EPOR ab10653 ELISA, WB polyclonal Unconjugated ABR-Affinity Mouse Anti-Human EPOR Monoclonal Antibody, EPOR MA1-51823 WB, ELISA Monoclonal BioReagents Unconjugated, Clone 3D10 Abnova Mouse Anti-Human EPOR Monoclonal Antibody, EPOR H00002057- WB, Capture Monoclonal Corporation Unconjugated, Clone 3D10 M01 ELISA Abcam Goat Anti-Human EPO Receptor Polyclonal EPOR ab27497 ELISA, WB Polyclonal Antibody, Unconjugated Abcam Mouse Anti-Human EPO Receptor Monoclonal EPOR ab56310 WB Monoclonal Antibody, Unconjugated, Clone MM-0031-6G7 ABR-Affinity Mouse Anti-Human EPOR Polyclonal Antibody, EPOR PA1-51822 WB Polyclonal BioReagents Unconjugated IMGENEX Goat Anti-Human EPOR Polyclonal Antibody, EPOR IMG-3771 WB, ELISA Polyclonal Unconjugated Lifespan Rabbit Anti-Human Erythropoietin Receptor EPOR LS-C6720 ELISA Polyclonal Biosciences (EPOR) Polyclonal Antibody, Unconjugated GeneTex Mouse Anti-Human EPOR Monoclonal Antibody, EPOR GTX91710 ELISA, WB Monoclonal Unconjugated, Clone 3D10 Lifespan Rabbit Anti-Human EPOR Polyclonal Antibody, EPOR LS-C6719- ELISA Polyclonal Biosciences Unconjugated 100 Novus Biologicals Mouse Anti-Human EPOR Polyclonal Antibody, EPOR H00002057- Polyclonal Unconjugated A01 Novus Biologicals Mouse Anti-Human EPOR Monoclonal Antibody, EPOR H00002057- ELISA, WB Monoclonal Unconjugated, Clone 3D10 M01 Lifespan Sheep Anti-Human Erythropoietin Receptor EPOR LS-C6718 ELISA, WB Polyclonal Biosciences (EPOR) Polyclonal Antibody, Unconjugated Lifespan Sheep Anti-Human Erythropoietin Receptor EPOR LS-C6716 Polyclonal Biosciences (EPOR) Polyclonal Antibody, Unconjugated Lifespan Sheep Anti-Human EPOR Polyclonal Antibody, EPOR LS-C6717-50 ELISA Polyclonal Biosciences Unconjugated Santa Cruz Rabbit Anti-Human EpoR (C-20) Polyclonal EpoR (C-20) sc-695 WB, IP, IF, ICH. Polyclonal Biotechnology, Inc. Antibody, Unconjugated Santa Cruz Rabbit Anti-EpoR (M-20) Polyclonal Antibody, EpoR (M-20) sc-697 WB, IP, IF, ICH. Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Rabbit Anti-EpoR Polyclonal Antibody, EpoR sc-5624 WB, IP, IF, ICH. Polyclonal Biotechnology, Inc. Unconjugated EPH-B4 Abgent Rabbit Anti-EPH-B4 C-term RB1659-1660 EPH-B4 C-term AP7625a ELISA; IHC. Polyclonal Polyclonal Antibody, Unconjugated ABR-Affinity Rabbit Anti-Human EPH-B4 Polyclonal Antibody, EPH-B4 PA1-24241 WB Polyclonal BioReagents Unconjugated ABR-Affinity Mouse Anti-Human EPH-B4 Monoclonal Antibody, EPH-B4 MA1-51815 ELISA Monoclonal BioReagents Unconjugated, Clone 1D1 AbD Serotec Human Anti-Human EPH-B4 Monoclonal EPH-B4 HCA001 IHC, WB, ELISA Monoclonal Antibody, Unconjugated, Clone 1327 AbD Serotec Human Anti-Human EPH-B4 Monoclonal EPH-B4 HCA025 IHC, WB, ELISA Monoclonal Antibody, Unconjugated, Clone 3934 Invitrogen Anti-EPH-B4 receptor Monoclonal Antibody, EPH-B4 35-2900 WB, ELISA Monoclonal Unconjugated, Clone 3D7F8 GeneTex Rabbit Anti-EPH-B4 Polyclonal Antibody, EPH-B4 GTX77656 WB Polyclonal Unconjugated Invitrogen Mouse Anti-EPH-B4 Receptor Monoclonal EPH-B4 182394 IHC(FFPE) Monoclonal Antibody,, Clone 3D7G8 Invitrogen Anti-Eph Receptor Sampler Pack Antibody, EPH-B4 901100 ELISA Monoclonal GeneTex Mouse Anti-Human EPH-B4 Monoclonal Antibody, EPH-B4 GTX91629 Unconjugated, Clone 1D1 Invitrogen Mouse Anti-Human EPH-B4 Receptor Monoclonal EPH-B4 371800 WB ELISA IP, Monoclonal Antibody,, Clone 3D7G8 IHC Novus Biologicals Mouse Anti-Human EPH-B4 Monoclonal Antibody, EPH-B4 H00002050- Monoclonal Unconjugated, Clone 1D1 M01 R&D Systems Goat Anti-Human EPH-B4 Polyclonal Antibody, EPH-B4 AF3038 FC, IHC, WB Polyclonal Unconjugated Raybiotech, Inc. Human Anti-Human EPH-B4, (packaged with EPH-B4 DS-MB- Monoclonal HRP-Conjugated Secondary Antibody); 01224 Monoclonal Antibody, Unconjugated R&D Systems Rat Anti-Human EPH-B4 Monoclonal Antibody, EPH-B4 MAB3038 FC, IHC, WB Monoclonal Unconjugated, Clone 395810 Santa Cruz Goat Anti-EPH-B4 Polyclonal Antibody, EPH-B4 sc-7284 WB, IF Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Goat Anti-EPH-B4 Polyclonal Antibody, EPH-B4 sc-7285 WB, IF Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Rabbit Anti-EPH-B4 Polyclonal Antibody, EPH-B4 sc-5536 WB, IF Polyclonal Biotechnology, Inc. Unconjugated Raybiotech, Inc. Human Anti-Human EPH-B4, (packaged with EPH-B4 DS-MB- Monoclonal HRP-Conjugated Secondary Antibody); 01225 Monoclonal Antibody, Unconjugated EFNA1 Invitrogen Anti-Ephrin A1 Polyclonal Antibody, EFNA1 34-3300 Polyclonal Unconjugated, Clone ZMD.39 Novus Biologicals Mouse Anti-Human EFNA1 Monoclonal Antibody, EFNA1 H00001942- Monoclonal Unconjugated, Clone 3C6 M01 Santa Cruz Rabbit Anti-ephrin-A1 (V-18) Polyclonal Antibody, EFNA1 (V-18) sc-911 WB, IF Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Rabbit Anti-ephrin-A1 Polyclonal Antibody, EFNA1 sc-20719 WB, IP, IF Polyclonal Biotechnology, Inc. Unconjugated Abcam Rabbit Anti-Human Ephrin A1 Receptor Polyclonal EFNA1 ab37857 ELISA, IHC, WB Polyclonal Antibody, Unconjugated GeneTex Mouse Anti-Human EFNA1 Monoclonal Antibody, EFNA1 GTX91614 Monoclonal Unconjugated, Clone 3C7 EFNB2 Novus Biologicals Mouse Anti-Human EFNB2 Polyclonal Antibody, EFNB2 H00001948- Polyclonal Unconjugated A01 Santa Cruz Rabbit Anti-ephrin-B2 (P-20) Polyclonal Antibody, EFNB2 (P-20) sc-1010 WB, IF Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Goat Anti-ephrin-B2 Polyclonal Antibody, EFNB2 sc-19227 WB, IF, IP Polyclonal Biotechnology, Inc. Unconjugated Santa Cruz Rabbit Anti-ephrin-B2 Polyclonal Antibody, EFNB2 sc-15397 WB, IF, IP Polyclonal Biotechnology, Inc. Unconjugated

[0147] In one aspect there is provided a method for assessing tumour tissue for expression of EPH-B4 and/or Ephrin A1, comprising: (a) isolating a tissue sample from an individual who is receiving or shall receive erythropoietin, (b) determining the level of expression of the EPH-B4 and/or Ephrin A1, (c) correlating the presence of these component gene expression products to a negative physiological response to the treatment with erythropoietin. In one embodiment, the level of expression of the component genes (mRNA) is determined by a molecular biological technique selected from the group consisting of PCR, QPCR, R-PCR, gene expression microarray analysis, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), to capillary electrophoresis with laser induced fluorescence (CE-LIF). In another, the individual is a cancer patient who is to be treated with erythropoietin or is being treated with erythropoietin. In one example, the presence of EPH-B4 and/or Ephrin A1 gene expression products is indicative of poorer loco-regional tumor control and poorer patient survival upon treatment with erythropoietin. In another, the presence of a higher level of EPH-B4 and/or is Ephrin A1 gene expression products is indicative of poorer loco-regional tumour control and poorer patient survival upon treatment with erythropoietin. In some embodiments, the means for testing for the presence of the gene expression products are a protein array or binding to a mass microbalance instrument. In others, the determination of the presence of the EPH-B4 and/or Ephrin A1 gene products is done by detecting the respective proteins with an immunoassay procedure, where the immunoassay procedure is selected from the group of immunoprecipitation, enzyme immunoassay (EIA), radioimmunoassay (RIA) or fluorescent immunoassay, a chemiluminescent assay, an agglutination assay, nephelometric assay, turbidimetric assay, a Western blot, a competitive immunoassay, a noncompetitive immunoassay, a homogeneous immunoassay a heterogeneous immunoassay, a bioassay and a reporter-assay. In one example, the immunoassay is an ELISA. In another embodiment, the tissue sample may be selected from the cancerous tissue or circulating cells derived from same or from a group of biological tissues and fluids such as blood, lymph, urine, cerebral fluid.

[0148] In another aspect, a prognostic method is provided to stratify patients having a tumour as suitable (EPH-B4 and/or Ephrin A1) or non-suitable (EPH-B4 and/or Ephrin A1) for EPO treatment, comprising: (a) isolating a tissue sample from an individual who is receiving or shall receive erythropoietin; (b) determining the level of expression of the EPH-B4 and/or Ephrin A1 gene(s) component, a EPH-B4 and/or Ephrin A1 from the isolated tissue; and (c) correlating the presence of EPH-B4 and/or Ephrin A1 component gene expression products to a negative physiological response to the treatment with erythropoietin. In one embodiment, the level of expression of EPH-B4 and/or Ephrin A1 component genes is determined by a molecular biological technique selected from the group consisting of PCR, QPCR, R-PCR, gene expression microarray analysis, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), capillary electrophoresis with laser induced fluorescence (CE-LIF). In another, the determination of the presence of the EPH-B4 and/or Ephrin A1 gene products is done by detecting the respective protein with an immunoassay procedure, where the immunoassay procedure is selected from the group of ELISA, immunoprecipitation, enzyme immunoassay (ETA), radioimmunoassay (RIA) or fluorescent immunoassay, a chemiluminescent assay, an agglutination assay, nephelometric assay, turbidimetric assay, a Western blot, a competitive immunoassay, a noncompetitive immunoassay, a homogeneous immunoassay a heterogeneous immunoassay, a bioassay and a reporter-assay such as a luciferase-assay. The tissue sample can be selected from the cancerous tissue or circulating is cells derived from same, or from a group of biological tissues and fluids such as blood, lymph, urine, cerebral fluid.

[0149] In another aspect, a method is provided for imaging tumour tissue that is susceptible to enhanced survival in response to EPO treatment, comprising administering an anti-EPH-B4 and/or anti-Ephrin A1 antibody or EPH-B4 and/or Ephrin A1 binding peptide linked to a radio-ligand or other imaging agent, and measuring for tissue distribution and location of the radio-ligand or other imaging agent. In one embodiment, the anti-EPH-B4 and/or anti-Ephrin A1 antibody is a monoclonal or polyclonal antibody selected from the group of antibodies listed in Table 6.

[0150] In another aspect, a method is provided for designing a compound which interferes with NEPOR's survival promoting activity, comprising: (a) providing the molecular makeup of the NEPOR species and providing amino acid sequences of a component NEPOR polypeptides; (b) using software comprised by the digital computer to design a chemical compound/protein construct which is predicted to bind to NEPOR; and

[0151] (c) optionally designing protein constructs which mimic NEPOR in its dimerised/multimerised state (e.g. Fc constructs).

[0152] A method also is provided for identifying compounds that modulate NEPOR's tissue protective signalling activity, comprising (a) contacting a test compound with the NEPOR receptor complex; (b) measuring the level of tissue protective activity initiated by NEPOR activation; (c) identifying a test compound which increases or decreases the level of tissue protective NEPOR complex activity; (d) assaying the identified therapeutics for tissue protective activity mediated via NEPOR; and (e) assaying the identified therapeutics for NEPOR inhibitory activity. In one embodiment, the tissue protective NEPOR receptor complex activity is measured by measuring the binding of the test compound to the NEPOR receptor complex. In another, the test compound is labelled and binding of the labelled test compound to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the test compound. The tissue protective NEPOR receptor complex activity can be measured by measuring the binding of the test compound to the tissue protective NEPOR receptor complex.

[0153] In another aspect, a method is provided for identifying compounds that modulate NEPOR's tissue protective signalling activity, comprising (a) contacting a test compound with the NEPOR receptor complex expressing cell; (b) measuring the level of tissue protective activity initiated by NEPOR activation in the cell; (c) identifying a test compound which increases or decreases the level of tissue protective NEPOR complex activity in a cell; (d) assaying the identified compounds for tissue protective activity mediated via NEPOR; and (e) assaying the identified therapeutics for NEPOR inhibitory activity. In one embodiment, the assay in step (d) is a tissue protective NEPOR receptor complex activity is measured by a cell proliferation/differentiation assay. In one example, the cells in the cell proliferentiation/differentiation assay are recombinantly engineered to express EPH-B4, and/or EPOR, and/or Ephrin A1. In another, the cells endogenously expresses an EPO receptor and are transformed with a nucleic acid comprising a nucleotide sequence that (i) is operably linked to a promoter, and (ii) encodes either EPH-B4 and/or Ephrin A1. In another example, the cells endogenously express EPH-B4 and/or Ephrin A1 and are transformed with a nucleic acid comprising a nucleotide sequence that (i) is operably linked to a promoter, and (ii) encodes an EPO receptor polypeptide.

[0154] In one aspect, a method is provided for identifying a compound that modulates the interaction between a tissue protective NEPOR receptor complex and a tissue protective NEPOR receptor complex ligand, comprising: (a) contacting a tissue protective NEPOR receptor complex with one or more test compounds; and (b) measuring the tissue protective NEPOR receptor complex activity, whereby if the activity measured in (b) differs from the tissue protective NEPOR receptor complex activity in the absence of the one or more test compounds, then a compound that modulates the interaction between the tissue protective NEPOR receptor complex and the tissue protective NEPOR receptor complex ligand is identified. In one embodiment, the tissue protective NEPOR receptor complex activity is measured by cell proliferation or cell differentiation. In another, the tissue protective NEPOR receptor complex activity measured is the ability of the tissue protective NEPOR receptor complex to interact with a tissue protective NEPOR receptor complex ligand. In another, the step of assaying the identified compound for tissue protective activity comprises detecting the presence of nucleolin in the cell. In some embodiments, the step of assaying the identified compound for tissue protective activity comprises detecting or measuring an increased level of activity of neuroglobin or cytoglobin in a cell. In others, the tissue protective NEPOR receptor complex is in solution. In another the tissue protective NEPOR receptor complex is in a cell. In some aspects, the compound inhibits the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex, while in others the compound enhances the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex. The tissue protective NEPOR receptor complex contacted in step (a) can be on a cell surface or on an isolated cell membrane. In some embodiments, the tissue protective NEPOR receptor complex activity is compared to EPOR receptor activation to identify NEPOR specific compounds. In some embodiments, the tissue protective NEPOR receptor complex is immobilized to a solid surface. In one example, the solid surface is a microtiter dish, and in another it is a chip.

[0155] In another aspect, there is provided a method for identifying a compound that binds a tissue protective NEPOR receptor complex, comprising: (a) contacting a test compound with a ligand-binding tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; and (b) contacting a test compound with a ligand-binding EPOR receptor complex fragment comprising at least two EPO receptor extracellular domains fused to an Fc fragment attached to a solid support (c) removing unbound test compounds from the solid supports; (d) identifying the compound attached to the tissue protective NEPOR receptor complex fragment, but not the EPOR receptor complex (and vice versa), whereby a compound bound to the solid support is identified as a compound that binds specifically to a tissue protective NEPOR receptor complex or a compound that binds specifically to an EPOR receptor complex.

[0156] In another aspect, a method is provided for identifying a compound that binds a tissue protective NEPOR receptor complex, comprising: (a) contacting a test compound with a ligand-binding tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; (b) removing unbound test compounds from the solid supports; (c) identifying the compound attached to the tissue protective NEPOR receptor complex fragment, whereby a compound bound to the solid support is identified as a compound that binds specifically to a tissue protective NEPOR receptor complex.

[0157] In another aspect, there is provided a method for identifying a compound that binds to a tissue protective NEPOR receptor complex, comprising: (a) contacting a tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support with (i) a tissue protective NEPOR receptor complex ligand attached to a first label and (ii) an equivalent amount of a test compound attached to a second label under conditions conducive to binding; (b) removing unbound material from the tissue protective NEPOR receptor complex; and (c) detecting the level of the first and second labels wherein if the second label is present the compound binds the complex and if the level of the first label decreases relative to the level of the first label where the labelled ligand is contacted with a tissue protective NEPOR receptor complex under conditions conducive to binding in the absence of a test compound after removal of unbound material, then a compound that binds to a tissue protective NEPOR receptor complex is identified.

[0158] In another aspect, a method is provided for identifying a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to a tissue protective NEPOR receptor complex, comprising: (a) contacting a tissue protective NEPOR receptor complex ligand with a tissue protective NEPOR receptor complex fragment comprising at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor extracellular domain and at least one EPO receptor or EPH-B4 receptor or Ephrin A1 receptor, extracellular domain fused to an Fc fragment attached to a solid support; in the presence of one or more test compounds under conditions conducive to binding; and (b) measuring the amount of tissue protective NEPOR receptor complex ligated bound to the tissue protective NEPOR receptor complex; whereby if the amount of bound tissue protective NEPOR receptor complex ligand measured in (b) differs from the amount of bound tissue protective NEPOR receptor complex ligand measured in the absence of the one or more test compounds, then a compound that modulates the binding of a tissue protective NEPOR receptor complex ligand to the tissue protective NEPOR receptor complex is identified. In one embodiment, the amount of bound tissue protective NEPOR receptor complex ligand is measured using a tissue protective NEPOR receptor complex ligand-specific antibody. In another, the tissue protective NEPOR receptor complex ligand is labelled and binding of the tissue protective NEPOR receptor complex ligand to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the tissue protective NEPOR receptor complex ligand. In one aspect, the tissue protective NEPOR receptor complex ligand is labelled and binding of the labelled ligand to the tissue protective NEPOR receptor complex is measured by detecting the label attached to the tissue protective NEPOR receptor complex ligand. In one example, the label is fluorescent. In another embodiment, the test compound is an antibody specific for the tissue protective NEPOR receptor complex. In another, the test compound is a small molecule or a peptide or a member of a library. In one embodiment, the tissue protective NEPOR receptor complex ligand is EPO, or derivatives thereof. In some aspects, the compound binds the tissue protective NEPOR receptor complex. In others, the compound binds the tissue protective NEPOR receptor complex ligand. In some embodiments, the tissue protective NEPOR receptor complex activity is compared to EPOR receptor activation to identify NEPOR specific compounds.

[0159] In one aspect, a method is provided for identifying a compound that modulates a tissue protective activity in a mammal, comprising: (a) administering the compound to a first animal immediately following infliction of an injury, wherein the first animal endogenously expresses a tissue protective NEPOR receptor complex; and (b) administering the compound to a second animal immediately following infliction of the same injury as in step (a), wherein the second animal is deficient in expression of a tissue protective NEPOR receptor complex or components thereof; such that if recovery from the injury differs in the animal of step (a) as compared to the animal of step (b), a compound that modulates a tissue protective activity is identified.

[0160] In another aspect, there is provided a method for treating the negative patient outcomes associated with EPO stimulated NEPOR function, involving the co-administration of EPO with an inhibitor of NEPOR activity. In one embodiment, the method comprises administering an effective amount of anti-NEPOR antibody from claim 1, in combination with EPO, whereby such combinations permits haematopoietic signalling whilst switching off NEPOR signalling and thus EPO mediated cell survival signals on tumour cells. In another, the method further comprises administering an effective amount of EPHB4 tyrosine kinase inhibitor in combination with EPO, whereby such combinations permits haematopoietic signalling whilst switching off NEPOR signalling and thus EPO mediated cell survival signals on tumour cells. In another, the method further comprises administering an effective amount of anti-NEPOR siRNA's, in combination with EPO, whereby such combinations permits haematopoietic signalling whilst switching off NEPOR signalling and thus EPO mediated cell survival signals on tumour cells.

[0161] In another aspect, a method is provided for decreasing the survival of tumour cells or tissues in a human comprising administering a therapeutically effective amount of a compound that modulates the activity of a tissue protective NEPOR receptor complex to a human in need thereof, wherein said decreased survival of cancer cells/tissues results in the decrease of tumour growth and/or an increase in patient survival, with the proviso that the compound is an EPO derivative and not a wild-type EPO.

[0162] In one aspect, there is provided a method for modulating cell survival in NEPOR positive tissue comprising administering an EPO mutants and peptides selected from the group consisting of peptides from each of SEQ ID NO. 17 through SEQ ID NO. 212.

[0163] In another, a method is provided for modulating cell survival in NEPOR positive tissue comprising administering an effective amount of an EPO chimera's, comprising an ephrin receptor ligand binding domain selected from the group consisting of SEQ ID NO.215, and SEQ ID NO. 216. In one embodiment, the compound is an antibody specific for the tissue protective NEPOR receptor complex. In another, the compound is an antibody is specific for a tissue protective NEPOR receptor complex ligand. In another, the compound is a small molecule, peptide, or a member of a library. In another, the compound binds to the tissue protective NEPOR receptor complex. In another, the compound decreases the activity of the tissue protective NEPOR receptor complex. In another, the compound is administered in conjunction with an EPO. In another embodiment, the disease or disorder is a cancer including, head and neck cancer, breast cancer, liver cancer, colorectal cancer, small intestine cancer, leukemia, prostate cancer, lung cancer, ovarian cancer, pancreatic cancer, endometrial cancer, stomach cancer, non-Hodgkin lymphoma, kidney cancer, Renal cell carcinoma (RCC), malignant melanoma, gallbladder cancer, bladder cancer, vulvar cancer, Penile cancer, testicular cancer, thymus cancer, Kaposi's sarcoma, eye cancer, adrenal gland cancer, brain cancer, cervical cancer, appendix cancer, adenoid cancer, bile duct cancer, urethral cancer, spinal cancer, Ewing's family of tumors, extragonal germ cell cancer, extra hepatic bile duct cancer, fallopian tube cancer, soft tissue cancers, bone cancer, Hodgkin's lymphoma, anal cancer, malignant mesothelioma, vaginal cancer skin cancer, central nervous system cancer (craniopharyngioma), pleuropulmonary blastoma, nasal cavity and paranasal sinus cancer transitional cell cancer of renal pelvis and ureter, pituitary gland cancer, squamous cell carcinoma of the head and neck (HNSCC), prostate cancer, colorectal cancer, lung cancer, brain cancer, bladder cancer, and salivary gland cancer. In one embodiment, the cancer comprises cancer cells expressing the tissue protective NEPOR receptor complex. In another the cancer is metastatic cancer. In another, the cancer is an angiogenesis-dependent cancer.

[0164] In another aspect, there is provided a method for treating a patient suffering from an angiogenesis-associated disease, comprising administering to the patient a compound identified by the inventive methods.

[0165] In another aspect, there is provided siRNA which is specific for EPH-B4 for use in treating a cancer and/or tumor patient that is receiving or will receive Erythropoietin.

EXAMPLES

Example 1

[0166] A variety of sequence analysis approaches were pursued, including the search for homologues of the EPO binding domain from EPOR, a domain analysis based method combined with text-mining, and EPO homology analysis followed by text-mining of resultant hits. Only that part of the human proteome exposed to the extracellular environment was investigated. This allowed a focus on homologies that were significant, though possibly overlooked within the context of a complete proteome analysis. This formed the XtraCell database. The XtraCell database performed a signal peptide and transmembrane prediction for the entire human proteome. All proteins possessing at least one of these features were stored in a first version of the extracellular database. Given that not all extracellular proteins actually possess either of these features, there was extracted a list of protein domains specific to the extracellular environment from a SMART (Simple Modular Architecture Research Tool--.SMART is a well-known protein domain database with a strong bias towards domains contained in signalling proteins.) These were then screened against the human proteome using the HMMER algorithm. HMMER is a freely distributable implementation of profile HMM software for protein sequence analysis--Profile hidden Markov models (profile HMMs) can be used to do sensitive database searching using statistical descriptions of a sequence family's consensus. All hits were added to the XtraCell database and the dataset made non-redundant. A final version of the XtraCell database was established for the purpose of these EPO specific analyses.

Example 2

[0167] This example illustrates a domain-based approach coupled with a text-mining and genome-wide analysis. The operating theory was that any novel EPO receptor involved in mediating EPO's neuroprotective effect might also possess the two membrane proximal fibronectin 3 (FN3) domains (as found in EPOR), whilst at the same time being hypoxia inducible. Such conserved domain architecture is compatible with both a heterodimeric complex containing EPOR and/or an independent hypoxia inducible homodimeric receptor. All proteins containing two membrane proximal FN3 domains from the human proteome (84 in all) were extracted and asked whether there was any evidence for their role in response to low oxygen conditions/ischaemia. (See FIG. 4) The latter analysis was performed using a text-mining approach that encompasses the use of comprehensive protein synonyms, and concepts such as hypoxia and ischaemia. Of the 84 proteins containing the 2FN3-TM domain composition, only four showed evidence for mediating response to low oxygen conditions: EPH-B4, IL6RB, TIE1 and GM-CSF. Apart from EPH-B4, the cellular role of each of these proteins has been studied and an important role in response to hypoxia established.

[0168] Direct examination of the EPH-B4 locus revealed that it directly juxtaposes the EPO locus, albeit on the opposite strand. (See FIG. 5) This close genomic association was conserved in all vertebrate genomes examined. The need for immediate response of cells to low oxygen conditions and thus the need to co-transcribe/-translate key effector molecules was seen. Moreover, such genomic co-localisation of functionally associated molecules is seen for other receptor:ligand partners (e.g. MST1 and its receptor MST1R: see worldwide web at ensembl.org/Homo_sapiens/contigview?gene=OTTHUMG00000136237; db=vega).

[0169] To examine this possibility in greater detail, we analysed the promoter, 5' UTR and 3' UTR regions of EPO, EPHB4 and EPOR in search of hypoxia inducible factor binding sites. Here we utilised the `match` algorithm from Genomatix, searching for strict conservation of the core binding site residues and at least 90% conservation of non-core residues. We found that the EPO and EPH-B4 loci possessed numerous hypoxia-inducible transcription factor binding sites. In contrast, the EPOR gene regulatory regions were found to be complete devoid of such HIF-1 binding sites, again hinting at a possible role for EPHB4 as a hypoxia inducible EPO receptor. (See FIG. 6)

Example 3

[0170] This example shows the homology-based approach using human extra-cellular database. Here we sought to directly identify regions of EPO binding activity in other proteins, by direct comparison to the EPO binding domain of EPOR. The region of EPOR responsible for EPO binding was thus extracted and used to identify homologies with proteins of the XtraCellDB. This specially developed database holds distinct advantages in that all homologies identified are to human extracellular proteins, thus avoiding the need to assess spurious homologies to irrelevant intracellular species. Analysis of resultant homologues revealed a striking homology to the Ephrin A1 protein, within the top four hits. Given what we had learned about EPH-B4's possible role in EPO signalling we decided to assess this homology in greater detail using the Swiss-model protein structure package. Here we employed information derived from the co-crystal structure of Ephrin A5 in association with EphB2 and compared it to EPO:EPOR co-crystal information. Conservation of key residues in structurally aligned positions allowed us to conclude a firm structural basis for association between Ephrin A1 and EPO. Moreover, the realisation that both EphrinA1 and EPHB4 possess a putative affinity for EPO, suggests a more exciting functional context for ephrin biology than heretofore recognised (See FIG. 7).

Example 4

[0171] This example provides wet lab or in vivo data that validates the bioinformatics analysis provide in Examples 1-3 herein. In vivo validation of EPH-B4's role in EPO signalling has focussed on the neuroprotective aspect of EPO's function, with a bias towards the hypothesis that EPH-B4 and EPOR are heterodimeric partners. The following table lists the validation experiments for which data are available (see Table 7).

TABLE-US-00018 TABLE 7 LAB-BASED validation experiments Method Goal Result Immuno- To assess the expression Precipitation stainings on adult rodent brain histochemistry of EPHB4 protein in brain showed that EPHB4 was expressed in adult and how it relates to neurons in the same pattern as EPO receptor. EPOR expression. Staining in hippocampus showed co-expression of EPOR and EPHB4 (See FIG. 8). Strikingly, the staining was restricted to particular cells within the field of tissue. Co-IP Exogenous expression of Positive. Use of EPOR antibody successfully EPOR/EPHB4 in COS Co-IP's EPHB4 protein. cells. Co-ip with EpoR- and EphB4-antibodies => WB analysis.

[0172] Immunohistochemistry.

[0173] For immunofluorescence, sections of paraffin-embedded rat brain tissues (2 .mu.m) were deparaffinated and microwaved (citrate buffer at 600 W for 15 min). Afterwards, sections were incubated simultaneously with the EpoR antiserum (1:200; sc-697, Santa Cruz Biotechnology) and the EphB4 antibody (1:100; AF446, R&D Systems) at 4.degree. C. over night. After adding a biotinylated anti-goat secondary antibody (1:200; Dianova), sections were incubated with Streptavidin-coupled Alexa Fluor 555 (1:200; Invitrogen, Karlsruhe, Germany) and a FITC-coupled anti-rabbit secondary antibody (1:200; Dianova). The nuclei were counterstained with Hoechst 33342 (1:10,000; Molecular Probes). Controls for the stainings included omission of primary antibodies, fluorophor swapping, and single-fluorescence stainings. Images were obtained with an Olympus IX-81 microscope with narrow-bandwidth monochromator excitation (Polychrome IV, Till Photonics, Grafelfing, Germany) and appropriate filters.

[0174] Both EPHB4 and EPOR displayed a striking co-localisation when assessed in rat brain tissue sections. Without being bound by theory, this co-expression suggests functional coupling of both receptors.

[0175] Co-Immunoprecipitation.

[0176] The principle of an immunoprecipitation is an antibody (monoclonal or polyclonal) against a specific target antigen is allowed to form an immune complex with that target in a sample, such as a cell lysate. The immune complex is then captured on a solid support to which either Protein A or Protein G has been immobilized (Protein A or G binds to the antibody, which is bound to its antigen). The process of capturing this complex from the solution is referred to as precipitation. Any proteins not "precipitated" by the immobilized Protein A or G support are washed away. Finally, components of the bound immune complex (both antigen and antibody) are eluted from the support and analyzed by SDS-PAGE (gel electrophoresis), often followed by Western blot detection to verify the identity of the antigen.

[0177] Traditional immunoprecipitation involves the following steps:

1. Form the antigen-antibody complex (immune complex) by incubating specific antibody with the antigen-containing sample for 1 hour to several hours. 2. Capture the immune complex on an immobilized Protein A or Protein G agarose gel support by incubation for 0.5-2 hours. 3. Remove any non-bound protein (non-immune complex sample components) from the precipitated complex by washing gel support with additional sample buffer. 4. Boil gel support in reducing SDS-PAGE sample loading buffer. 5. Recover sample eluted in loading buffer from gel support and analyze by SDS-PAGE. 6. Perform Western blot analysis, probing with antigen-specific antibody.

[0178] In a co-immunoprecipitation the target antigen precipitated by the antibody "co-precipitates" a binding partner/protein complex from a lysate, that is, the interacting protein is bound to the target antigen, which becomes bound by the antibody that becomes captured on the Protein A or G gel support. The assumption that is usually made when associated proteins are co-precipitated is that these proteins are related to the function of the target antigen at the cellular level.

[0179] Assessment of a putative EPHB4:EPOR association using co-immmunoprecipitation showed that both proteins were physically associated. Here, FLAG-tagged EPOR was co-expressed with EPH-B4 in COS cells and then immunoprecipitated using an a-FLAG antibody. As can be seen from FIG. 9, EPHB4 was shown to co-immunoprecipitate in these experiments.

[0180] Human Fc Antibody Constructs.

[0181] The Fc conjugate approach is most appropriate when dealing with dimeric cell surface receptors. Here the extracellular portion of EPHB4/EPOR can be fused to an Fc fragment. This method has advantages due to its in vivo (therapeutic) viability and the fact that it optimally mimics the dimerised receptor state. FIG. 10 highlights the Human constructs that can be used to show EPHB4's/EphrinA1's affinity for EPO.

[0182] One of two alternatives can assay the interaction of the Fc constructs with EPO, including, for example, a protein array approach or a surface plasmon resonance analysis.

Example 5

Further In Vitro and In Vivo Validation of NEPORs Role in Mediating EPO Function

[0183] In these experiments we sought to determine the response to erythropoietin (EPO) treatment in a panel of ovarian cancer cell lines. This would be mediated by the expression of erythropoietin receptor (EPO as well as two receptors that potentially may be able to activate signaling pathways in response to EPO binding, EPH-B4 and Ephrin A1. It was first necessary to characterize the expression of these receptors in a panel of ovarian cancer cell lines. First we collected RNA from each cell line and reverse transcribed them into cDNA. Using specific primers for each receptor we analyzed their RNA expression. As evident in the figures the expression of EPOR and EPH-B4 RNA is different in different cell lines suggesting changes in transcriptional regulation during tumorigenesis no significant changes were seen in the EphrinA1. It was then necessary to determine protein expression of these receptors in the panel. Again we see significant differences in the expression of the EPOR and EPH-B4 receptors though they do not coincide with the RNA expression suggesting there is changes in post transcriptional regulation of these receptors in the cell lines. We then categorized these expression changes particularly with regard to the EPOR and EPH-B4 to then analyze the response to EPO treatment. We analyzed their response to chemotherapy in conjunction with EPO. We found that particularly in the HeyA8 ovarian cancer cell line that EPO was able to abrogate the apoptosis induced by docetaxel. It was then necessary to analyze the activation of signaling pathways known to be activated by these receptors in response to EPO treatment. Three cell lines were starved for two hours to isolate their response to EPO. Cell lines with higher expression (HeyA8 and HeyA8 MDR) of the EPOR demonstrated activation of the MAPK/ERK pathway while cell lines that expressed higher EPH-B4 (SKOV3ip1) demonstrated increased activation of the AKT and STAT5b signaling pathways. We then sought to determine a EPO dose that optimized its tumor promoting effect in vivo. Female nude mice were injected i.p. with HeyA8 MDR (positive for both EPOR and EPH-B4). At day eight the mice were treated with increasing doses of EPO (10, 50, 100 U) every two days. Treatment continued until tumors became evident, the mice were then sacrificed and the tumor weight was determined. We saw an increase in tumor weight as compared to control in the mice treated with 10 and 50 U EPO. The differential expression of EphB4 in cell lines as well as the activation of particular signaling pathways suggested that it would also mediate the tumor promoting effect in vivo. To determine this we again injected mice with HeyA8 MDR cell lines i.p. At day eight we began treatment with EPO (50 U 3.times. week) in conjunction with siRNA specific to EPH-B4 [sense: (SEQ ID NO: 266) 5'CAGCCAAUAGCCACUCUAA3'; antisense: (SEQ ID NO: 267) 5'UUAGAGUGGCUAUUGGCUG3']. As previously described EphB4 siRNA was able decrease tumor growth alone. Moreover, EPH-B4 siRNA also completely abrogated the EPO induced tumor growth.

Example 6

[0184] To further validate that EPHB4 is a novel EPO receptor a co-immunoprecipitation experiment was conducted using an anti-EPHB4 antibody to immuno-precipitate EPHB4 from cellular lysate.

[0185] In particular, cells (HeyA8 MDR and A2780cp20) were grown in RPMI-1640 supplemented with 15% fetal calf serum and gentamycin. At 70% confluency, the cells were treated with Epo (50U/ml) for 15 and 30 minutes. In addition, one group of cells were exposed to MG132 (10 .mu.M) for 30 minutes. Cell lysates were prepared after washing twice with cold-PBS and incubated in modified radioimmunoprecipitation assay buffer (RIPA). Protein concentrations were determined using a BCA Protein Assay Reagent kit (Pierce Biotechnology, Rockford, Ill.). For immunoprecipitation, 500 .mu.g of cell lysate was incubated with 6 .mu.l of primary antibody (EphB4-Abcam) overnight at 4.degree. C. Protein A Sepharose beads were added, and the mixture was incubated for 3 hours at 4.degree. C. Laemilli buffer was added to dislodge complexes from beads, and beads were separated by centrifugation at 3,500 g for 5 minutes at 4.degree. C. The supernatant were then used for immunoblot analysis. Supernatants were subjected to 8% SDS-PAGE separation. Samples transferred to a nitrocellulose membrane electrophoresis (Bio-Rad Laboratories, Hercules, Calif.) were incubated with EphB4 (Abeam Co.) and Epo (R & D Systems) antibodies overnight at 4.degree. C., detected with horseradish peroxidase (HRP)-conjugated anti-mouse/rabbit IgG (Amersham, Piscataway, N.J.), and developed using enhanced chemiluminescence detection kit (Pierce Biotechnology).

[0186] The results are provided in FIG. 19. The data clearly demonstrates a direct association between EPHB4 and EPO, suggesting tight functional coupling of both proteins.

Example 7

[0187] Radiolabelled EPO is capable of binding to independent cell-lines to various degrees. The capacity of EPHB4 to mediate such binding was investigated in three different cell-lines with varying degrees of EPHB4 and EPOR expression.

[0188] Cells (HeyA8, HeyA8 MDR and A2780cp20) were grown in RPMI-1640 supplemented with 15% fetal calf serum and gentamycin. Cells were transiently transfected with control siRNA [sense: 5'UUCUCCGAACGUUGUCACGU3' (SEQ ID NO: 264); antisense: 5'ACGUGACACGUUCGGAGAA3' (SEQ ID NO: 265)], EphB4 siRNA [sense: 5'CAGCCAAUAGCCACUCUAA3' (SEQ ID NO: 266); antisense: 5'UUAGAGUGGCUAUUGGCUG3' (SEQ ID NO: 267)] or EpoR siRNA[sense: 5'CCGAAGAGCUUCUGUGCUU3' (SEQ ID NO: 262); antisense: 5'AAGCACAGAAGCUCUUCGG3' (SEQ ID NO: 263)]. After 72 hours, the cells were detached with 0.1% EDTA. 1.times.10.sup.6 cells were diluted in 80 .mu.l of binding buffer (MEM+20 mM HEpes, Ph 7.4, 0.1% BSA). They were incubated with 7.5 mM .sup.125I-Epo at room temperature for 2.5 hours. Non-specific binding was determined by exposing the cells to 7.5 mM .sup.125I-Epo and cold-Epo (.times.200). The cells were washed with PBS and resuspended in cushion buffer (10% BSA in PBS). After centrifugation, the tubes were frozen in dry ice, and the pellet clipped and placed in scintillation fluid. Total binding was calculated by subtracting non-specific from total binding.

[0189] The results, provided in FIG. 20, demonstrate that EPHB4 is indeed responsible for the bulk of EPO binding in certain cell types (e.g. A2780cp20).

Example 8

[0190] To demonstrate that EPHB4 is responsible for mediating tumour cell survival and reduced patient outcome in response to EPO treatment, immunohistochemical analysis of EphB4 and EpoR was conducted on tumour samples from 71 patients with high grade and high stage epithelial ovarian cancer. All patients were previously treated with surgery followed by taxane-platinum chemotherapy and EPO therapy.

[0191] Specifically, immunohistochemical analysis of EphB4 and EpoR was conducted on 4 .mu.m-thick formalin-fixed paraffin-embedded epithelial ovarian cancer specimens. Slides were deparaffinized with xylene and decreasing concentrations of ethanol and rehydrated with PBS. Antigen retrival for EphB4 was performed using 1.times. Diva Decloaker (Biocare Medical, Concord, Calif.) under steam for 40 minutes followed by a 20 minute cool down at room temperature. Antigen retrival for EpoR was performed using 1.times. Borg Decloaker (Biocare Medical) under heat (125.degree. C.) and pressure for 4 minutes followed by a 60 minute cool down at room temperature. Following antigen retrival, all sections were washed with PBS. Endogenous peroxidases were blocked with 3% hydrogen peroxide in PBS for 12 minutes at room temperature followed by nonspecific protein blocking with either 5% BSA in TBST for 10 minutes at room temperature for EphB4 or 5% normal horse serum for 20 minutes at room temperature for EpoR. Sections were then incubated with primary antibody to EphB4 (mouse monoclonal anti-human, 1:500 dilution, Abcam, Cambridge, Mass.) or EpoR (biotinylated mouse monoclonal anti-human, 1:25 dilution, R&D Systems, Minneapolis, Minn.) in the respectively blocking solution overnight at 4.degree. C. Secondary amplification was performed using either the MACH4 polymer detection system (EphB4: Biocare Medical) or the 4plus Streptavidin AP label (EpoR: Biocare Medical). Visualization was achieved with 3,3'-diaminobezidine (DAB; Open Biosystems, Huntsville, Ala.). Slides were counterstained with Gill No. 2 hematoxylin (Sigma-Aldrich, St. Louis, Mo.), washed with PBS for 1 minute and mounted with Universal Mount (Research Genetics, Huntsville, Ala.). Clinical samples were scored for staining with the EphB4 and EpoR antibodies by a board-certified pathologist who was blinded to the clinical outcome of the patients. EphB4 and EpoR expression was determined semi-quantitatively by assessing the distribution of the positive cells and the staining intensity in the tumor cells. The distribution of positive cells was rated as follows: 0 points, no staining; 1 point, focal or <25%; 2 points, 25-50%, 3 points, 50-75%; 4 points, 75-100%. The staining intensity was rated as focal or weak (1 point), moderate (2 points) or heavy (3 points). Points for intensity and distribution were multiplied, and an overall score ranging from 0 to 12 was assigned. An overall score <3 was deemed negative and >3 positive.

[0192] The results are depicted in FIGS. 21-23. Overexpression of EPHB4, but not EPOR was found to correlate with poorer clinical outcome in response to EPO treatment. High levels of EPHB4 expression with low levels of EPOR showed the worst median survival (2.53 years), while low levels of EPHB4 and high levels of EPOR showed the best median survival (7.67 years). The data supports the need for a theranostic test to assess EPHB4 expression prior to, and/or during, an EPO treatment regimen.

Sequence CWU 1

1

2671508PRTHomo sapiens 1Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys 1 5 10 15 Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30 Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45 Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60 Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser 65 70 75 80 Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95 Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110 Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125 Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140 Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly 145 150 155 160 His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175 His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190 Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205 Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220 Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val 225 230 235 240 Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255 Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270 Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285 Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300 Gln Leu Trp Leu Tyr Gln Asn Asp Gly Cys Leu Trp Trp Ser Pro Cys 305 310 315 320 Thr Pro Phe Thr Glu Asp Pro Pro Ala Ser Leu Glu Val Leu Ser Glu 325 330 335 Arg Cys Trp Gly Thr Met Gln Ala Val Glu Pro Gly Thr Asp Asp Glu 340 345 350 Gly Pro Leu Leu Glu Pro Val Gly Ser Glu His Ala Gln Asp Thr Tyr 355 360 365 Leu Val Leu Asp Lys Trp Leu Leu Pro Arg Asn Pro Pro Ser Glu Asp 370 375 380 Leu Pro Gly Pro Gly Gly Ser Val Asp Ile Val Ala Met Asp Glu Gly 385 390 395 400 Ser Glu Ala Ser Ser Cys Ser Ser Ala Leu Ala Ser Lys Pro Ser Pro 405 410 415 Glu Gly Ala Ser Ala Ala Ser Phe Glu Tyr Thr Ile Leu Asp Pro Ser 420 425 430 Ser Gln Leu Leu Arg Pro Trp Thr Leu Cys Pro Glu Leu Pro Pro Thr 435 440 445 Pro Pro His Leu Lys Tyr Leu Tyr Leu Val Val Ser Asp Ser Gly Ile 450 455 460 Ser Thr Asp Tyr Ser Ser Gly Asp Ser Gln Gly Ala Gln Gly Gly Leu 465 470 475 480 Ser Asp Gly Pro Tyr Ser Asn Pro Tyr Glu Asn Ser Leu Ile Pro Ala 485 490 495 Ala Glu Pro Leu Pro Pro Ser Tyr Val Ala Cys Ser 500 505 2987PRTHomo sapiens 2Met Glu Leu Arg Val Leu Leu Cys Trp Ala Ser Leu Ala Ala Ala Leu 1 5 10 15 Glu Glu Thr Leu Leu Asn Thr Lys Leu Glu Thr Ala Asp Leu Lys Trp 20 25 30 Val Thr Phe Pro Gln Val Asp Gly Gln Trp Glu Glu Leu Ser Gly Leu 35 40 45 Asp Glu Glu Gln His Ser Val Arg Thr Tyr Glu Val Cys Asp Val Gln 50 55 60 Arg Ala Pro Gly Gln Ala His Trp Leu Arg Thr Gly Trp Val Pro Arg 65 70 75 80 Arg Gly Ala Val His Val Tyr Ala Thr Leu Arg Phe Thr Met Leu Glu 85 90 95 Cys Leu Ser Leu Pro Arg Ala Gly Arg Ser Cys Lys Glu Thr Phe Thr 100 105 110 Val Phe Tyr Tyr Glu Ser Asp Ala Asp Thr Ala Thr Ala Leu Thr Pro 115 120 125 Ala Trp Met Glu Asn Pro Tyr Ile Lys Val Asp Thr Val Ala Ala Glu 130 135 140 His Leu Thr Arg Lys Arg Pro Gly Ala Glu Ala Thr Gly Lys Val Asn 145 150 155 160 Val Lys Thr Leu Arg Leu Gly Pro Leu Ser Lys Ala Gly Phe Tyr Leu 165 170 175 Ala Phe Gln Asp Gln Gly Ala Cys Met Ala Leu Leu Ser Leu His Leu 180 185 190 Phe Tyr Lys Lys Cys Ala Gln Leu Thr Val Asn Leu Thr Arg Phe Pro 195 200 205 Glu Thr Val Pro Arg Glu Leu Val Val Pro Val Ala Gly Ser Cys Val 210 215 220 Val Asp Ala Val Pro Ala Pro Gly Pro Ser Pro Ser Leu Tyr Cys Arg 225 230 235 240 Glu Asp Gly Gln Trp Ala Glu Gln Pro Val Thr Gly Cys Ser Cys Ala 245 250 255 Pro Gly Phe Glu Ala Ala Glu Gly Asn Thr Lys Cys Arg Ala Cys Ala 260 265 270 Gln Gly Thr Phe Lys Pro Leu Ser Gly Glu Gly Ser Cys Gln Pro Cys 275 280 285 Pro Ala Asn Ser His Ser Asn Thr Ile Gly Ser Ala Val Cys Gln Cys 290 295 300 Arg Val Gly Tyr Phe Arg Ala Arg Thr Asp Pro Arg Gly Ala Pro Cys 305 310 315 320 Thr Thr Pro Pro Ser Ala Pro Arg Ser Val Val Ser Arg Leu Asn Gly 325 330 335 Ser Ser Leu His Leu Glu Trp Ser Ala Pro Leu Glu Ser Gly Gly Arg 340 345 350 Glu Asp Leu Thr Tyr Ala Leu Arg Cys Arg Glu Cys Arg Pro Gly Gly 355 360 365 Ser Cys Ala Pro Cys Gly Gly Asp Leu Thr Phe Asp Pro Gly Pro Arg 370 375 380 Asp Leu Val Glu Pro Trp Val Val Val Arg Gly Leu Arg Pro Asp Phe 385 390 395 400 Thr Tyr Thr Phe Glu Val Thr Ala Leu Asn Gly Val Ser Ser Leu Ala 405 410 415 Thr Gly Pro Val Pro Phe Glu Pro Val Asn Val Thr Thr Asp Arg Glu 420 425 430 Val Pro Pro Ala Val Ser Asp Ile Arg Val Thr Arg Ser Ser Pro Ser 435 440 445 Ser Leu Ser Leu Ala Trp Ala Val Pro Arg Ala Pro Ser Gly Ala Val 450 455 460 Leu Asp Tyr Glu Val Lys Tyr His Glu Lys Gly Ala Glu Gly Pro Ser 465 470 475 480 Ser Val Arg Phe Leu Lys Thr Ser Glu Asn Arg Ala Glu Leu Arg Gly 485 490 495 Leu Lys Arg Gly Ala Ser Tyr Leu Val Gln Val Arg Ala Arg Ser Glu 500 505 510 Ala Gly Tyr Gly Pro Phe Gly Gln Glu His His Ser Gln Thr Gln Leu 515 520 525 Asp Glu Ser Glu Gly Trp Arg Glu Gln Leu Ala Leu Ile Ala Gly Thr 530 535 540 Ala Val Val Gly Val Val Leu Val Leu Val Val Ile Val Val Ala Val 545 550 555 560 Leu Cys Leu Arg Lys Gln Ser Asn Gly Arg Glu Ala Glu Tyr Ser Asp 565 570 575 Lys His Gly Gln Tyr Leu Ile Gly His Gly Thr Lys Val Tyr Ile Asp 580 585 590 Pro Phe Thr Tyr Glu Asp Pro Asn Glu Ala Val Arg Glu Phe Ala Lys 595 600 605 Glu Ile Asp Val Ser Tyr Val Lys Ile Glu Glu Val Ile Gly Ala Gly 610 615 620 Glu Phe Gly Glu Val Cys Arg Gly Arg Leu Lys Ala Pro Gly Lys Lys 625 630 635 640 Glu Ser Cys Val Ala Ile Lys Thr Leu Lys Gly Gly Tyr Thr Glu Arg 645 650 655 Gln Arg Arg Glu Phe Leu Ser Glu Ala Ser Ile Met Gly Gln Phe Glu 660 665 670 His Pro Asn Ile Ile Arg Leu Glu Gly Val Val Thr Asn Ser Met Pro 675 680 685 Val Met Ile Leu Thr Glu Phe Met Glu Asn Gly Ala Leu Asp Ser Phe 690 695 700 Leu Arg Leu Asn Asp Gly Gln Phe Thr Val Ile Gln Leu Val Gly Met 705 710 715 720 Leu Arg Gly Ile Ala Ser Gly Met Arg Tyr Leu Ala Glu Met Ser Tyr 725 730 735 Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Asn Ser Asn Leu 740 745 750 Val Cys Lys Val Ser Asp Phe Gly Leu Ser Arg Phe Leu Glu Glu Asn 755 760 765 Ser Ser Asp Pro Thr Tyr Thr Ser Ser Leu Gly Gly Lys Ile Pro Ile 770 775 780 Arg Trp Thr Ala Pro Glu Ala Ile Ala Phe Arg Lys Phe Thr Ser Ala 785 790 795 800 Ser Asp Ala Trp Ser Tyr Gly Ile Val Met Trp Glu Val Met Ser Phe 805 810 815 Gly Glu Arg Pro Tyr Trp Asp Met Ser Asn Gln Asp Val Ile Asn Ala 820 825 830 Ile Glu Gln Asp Tyr Arg Leu Pro Pro Pro Pro Asp Cys Pro Thr Ser 835 840 845 Leu His Gln Leu Met Leu Asp Cys Trp Gln Lys Asp Arg Asn Ala Arg 850 855 860 Pro Arg Phe Pro Gln Val Val Ser Ala Leu Asp Lys Met Ile Arg Asn 865 870 875 880 Pro Ala Ser Leu Lys Ile Val Ala Arg Glu Asn Gly Gly Ala Ser His 885 890 895 Pro Leu Leu Asp Gln Arg Gln Pro His Tyr Ser Ala Phe Gly Ser Val 900 905 910 Gly Glu Trp Leu Arg Ala Ile Lys Met Gly Arg Tyr Glu Glu Ser Phe 915 920 925 Ala Ala Ala Gly Phe Gly Ser Phe Glu Leu Val Ser Gln Ile Ser Ala 930 935 940 Glu Asp Leu Leu Arg Ile Gly Val Thr Leu Ala Gly His Gln Lys Lys 945 950 955 960 Ile Leu Ala Ser Val Gln His Met Lys Ser Gln Ala Lys Pro Gly Thr 965 970 975 Pro Gly Gly Thr Gly Gly Pro Ala Pro Gln Tyr 980 985 3205PRTHomo sapiens 3Met Glu Phe Leu Trp Ala Pro Leu Leu Gly Leu Cys Cys Ser Leu Ala 1 5 10 15 Ala Ala Asp Arg His Thr Val Phe Trp Asn Ser Ser Asn Pro Lys Phe 20 25 30 Arg Asn Glu Asp Tyr Thr Ile His Val Gln Leu Asn Asp Tyr Val Asp 35 40 45 Ile Ile Cys Pro His Tyr Glu Asp His Ser Val Ala Asp Ala Ala Met 50 55 60 Glu Gln Tyr Ile Leu Tyr Leu Val Glu His Glu Glu Tyr Gln Leu Cys 65 70 75 80 Gln Pro Gln Ser Lys Asp Gln Val Arg Trp Gln Cys Asn Arg Pro Ser 85 90 95 Ala Lys His Gly Pro Glu Lys Leu Ser Glu Lys Phe Gln Arg Phe Thr 100 105 110 Pro Phe Thr Leu Gly Lys Glu Phe Lys Glu Gly His Ser Tyr Tyr Tyr 115 120 125 Ile Ser Lys Pro Ile His Gln His Glu Asp Arg Cys Leu Arg Leu Lys 130 135 140 Val Thr Val Ser Gly Lys Ile Thr His Ser Pro Gln Ala His Asp Asn 145 150 155 160 Pro Gln Glu Lys Arg Leu Ala Ala Asp Asp Pro Glu Val Arg Val Leu 165 170 175 His Ser Ile Gly His Ser Ala Ala Pro Arg Leu Phe Pro Leu Ala Trp 180 185 190 Thr Val Leu Leu Leu Pro Leu Leu Leu Leu Gln Thr Pro 195 200 205 4333PRTHomo sapiens 4Met Ala Val Arg Arg Asp Ser Val Trp Lys Tyr Cys Trp Gly Val Leu 1 5 10 15 Met Val Leu Cys Arg Thr Ala Ile Ser Lys Ser Ile Val Leu Glu Pro 20 25 30 Ile Tyr Trp Asn Ser Ser Asn Ser Lys Phe Leu Pro Gly Gln Gly Leu 35 40 45 Val Leu Tyr Pro Gln Ile Gly Asp Lys Leu Asp Ile Ile Cys Pro Lys 50 55 60 Val Asp Ser Lys Thr Val Gly Gln Tyr Glu Tyr Tyr Lys Val Tyr Met 65 70 75 80 Val Asp Lys Asp Gln Ala Asp Arg Cys Thr Ile Lys Lys Glu Asn Thr 85 90 95 Pro Leu Leu Asn Cys Ala Lys Pro Asp Gln Asp Ile Lys Phe Thr Ile 100 105 110 Lys Phe Gln Glu Phe Ser Pro Asn Leu Trp Gly Leu Glu Phe Gln Lys 115 120 125 Asn Lys Asp Tyr Tyr Ile Ile Ser Thr Ser Asn Gly Ser Leu Glu Gly 130 135 140 Leu Asp Asn Gln Glu Gly Gly Val Cys Gln Thr Arg Ala Met Lys Ile 145 150 155 160 Leu Met Lys Val Gly Gln Asp Ala Ser Ser Ala Gly Ser Thr Arg Asn 165 170 175 Lys Asp Pro Thr Arg Arg Pro Glu Leu Glu Ala Gly Thr Asn Gly Arg 180 185 190 Ser Ser Thr Thr Ser Pro Phe Val Lys Pro Asn Pro Gly Ser Ser Thr 195 200 205 Asp Gly Asn Ser Ala Gly His Ser Gly Asn Asn Ile Leu Gly Ser Glu 210 215 220 Val Ala Leu Phe Ala Gly Ile Ala Ser Gly Cys Ile Ile Phe Ile Val 225 230 235 240 Ile Ile Ile Thr Leu Val Val Leu Leu Leu Lys Tyr Arg Arg Arg His 245 250 255 Arg Lys His Ser Pro Gln His Thr Thr Thr Leu Ser Leu Ser Thr Leu 260 265 270 Ala Thr Pro Lys Arg Ser Gly Asn Asn Asn Gly Ser Glu Pro Ser Asp 275 280 285 Ile Ile Ile Pro Leu Arg Thr Ala Asp Ser Val Phe Cys Pro His Tyr 290 295 300 Glu Lys Val Ser Gly Asp Tyr Gly His Pro Val Tyr Ile Val Gln Glu 305 310 315 320 Met Pro Pro Gln Ser Pro Ala Asn Ile Tyr Tyr Lys Val 325 330 51849DNAHomo sapiens 5acttagaggc gcctggtcgg gaagggcctg gtcagctgcg tccggcggag gcagctgctg 60acccagctgt ggactgtgcc gggggcgggg gacggagggg caggagccct gggctccccg 120tggcgggggc tgtatcatgg accacctcgg ggcgtccctc tggccccagg tcggctccct 180ttgtctcctg ctcgctgggg ccgcctgggc gcccccgcct aacctcccgg accccaagtt 240cgagagcaaa gcggccttgc tggcggcccg ggggcccgaa gagcttctgt gcttcaccga 300gcggttggag gacttggtgt gtttctggga ggaagcggcg agcgctgggg tgggcccggg 360caactacagc ttctcctacc agctcgagga tgagccatgg aagctgtgtc gcctgcacca 420ggctcccacg gctcgtggtg cggtgcgctt ctggtgttcg ctgcctacag ccgacacgtc 480gagcttcgtg cccctagagt tgcgcgtcac agcagcctcc ggcgctccgc gatatcaccg 540tgtcatccac atcaatgaag tagtgctcct agacgccccc gtggggctgg tggcgcggtt 600ggctgacgag agcggccacg tagtgttgcg ctggctcccg ccgcctgaga cacccatgac 660gtctcacatc cgctacgagg tggacgtctc ggccggcaac ggcgcaggga gcgtacagag 720ggtggagatc ctggagggcc gcaccgagtg tgtgctgagc aacctgcggg gccggacgcg 780ctacaccttc gccgtccgcg cgcgtatggc tgagccgagc ttcggcggct tctggagcgc 840ctggtcggag cctgtgtcgc tgctgacgcc tagcgacctg gaccccctca tcctgacgct 900ctccctcatc ctcgtggtca tcctggtgct gctgaccgtg ctcgcgctgc tctcccaccg 960ccgggctctg aagcagaaga tctggcctgg catcccgagc ccagagagcg agtttgaagg 1020cctcttcacc acccacaagg gtaacttcca gctgtggctg taccagaatg atggctgcct 1080gtggtggagc ccctgcaccc ccttcacgga ggacccacct gcttccctgg aagtcctctc 1140agagcgctgc tgggggacga tgcaggcagt ggagccgggg acagatgatg agggccccct 1200gctggagcca gtgggcagtg agcatgccca ggatacctat ctggtgctgg acaaatggtt 1260gctgccccgg aacccgccca gtgaggacct cccagggcct ggtggcagtg tggacatagt 1320ggccatggat gaaggctcag aagcatcctc ctgctcatct gctttggcct cgaagcccag 1380cccagaggga gcctctgctg ccagctttga gtacactatc ctggacccca gctcccagct 1440cttgcgtcca tggacactgt gccctgagct gccccctacc ccaccccacc taaagtacct 1500gtaccttgtg gtatctgact ctggcatctc aactgactac agctcagggg actcccaggg 1560agcccaaggg ggcttatccg atggccccta ctccaaccct tatgagaaca gccttatccc 1620agccgctgag cctctgcccc ccagctatgt ggcttgctct taggacacca ggctgcagat 1680gatcagggat ccaatatgac tcagagaacc

agtgcagact caagacttat ggaacaggga 1740tggcgaggcc tctctcagga gcaggggcat tgctgatttt gtctgcccaa tccatcctgc 1800tcaggaaacc acaaccttgc agtattttta aatatgtata gtttttttg 184964369DNAHomo sapiens 6ttccagcgca gctcagcccc tgcccggccc ggcccgcccg gctccgcgcc gcagtctccc 60tccctcccgc tccgtccccg ctcgggctcc caccatcccc gcccgcgagg agagcactcg 120gcccggcggc gcgagcagag ccactccagg gaggggggga gaccgcgagc ggccggctca 180gcccccgcca cccggggcgg gaccccgagg ccccggaggg accccaactc cagccacgtc 240ttgctgcgcg cccgcccggc gcggccactg ccagcacgct ccgggcccgc cgcccgcgcg 300cgcggcacag acgcggggcc acacttggcg ccgccgcccg gtgccccgca cgctcgcatg 360ggcccgcgct gagggccccg acgaggagtc ccgcgcggag tatcggcgtc cacccgccca 420gggagagtca gacctggggg ggcgagggcc ccccaaactc agttcggatc ctacccgagt 480gaggcggcgc catggagctc cgggtgctgc tctgctgggc ttcgttggcc gcagctttgg 540aagagaccct gctgaacaca aaattggaaa ctgctgatct gaagtgggtg acattccctc 600aggtggacgg gcagtgggag gaactgagcg gcctggatga ggaacagcac agcgtgcgca 660cctacgaagt gtgtgacgtg cagcgtgccc cgggccaggc ccactggctt cgcacaggtt 720gggtcccacg gcggggcgcc gtccacgtgt acgccacgct gcgcttcacc atgctcgagt 780gcctgtccct gcctcgggct gggcgctcct gcaaggagac cttcaccgtc ttctactatg 840agagcgatgc ggacacggcc acggccctca cgccagcctg gatggagaac ccctacatca 900aggtggacac ggtggccgcg gagcatctca cccggaagcg ccctggggcc gaggccaccg 960ggaaggtgaa tgtcaagacg ctgcgtctgg gaccgctcag caaggctggc ttctacctgg 1020ccttccagga ccagggtgcc tgcatggccc tgctatccct gcacctcttc tacaaaaagt 1080gcgcccagct gactgtgaac ctgactcgat tcccggagac tgtgcctcgg gagctggttg 1140tgcccgtggc cggtagctgc gtggtggatg ccgtccccgc ccctggcccc agccccagcc 1200tctactgccg tgaggatggc cagtgggccg aacagccggt cacgggctgc agctgtgctc 1260cggggttcga ggcagctgag gggaacacca agtgccgagc ctgtgcccag ggcaccttca 1320agcccctgtc aggagaaggg tcctgccagc catgcccagc caatagccac tctaacacca 1380ttggatcagc cgtctgccag tgccgcgtcg ggtacttccg ggcacgcaca gacccccggg 1440gtgcaccctg caccacccct ccttcggctc cgcggagcgt ggtttcccgc ctgaacggct 1500cctccctgca cctggaatgg agtgcccccc tggagtctgg tggccgagag gacctcacct 1560acgccctccg ctgccgggag tgccgacccg gaggctcctg tgcgccctgc gggggagacc 1620tgacttttga ccccggcccc cgggacctgg tggagccctg ggtggtggtt cgagggctac 1680gtcctgactt cacctatacc tttgaggtca ctgcattgaa cggggtatcc tccttagcca 1740cggggcccgt cccatttgag cctgtcaatg tcaccactga ccgagaggta cctcctgcag 1800tgtctgacat ccgggtgacg cggtcctcac ccagcagctt gagcctggcc tgggctgttc 1860cccgggcacc cagtggggct gtgctggact acgaggtcaa ataccatgag aagggcgccg 1920agggtcccag cagcgtgcgg ttcctgaaga cgtcagaaaa ccgggcagag ctgcgggggc 1980tgaagcgggg agccagctac ctggtgcagg tacgggcgcg ctctgaggcc ggctacgggc 2040ccttcggcca ggaacatcac agccagaccc aactggatga gagcgagggc tggcgggagc 2100agctggccct gattgcgggc acggcagtcg tgggtgtggt cctggtcctg gtggtcattg 2160tggtcgcagt tctctgcctc aggaagcaga gcaatgggag agaagcagaa tattcggaca 2220aacacggaca gtatctcatc ggacatggta ctaaggtcta catcgacccc ttcacttatg 2280aagaccctaa tgaggctgtg agggaatttg caaaagagat cgatgtctcc tacgtcaaga 2340ttgaagaggt gattggtgca ggtgagtttg gcgaggtgtg ccgggggcgg ctcaaggccc 2400cagggaagaa ggagagctgt gtggcaatca agaccctgaa gggtggctac acggagcggc 2460agcggcgtga gtttctgagc gaggcctcca tcatgggcca gttcgagcac cccaatatca 2520tccgcctgga gggcgtggtc accaacagca tgcccgtcat gattctcaca gagttcatgg 2580agaacggcgc cctggactcc ttcctgcggc taaacgacgg acagttcaca gtcatccagc 2640tcgtgggcat gctgcggggc atcgcctcgg gcatgcggta ccttgccgag atgagctacg 2700tccaccgaga cctggctgct cgcaacatcc tagtcaacag caacctcgtc tgcaaagtgt 2760ctgactttgg cctttcccga ttcctggagg agaactcttc cgatcccacc tacacgagct 2820ccctgggagg aaagattccc atccgatgga ctgccccgga ggccattgcc ttccggaagt 2880tcacttccgc cagtgatgcc tggagttacg ggattgtgat gtgggaggtg atgtcatttg 2940gggagaggcc gtactgggac atgagcaatc aggacgtgat caatgccatt gaacaggact 3000accggctgcc cccgccccca gactgtccca cctccctcca ccagctcatg ctggactgtt 3060ggcagaaaga ccggaatgcc cggccccgct tcccccaggt ggtcagcgcc ctggacaaga 3120tgatccggaa ccccgccagc ctcaaaatcg tggcccggga gaatggcggg gcctcacacc 3180ctctcctgga ccagcggcag cctcactact cagcttttgg ctctgtgggc gagtggcttc 3240gggccatcaa aatgggaaga tacgaagaaa gtttcgcagc cgctggcttt ggctccttcg 3300agctggtcag ccagatctct gctgaggacc tgctccgaat cggagtcact ctggcgggac 3360accagaagaa aatcttggcc agtgtccagc acatgaagtc ccaggccaag ccgggaaccc 3420cgggtgggac aggaggaccg gccccgcagt actgacctgc aggaactccc caccccaggg 3480acaccgcctc cccattttcc ggggcagagt ggggactcac agaggccccc agccctgtgc 3540cccgctggat tgcactttga gcccgtgggg tgaggagttg gcaatttgga gagacaggat 3600ttgggggttc tgccataata ggaggggaaa atcacccccc agccacctcg gggaactcca 3660gaccaagggt gagggcgcct ttccctcagg actgggtgtg accagaggaa aaggaagtgc 3720ccaacatctc ccagcctccc caggtgcccc cctcaccttg atgggtgcgt tcccgcagac 3780caaagagagt gtgactccct tgccagctcc agagtggggg ggctgtccca gggggcaaga 3840aggggtgtca gggcccagtg acaaaatcat tggggtttgt agtcccaact tgctgctgtc 3900accaccaaac tcaatcattt ttttcccttg taaatgcccc tcccccagct gctgccttca 3960tattgaaggt ttttgagttt tgtttttggt cttaattttt ctccccgttc cctttttgtt 4020tcttcgtttt gtttttctac cgtccttgtc ataactttgt gttggaggga acctgtttca 4080ctatggcctc ctttgcccaa gttgaaacag gggcccatca tcatgtctgt ttccagaaca 4140gtgccttggt catcccacat ccccggaccc cgcctgggac ccccaagctg tgtcctatga 4200aggggtgtgg ggtgaggtag tgaaaagggc ggtagttggt ggtggaaccc agaaacggac 4260gccggtgctt ggaggggttc ttaaattata tttaaaaaag taactttttg tataaataaa 4320agaaaatggg acgtgtccca gctccagggg taaaaaaaaa aaaaaaaaa 436971590DNAHomo sapiens 7gccagatctg tgagcccagc gctgactgcg ccgcggagaa agccagtggg aacccagacc 60cataggagac ccgcgtcccc gctcggcctg gccaggcccc gcgctatgga gttcctctgg 120gcccctctct tgggtctgtg ctgcagtctg gccgctgctg atcgccacac cgtcttctgg 180aacagttcaa atcccaagtt ccggaatgag gactacacca tacatgtgca gctgaatgac 240tacgtggaca tcatctgtcc gcactatgaa gatcactctg tggcagacgc tgccatggag 300cagtacatac tgtacctggt ggagcatgag gagtaccagc tgtgccagcc ccagtccaag 360gaccaagtcc gctggcagtg caaccggccc agtgccaagc atggcccgga gaagctgtct 420gagaagttcc agcgcttcac acctttcacc ctgggcaagg agttcaaaga aggacacagc 480tactactaca tctccaaacc catccaccag catgaagacc gctgcttgag gttgaaggtg 540actgtcagtg gcaaaatcac tcacagtcct caggcccatg acaatccaca ggagaagaga 600cttgcagcag atgacccaga ggtgcgggtt ctacatagca tcggtcacag tgctgcccca 660cgcctcttcc cacttgcctg gactgtgctg ctccttccac ttctgctgct gcaaaccccg 720tgaaggtgta tgccacacct ggccttaaag agggacaggc tgaagagagg gacaggcact 780ccaaacctgt cttggggcca ctttcagagc ccccagccct gggaaccact cccaccacag 840gcataagcta tcacctagca gcctcaaaac gggtcagtat taaggttttc aaccggaagg 900aggccaacca gcccgacagt gccatcccca ccttcacctc ggagggatgg agaaagaagt 960ggagacagtc ctttcccacc attcctgcct ttaagccaaa gaaacaagct gtgcaggcat 1020ggtcccttaa ggcacagtgg gagctgagct ggaaggggcc acgtggatgg gcaaagcttg 1080tcaaagatgc cccctccagg agagagccag gatgcccaga tgaactgact gaaggaaaag 1140caagaaacag tttcttgctt ggaagccagg tacaggagag gcagcatgct tgggctgacc 1200cagcatctcc cagcaagacc tcatctgtgg agctgccaca gagaagtttg tagccaggta 1260ctgcattctc tcccatcctg gggcagcact ccccagagct gtgccagcag gggggctgtg 1320ccaacctgtt cttagagtgt agctgtaagg gcagtgccca tgtgtacatt ctgcctagag 1380tgtagcctaa agggcagggc ccacgtgtat agtatctgta tataagttgc tgtgtgtctg 1440tcctgatttc tacaactgga gtttttttat acaatgttct ttgtctcaaa ataaagcaat 1500gtgttttttc ggacatgctt ttctgccact ccatattaaa acatatgacc attgagtccc 1560tgctaaaaaa aaaaaaaaaa aaaaaaaaaa 159084335DNAHomo sapiens 8gcgcggagct gggagtggct tcgccatggc tgtgagaagg gactccgtgt ggaagtactg 60ctggggtgtt ttgatggttt tatgcagaac tgcgatttcc aaatcgatag ttttagagcc 120tatctattgg aattcctcga actccaaatt tctacctgga caaggactgg tactataccc 180acagatagga gacaaattgg atattatttg ccccaaagtg gactctaaaa ctgttggcca 240gtatgaatat tataaagttt atatggttga taaagaccaa gcagacagat gcactattaa 300gaaggaaaat acccctctcc tcaactgtgc caaaccagac caagatatca aattcaccat 360caagtttcaa gaattcagcc ctaacctctg gggtctagaa tttcagaaga acaaagatta 420ttacattata tctacatcaa atgggtcttt ggagggcctg gataaccagg agggaggggt 480gtgccagaca agagccatga agatcctcat gaaagttgga caagatgcaa gttctgctgg 540atcaaccagg aataaagatc caacaagacg tccagaacta gaagctggta caaatggaag 600aagttcgaca acaagtccct ttgtaaaacc aaatccaggt tctagcacag acggcaacag 660cgccggacat tcggggaaca acatcctcgg ttccgaagtg gccttatttg cagggattgc 720ttcaggatgc atcatcttca tcgtcatcat catcacgctg gtggtcctct tgctgaagta 780ccggaggaga cacaggaagc actcgccgca gcacacgacc acgctgtcgc tcagcacact 840ggccacaccc aagcgcagcg gcaacaacaa cggctcagag cccagtgaca ttatcatccc 900gctaaggact gcggacagcg tcttctgccc tcactacgag aaggtcagcg gggactacgg 960gcacccggtg tacatcgtcc aggagatgcc cccgcagagc ccggcgaaca tttactacaa 1020ggtctgagag ggaccctggt ggtacctgtg ctttcccaga ggacacctaa tgtcccgatg 1080cctcccttga gggtttgaga gcccgcgtgc tggagaattg actgaagcac agcaccgggg 1140gagagggaca ctcctcctcg gaagagcccg tcgcgctgga cagcttacct agtcttgtag 1200cattcggcct tggtgaacac acacgctccc tggaagctgg aagactgtgc agaagacgcc 1260cattcggact gctgtgccgc gtcccacgtc tcctcctcga agccatgtgc tgcggtcact 1320caggcctctg cagaagccaa gggaagacag tggtttgtgg acgagagggc tgtgagcatc 1380ctggcaggtg ccccaggatg ccacgcctgg aagggccggc ttctgcctgg ggtgcatttc 1440ccccgcagtg cataccggac ttgtcacacg gacctcgggc tagttaaggt gtgcaaagat 1500ctctagagtt tagtccttac tgtctcactc gttctgttac ccagggctct gcagcacctc 1560acctgagacc tccactccac atctgcatca ctcatggaac actcatgtct ggagtcccct 1620cctccagccg ctggcaacaa cagcttcagt ccatgggtaa tccgttcata gaaattgtgt 1680ttgctaacaa ggtgcccttt agccagatgc taggctgtct gcgaagaagg ctaggagttc 1740atagaaggga gtggggctgg ggaaagggct ggctgcaatt gcagctcact gctgctgcct 1800ctgaaacaga aagttggaaa ggaaaaaaga aaaaagcaat taggtagcac agcactttgg 1860ttttgctgag atcgaagagg ccagtaggag acacgacagc acacacagtg gattccagtg 1920catggggagg cactcgctgt tatcaaatag cgatgtgcag gaagaaaagc ccctcttcat 1980tccggggaac aaagacgggt attgttggga aaggaacagg cttggaggga agggagaaag 2040taggccgctg atgatatatt cgggcaggac tgttgtggta ctggcaataa gatacacagc 2100tccgagctgt aggagagtcg gtctgctttg gatgattttt taagcagact cagctgctat 2160acttatcaca ttttattaaa cacagggaaa gcatttagga gaatagcaga gagccaaatc 2220tgacctaaaa gttgaaaagc caaaggtcaa acaggctgta attccatcat catcgttgtt 2280attaaagaat ccttatctat aaaaggtagg tcagatcccc ctccccccag gttcctcctt 2340cccctcccga ttgagcctta cgacactttg gtttatgcgg tgctgtccgg gtgccagggc 2400tgcagggtcg gtactgatgg aggctgcagc gcccggtgct ctgtgtcaag gtgaagcaca 2460tacggcagac ctcttagagt ccttaagacg gaagtaaatt atgatgtcca gggggagaag 2520gaagatagga cgtatttata ataggtatat agaacacaag ggatataaaa tgaaagattt 2580ttactaatat atattttaag gttgcacaca gtacacacca gaagatgtga aattcatttg 2640tggcaattaa gtggtcccaa tgctcagcgc ttaaaaaaac aaattggaca gctacttctg 2700ggaaaaacaa catcattcca aaaagaacaa taatgagagc aaatgcaaaa ataaccaagt 2760cctccgaagg catctcacgg aaccgtagac taggaagtac gagccccaca gagcaggaag 2820ccgatgtgac tgcatcatat atttaacaat gacaagatgt tccggcgttt atttctgcgt 2880tgggttttcc cttgccttat gggctgaagt gttctctaga atccagcagg tcacactggg 2940ggcttcaggt gacgatttag ctgtggctcc ctcctcctgt cctcccccgc accccctccc 3000ttctgggaaa caagaagagt aaacaggaaa cctacttttt atgtgctatg caaaatagac 3060atctttaaca tagtcctgtt actatggtaa cactttgctt tctgaattgg aagggaaaaa 3120aaatgtagcg acagcatttt aaggttctca gacctccagt gagtacctgc aaaaatgagt 3180tgtcacagaa attatgatcc tctatttcct gaacctggaa atgatgttgg tccaaagtgc 3240gtgtgtgtat gtgtgagtgg gtgcgtggta tacatgtgta catatatgta taatatatat 3300ctacaatata tattatatat atctatatca tatttctgtg gagggttgcc atggtaacca 3360gccacagtac atatgtaatt ctttccatca ccccaacctc tcctttctgt gcattcatgc 3420aagagtttct tgtaagccat cagaagttac ttttaggatg ggggagaggg gcgagaaggg 3480gaaaaatggg aaatagtctg attttaatga aatcaaatgt atgtatcatc agttggctac 3540gttttggttc tatgctaaac tgtgaaaaat cagatgaatt gataaaagag ttccctgcaa 3600ccaattgaaa agtgttctgt gcgtctgttt tgtgtctggt gcagaatatg acaatctacc 3660aactgtccct ttgtttgaag ttggtttagc tttggaaagt tactgtaaat gccttgcttg 3720tatgatcgtc cctggtcacc cgactttgga atttgcacca tcatgtttca gtgaagatgc 3780tgtaaatagg ttcagatttt actgtctatg gatttggggt gttacagtag ccttattcac 3840ctttttaata aaaatacaca tgaaaacaag aaagaaatgg cttttcttac ccagattgtg 3900tacatagagc aatgttggtt ttttataaag tctaagcaag atgttttgta taaaatctga 3960attttgcaat gtatttagct acagcttgtt taacggcagt gtcattcccc tttgcactgt 4020aatgaggaaa aaatggtata aaaggttgcc aaattgctgc atatttgtgc cgtaattatg 4080taccatgaat atttatttaa aatttcgttg tccaatttgt aagtaacaca gtattatgcc 4140tgagttataa atattttttt ctttctttgt tttattttaa tagcctgtca taggttttaa 4200atctgcttta gtttcacatt gcagttagcc ccagaaaatg aaatccgtga agtcacattc 4260cacatctgtt tcaaactgaa tttgttctta aaaaaataaa atattttttt cctatggaaa 4320aaaaaaaaaa aaaaa 43359642DNAHomo sapiens 9agcaaagcgg ccttgctggc ggcccggggg cccgaagagc ttctgtgctt caccgagcgg 60ttggaggact tggtgtgttt ctgggaggaa gcggcgagcg ctggggtggg cccgggcaac 120tacagcttct cctaccagct cgaggatgag ccatggaagc tgtgtcgcct gcaccaggct 180cccacggctc gtggtgcggt gcgcttctgg tgttcgctgc ctacagccga cacgtcgagc 240ttcgtgcccc tagagttgcg cgtcacagca gcctccggcg ctccgcgata tcaccgtgtc 300atccacatca atgaagtagt gctcctagac gcccccgtgg ggctggtggc gcggttggct 360gacgagagcg gccacgtagt gttgcgctgg ctcccgccgc ctgagacacc catgacgtct 420cacatccgct acgaggtgga cgtctcggcc ggcaacggcg cagggagcgt acagagggtg 480gagatcctgg agggccgcac cgagtgtgtg ctgagcaacc tgcggggccg gacgcgctac 540accttcgccg tccgcgcgcg tatggctgag ccgagcttcg gcggcttctg gagcgcctgg 600tcggagcctg tgtcgctgct gacgcctagc gacctggacc cc 64210651DNAHomo sapiens 10ccttcggctc cgcggagcgt ggtttcccgc ctgaacggct cctccctgca cctggaatgg 60agtgcccccc tggagtctgg tggccgagag gacctcacct acgccctccg ctgccgggag 120tgccgacccg gaggctcctg tgcgccctgc gggggagacc tgacttttga ccccggcccc 180cgggacctgg tggagccctg ggtggtggtt cgagggctac gtcctgactt cacctatacc 240tttgaggtca ctgcattgaa cggggtatcc tccttagcca cggggcccgt cccatttgag 300cctgtcaatg tcaccactga ccgagaggta cctcctgcag tgtctgacat ccgggtgacg 360cggtcctcac ccagcagctt gagcctggcc tgggctgttc cccgggcacc cagtggggct 420gtgctggact acgaggtcaa ataccatgag aagggcgccg agggtcccag cagcgtgcgg 480ttcctgaaga cgtcagaaaa ccgggcagag ctgcgggggc tgaagcgggg agccagctac 540ctggtgcagg tacgggcgcg ctctgaggcc ggctacgggc ccttcggcca ggaacatcac 600agccagaccc aactggatga gagcgagggc tggcgggagc agctggccct g 65111417DNAHomo sapiens 11ctggccgctg ctgatcgcca caccgtcttc tggaacagtt caaatcccaa gttccggaat 60gaggactaca ccatacatgt gcagctgaat gactacgtgg acatcatctg tccgcactat 120gaagatcact ctgtggcaga cgctgccatg gagcagtaca tactgtacct ggtggagcat 180gaggagtacc agctgtgcca gccccagtcc aaggaccaag tccgctggca gtgcaaccgg 240cccagtgcca agcatggccc ggagaagctg tctgagaagt tccagcgctt cacacctttc 300accctgggca aggagttcaa agaaggacac agctactact acatctccaa acccatccac 360cagcatgaag accgctgctt gaggttgaag gtgactgtca gtggcaaaat cactcac 41712426DNAHomo sapiens 12tccaaatcga tagttttaga gcctatctat tggaattcct cgaactccaa atttctacct 60ggacaaggac tggtactata cccacagata ggagacaaat tggatattat ttgccccaaa 120gtggactcta aaactgttgg ccagtatgaa tattataaag tttatatggt tgataaagac 180caagcagaca gatgcactat taagaaggaa aatacccctc tcctcaactg tgccaaacca 240gaccaagata tcaaattcac catcaagttt caagaattca gccctaacct ctggggtcta 300gaatttcaga agaacaaaga ttattacatt atatctacat caaatgggtc tttggagggc 360ctggataacc aggagggagg ggtgtgccag acaagagcca tgaagatcct catgaaagtt 420ggacaa 42613214PRTHomo sapiens 13Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu Glu Leu Leu Cys 1 5 10 15 Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp Glu Glu Ala Ala 20 25 30 Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser Tyr Gln Leu Glu 35 40 45 Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala Pro Thr Ala Arg 50 55 60 Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala Asp Thr Ser Ser 65 70 75 80 Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser Gly Ala Pro Arg 85 90 95 Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu Leu Asp Ala Pro 100 105 110 Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly His Val Val Leu 115 120 125 Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser His Ile Arg Tyr 130 135 140 Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser Val Gln Arg Val 145 150 155 160 Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser Asn Leu Arg Gly 165 170 175 Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met Ala Glu Pro Ser 180 185 190 Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val Ser Leu Leu Thr 195 200 205 Pro Ser Asp Leu Asp Pro 210 14217PRTHomo sapiens 14Pro Ser Ala Pro Arg Ser Val Val Ser Arg Leu Asn Gly Ser Ser Leu 1 5 10 15 His Leu Glu Trp Ser Ala Pro Leu Glu Ser Gly Gly Arg Glu Asp Leu 20 25 30 Thr Tyr Ala Leu Arg Cys Arg Glu Cys Arg Pro Gly Gly Ser Cys Ala 35 40 45 Pro Cys Gly Gly Asp Leu Thr Phe Asp Pro Gly Pro Arg Asp Leu Val 50 55 60 Glu Pro Trp Val Val Val Arg Gly Leu Arg Pro Asp Phe Thr Tyr Thr 65 70 75 80 Phe Glu Val Thr Ala Leu Asn Gly Val Ser Ser Leu Ala Thr Gly Pro 85 90 95 Val Pro Phe Glu Pro Val Asn Val Thr Thr Asp Arg Glu Val Pro Pro 100 105 110 Ala

Val Ser Asp Ile Arg Val Thr Arg Ser Ser Pro Ser Ser Leu Ser 115 120 125 Leu Ala Trp Ala Val Pro Arg Ala Pro Ser Gly Ala Val Leu Asp Tyr 130 135 140 Glu Val Lys Tyr His Glu Lys Gly Ala Glu Gly Pro Ser Ser Val Arg 145 150 155 160 Phe Leu Lys Thr Ser Glu Asn Arg Ala Glu Leu Arg Gly Leu Lys Arg 165 170 175 Gly Ala Ser Tyr Leu Val Gln Val Arg Ala Arg Ser Glu Ala Gly Tyr 180 185 190 Gly Pro Phe Gly Gln Glu His His Ser Gln Thr Gln Leu Asp Glu Ser 195 200 205 Glu Gly Trp Arg Glu Gln Leu Ala Leu 210 215 15139PRTHomo sapiens 15Leu Ala Ala Ala Asp Arg His Thr Val Phe Trp Asn Ser Ser Asn Pro 1 5 10 15 Lys Phe Arg Asn Glu Asp Tyr Thr Ile His Val Gln Leu Asn Asp Tyr 20 25 30 Val Asp Ile Ile Cys Pro His Tyr Glu Asp His Ser Val Ala Asp Ala 35 40 45 Ala Met Glu Gln Tyr Ile Leu Tyr Leu Val Glu His Glu Glu Tyr Gln 50 55 60 Leu Cys Gln Pro Gln Ser Lys Asp Gln Val Arg Trp Gln Cys Asn Arg 65 70 75 80 Pro Ser Ala Lys His Gly Pro Glu Lys Leu Ser Glu Lys Phe Gln Arg 85 90 95 Phe Thr Pro Phe Thr Leu Gly Lys Glu Phe Lys Glu Gly His Ser Tyr 100 105 110 Tyr Tyr Ile Ser Lys Pro Ile His Gln His Glu Asp Arg Cys Leu Arg 115 120 125 Leu Lys Val Thr Val Ser Gly Lys Ile Thr His 130 135 16142PRTHomo sapiens 16Ser Lys Ser Ile Val Leu Glu Pro Ile Tyr Trp Asn Ser Ser Asn Ser 1 5 10 15 Lys Phe Leu Pro Gly Gln Gly Leu Val Leu Tyr Pro Gln Ile Gly Asp 20 25 30 Lys Leu Asp Ile Ile Cys Pro Lys Val Asp Ser Lys Thr Val Gly Gln 35 40 45 Tyr Glu Tyr Tyr Lys Val Tyr Met Val Asp Lys Asp Gln Ala Asp Arg 50 55 60 Cys Thr Ile Lys Lys Glu Asn Thr Pro Leu Leu Asn Cys Ala Lys Pro 65 70 75 80 Asp Gln Asp Ile Lys Phe Thr Ile Lys Phe Gln Glu Phe Ser Pro Asn 85 90 95 Leu Trp Gly Leu Glu Phe Gln Lys Asn Lys Asp Tyr Tyr Ile Ile Ser 100 105 110 Thr Ser Asn Gly Ser Leu Glu Gly Leu Asp Asn Gln Glu Gly Gly Val 115 120 125 Cys Gln Thr Arg Ala Met Lys Ile Leu Met Lys Val Gly Gln 130 135 140 17138PRTHomo sapiens 17Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Arg Val Gly Gln Gln Ala 20 25 30 Val Glu Val Trp Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg 35 40 45 Gly Gln Ala Leu Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln 50 55 60 Leu His Val Asp Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu 65 70 75 80 Leu Arg Ala Leu Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala 85 90 95 Ala Ser Ala Ala Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys 100 105 110 Leu Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr 115 120 125 Thr Gly Glu Ala Cys Arg Thr Gly Asp Arg 130 135 18147PRTHomo sapiens 18Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Leu Leu Val Asn 50 55 60 Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp Lys Ala Val 65 70 75 80 Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu Gly Ala Gln 85 90 95 Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala Pro Leu Arg 100 105 110 Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn 115 120 125 Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr 130 135 140 Gly Asp Arg 145 19137PRTHomo sapiens 19Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Leu Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu 50 55 60 His Val Asp Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu 65 70 75 80 Arg Ala Leu Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala 85 90 95 Ser Ala Ala Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu 100 105 110 Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr 115 120 125 Gly Glu Ala Cys Arg Thr Gly Asp Arg 130 135 20135PRTHomo sapiens 20Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Pro Trp Glu Pro Leu Gln Leu His Val 50 55 60 Asp Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala 65 70 75 80 Leu Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala 85 90 95 Ala Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg 100 105 110 Val Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu 115 120 125 Ala Cys Arg Thr Gly Asp Arg 130 135 2160PRTHomo sapiens 21Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Pro Gly Val Gly Gln Leu 35 40 45 Phe Pro Ala Val Gly Ala Pro Ala Ala Ala Cys Gly 50 55 60 2241PRTHomo sapiens 22Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Asn His Cys 35 40 2326PRTHomo sapiens 23Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr 20 25 2426PRTHomo sapiens 24Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Ala Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr 20 25 2526PRTHomo sapiens 25Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Glu Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr 20 25 2616PRTHomo sapiens 26Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 276PRTHomo sapiens 27Ala Pro Pro Arg Leu Ile 1 5 28106PRTHomo sapiens 28Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Ile 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Val Thr Met Gly Cys Ala Glu Gly 20 25 30 Pro Arg Leu Ser Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Lys Glu Leu Met Ser Pro Pro Asp Thr 50 55 60 Thr Pro Pro Ala Pro Leu Arg Thr Leu Thr Val Asp Thr Phe Cys Lys 65 70 75 80 Leu Phe Arg Val Tyr Ala Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr 85 90 95 Thr Gly Glu Val Cys Arg Arg Gly Asp Arg 100 105 29153PRTHomo sapiens 29Ala Pro Pro Arg Leu Ile Cys Glu Ala Glu Asn Ile Thr Thr Gly Cys 1 5 10 15 Ala Glu His Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys 20 25 30 Val Asn Phe Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val 35 40 45 Glu Val Trp Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly 50 55 60 Gln Ala Leu Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu 65 70 75 80 His Val Asp Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu 85 90 95 Arg Ala Leu Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala 100 105 110 Ser Ala Ala Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu 115 120 125 Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr 130 135 140 Gly Glu Ala Cys Arg Thr Gly Asp Arg 145 150 30162PRTHomo sapiens 30Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Phe Tyr Ala Trp Lys 35 40 45 Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp Gln Gly Leu Ala 50 55 60 Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu Leu Val Asn Ser 65 70 75 80 Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp Lys Ala Val Ser 85 90 95 Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu Gly Ala Gln Lys 100 105 110 Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala Pro Leu Arg Thr 115 120 125 Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn Phe 130 135 140 Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr Gly 145 150 155 160 Asp Arg 31158PRTHomo sapiens 31Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Leu Thr Thr Leu Leu Arg Ala Leu Gly Ala Gln Lys Glu Ala Ile Ser 100 105 110 Pro Pro Asp Ala Ala Ser Ala Ala Pro Leu Arg Thr Ile Thr Ala Asp 115 120 125 Thr Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys 130 135 140 Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr Gly Asp Arg 145 150 155 32158PRTHomo sapiens 32Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Gly Lys 130 135 140 Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr Gly Asp Arg 145 150 155 33160PRTHomo sapiens 33Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 34159PRTHomo sapiens 34Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu 145 150 155 35158PRTHomo sapiens 35Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35

40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly 145 150 155 36157PRTHomo sapiens 36Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr 145 150 155 37156PRTHomo sapiens 37Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr 145 150 155 38155PRTHomo sapiens 38Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu 145 150 155 39154PRTHomo sapiens 39Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys 145 150 40153PRTHomo sapiens 40Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu 145 150 41152PRTHomo sapiens 41Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys 145 150 42151PRTHomo sapiens 42Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly 145 150 43150PRTHomo sapiens 43Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg 145 150 44149PRTHomo sapiens 44Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu 145 45148PRTHomo sapiens 45Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe 145 46147PRTHomo sapiens 46Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn 145 47146PRTHomo sapiens 47Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser 145 48145PRTHomo sapiens 48Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr 145 49144PRTHomo sapiens 49Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 50143PRTHomo sapiens 50Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile

Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg 130 135 140 51142PRTHomo sapiens 51Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe 130 135 140 52141PRTHomo sapiens 52Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu 130 135 140 53140PRTHomo sapiens 53Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys 130 135 140 54139PRTHomo sapiens 54Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg 130 135 55138PRTHomo sapiens 55Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe 130 135 56137PRTHomo sapiens 56Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr 130 135 57136PRTHomo sapiens 57Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp 130 135 58135PRTHomo sapiens 58Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala 130 135 59134PRTHomo sapiens 59Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr 130 60133PRTHomo sapiens 60Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile 130 61132PRTHomo sapiens 61Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr 130 62131PRTHomo sapiens 62Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg 130 63130PRTHomo sapiens 63Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu 130 64129PRTHomo sapiens 64Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro 65128PRTHomo sapiens 65Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 66127PRTHomo sapiens 66Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala 115 120

125 67126PRTHomo sapiens 67Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser 115 120 125 68125PRTHomo sapiens 68Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala 115 120 125 69124PRTHomo sapiens 69Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala 115 120 70123PRTHomo sapiens 70Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp 115 120 71122PRTHomo sapiens 71Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro 115 120 72121PRTHomo sapiens 72Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro 115 120 73120PRTHomo sapiens 73Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser 115 120 74119PRTHomo sapiens 74Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile 115 75118PRTHomo sapiens 75Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala 115 76117PRTHomo sapiens 76Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu 115 77116PRTHomo sapiens 77Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys 115 78115PRTHomo sapiens 78Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln 115 79114PRTHomo sapiens 79Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala 80113PRTHomo sapiens 80Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly 81112PRTHomo sapiens 81Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 82111PRTHomo sapiens 82Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala 100 105 110 83110PRTHomo sapiens 83Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg 100 105 110 84109PRTHomo sapiens 84Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu 100 105 85108PRTHomo sapiens 85Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu 100 105 86107PRTHomo sapiens 86Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5

10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr 100 105 87106PRTHomo sapiens 87Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr 100 105 88105PRTHomo sapiens 88Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu 100 105 89104PRTHomo sapiens 89Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser 100 90103PRTHomo sapiens 90Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg 100 91102PRTHomo sapiens 91Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu 100 92101PRTHomo sapiens 92Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly 100 93100PRTHomo sapiens 93Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser 100 9499PRTHomo sapiens 94Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val 9598PRTHomo sapiens 95Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala 9697PRTHomo sapiens 96Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys 9796PRTHomo sapiens 97Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 9895PRTHomo sapiens 98Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val 85 90 95 9994PRTHomo sapiens 99Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His 85 90 10093PRTHomo sapiens 100Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu 85 90 10192PRTHomo sapiens 101Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln 85 90 10291PRTHomo sapiens 102Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu 85 90 10390PRTHomo sapiens 103Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro 85 90 10489PRTHomo sapiens 104Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu 85 10588PRTHomo sapiens 105Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp 85 10687PRTHomo sapiens 106Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro 85 10786PRTHomo sapiens 107Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln 85 10885PRTHomo sapiens 108Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser 85 10984PRTHomo sapiens 109Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25

30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser 11083PRTHomo sapiens 110Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn 11182PRTHomo sapiens 111Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val 11281PRTHomo sapiens 112Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu 11380PRTHomo sapiens 113Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 11479PRTHomo sapiens 114Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala 65 70 75 11578PRTHomo sapiens 115Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln 65 70 75 11677PRTHomo sapiens 116Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly 65 70 75 11776PRTHomo sapiens 117Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg 65 70 75 11875PRTHomo sapiens 118Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu 65 70 75 11974PRTHomo sapiens 119Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val 65 70 12073PRTHomo sapiens 120Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala 65 70 12172PRTHomo sapiens 121Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu 65 70 12271PRTHomo sapiens 122Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser 65 70 12370PRTHomo sapiens 123Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu 65 70 12469PRTHomo sapiens 124Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu 65 12568PRTHomo sapiens 125Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala 65 12667PRTHomo sapiens 126Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu 65 12766PRTHomo sapiens 127Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly 65 12865PRTHomo sapiens 128Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln 65 12964PRTHomo sapiens 129Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 13063PRTHomo sapiens 130Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val 50 55 60 13162PRTHomo sapiens 131Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu 50 55 60 13261PRTHomo sapiens 132Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val 50 55 60 13360PRTHomo sapiens 133Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala 50 55 60 13459PRTHomo sapiens 134Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln 50 55 13558PRTHomo sapiens 135Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln 50 55 13657PRTHomo sapiens 136Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly 50 55 13756PRTHomo sapiens 137Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val 50 55 13855PRTHomo sapiens 138Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu 50 55 13954PRTHomo sapiens 139Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met 50 14053PRTHomo sapiens 140Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg 50 14152PRTHomo sapiens 141Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn

Phe 35 40 45 Tyr Ala Trp Lys 50 14251PRTHomo sapiens 142Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp 50 14350PRTHomo sapiens 143Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala 50 14449PRTHomo sapiens 144Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr 14548PRTHomo sapiens 145Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 14647PRTHomo sapiens 146Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn 35 40 45 14746PRTHomo sapiens 147Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val 35 40 45 14845PRTHomo sapiens 148Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys 35 40 45 14944PRTHomo sapiens 149Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr 35 40 15043PRTHomo sapiens 150Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp 35 40 15142PRTHomo sapiens 151Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro 35 40 15241PRTHomo sapiens 152Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val 35 40 15340PRTHomo sapiens 153Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr 35 40 15439PRTHomo sapiens 154Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile 35 15538PRTHomo sapiens 155Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn 35 15637PRTHomo sapiens 156Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu 35 15736PRTHomo sapiens 157Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn 35 15835PRTHomo sapiens 158Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu 35 15934PRTHomo sapiens 159Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser 16033PRTHomo sapiens 160Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys 16132PRTHomo sapiens 161Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 16231PRTHomo sapiens 162Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu 20 25 30 16330PRTHomo sapiens 163Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala 20 25 30 16429PRTHomo sapiens 164Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys 20 25 16528PRTHomo sapiens 165Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly 20 25 16627PRTHomo sapiens 166Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr 20 25 16726PRTHomo sapiens 167Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr 20 25 16825PRTHomo sapiens 168Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile 20 25 16924PRTHomo sapiens 169Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn 20 17023PRTHomo sapiens 170Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu 20 17122PRTHomo sapiens 171Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala 20 17221PRTHomo sapiens 172Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu 20 17320PRTHomo sapiens 173Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys 20 17419PRTHomo sapiens 174Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala 17518PRTHomo sapiens 175Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu 17617PRTHomo sapiens 176Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu 17716PRTHomo sapiens 177Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 17815PRTHomo sapiens 178Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr 1 5 10 15 17914PRTHomo sapiens 179Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg 1 5 10 18013PRTHomo sapiens 180Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu 1 5 10 18112PRTHomo sapiens 181Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu 1 5 10 18211PRTHomo sapiens 182Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val 1 5 10 18310PRTHomo sapiens 183Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg 1 5 10 1849PRTHomo sapiens 184Ala Pro Pro Arg Leu Ile Cys Asp Ser 1 5 1858PRTHomo sapiens 185Ala Pro Pro Arg Leu Ile Cys Asp 1 5 1867PRTHomo sapiens 186Ala Pro Pro Arg Leu Ile Cys 1 5 187166PRTHomo sapiens 187Ala Pro Pro Arg Leu Ile Cys Ala Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 188166PRTHomo sapiens 188Ala Pro Pro Arg Leu Ile Cys Arg Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 189166PRTHomo sapiens 189Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Ala Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 190166PRTHomo sapiens 190Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Glu Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 191166PRTHomo sapiens 191Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Ala Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155

160 Cys Arg Thr Gly Asp Arg 165 192166PRTHomo sapiens 192Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Glu Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 193166PRTHomo sapiens 193Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Ala Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 194166PRTHomo sapiens 194Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Ala Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 195166PRTHomo sapiens 195Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Glu Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 196166PRTHomo sapiens 196Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Ala Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 197166PRTHomo sapiens 197Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Ala Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 198166PRTHomo sapiens 198Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Ala Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 199166PRTHomo sapiens 199Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Glu Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 200166PRTHomo sapiens 200Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ala Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 201166PRTHomo sapiens 201Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Ala Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 202166PRTHomo sapiens 202Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Glu Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 203166PRTHomo sapiens 203Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ala Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 204166PRTHomo sapiens 204Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Ala Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 205166PRTHomo sapiens 205Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu

Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Glu Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 206166PRTHomo sapiens 206Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Ala Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 207166PRTHomo sapiens 207Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Ala Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 208166PRTHomo sapiens 208Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Glu Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 20927PRTHomo sapiens 209Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr 20 25 21021PRTHomo sapiens 210Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe Tyr Ala Trp 1 5 10 15 Lys Arg Met Glu Val 20 21127PRTHomo sapiens 211Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp Lys Ala 1 5 10 15 Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu 20 25 21218PRTHomo sapiens 212Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys Leu 1 5 10 15 Lys Leu 213193PRTHomo sapiens 213Met Gly Val His Glu Cys Pro Ala Trp Leu Trp Leu Leu Leu Ser Leu 1 5 10 15 Leu Ser Leu Pro Leu Gly Leu Pro Val Leu Gly Ala Pro Pro Arg Leu 20 25 30 Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu Leu Glu Ala Lys Glu 35 40 45 Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His Cys Ser Leu Asn Glu 50 55 60 Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe Tyr Ala Trp Lys Arg 65 70 75 80 Met Glu Val Gly Gln Gln Ala Val Glu Val Trp Gln Gly Leu Ala Leu 85 90 95 Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu Leu Val Asn Ser Ser 100 105 110 Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp Lys Ala Val Ser Gly 115 120 125 Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu Gly Ala Gln Lys Glu 130 135 140 Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala Pro Leu Arg Thr Ile 145 150 155 160 Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn Phe Leu 165 170 175 Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr Gly Asp 180 185 190 Arg 214181PRTHomo sapiens 214Glu Glu Thr Leu Leu Asn Thr Lys Leu Glu Thr Ala Asp Leu Lys Trp 1 5 10 15 Val Thr Phe Pro Gln Val Asp Gly Gln Trp Glu Glu Leu Ser Gly Leu 20 25 30 Asp Glu Glu Gln His Ser Val Arg Thr Tyr Glu Val Cys Asp Val Gln 35 40 45 Arg Ala Pro Gly Gln Ala His Trp Leu Arg Thr Gly Trp Val Pro Arg 50 55 60 Arg Gly Ala Val His Val Tyr Ala Thr Leu Arg Phe Thr Met Leu Glu 65 70 75 80 Cys Leu Ser Leu Pro Arg Ala Gly Arg Ser Cys Lys Glu Thr Phe Thr 85 90 95 Val Phe Tyr Tyr Glu Ser Asp Ala Asp Thr Ala Thr Ala Leu Thr Pro 100 105 110 Ala Trp Met Glu Asn Pro Tyr Ile Lys Val Asp Thr Val Ala Ala Glu 115 120 125 His Leu Thr Arg Lys Arg Pro Gly Ala Glu Ala Thr Gly Lys Val Asn 130 135 140 Val Lys Thr Leu Arg Leu Gly Pro Leu Ser Lys Ala Gly Phe Tyr Leu 145 150 155 160 Ala Phe Gln Asp Gln Gly Ala Cys Met Ala Leu Leu Ser Leu His Leu 165 170 175 Phe Tyr Lys Lys Cys 180 215199PRTHomo sapiens 215Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Leu 115 120 125 Thr Pro Ala Trp Met Glu Asn Pro Tyr Ile Lys Val Asp Thr Val Ala 130 135 140 Ala Glu His Leu Thr Arg Lys Arg Pro Gly Ala Glu Ala Thr Gly Lys 145 150 155 160 Val Asn Val Lys Thr Leu Arg Leu Gly Pro Leu Ser Lys Ala Gly Phe 165 170 175 Tyr Leu Ala Phe Gln Asp Gln Gly Ala Cys Met Ala Leu Leu Ser Leu 180 185 190 His Leu Phe Tyr Lys Lys Cys 195 216206PRTHomo sapiens 216Glu Glu Thr Leu Leu Asn Thr Lys Leu Glu Thr Ala Asp Leu Lys Trp 1 5 10 15 Val Thr Phe Pro Gln Val Asp Gly Gln Trp Glu Glu Leu Ser Gly Leu 20 25 30 Asp Glu Glu Gln His Ser Val Arg Thr Tyr Glu Val Cys Asp Val Gln 35 40 45 Arg Ala Pro Gly Gln Ala His Trp Leu Arg Thr Gly Trp Val Pro Arg 50 55 60 Arg Gly Ala Val His Val Tyr Ala Thr Leu Arg Phe Thr Met Leu Glu 65 70 75 80 Cys Leu Ser Leu Pro Arg Ala Gly Arg Ser Cys Lys Glu Thr Phe Thr 85 90 95 Val Phe Tyr Tyr Glu Ser Asp Ala Asp Thr Ala Thr Ala Leu Ser Glu 100 105 110 Ala Val Leu Arg Gly Gln Ala Leu Leu Val Asn Ser Ser Gln Pro Trp 115 120 125 Glu Pro Leu Gln Leu His Val Asp Lys Ala Val Ser Gly Leu Arg Ser 130 135 140 Leu Thr Thr Leu Leu Arg Ala Leu Gly Ala Gln Lys Glu Ala Ile Ser 145 150 155 160 Pro Pro Asp Ala Ala Ser Ala Ala Pro Leu Arg Thr Ile Thr Ala Asp 165 170 175 Thr Phe Arg Lys Leu Phe Arg Val Tyr Ser Asn Phe Leu Arg Gly Lys 180 185 190 Leu Lys Leu Tyr Thr Gly Glu Ala Cys Arg Thr Gly Asp Arg 195 200 205 21720DNAHomo sapiens 217atggaggcct cgctcagaaa 2021820DNAHomo sapiens 218tacctgaagg tcaggcgaac 2021921RNAHomo sapiens 219ggugaauguc aagacgcugu u 2122021RNAHomo sapiens 220cagcgucuug acauucaccu u 2122121RNAHomo sapiens 221aguuaauauc aagacgcugu u 2122221RNAHomo sapiens 222cagcgucuug auauuaacuu u 2122321DNAArtificial SequenceDescription of Combined DNA/RNA Molecule Synthetic oligonucleotide 223cgcugacccu gaaguucatu u 2122421RNAArtificial SequenceDescription of Artificial Sequence Synthetic oligonucleotide 224augaacuuca gggucagcgu u 2122516DNAHomo sapiens 225cgcggagatg ggggtg 1622617DNAHomo sapiens 226acagatgacc aggtgtg 1722710DNAHomo sapiens 227ttccagcgca 1022815DNAHomo sapiens 228ggcgccatgg agctc 1522916DNAHomo sapiens 229gcagtactga cctgca 1623014DNAHomo sapiens 230ccagctccag gggt 1423115DNAHomo sapiens 231tgtatcatgg accac 1523216DNAHomo sapiens 232ttgctcttag gacacc 1623314DNAHomo sapiens 233tgtatagttt tttt 14234107PRTHomo sapiens 234Leu Glu Pro Val Ser Trp Ser Ser Leu Asn Pro Lys Phe Leu Ser Gly 1 5 10 15 Lys Gly Leu Val Ile Tyr Pro Lys Ile Gly Asp Lys Leu Asp Ile Ile 20 25 30 Cys Pro Arg Ala Glu Ala Gly Arg Pro Tyr Glu Tyr Tyr Lys Leu Tyr 35 40 45 Leu Val Arg Pro Glu Gln Ala Ala Ala Cys Ser Thr Val Leu Asp Pro 50 55 60 Asn Val Leu Val Thr Cys Asn Arg Pro Glu Gln Glu Ile Arg Phe Thr 65 70 75 80 Ile Lys Phe Gln Glu Phe Ser Pro Asn Tyr Met Gly Leu Glu Phe Lys 85 90 95 Lys His His Asp Tyr Tyr Ile Thr Ser Thr Ser 100 105 235109PRTHomo sapiens 235Leu Glu Pro Ile Tyr Trp Asn Ser Ser Asn Ser Lys Phe Leu Pro Gly 1 5 10 15 Gln Gly Leu Val Leu Tyr Pro Gln Ile Gly Asp Lys Leu Asp Ile Ile 20 25 30 Cys Pro Lys Val Asp Ser Lys Thr Val Gly Gln Tyr Glu Tyr Tyr Lys 35 40 45 Val Tyr Met Val Asp Lys Asp Gln Ala Asp Arg Cys Thr Ile Lys Lys 50 55 60 Glu Asn Thr Pro Leu Leu Asn Cys Ala Lys Pro Asp Gln Asp Ile Lys 65 70 75 80 Phe Thr Ile Lys Phe Gln Glu Phe Ser Pro Asn Leu Trp Gly Leu Glu 85 90 95 Phe Gln Lys Asn Lys Asp Tyr Tyr Ile Ile Ser Thr Ser 100 105 236119PRTHomo sapiens 236Val Ala Asp Arg Tyr Ala Val Tyr Trp Asn Ser Ser Asn Pro Arg Phe 1 5 10 15 Gln Arg Gly Asp Tyr His Ile Asp Val Cys Ile Asn Asp Tyr Leu Asp 20 25 30 Val Phe Cys Pro His Tyr Glu Asp Ser Val Pro Glu Asp Lys Thr Glu 35 40 45 Arg Tyr Val Leu Tyr Met Val Asn Phe Asp Gly Tyr Ser Ala Cys Asp 50 55 60 His Thr Ser Lys Gly Phe Lys Arg Trp Glu Cys Asn Arg Pro His Ser 65 70 75 80 Pro Asn Gly Pro Leu Lys Phe Ser Glu Lys Phe Gln Leu Phe Thr Pro 85 90 95 Phe Ser Leu Gly Phe Glu Phe Arg Pro Gly Arg Glu Tyr Phe Tyr Ile 100 105 110 Ser Ser Ala Ile Pro Asp Asn 115 237123PRTHomo sapiens 237Arg Tyr Ala Val Tyr Trp Asn Arg Ser Asn Pro Arg Phe His Ala Gly 1 5 10 15 Ala Gly Asp Asp Gly Gly Gly Tyr Thr Val Glu Val Ser Ile Asn Asp 20 25 30 Tyr Leu Asp Ile Tyr Cys Pro His Tyr Gly Ala Pro Leu Pro Pro Ala 35 40 45 Glu Arg Met Glu His Tyr Val Leu Tyr Met Val Asn Gly Glu Gly His 50 55 60 Ala Ser Cys Asp His Arg Gln Arg Gly Phe Lys Arg Trp Glu Cys Asn 65 70 75 80 Arg Pro Ala Ala Pro Gly Gly Pro Leu Lys Phe Ser Glu Lys Phe Gln 85 90 95 Leu Phe Thr Pro Phe Ser Leu Gly Phe Glu Phe Arg Pro Gly His Glu 100 105 110 Tyr Tyr Tyr Ile Ser Ala Thr Pro Pro Asn Ala 115 120 238114PRTHomo sapiens 238Arg His Val Val Tyr Trp Asn Ser Ser Asn Pro Arg Leu Leu Arg Gly 1 5 10 15 Asp Ala Val Val Glu Leu Gly Leu Asn Asp Tyr Leu Asp Ile Val Cys 20 25 30 Pro His Tyr Glu Gly Pro Gly Pro Pro Glu Gly Pro Glu Thr Phe Ala 35 40 45 Leu Tyr Met Val Asp Trp Pro Gly Tyr Glu Ser Cys Gln Ala Glu Gly 50 55 60 Pro Arg Ala Tyr Lys Arg Trp Val Cys Ser Leu Pro Phe Gly His Val 65 70 75 80 Gln Phe Ser Glu Lys Ile Gln Arg Phe Thr Pro Phe Ser Leu Gly Phe 85 90 95 Glu Phe Leu Pro Gly Glu Thr Tyr Tyr Tyr Ile Ser Val Pro Thr Pro 100 105 110 Glu Ser

239117PRTHomo sapiens 239Arg His Thr Val Phe Trp Asn Ser Ser Asn Pro Lys Phe Arg Asn Glu 1 5 10 15 Asp Tyr Thr Ile His Val Gln Leu Asn Asp Tyr Val Asp Ile Ile Cys 20 25 30 Pro His Tyr Glu Asp His Ser Val Ala Asp Ala Ala Met Glu Gln Tyr 35 40 45 Ile Leu Tyr Leu Val Glu His Glu Glu Tyr Gln Leu Cys Gln Pro Gln 50 55 60 Ser Lys Asp Gln Val Arg Trp Gln Cys Asn Arg Pro Ser Ala Lys His 65 70 75 80 Gly Pro Glu Lys Leu Ser Glu Lys Phe Gln Arg Phe Thr Pro Phe Thr 85 90 95 Leu Gly Lys Glu Phe Lys Glu Gly His Ser Tyr Tyr Tyr Ile Ser Lys 100 105 110 Pro Ile His Gln His 115 240105PRTHomo sapiens 240Asp Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro 1 5 10 15 Glu Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe 20 25 30 Trp Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe 35 40 45 Ser Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln 50 55 60 Ala Pro Thr Ala Arg Phe Trp Cys Ser Leu Pro Thr Ala Asp Thr Ser 65 70 75 80 Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser Gly Ala Pro 85 90 95 Arg Tyr His Arg Val Ile His Ile Asn 100 105 241166PRTArtificial SequenceDescription of Artificial Sequence Synthetic consensus sequence 241Ala Pro Pro Arg Leu Ile Cys Asp Ser Arg Val Leu Glu Arg Tyr Leu 1 5 10 15 Leu Glu Ala Lys Glu Ala Glu Asn Ile Thr Thr Gly Cys Ala Glu His 20 25 30 Cys Ser Leu Asn Glu Asn Ile Thr Val Pro Asp Thr Lys Val Asn Phe 35 40 45 Tyr Ala Trp Lys Arg Met Glu Val Gly Gln Gln Ala Val Glu Val Trp 50 55 60 Gln Gly Leu Ala Leu Leu Ser Glu Ala Val Leu Arg Gly Gln Ala Leu 65 70 75 80 Leu Val Asn Ser Ser Gln Pro Trp Glu Pro Leu Gln Leu His Val Asp 85 90 95 Lys Ala Val Ser Gly Leu Arg Ser Leu Thr Thr Leu Leu Arg Ala Leu 100 105 110 Gly Ala Gln Lys Glu Ala Ile Ser Pro Pro Asp Ala Ala Ser Ala Ala 115 120 125 Pro Leu Arg Thr Ile Thr Ala Asp Thr Phe Arg Lys Leu Phe Arg Val 130 135 140 Tyr Ser Asn Phe Leu Arg Gly Lys Leu Lys Leu Tyr Thr Gly Glu Ala 145 150 155 160 Cys Arg Thr Gly Asp Arg 165 24221RNAHomo sapiens 242caauagccac ucuaacaccu u 2124321RNAHomo sapiens 243gguguuagag uggcuauugu u 2124421RNAHomo sapiens 244ggggcccguc ccauuugagu u 2124521RNAHomo sapiens 245cucaaauggg acgggccccu u 2124621RNAHomo sapiens 246cugaucugaa gugggugacu u 2124721RNAHomo sapiens 247gucacccacu ucagaucagu u 2124821RNAHomo sapiens 248aagacccuaa ugaggcuguu u 2124921RNAHomo sapiens 249acagccucau uagggucuuu u 2125021RNAHomo sapiens 250ucgaugucuc cuacgucaau u 2125121RNAHomo sapiens 251uugacguagg agacaucgau u 2125221RNAHomo sapiens 252auugaagagg ugauuggugu u 2125321RNAHomo sapiens 253caccaaucac cucuucaauu u 2125421RNAHomo sapiens 254ggaguuacgg gauugugauu u 2125521RNAHomo sapiens 255aucacaaucc cguaacuccu u 2125621RNAHomo sapiens 256gguacuaagg ucuacaucgu u 2125721RNAHomo sapiens 257cgauguagac cuuaguaccu u 2125821RNAHomo sapiens 258guccugacuu caccuauacu u 2125921RNAHomo sapiens 259guauagguga agucaggacu u 2126021RNAHomo sapiens 260ugccgcgucg gguacuuccu u 2126121RNAHomo sapiens 261ggaaguaccc gacgcggcau u 2126219RNAHomo sapiens 262ccgaagagcu ucugugcuu 1926319RNAHomo sapiens 263aagcacagaa gcucuucgg 1926420RNAArtificial SequenceDescription of Artificial Sequence Synthetic oligonucleotide 264uucuccgaac guugucacgu 2026519RNAArtificial SequenceDescription of Artificial Sequence Synthetic oligonucleotide 265acgugacacg uucggagaa 1926619RNAHomo sapiens 266cagccaauag ccacucuaa 1926719RNAHomo sapiens 267uuagaguggc uauuggcug 19

Claims

1.-21. (canceled)

22. A method of determining whether a patient is suitable for erythropoietin (EPO) therapy, comprising (A) isolating a tissue sample from said patient; (B) determining the level of expression of EPH-B4 in said sample; and (C) correlating a presence of EPH-B4 expression to a negative physiological response to EPO therapy.

23. The method of claim 22, wherein the level of expression is determined by measuring the amount of EPH-B4 protein in said sample.

24. The method of claim 23, wherein the presence of EPH-B4 expression is defined by the percentage of cells in said sample showing detectable levels of EPH-B4 protein and the concentration of EPH-B4 protein in said cells.

25. The method of claim 23, wherein the presence of EPH-B4 expression is defined by the formula P.times.C wherein P is the percentage of cells in said sample showing detectable levels of EPH-B4 protein and C is the relative concentration of EPH-B4 protein in said cells, wherein a score of 0, 1, 2, 3 or 4 is assigned to a sample comprising a percentage of cells showing detectable levels of EPH-B4 protein of, respectively, 0%, <25%, 25-50%, 50-75% and 75-100%, wherein a score of 1, 2 or 3 is assigned to relative concentrations of EPH-B4 protein of, respectively, weak, moderate and heavy, and wherein a resulting product of >3 denotes EPH-B4 expression in the sample.

26. The method of claim 23, wherein the level of expression is determined by a technique selected from the group consisting of immunoassay, chemiluminescent assay, nephelometric assay, turbidimetric assay, bioassay and reporter-assay.

27. The method of claim 23, wherein the level of expression is determined by an immunoassay selected from the group consisting of enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, enzyme immunoassay (EIA), radioimmunoassay (RIA), fluorescent immunoassay, Western blot, competitive immunoassay, noncompetitive immunoassay, homogeneous immunoassay and heterogeneous immunoassay.

28. The method of claim 23, wherein the level of expression is determined by ELISA.

29. The method of claim 23, wherein said level of expression is determined by immunohistochemistry.

30. The method of claim 22, wherein the level of expression is determined by measuring the amount of EPH-B4 mRNA in said sample.

31. The method of claim 29, wherein the level of expression is determined by a technique selected from the group consisting of PCR, QPCR, R-PCR, gene expression microarray analysis, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), capillary electrophoresis with laser induced fluorescence (CE-LIF), and RT-PCR.

32. The method of claim 22, wherein the level of expression is determined by RT-PCR.

33. The method of claim 22, wherein the tissue sample is selected from cancerous tissue or circulating cells derived from same.

34. The method of claim 22, wherein the tissue sample is selected from blood, lymph, urine or cerebral fluid.

35. The method of claim 22, further comprising determining the level of expression in said sample of at least one of Ephrin A1 or EPOR.

36. The method of claim 22, further comprising determining the level of expression of Ephrin A1 in said sample.

37. The method of claim 22, wherein said patient is a cancer patient considering EPO therapy.

38. The method of claim 37, wherein said negative physiological response includes the proliferation of cancer cells of said patient.

39. A method of enhancing the effectiveness of EPO therapy in a patient, comprising administering to said patient, in conjunction with EPO therapy, an siRNA specific for EPH-B4.

40. The method of claim 39, wherein the siRNA is selected from the group of nucleic acid duplexes consisting of SEQ ID NO: 242 and SEQ ID NO: 243; SEQ ID NO: 244 and SEQ ID NO: 245; SEQ ID NO: 246 and SEQ ID NO: 247; SEQ ID NO: 248 and SEQ ID NO: 249; SEQ ID NO: 250 and SEQ ID NO: 251; SEQ ID NO: 252 and SEQ ID NO: 253; SEQ ID NO: 254 and SEQ ID NO: 255; SEQ ID NO: 256 and SEQ ID NO: 257; SEQ ID NO: 258 and SEQ ID NO: 259; SEQ ID NO: 260 and SEQ ID NO: 261; and SEQ ID NO: 266 and SEQ ID NO: 267.

41. The method of claim 39, wherein the siRNA is a duplex of SEQ ID NO: 219 and SEQ ID NO: 220.