MODIFIED UROKINASE-TYPE PLASMINOGEN ACTIVATOR POLYPEPTIDES AND METHODS OF USE

Abstract

u-PA polypeptides and fusion proteins containing u-PA polypeptides are provided. The u-PA polypeptides are modified to have altered activity and / or specificity such that they cleave a complement protein, such as complement protein C3, thereby inhibiting complement activation. The modified u-PA polypeptides and fusion proteins that inhibit complement activation can be used to treat diseases and conditions that are mediated by complement activation, or in which complement activation plays a role. These disorders include ischemic and reperfusion disorders, including myocardial infarction and stroke, sepsis, autoimmune diseases, diabetic retinopathy, age-related macular degeneration, organ transplant rejection, inflammatory diseases, and diseases with an inflammatory component.

Description

PLASMINOGEN ACTIVATOR POLYPEPTIDES, UROKINASE TYPE, MODIFIED AND METHODS OF USE RELATED REQUESTS Priority benefit is claimed from United States Provisional Application No. 62 / 786,302, entitled "PLASMINOGEN ACTIVATOR POLYPEPTIDES, UROCINASE TYPE, MODIFIED AND METHODS OF USE", filed December 28, 2018, by inventors Edwin L. Madison, Christopher Thanos, Mikhail Popkov, Vanessa Soros, and Kimberly Tipton, and the applicant Catalyst Biosciences, Inc. Where permitted, the subject matter of this application is incorporated by reference in its entirety. INCORPORATION BY REFERENCE OF ELECTRONICALLY PROVIDED SEQUENCE LISTING An electronic version of the Sequence Listing is presented, the contents of which are incorporated herein by reference in their entirety. The electronic file was created on December 26, 2019, is 2,294 kilobytes in size, and is titled 4940seqPC1 .txt. FIELD OF THE INVENTION Modified u-PA polypeptides are provided that cleave a complement protein, thereby inhibiting complement activation. By virtue of this inhibition, the modified u-PA polypeptides can be used for the treatment of diseases and conditions mediated by complement or in which complement activation plays a role. These diseases and conditions include, but are not limited to, ophthalmic indications, including macular degeneration, such as age-related macular degeneration (AMD) and Stargardt disease, delayed renal graft function (DGF), ischemic and reperfusion disorders. , including myocardial infarction and stroke, sepsis, autoimmune diseases, inflammatory diseases, and diseases with an inflammatory component, including Alzheimer's disease and other neurodegenerative disorders. BACKGROUND OF THE INVENTION The complement (C) system is part of the immune system and plays a role in eliminating invading pathogens and initiating the inflammatory response. The human and other mammalian complement system comprises more than 30 soluble and membrane-bound proteins that participate in an ordered sequence of reactions that result in complement activation. The blood complement system has a wide variety of functions associated with a broad spectrum of host defense mechanisms including antimicrobial and antiviral actions. Products derived from the activation of C components include the non-self-recognition molecules C3b, C4b and PbQynn / i znz / E / v C5b, as well as C3a, C4a, and C5a anaphylatoxins that influence a variety of cellular immune responses. These anaphylatoxins also act as pro-inflammatory agents. The complement system is composed of an arrangement of enzymes and nonenzymatic proteins and receptors. Complement activation occurs by one of three primary modes known as the classical pathway, the alternative pathway, and the "lectin" pathway (see Figure 1). Usually, complement is activated or triggered by 1 of these 3 pathways, which, as shown in Figure 1, converge in the activation of 03. In a fourth mechanism of complement activation, referred to as the intrinsic pathway, serine proteases Associated with the fibrinolytic / coagulation cascade, they activate the complement system directly through cleavage of 03 or 05, independently of the classical, alternative, and lectin pathways. These pathways can be distinguished by the process that initiates complement activation. The classical pathway is initiated by antibody-antigen complexes or aggregated forms of immunoglobulins; the alternative pathway is initiated by recognition of structures on microbial and cell surfaces; and the lectin pathway, which is an antibody-independent pathway, is initiated by the binding of mannan-binding lectin (MBL, also designated mannose-binding protein) to carbohydrates, such as those displayed on the surface of bacteria. or viruses. Activation of the cascades results in the production of complexes involved in proteolysis or cell lysis and peptides involved in opsonization, anaphylaxis, and chemotaxis. The complement cascade, which is a central component of an animal's immune response, is an irreversible cascade. Numerous protein cofactors regulate the process. Inappropriate regulation, usually inappropriate activation, of the process may be a facet of, or may present in, a variety of disorders involving inappropriate inflammatory and immune responses, such as those seen in acute and chronic inflammatory diseases and other conditions involving an immune response. inappropriate. These diseases and disorders include autoimmune diseases, such as rheumatoid arthritis and lupus, cardiac disorders, and other inflammatory diseases, such as sepsis and ischemia-reperfusion injury. Due to the involvement of complement pathways in a variety of diseases and conditions, components of complement pathways are targets for therapeutic intervention, particularly for inhibition of the pathway. Examples of these therapeutic agents include synthetic and natural small molecule therapeutics, antibody inhibitors, and recombinant soluble forms of membrane complement regulators. There are limitations to the strategies for preparing these therapeutics. The small molecules have short half-lives in vivo and must be infused cboynn / i znz / B / v continuously to maintain complement inhibition, thus limiting their role, especially in chronic disease. Therapeutic antibodies can result in an immune response in a subject and, therefore, can lead to complications in treatment, particularly in treatments designed to modulate immune responses. Therefore, there is a need for therapeutics for the treatment of complement-mediated diseases and diseases in which complement activation plays a role. These include acute and chronic inflammatory diseases. Accordingly, among the objectives herein, it is an objective to provide these therapeutics to target or focus on the activation of the complement cascade and to provide therapeutics and methods of treating diseases. BRIEF DESCRIPTION OF THE INVENTION Modified urokinase-like plasminogen activator polypeptides (u-PA) are provided that include amino acid insertions, deletions, and / or replacements in the protease domain that result in increased cleavage activity in complement protein C3 as compared to with the wild-type u-PA protease domain (where the protease domain may include replacement of free Cys with Ser to reduce / eliminate aggregation). Modified u-PA polypeptides are any that comprise the protease domain, such as full-length activated protease, zymogenic forms thereof, and fusion proteins containing a modified u-PA polypeptide and a property-conferring fusion partner. or pharmacological activity. The modified u-PA polypeptides and fusion proteins containing the modified u-PA polypeptides, when in active form, inhibit complement activation. In particular, these fusion polypeptides and proteins cleave C3 whereby C3 activity is inhibited or eliminated. Modifications, including amino acid deletions, replacements, and insertions, provided herein are in the protease domain. The modified u-PA polypeptides include or are the protease domains. Modified u-PA polypeptides can further include post-translational and other modifications to a primary amino acid sequence, such as conjugation or binding to other polypeptides, and portions that alter properties, such as serum half-life, and resistance to endogenous protease. These modifications include, but are not limited to, binding to albumin, binding to the multimerization domain(s), and PEGylation. Thus, modified u-PA polypeptides can be modified by PEGylation, albumination, farnisylation, carboxylation, hydroxylation, phosphorylation, and other polypeptide modifications, known in the art. Among the modifications is the replacement of a free cisterna, in the cboynn / i znz / B / v zymogen, such as 0122, by chymotrypsin numbering, with serine or alanine, to reduce aggregation, particularly upon in vitro expression. This replacement is optional and is not necessarily included in polypeptides that will be pegylated or expressed in vivo. Modified u-PA polypeptides inactivate complement protein 03 by cleavage. Modified u-PA polypeptides cleave 03 to thereby inhibit complement activation, cleave 03 at a site, such as the active site, that inactivates or inhibits the activity of 03 to thereby inhibit complement activation. The modified uPA polypeptides provided herein were selected and designed to cleave within QHARASHLG, and in particular where Ρ1-ΡΓ is RA (QHARjASHL; see SEQ ID NO:47 residues 737-744, where cleavage is between residues 740 and 741). As a result, these modified u-PA polypeptides can be used as therapeutics to treat disorders, diseases and / or conditions in which complement activation plays a role such that inhibition thereof can treat the disorders, diseases and / or or conditions. Modified u-PA polypeptides may also have reduced activity for a natural substrate, such as plasminogen, compared to a wild-type uPA or compared to one that has only the C122S replacement, by chymotrypsin numbering. Among the diseases and conditions for which the modified u-PA polypeptides are any 03-mediated or complement-mediated or involved diseases and conditions. These include ophthalmic disorders, such as age-related macular degeneration (AMD) and diabetic retinopathies, and organ rejection, such as delayed renal graft function (DGF), as well as other diseases, disorders, and conditions that can be treated by inhibition of complement activation. AMD is treated by administration to the vitreous humor, such as by intravitreal injection or intraretinal or subretinal injection, and DGF is treated by intravenous or other systemic administration. The modified u-PA polypeptides and fusion proteins can be further modified, such as by PEGylation, to enhance or improve or impart desirable pharmacological properties, including increased half-life and / or decreased immunogenicity. Other diseases and conditions include, for example, rheumatoid arthritis (RA), eye diseases, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory injury ( HF), Alzheimer's disease (AD), ischemia-perfusion injury, atypical hemolytic uremic syndrome (aHUS), and complement 3 glomerulopathy (C3G). Unmodified u-PA polypeptides include precursor forms, mature forms, the catalytic domain and catalytically active forms thereof, and also proteins PbQynn / i znz / B / v fusion, such as those described in Examples 14-16. Examples of unmodified u-PA polypeptides are those whose sequences are set forth in SEQ ID NO.: 1-6. Unmodified u-PA polypeptides include those in which the free cysteine ​​in the catalytic domain (corresponding to C122 by chymotrypsin numbering) is replaced by another amino acid, such as S or A, particularly S, which does not alter the catalytic activity, but decreases polypeptide aggregation. It is understood that all modified uPA polypeptides may include a replacement, generally S, at the residue corresponding to 0122 by chymotrypsin numbering. Among the modified urokinase-like plasminogen activator (u-PA) polypeptides provided herein are those that contain one or more amino acid modifications selected from replacements corresponding to R35Q, H37Y, V41R, V41 L, Y40Q, D60aP , L97bA, T97al, and H99Q, and conservative amino acid modifications thereto, whereby the modified u-PA polypeptide has greater activity / specificity for a complement protein compared to the unmodified active form of the u-PA polypeptide, where: amino acid modifications are selected from among replacements, insertions, and deletions; the corresponding residues can be determined by alignment with the mature form of u-PA; the modified u-PA polypeptide cleaves a complement protein to thereby inhibit or reduce complement activation compared to unmodified u-PA polypeptide that does not contain the amino acid modifications; residues are numbered by chymotrypsin numbering; Unmodified u-PA polypeptide comprises the sequence set forth in any of SEQ ID NO: 1-6 (wild-type full-length human u-PA, wild-type (catalytic) protease domain u-PA), mature u-PA wild-type, full-length u-PA with the C122S replacement, protease domain u-PA with the C122S replacement, and mature u-PA with the C122S replacement) and catalytically active fragment thereof including amino acid replacement(s). Conservative modifications are selected from R35Y, W, F, or N; H37 R, Q, E, W or F, V41K, D60aS, T97aD, L or V, L97bG or S and H99N, by chymotrypsin numbering. In particular, among these modified urokinase-like plasminogen activator (uPA) polypeptides are those containing one or more amino acid modifications selected from replacements corresponding to R35Q, H37Y, V41R, V41L, Y40Q, D60aP, L97bA, T97al and H99Q. Modified u-PA polypeptides have reduced activity and / or specificity for cleavage of a substrate sequence in plasminogen. The complement protein for which polypeptides have the highest specificity / activity is C3; the inactive cleavage C3. Example cboynn / i znz / B / v cleavage sites are within the C3 active site. Among the modified u-PA polypeptides are those that have greater activity for C3 cleavage that is at least 3-fold greater than the unmodified u-PA polypeptide of SEQ ID NO: 5 (protease domain with the C122S replacement). . Modified u-PA polypeptides include those containing the H37Y replacement, such as the H37Y / V38E replacements. Modified u-PA polypeptides include those containing the R35Y / H37K or R35Q / H37K replacements, such as those comprising the R35Y / H37K / V38E or R35Q / H37K / V38E replacements. Also provided are modified u-PA polypeptides, including those described above, which also contain the L97bA and / or R35Q, and / or H99Q, and / or D60aP, and / or T97al replacement. The modified u-PA polypeptide may further include the amino acid replacement corresponding to T39Y, T39W, T39F, such as T39Y, or selected conservative replacements of T39M or T39L. Other modified u-PA polypeptides include or further include the R35Q / H37Y and / or V38E / V41R / Y149R amino acid replacements. Other modified u-PA polypeptides are those comprising the V41R modification, such as modified u-PA polypeptides comprising the V38E / V41 R modifications, including those that further comprise a replacement at one or more of the R35 positions. , H37 and V38. These include modified u-PA polypeptide in which the replacement in V38 is E, such as, for example, modified u-PA polypeptides comprising R35Y / H37S / V38E / V41R, H37Y / V38E, and other combinations of contributing residues. to 03 cleavage and / or stability, such as in a body fluid. Among the modified u-PA polypeptides provided herein are those with an ED5o for 03 inactivation cleavage of less than either 100 nM, or 50 nM, or 30 nM, or 25 nM in an in vitro assay. Examples of these are those set forth in Table 14, where the ED5o is 100 nM or less, or those set forth in Table 14, where the ED5o is less than 50 nM, or those set forth in Table 14, where the ED5o is less than 30 nM, or those set forth in Table 14, where the ED5o is less than 25 nM. Example of an assay to assess ED5o is one that involves incubating the substrate complement human C3 protein with various concentrations of each modified protease for 1 hour at 37SC to determine ED5o- In particular, modified u-PA polypeptides are any that cleaves C3 with an ED5ode of 50 nM or less. Unmodified u-PA polypeptides may consist of the amino acid sequence set forth in any of SEQ ID NO:1-6 or may include additional modifications, including additional insertions and deletions. Any of the replacements, cboynn / i znz / B / v insertions or deletions herein can be included in the unmodified u-PA polypeptides, such as the protease domain, particularly the protease domain of SEQ ID NO:5. . The modified u-PA polypeptide can have at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the polypeptides of any of SEQ ID NO.: 1-6 or a catalytically active portion thereof. Modified u-PA polypeptides may contain 1 or up to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acid replacements, insertions, or deletions. , as compared to the unmodified u-PA polypeptide of any of SEQ ID NO.: 1-6 or a catalytically active portion thereof. Thus, modified u-PA polypeptides containing the V41R, or H37Y, or L97bA, or R35Q, or H99Q, or D60aP, or T97al modification or combinations thereof are provided. Any of the modified u-PA polypeptides may further contain the amino acid replacement corresponding to T39Y, T39W, T39F or conservative replacements thereof selected from T39M or T39L. In particular, the modified u-PA polypeptides may additionally contain the T39Y amino acid replacement, such as the T39Y / V41R combination, and up to 12 or 13 additional modifications, as well as the optional C122S. Any of the modified u-PA polypeptides may further contain the V38E amino acid replacement, and may further contain one or more of the R35Q, Y60bQ and / or Y149R amino acid modifications. Any of the modified uPA polypeptides may further contain the R37aE or R37aS amino acid modification. Therefore, the modified u-PA polypeptides provided herein may contain the replacements R35Q / H37Y / T39Y / V41R or R35Q / H37Y / T39Y / V41R / C122S. Any of the modified u-PA polypeptides may contain the replacement corresponding to H99Q. Modified u-PA polypeptides provided herein include those containing the amino acid modifications R35Q / H37Y / T39Y / V41 R / L97bA / H99Q / C122S or R35Q / H37Y / T39Y / V41 R / L97bA / H99Q, or T39Y / V41 R / Y60bQ / L97bA / H99Q or T39Y / V41 R / Y60bQ / L97bA / H99Q / C122S or T39Y / V41 R / D60aP / L97bA / H99Q / C122S or T39Y / V41 R / D60aP / L97bA / H99Q / C1 22S. Also among the modified u-PA polypeptides provided herein are those containing the amino acid modifications corresponding to Y40Q / V41 L / L97bA / C122S or Y40Q / V41 R / L97bA / C122S or Y40Q / V41 L / L97bA or Y40Q / V41R / L97bA or R37aSA / 41R / L97bG / H99Q or R37aS / V41 R / L97bG / H99Q / C122S or T39Y / V41 L / L97bA / H99Q / C122S or T39Y / V41 R / L97bA / H99Q / C122S. Modified u-PA polypeptides include those containing the modifications: cboynn / i znz / B / v R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97b A / H99Q / Y149R. Modified u-PA polypeptides containing the amino acid modifications are provided, including polypeptides with the modifications: H37Y / R37aE / V38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / R37aE / V38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37YA / 38E / T39YA / 41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aE / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aE / V38E / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / T97al / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / C122S / Y149R; o R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S o R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aA / Y60bP / T97al / L97bA / H99Q / C122S / Y149R or R35L / H37D / R37aS / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R or R35M / H37G / R37aD / V38E / T39W / V41 R / D60aP / Y60bD / T 97al / L97bA / H99Q / C122S / Y149R or R35Q / H37G / R37aP / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R or R35A / H37G / R37aEA / 38E / T39FA / 41 R / D60a E / Y60bP / T97al / L97bA / H99Q / C122S / Y149R OR R35Q / H37S / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37T / R37aP / V38E / T3 9Y / V41R / D60aE / Y60bD / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37G / R37aE / V38E / T39H / V41 R / D60aP / Y60bA / T97al / L97bA / H99Q / C122S / Y149R or R35W / H37D / R37aS / V38E / T39Y / V41 R / D60aE / Y60bS / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37G / R37aE / V38E / T39Y / V41 R / D60aP / Y60bT / T97al / L97bA / H99Q / C122S / Y149R or R 35W / H37P / R37aN / V38E / T39Y / V41R / D60aP / Y60bL / D97T / T97aE / L97bG / A98S / H99L / C1 22S CbQjnn / l 7Π7 / Β / Υ R35W / H37P / R37aN / V38E / T39Y / V41K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 A or R35Y / H37V / R37aW / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y151L / Q192 To R35W / H37P / R37aN / V38E / T39Y / V41 K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151 L / Q192 To each without replacement at 0122. Examples of these modified deu-PA polypeptides are those containing the modifications R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R. Examples of these polypeptides are those whose sequences are set forth in any of SEQ ID NO.: 8-44 and 987, such as 21 and 39-44, as well as full-length modified u-PA polypeptides and precursors containing the polypeptides. whose sequences are set forth in SEQ ID NO.: 8-44 and catalytically active portions thereof. It is also understood that in any of the modified u-PA polypeptides provided herein, the Cys at residue 122, via chymotrypsin numbering, may be substituted with Ser or Cys may remain. The Cys is retained for embodiments in which the polypeptide, including fusion proteins, containing the modified u-PA protease domain is intended for use as a two-chain form in which free C122 forms a disulfide bond with another free Cys in the polypeptide, or the Cys is modified, such as by PEGylation. In all of the embodiments described herein, position 122 can be Cys or Ser. One skilled in the art can select the appropriate residue depending on the intended use. The unmodified u-PA polypeptide comprises the protease domain of any of SEQ ID NO: 1-6, or a catalytically active part thereof, which includes or contains only the protease domain of SEQ ID NO: 2 or SEQ ID NO:5. The modified u-PA polypeptide may contain additional modifications, including post-translational modifications, modifications that introduce or eliminate a glycosylation site, modification, such as binding or conjugation to a polymer, such as a PEG to increase serum half-life, and / or reduce immunogenicity or both. In particular, any and all of the modified u-PA polypeptides described and provided herein can be PEGylated. Also provided are fusion proteins containing the modified u-PA polypeptides provided herein, such as a fusion with an Fc domain, or a targeting agent specific for a targeted cell or antigen. Among the modified u-PA polypeptides and fusion proteins provided herein are those that have greater than 50% or 80% stability after cboynn / i znz / B / v incubation in PBS or in a body fluid, such as as aqueous humor or serum for 7 days. Also among modified u-PA polypeptides are those which, when in active form, have at least 100-fold decreased activity on plasmin as compared to a corresponding form of unmodified u-PA polypeptide. Also among the modified u-PA polypeptides and fusion proteins provided herein are those that have an ED5o for cleavage by C3 inactivation of less than either 100 nM, or 50 nM, or 30 nM, or 25 nM, or 15 nM. nM or 10 nM in an in vitro assay, such as any exemplified in the Examples herein. These include polypeptides that contain or are the protease domains set forth in Table 14, which lists numerous mutation strands and the ED5o for protease domains of modified u-PA polypeptides that exhibit the ED5o titrated as described in Example 2. Modified u-PA polypeptides and fusion polypeptides having an ED5o of 100 nM or less, less than 50 nM, less than 30 nM, less than 25 nM, less than 15 nM, less than 10 nM, are among those which can be used as protease domains, or in longer u-PA forms and / or in fusion proteins as described herein. Conjugate proteins are provided, including fusion proteins that contain a modified u-PA polypeptide or a catalytically active portion of any of the modified u-PA polypeptides fused to a non-proteinaceous polypeptide or portion thereof. Non-protease polypeptides are provided such as those that include a multimerization domain, such as an Fe domain, a polypeptide, such as albumin, that increases stability in serum, or a protein transduction domain (PTD). As discussed above, all modifications can be in the unmodified polypeptides whose sequences are set forth in any of SEQ ID NO.: 1-6 and catalytically active portions thereof. Polypeptides include those in which the unmodified polypeptide has the sequence set forth in SEQ ID NO:5 (the protease domain with the C122S replacement). Fusion proteins containing the modified u-PA polypeptides provided herein and additional polypeptides, such as serum albumin, multimerization domains, signal sequences, and other trafficking sequences and tags are also provided for ease of expression and isolation. Fusion proteins can also include activation sequences to activate the u-PA portions. Active forms of the fusion proteins are produced upon expression and removal of signal sequences, and any other signal processing and trafficking to result in active fusion proteins that cleave C3. Active forms of fusion proteins include 2-chain activated forms and also dimers, such as those resulting from the inclusion of a multimerization domain. PbQynn / i znz / B / v Fusion proteins include those that contain a modified u-PA polypeptide or a catalytically active portion of a modified u-PA polypeptide, such as those in Table 14, that is fused to a non-proteinaceous polypeptide or portion. of the same. Fusion proteins can also include activation sequences and, prior to processing, signal sequences and other traffic signals. Non-protease polypeptides include, but are not limited to, any known to those of skill in the art to confer a desirable pharmaceutical property or activity, a multilinking domain, such as an Fc, a protein transduction domain (PTD), a hyaluronic acid binding domain (HABD), an antibody for directing or targeting a particular antigen. Fusion proteins can also include activation sequences, such as a native u-PA activation sequence or a funna activation sequence. Examples of furin activation sequences are those that are or comprise QSGQKTLRRRKR (SEQ ID NO:996) or QCGQKTLRRRKR (SEQ ID NO:995) or QSGQKTLRRRKR (SEQ ID NO:1044) or a furin activation site having at least least 98% sequence identity therewith. For example, fusion proteins comprising any of the modified u-PA polypeptides as described or provided herein, and also include, prior to processing or activation, a signal sequence and the modified u-PA polypeptide. or a catalytically active portion thereof. Signal sequences to code for secretion of the fusion proteins include, for example, a II-2, u-PA or IgGK signal sequence. Fusion proteins can include a fusion partner, such as a multimerization domain, or a polypeptide that increases serum half-life, or one that confers another desirable pharmacological property or activity. Examples thereof are an albumin, or Fc domain, or a single chain antibody or other antigen-binding fragment of an antibody, or a hyaluronic acid-binding domain (HABD). Exemplary fusion partners include, but are not limited to, tumor necrosis factor-stimulated gene 6 (TSG-6); HSA, IgG Fc, an antibody or antigen-binding fragment thereof, such as an anti-collagen type II antibody scFv fragment or an anti-VEGFR antibody or fragment thereof. Fusion proteins can also include an activation sequence so that the resulting u-PA-containing fusion protein is in an active form, such as a two-chain form. The activation sequences can contain or be modified to contain a cysteine, which can form a disulfide bond with a free Cys, such as 0122, in the modified u-PA polypeptide, whereby, upon activation, the resulting activated polypeptide contains two strings. Example cboynn / i znz / B / v activation sequences are a u-PA activation sequence and a furin activation sequence, and modified forms thereof, this activation sequence having the sequence set forth in either of SEQ ID NO:995-998, 1041, and 1044 or a sequence having at least 98% or 99% sequence identity thereto. Exemplary fusion proteins are those that contain an activation sequence, a modified u-PA polypeptide, and HSA, such as any comprising the amino acid sequence set forth in any of SEQ ID NO: 1014, 1015, 1016, 1019 and 1040 or a modified form thereof having at least 95%, 96%, 97%, 98%, 99% sequence identity (and containing the sequence modifications in the u-PA portion). For use in methods of treatment, fusion proteins generally do not contain the signal sequence. For use in gene therapy methods, the nucleic acid may encode the signal sequence. Such fusion proteins are provided, such as those containing the amino acid sequence set forth in any of SEQ ID Nos: 1004-1019 and 1034-1040 or any having at least 95%, 96%, 97%, 98%, 99% sequence identity (and containing the sequence modifications of the u-PA portion). Examples of fusion proteins are those having the amino acid sequence set forth in SEQ ID NQ:1015 or 1019. In particular, the signal sequence is removed prior to use or upon expression in vivo or when produced in vitro. These include those that are in the activated two-chain form containing an A chain and a B chain. For example, fusion proteins, where the B chain begins at residues IIGG of the modified u-PA polypeptide and ends at the C-terminus of the fusion protein, such as those containing a modified u-PA polypeptide and HSA, those containing the amino acid sequence set forth in any of SEQ ID Nos. 1005, 1011, 1014, 1015 and 1036, but lacking the signal sequence. Examples of fusion proteins in activated form is a fusion protein containing an A chain from residues 21-178 and a B chain from residues 179- to the C-terminus of the protein with a disulfide bond between residues 168-299. These are also understood to include fusion proteins having at least 95%, 96%, 97%, 98%, 99% sequence identity (and containing the sequence modifications of the u-PA portion). For example, a fusion protein is provided that contains an A chain and a B chain, where the A chain consists of residues 21-178 of SEQ ID No. 1015, and the B chain consists of residues 179-1022; and chains A and B are linked via a disulfide bridge between C168 and C299 of SEQ ID NO:1015. Other fusion proteins provided herein contain multimerization domains such that, upon processing, they form multimers, such as dimers that are formed through the interaction of complementary multimerization domains, such as cboynn / i znz / B / v as Fe domains. Also provided are combinations, which may be packaged as a kit, containing a first composition containing a modified u-PA polypeptide, including, as in all embodiments, fusion proteins, particularly those in activated form, or a plurality thereof, and a second composition containing a second agent or agents for treating a complement-mediated disease or disorder. The second agent or agents, for example, may be one or more anti-inflammatory or anticoagulant agents. Examples of these agents are one or more anti-inflammatory agents selected from one or more nonsteroidal anti-inflammatory drugs (NSAIDs), antimetabolite, corticosteroid, analgesic, cytotoxic agent, inhibitor of proinflammatory cytokines, anti-inflammatory cytokines, agents directed at or with specificity to B cells, compounds directed at T antigens, adhesion molecule blockers, chemokine receptor antagonists, kinase inhibitors , PPAR-γ (gamma) ligands, complement inhibitors, heparin, warfarin, acenocoumarol, phenindione, EDTA, citrate, oxalate, argatroban, lepirudin, bivalirudin, and ximelagatran. Nucleic acid molecules encoding any of the modified u-PA polypeptides and fusion proteins provided herein are provided. Vectors containing these nucleic acid molecules and encoding the modified u-PA polypeptides are also provided. Vectors include prokaryotic vectors and eukaryotic vectors, including mammalian and insect vectors, such as a baculovirus vector, yeast vectors, such as Pichia and Saccharomyces, and viral vectors, such as a herpes simplex virus vector, or vaccinia virus vector, AAV vector, adenoviral vector, or retroviral vector. The vectors may be expression vectors for the production of the vectors and / or modified u-PA polypeptides, such as adenoviruses and AAV viruses, particularly those with tropism for tissue of interest, such as liver or the eye, for gene therapy. Methods are provided for producing the modified u-PA polypeptides by culturing a cell containing a vector or nucleic acid encoding a modified u-PA polypeptide or fusion protein under conditions in which the vector is expressed, and, optionally isolating or recovering the expressed modified u-PA polypeptide. Isolated cells and cell cultures containing the nucleic acid molecules or vectors are also provided. The cells may be non-human cells or human cell cultures, but do not include any zygotes or cells that develop into a human. Cells include mammalian cells and bacterial cells, including, but not limited to, bacterial cells, such as E. coli, CHO, Balb / 3T3, HeLa, MT2, mouse NSO, BHK, insect cells, yeast cells, and other cells used routinely for cboynn / i znz / E / v recombinant expression of polypeptides. Methods for producing the modified u-PA polypeptide include culturing the cells under conditions in which the encoded modified u-PA polypeptide is expressed and optionally isolating or purifying the modified u-PA polypeptide. In general, modified u-PA polypeptides and conjugates thereof, such as fusion proteins, are produced in cells that glycosylate the proteins. Isolated modified u-PA polypeptides can be further modified, such as by PEGylation. Pharmaceutical compositions containing the modified polypeptides and fusion proteins and / or the nucleic acids and / or the vectors are also provided. Uses of the pharmaceutical compositions, nucleic acids or polypeptides of u-PA modified to inhibit complement activation are provided to thereby treat a disease or disorder mediated by complement activation or in which complement activation plays a role in the etiology. or underlying etiology of the disease or disorder. In particular, uses of the nucleic acid molecules and / or gene therapy vectors are provided for treating those diseases, disorders and conditions, mediated by or involving complement activation, where inhibition of complement activation affects treatment or improvement of the disease or condition. Methods of treating a disease or condition mediated by or involving complement activation by administration of the vectors or administration of the nucleic acid molecules are also provided. In particular, diseases, disorders and conditions are those in which the inactivation of C3 so as to inhibit or reduce complement activation effects of treatment. Complement-mediated diseases, disorders, or conditions, or diseases, disorders, and conditions in which complement activation plays a role in the etiology or underlying etiology, include, but are not limited to, any inflammatory disorder, sepsis, rheumatoid arthritis ( RA), eye or ophthalmic disease, cardiovascular disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory injury (IC), Alzheimer's disease ( AD), ischemia-reperfusion injury, atypical hemolytic uremic syndrome (aHUS), complement glomerulopathy 3 (C3G), and organ transplant rejection, particularly delayed organ transplant rejection. Particular diseases and disorders include ocular or ophthalmic disorders, such as macular degeneration or diabetic retinopathy, or inflammation due to a transplanted organ. Diseases, disorders and conditions include age-related macular degeneration (AMD) and delayed kidney graft function (DGF). CbQjnn / l 7Π7 / Ε / Υ Methods for inhibiting complement activation are provided. The methods are performed by contacting a modified u-PA polypeptide with complement protein 03, whereby complement protein 03 is cleaved such that complement activation is reduced or inhibited. Contacting can be effected in vitro, but is generally in vivo, by administration of the modified u-PA polypeptide to a subject in whom complement inactivation or reduction is desired. Administration can be systemic, such as parenteral, including intravenous, or locally, such as by contacting an affected tissue, such as the eye. Administration to the eye includes by drops, by binding of the modified u-PA polypeptide to a protein transduction domain, or by intravitreal injection, intraretinal or subretinal injection, or other such method. For diseases and conditions, such as DGF, administration can be effected by intravenous administration. Other methods include subcutaneous and transdermal administration. The methods and uses include the treatment of any disease, disorder, or condition where inhibition of complement activation leads to a reduction of inflammatory symptoms associated with a complement-mediated disease or disorder selected from an inflammatory disorder, a neurodegenerative disorder, an ophthalmic disorder and a cardiovascular disorder. These include, but are not limited to, inflammatory diseases, conditions and disorders, sepsis, rheumatoid arthritis (RA), eye disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease , immune complex-mediated (IC) acute inflammatory injury, atypical hemolytic uremic syndrome (aHUS), complement 3 glomerulopathy (C3G), Alzheimer's disease (AD), ophthalmic disorders such as AMD and diabetic retinopathies, and ischemia-injury reperfusion. Ischemia-reperfusion injury may involve or be caused by an event or treatment selected from myocardial infarction (MI), stroke, angioplasty, coronary artery bypass grafting, cardiopulmonary bypass (CPB), and hemodialysis or treatment of an subject. Treatment with the modified u-PA polypeptide is effected prior to treatment of a subject. Treatments include organ transplantation. The disease, disorder or condition includes ophthalmic conditions or is an ocular disease or is rejection or inflammation due to a transplanted organ, such as diabetic retinopathy or macular degeneration. In particular, age-related macular degeneration (AMD) treatment methods are provided, as well as delayed renal graft function (DGF) treatment methods. Treatment can be effected intravenously or subcutaneously or locally, such as by injection of the modified u-PA polypeptide into the eye. Included is intravitreal or intraretinal, subretinal injection or attachment of the cboynn / i znz / B / v modified u-PA polypeptide to a protein transduction domain to facilitate transduction into the vitreous. The modified u-PA polypeptide can be linked or conjugated with moieties that effect targeting of the polypeptide to a particular organ or tissue, or that increase serum half-life or reduce immunogenicity, such as PEGylation and / or binding to an Fe domain. or to an antibody or antigen-binding portion thereof. Therefore, methods are provided for treating a subject with a complement-mediated disorder or condition, or one in which complement activation plays a role in that disorder or condition, by administration of a modified u-PA polypeptide provided in the present. These uses of the modified u-PA polypeptides and fusion proteins provided herein are also provided. Modified u-PA polypeptides and fusion protein effect treatment can be used for this treatment because they cleave complement C3 protein to thereby inhibit or reduce complement activation. Inhibition of complement activation leads to a reduction in inflammatory symptoms associated with a complement-mediated disorder, disease, or condition involving an inflammatory response, which leads to a reduction in inflammatory symptoms associated with a disease, condition, or disorder complement-mediated selected from an inflammatory disorder, a neurodegenerative disorder and a cardiovascular disorder. These include ophthalmic conditions, such as diabetic retinopathy and macular degeneration, and also delayed organ rejection, such as DGF. The dosage for the uses and methods and individual dosage formulations are provided herein. An individual dose can be determined empirically by the skilled medical professional and includes, for example, individual doses that are in the range of 0.1 mg to 1 mg for local administration and 0.1 mg to 10, 15, 20, 30 mg or more for systemic administration. , such as intravenous administration. The particular dosage depends on the particular disorder or disease or condition, the subject treated, the stage of the disease, the disorder or condition, the route of administration, the regimen, and the like. Doses can be repeated daily, every two, three, four, five, six, or seven days, at least twice a week, at least every two weeks, three weeks, four weeks, or longer intervals. The particular regimen and doses depend, for example, on the disorder treated, the mode of administration, and details, such as weight, of the subject. Determination thereof is within the skill of the skilled medical professional. Also provided are methods, uses and combinations and modified u-PA polypeptides and fusion proteins, where the modified u-PA polypeptide comprises the V41R or V41L, particularly V41R, such as V411 or R and V38E modification, and those that PbQynn / i znz / B / v contain H37Y / V38E. Examples of this modified u-PA polypeptide are modified u-PA polypeptides containing the modifications Y40Q / V41 R / L97bA or Y40QA / 41L / L97BA or R37aS / V41 R / L97bG / H99Q, or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R. Modifications are found in any unmodified u-PA polypeptide, including those set forth in any of SEQ ID NO.: 1-6, and catalytically active portions thereof including the residue corresponding to V41. Examples of these modified u-PA polypeptides are modified u-PA polypeptides comprising the amino acid residue sequence set forth in SEQ ID NO: 21 or 987 or any of SEQ ID Nos. 40-44 or 40. -44 without the modification at C122, by chymotrypsin numbering, and catalytically active portions thereof, and modified forms thereof, such as PEGylated forms, and fusion proteins and modified forms thereof. Also provided are methods of treating disorders, such as DGF, by intravenous administration of a modified u-PA polypeptide or fusion protein (in activated form) as described and provided herein, including modified u-PA polypeptides. comprising the amino acid residue sequence set forth in any of SEQ ID NO:21 and 40-44, and modified forms thereof, such as PEGylated forms. An individual dose can be empirically determined by the skilled medical professional and includes individual doses that are in the range of 0.1 mg to 1 mg. The dose depends on the subject, the severity or stage of the disease or disorder, such as DGF. The treatment can be repeated multiple times, such as two, three or four times a day, once a day, repeated every 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, weekly, bi-monthly or monthly. The modified u-PA polypeptide may be one comprising the replacements / insertions, by chymotrypsin numbering, R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; and by mature numbering R20Q / H22Y / R23E / V27E / T28YA / 30R / D50P / Y51Q / T911 / L92A / H94Q / C121S / Y148R. Example thereof is the protease domain-containing modified u-PA polypeptide set forth in SEQ ID NO:21 or a catalytically active portion thereof, or the protease domain-containing full-length or precursor forms, and forms modified forms thereof, such as PEGylated forms and fusion proteins. Administration can be effected by any suitable method, including intravenous, subcutaneous, transdermal, local, intramuscular, oral, and other systemic routes of administration. In general, the administered form of the modified u-PA polypeptides provided herein is an activated form, which is generally, depending on the protein components (see, for example, Example 15), a two-chain form. . cboynn / i znz / B / v Methods as described herein, as described above and below, include methods of treating an ophthalmic disorder or ocular disorder by administration of any of the modified u-PA polypeptides and modified forms thereof, such as forms PEGylated and fusion proteins, such as those containing a protein transduction domain, provided herein, to the eye. Ophthalmic disorders, diseases or conditions involving complement activation include diabetic retinopathies and macular degeneration, such as AMD. Dosage is as described above and includes single doses of 0.1 mg to 1 mg. Modified u-PA polypeptides include those containing the replacements R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R, Y40QA / 41 L / L97bA / C122S or Y40Q / V41 R / L97bA / C122S or Y40Q / V41 L / L97bA or Y40Q / V41 R / L97bA, and those contain the amino acid residue sequence set forth in any of SEQ ID NO:21 and 40-44 and catalytically active portions thereof, as well as modified forms thereof. The treatment can be repeated multiple times, such as once a day. Uses of the modified u-PA polypeptides and modified forms thereof for treatment of AMD or DGF are provided. Modified u-PA polypeptides include any described herein, including those containing the replacements R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R or Y40QA / 41 L / L97bA / C122S or Y40Q / V41 R / L97bA / C122S or Y40QA / 41 L / L97bA or Y40Q / V41 R / L97bA, and modified forms thereof that are PEGylated or that are fusion proteins as described at the moment. BRIEF DESCRIPTION OF THE FIGURES FIGURE 1 illustrates an overview of the classical, lectin, and alternative complement pathways and the activation of the terminal complement complex, the membrane attack complex (MAC). The figure depicts many of the more than 30 proteins that participate in the complement cascade, their action within the cascade, and, where appropriate, their points of convergence between complement pathways. For example, the three pathways converge in the generation of a C3-convertase, which cleaves C3 to form a C5-convertase that results in the formation of the MAC complex. The figure also depicts the generation of many of the complement cleavage products. FIGURES 2A-2B are schematic views of N-terminal u-PA fusion proteins. FIGURE 2A is a schematic view of N-terminal u-PA fusion proteins containing the N-terminus of the fusion partner (ie, Fe) to the cbQynn / i ζηζ / Β / γ catalytic domain of u-PA. An exemplary N-terminal fusion protein is set forth in SEQ ID N0:1004, containing human immunoglobulin light chain kappa (k) signal sequence, Fe (fusion partner), AGS (linker), activation sequence of u-PA and a modified u-PA catalytic domain. FIGURE 2B is a schematic view of N-terminal wild-type protein that does not contain a fusion partner. An N-terminal wild-type protein is set forth in SEQ ID NO: 1005, containing human immunoglobulin light chain kappa(k) signal sequence, the N-terminus of u-PA, activation sequence of u-PA, and a modified u-PA catalytic domain. FIGURES 3A-3C are schematic views of u-PAC-terminal fusion proteins. FIGURE 3A is a schematic view of u-PAC-terminal fusion proteins containing the C-terminal fusion partner to the u-PA catalytic domain where the fusion protein lacks an activation sequence N-terminal to the u-catalytic domain. -PA. An exemplary C-terminal fusion protein is set forth in SEQID NQ:1006, containing a human IL2 signal sequence (hlL2SP), a modified u-PA catalytic domain, a linker, and Fe (fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NQ:1007, which contains a human IL2 signal sequence (hlL2SP), a modified u-PA catalytic domain, a linker, and HSA (human serum albumin as fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NQ:1008, which contains a human IL2 signal sequence (hlL2SP), a modified u-PA catalytic domain, a linker, and a collagen-binding scFv. II (C2scFv) (fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NO:1009, which contains a human IL2 signal sequence (hlL2SP), a modified u-PA catalytic domain, a linker, and a HABD (binding domain). hyaluronic acid (fusion partner) Another exemplary C-terminal fusion protein is set forth in SEQ ID NQ:1012, which contains a human IL2 signal sequence (hlL2SP), the catalytic domain of wild-type u-PA, a linker and Fe (fusion partner).Another exemplary C-terminal fusion protein is set forth in SEQ ID NO:1013, which contains a human IL2 signal sequence (hlL2SP), the wild-type u-PA catalytic domain, a linker, and Fe (fusion partner). HSA (fusion partner) FIGURE 3B is a schematic view of C-terminal u-PA fusion proteins containing the C-terminal (ie, Fe or HSA) fusion partner to the u-PA catalytic domain. An exemplary C-terminal fusion is set forth in SEQ ID NQ:1010, which contains a human immunoglobulin light chain kappa (k) signal sequence, a furin activation site, a modified u-PA catalytic domain, a linker and Fe (fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NO:1016, which contains a human immunoglobulin light chain kappa (k) signal sequence, a furin activation sequence, a u-PA catalytic domain Modified PbQynn / i znz / B / v, a linker and HSA (fusion partner). FIGURE 30 is a schematic view of u-PA fusion proteins containing a fusion partner (ie, Fe or HSA) C-terminal to the u-PA catalytic domain and a fusion partner (ie, u-PA wild-type N-terminus) to the u-PA catalytic domain. An exemplary fusion protein is set forth in SEQ ID NO:1011, which contains a human immunoglobulin light chain kappa(k) signal sequence, the N-terminal domain of u-PA, a modified u-PA catalytic domain, a linker, and Fe (fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NO:1014, which contains a human immunoglobulin light chain kappa (k) signal sequence, the N-terminal region of u-PA, a furin activation site, a modified u-PA catalytic domain, a linker, and HSA (fusion partner). Another exemplary C-terminal fusion protein is set forth in SEQ ID NO:1015, which contains a human immunoglobulin light chain kappa (k) signal sequence, the u-PA N-terminal region, the u-PA activation sequence, a modified u-PA catalytic domain, a linker, and HSA (fusion partner). FIGURES 4A-4H are schematic views of the activated forms of the fusion proteins, where SPD refers to the señna-protease domain (the modified u-PA polypeptide protease domains provided herein; the N-terminus of u-PA generally refers to residues 1-178 of u-PA or any modified form thereof. FIGURE 4A is a schematic view of the SE fusion protein Q ID NO: 1010, which contains an Fe domain at the C-terminus of the u-PA protease domain (SEQ ID NO: 21) and a furin activation sequence, where the disulfide bond between the Fe domains forms a dimer FIGURE 4B is a schematic view of the fusion protein of SEQ ID NO: 1011, which contains an Fe domain at the C-terminus of the u-PA protease domain ( SEQ ID NO: 987), and the N-terminus of u-PA and the u-PA activation sequence at the N-terminus of the protein, where the disulfide bond between the Fe domains forms a dimer. FIGURE 4C is a schematic view of the fusion protein set forth in SEQ ID NO: 1036, containing an Fe domain at the C-terminus of the u-PA protease domain (SEQ ID NO: 987), and the N-terminus of u-PA and a furin activation sequence at the N-terminus of the fusion protein, where the disulfide bond between the Fe domains forms a dimer. FIGURE 4D is a schematic view of the fusion protein set forth in SEQ ID NO: 1014, containing HSA at the C-terminus of the u-PA protease domain (SEQ ID NO: 987), and the N-terminus of u-PA and a furin activation sequence at the N-terminus of the fusion protein. FIGURE 4E is a schematic view of the fusion protein set forth in SEQ ID NO: 1015, containing HSA at the C-terminus of the u-PA protease domain (SEQ ID NO: 987), and the N-terminus of u-PA and the u-PA activation sequence cboynn / i znz / B / v at the N-terminus of the fusion protein. FIGURE 4F is a schematic view of the fusion protein set forth in SEQ ID NO: 1016, containing HSA at the C-terminus of the u-PA protease domain (SEQ ID NO: 21) and a furin activation sequence N-terminal to the protease domain. FIGURE 4F is a schematic view of the fusion protein set forth in SEQ ID NO: 1017, containing HSA at the C-terminus of the u-PA protease domain (SEQ ID NO: 21) and a SUMO activation sequence N-terminal to the protease domain. FIGURE 4H is a schematic view of the fusion protein set forth in SEQ ID NO: 1018, containing an Fe domain at the C-terminus of the u-PA protease domain (SEQ ID NO: 21) and the N-terminus of u-PA and a SUMO activation sequence N-terminal to the protease domain, where a disulfide bond between the Fe domains forms a dimer. DETAILED DESCRIPTION OF THE INVENTION Scheme A. DEFINITIONS B. STRUCTURE AND FUNCTION OF u-PA I.Serine Proteases 2.Structure 3.Function / activity C. INHIBITION OF COMPLEMENT BY TARGETING TO C3 .Complement C3 protein and its role in complement initiation a.Classical pathway b.Alternative route c. Lectin pathway d.Complement-mediated effector functions Yo. Complement-mediated lysis: Membrane Attack Complex II. inflammation iii. IV chemotaxis opsonization v. Activation of the Humoral Immune Response 2. Structure and Function of C3 a.C3a b.C3b c. C3b inhibitors D. MODIFIED U-PA POLYPEPTIDES THAT CLEAVE C3 .Example modified u-PA polypeptides.Additional modifications cboynn / i znz / E / v a.Decreased immunogenicity b.Fe domain c . Conjugation with d polymers. Protein transduction domain E. ASSAYS TO ASSESS OR MONITOR u-PA ACTIVITY IN COMPLEMENT MEDIATED FUNCTIONS. Methods to assess u-PA activity in complement protein C3 function a.Detection of proteins i. SDS-PAGE analysis ii. Enzyme immunoassay iii. Radial immunodiffusion (RID) b. Hemolytic Assays c. Methods to determine cleavage sites. Methods to assess wild-type u-PA activity. a.Plasminogen cleavage b. Plasminogen Activation Assays c. u-PA-uPAR Binding Assays d. Selection of C3 ACC-AGR + ELISA Specificity assessment using peptide libraries .Specificity .Disease models F. METHODS OF PRODUCTION OF NUCLEIC ACIDS ENCODING MODIFIED U-PA POLYPEPTIDES THEREOF. Isolation or Preparation of Nucleic Acids Encoding u-PA Polypeptides. Generation of Mutant or Modified Nucleic Acids and Encoding Polypeptides. Vectors and Cells 4.Expression a.Prokaryotic cells b.Yeast cells c. Insects and insect cells d. Mammalian expression e. Plants cbQynn / i ζπζ / β / υ 5. Purification. Additional Modifications a.PEGylation b.Fusion proteins and other conjugates.Nucleic acid molecules G. COMPOSITIONS, FORMULATIONS AND DOSAGES Administration of modified u-PA polypeptides Administration of nucleic acids encoding modified u-PA polypeptides (gene therapy) H. THERAPEUTIC USES AND TREATMENT METHODS .Disease mediated by complement activation a. Rheumatoid arthritis b.Sepsis c. Multiple sclerosis d.Alzheimer's disease e. Ischemia-Reperfusion Injury f. Eye disorders Age-related macular degeneration (ADM) g. Organ transplantation and delayed graft function (DGF). Therapeutic Uses a.Inflammatory disease mediated by the immune system b. Neurodegenerative disease c. Cardiovascular diseases e. Organ transplant Delayed graft function (DGF). Combination therapy I. EXAMPLES A. DEFINITIONS Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by one skilled in the art to which the invention pertains. All patents, patent applications, published applications and publications, GENBANK sequences, websites, and other published materials referenced throughout the description herein, unless otherwise indicated, are incorporated herein. by reference in its entirety. In the event that there are multiple definitions for terms herein, those in cboynn / i znz / B / v in this section shall control. When referring to a URL or other identifier or address, it is understood that these identifiers may change and particular information on the Internet may come and go, but equivalent information is known and can be easily accessed, such as by searching the Internet and / or appropriate databases. The reference to it reveals the availability and public dissemination of that information. As used herein, cleavage refers to the cleavage of peptide bonds by a protease. The protease cleavage site motif involves N- and C-terminal residues to the cleaved linker (the primed and unprimed sides, respectively, with the protease cleavage site defined as... P3-P2-P1-P1'- P2'-P3'..., and the cleavage occurs between residues P1 and ΡΓ). In human C3, cleavage by a C3-convertase occurs between residues R and S (see residues 746-751 of SEQ ID NO: 47, cleavage between residues 748 and 749 in human C3) of C3: P3 P2 P1 ΡΓ P2' P3' Leu Ala Arg J, Ser Asn Leu Usually, the cleavage of a substrate in a biochemical pathway is either an activating cleavage or an inhibitory cleavage. An activating cleavage refers to the cleavage of a polypeptide from an inactive to an active form. This includes, for example, cleavage of a zymogen to an active enzyme. An activating cleavage is also a cleavage whereby a protein is cleaved into one or more proteins that have activity. For example, the complement system is an irreversible cascade of proteolytic cleavage events whose termination results in the formation of multiple effector molecules that stimulate inflammation, facilitate antigen phagocytosis, and directly lyse some cells. Therefore, the cleavage of C3 by a C3-convertase into C3a and C3b is an activation cleavage. In contrast, the modified u-PA polypeptides provided herein effect inhibitory cleavage of C3, such as by cleavage at the active site. As used herein, an inhibitory cleavage or inactivating cleavage is the cleavage of a protein into one or more degradation products that are non-functional. Inhibitory cleavage results in decreased or reduced activity of a protein. Usually, a reduction in a protein's activity reduces the pathway or process for which the protein is involved. In one example, cleavage of any one or more complement proteins that is an inhibitory cleavage results in concomitant reduction or inhibition of any one or more of the classical, lectin, or alternative functional pathways of complement. To be inhibitory, cleavage reduces activity by at least or at least about 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90 %, 95%, 99% or more compared to a natural form of the protein. The percent cleavage of a protein that is required for cleavage to be inhibitory PbQynn / i znz / B / v varies between proteins, but can be determined by assaying for a protein activity. As used herein, "complement activation" refers to activation of complement pathways, for example, complement activation refers to an increase in the functions or activities of any one or more of the complement pathways by a protease or an increase in the activity of any of the proteins in the complement pathway. Activation of complement can lead to complement-mediated cell lysis or can lead to cell or tissue destruction. Inappropriate activation of complement in the host tissue plays an important role in the pathology of many autoimmune and inflammatory diseases, and is also responsible for or associated with many disease states associated with bioincompatibility. It is understood that activation can mean an increase in existing activity, as well as the induction of new activity. Activation of complement can occur in vitro or in vivo. Examples of complement functions that can be assayed and described herein include hemolytic assays and assays to measure any one or more of the complement effector molecules, such as by SDS PAGE followed by Western blot or Coomassie Brilliant Blue staining or by ELISA. In some embodiments, complement activation is inhibited by a protease, such as a protease described herein, by 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 99%. or more compared to complement activity in the absence of a protease. As used herein, "inhibiting complement activation" or "complement inactivation" refers to the reduction or diminution of a complement-mediated function or activity of any one or more of the complement pathways by a protease or in the activity of any of the proteins in a pathway. A complement function or activity can be presented in vitro or in vivo. Examples of complement functions that can be assayed and described herein include hemolytic assays and assays to measure any one or more of the complement effector molecules, such as by SDS PAGE followed by Western blot or Coomassie Brilliant Blue staining or by ELISA. A protease can inhibit complement activation by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In other embodiments, a protease inhibits complement activation by 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 99% or more compared to complement activity in the absence of a protease. As used herein, a complement protein or "complement component" is a protein of the complement system that functions in the host's defense against infection and in the inflammatory process. Complement proteins include those that function in the classical pathway, those that function in the alternative cboynn / i znz / B / v pathway, and those that function in the lectin pathway. Complement proteins include proteases involved in complement pathways. As used herein, complement proteins include any of the "cleavage products" (also referred to as "fragments") that are formed after activation of the complement cascade. Also included among complement proteins are inactive or altered forms of complement proteins, such as ¡C3b and C3a-desArg. Thus, complement proteins include, but are not limited to: C1q, C1 r, C1s, C2, C3, C3a, C3b, C3c, C3dg, C3g, C3d, C3f, iC3, C3a-desArg, C4, C4a , C4b, iC4, C4a-desArg, C5, C5a, C5a-des-Arg, C6, C7, C8, C9, MASP-1, MASP-2, MBL, Factor B, Factor D, Factor H, Factor I, CR1 , CR2, CR3, CR4, properdin, C1lnh, C4bp, MCP, DAF, CD59 (MIRL), clusterin and HRF and allelic and species variants of any complement protein. As used herein, a natural form of a complement protein is one that can be isolated from an organism such as a vertebrate in the absence of complement activation and has not been intentionally modified by man in the laboratory. Examples of native complement proteins include C1q, C1r, C1s, C2, C3, C4, Factor B, Factor D, properdin, C5, C6, C7, C6, and C9. In general, native complement proteins are inactive and become active upon activation. Activation may require activation cleavage, maturation cleavage, and / or complexation with other proteins. An exception to this is Factor I and Factor D which have enzyme activity in their native form. In some examples, activation of a natural complement protein occurs after cleavage of the protein. For example, complement zymogens such as 03 are proteases that activate themselves by protease cleavage such that cleavage of 03 by 03 convertase C4b2b generates the active fragments C3a and C3b. In another example, cleavage of an inactive natural complement protein results in changes in the structural stability of a protein that result in activation of the protein. For example, 03 contains an internal thioester bond which in the native protein is stable, but can become highly reactive and activated following conformational changes resulting from protein cleavage. Therefore, the 03 cleavage products are biologically active. Activation of 03 can also occur spontaneously in the absence of cleavage. It is the spontaneous conversion of the thioester bond to natural 03 that is an initiating event of the alternative complement pathway. In another example, activation of a natural complement protein occurs after the release of a complex regulatory molecule that inhibits the activity of an otherwise active natural complement protein. For example, 01 inh binds to and inactivates 01 s and 01 r, unless they are in complex with 01 q. cboynn / i znz / B / v As used herein, maturation cleavage is a general term referring to any cleavage necessary for activation of a zymogen. This includes cleavage leading to a conformational change that results in activity (ie, activation cleavage). It also includes cleavage in which a critical binding site is exposed or a steric hindrance is exposed or an inhibitory segment is removed or moved. As used herein, "altered form" of a complement protein refers to a complement protein that is present in a non-natural form resulting from modifications in its molecular structure. For example, reaction of C3 of the thioester with water can occur in the absence of convertase cleavage, yielding an inactive hydrolyzed form of C3 called ¡C3. In another example, anaphylatoxins including C3a, C5a and C4a can be desarginized by carboxypeptidase N into more stable and less active forms. As used herein, a fragment or "cleavage product" of a complement protein is a region or segment of a complement protein that contains a portion of the polypeptide sequence of a native complement protein. A fragment of a complement protein usually results after activation of a complement cascade. In general, a fragment results from the proteolytic cleavage of a natural complement protein. For example, the complement protein C3 is cleaved enzymatically by a C3-convertase, resulting in two fragments: C3a which constitutes the N-terminal portion of C3; and C3b which constitutes the C-terminal portion and contains the serine protease site. A fragment of a complement protein also results from the proteolytic cleavage of another fragment of a complement protein. For example, C3b, a fragment generated from the cleavage of C3, is cleaved by Factor I to generate the ¡C3b and C3f fragments. In general, the cleavage products of complement proteins are biologically active products and function as cleavage effector molecules of the complement system. Thus, a complement protein fragment or portion includes cleavage products of complement proteins and also portions of the proteins that retain or exhibit at least one complement protein activity. As used herein, "cleavage effector molecules" or "cleavage effector proteins" refers to the active cleavage products generated as a result of the complement system activated enzyme cascade. A cleavage effector molecule, fragment, or cleavage product resulting from complement activation may contribute to any one or more complement-mediated functions or activities, including opsonization, anaphylaxis, cell lysis, and inflammation. Examples of cleavage or effector molecules include, but are not limited to, C3a, C3b, C4a, C4b, C5a, C5b-9 cboynn / i znz / B / v and Bb. The cleavage effector molecules of the complement system, by virtue of participation in the cascade, exhibit activities including stimulating inflammation, facilitating phagocytosis of antigen, and lysing some cells directly. Complement cleavage products promote or participate in the activation of complement pathways. As used herein, anaphylatoxins are cleavage effector proteins that trigger the degranulation or release of substances from mast cells or basophils, which participate in the inflammatory response, particularly as part of the defense against parasites. If the degranulation is too strong, it can cause allergic reactions. Anaphylatoxins include, for example, C3a, C4a, and C5a. Anaphylatoxins also indirectly mediate smooth muscle cell spasms (such as bronchospasms), increases in blood capillary permeability, and chemotaxis. As used herein, "chemotaxis" refers to the receptor-mediated movement of leukocytes toward a chemoattractant usually in the direction of increasing concentration thereof, such as in the direction of increasing concentration of an anaphylatoxin. As used herein, opsonization refers to the alteration of the surface of a pathogen or other particle so that it can be ingested by phagocytes. A protein that binds to or disrupts the surface of a pathogen is called an opsonin. Complement and antibody proteins opsonize extracellular bacteria for uptake and destruction by phagocytes such as neutrophils and macrophages. As used herein, cell lysis refers to the open rupture of a cell by destruction of its wall or membrane. Red blood cell hemolysis is a measure of cell lysis. As used herein, "complement protein C3" or "C3" refers to the complement protein C3 of the complement system that functions in host defense against infection and in the inflammatory process. Human complement C3 protein is a 1663 amino acid single chain pre-proprotein or zymogen set forth in SEQ ID NO:47 containing a 22 amino acid signal peptide (amino acids 1-22 of SEQ ID NO:47) and a tetra-arginine sequence (amino acids 678-671 of SEQ ID NO:47) that is removed by a furin-like enzyme resulting in a mature two-chain protein containing a beta chain (amino acids 23-667 of SEQ ID NO:47) and an alpha chain (amino acids 672-1663 of SEQ ID NO:47) linked by a disulfide bond between residues C559 and C816. Complement protein C3 is further activated by proteolytic cleavage by a C3-convertase (C4b2b or C3bBb) between amino acids 748 and 749 of SEQ ID NO:47 generating C3a anaphylatoxin and C3b opsonin. As used herein, a zymogen refers to a protein that is activated cboynn / i znz / B / v by proteolytic cleavage, including maturational cleavage, such as activation cleavage, and / or complexation with other proteins. and / or cofactors. A zymogen is an inactive precursor of a protein. These precursors are generally larger, although not necessarily larger, than the active form. Referring to u-PA or complement protein C3, zymogens are converted to active enzymes by specific cleavage, including catalytic and autocatalytic cleavage, or by binding of an activating cofactor, which generates an active enzyme. A zymogen, thus, is an enzymatically inactive protein that is converted to a proteolytic enzyme by the action of an activator. Cleavage can be effected autocatalytically. Several complement proteins are zymogens; they are inactive, but are cleaved and activated at the onset of the complement system after infection. Zymogens, in general, are inactive and can be converted to mature active polypeptides by catalytic or autocatalytic cleavage of the zymogen proregion. As used herein, a "pro-region", "propeptide" or "pro-sequence" refers to a region or segment of a protein that is cleaved to produce a mature protein. This may include segments that function to suppress enzyme activity by masking the catalytic machinery and thus preventing formation of the catalytic intermediate (ie, by sterically occluding the substrate binding site). A proregion is an amino acid sequence positioned at the amino terminus of a mature biologically active polypeptide and may be as small as a few amino acids or may be a multidomain structure. As used herein, an "activation sequence" refers to an amino acid sequence in a zymogen that is the site necessary for activation cleavage or maturation cleavage to form an active protease. Cleavage of an activation sequence can be catalyzed autocatalytically or via activation partners. Activation cleavage is a type of maturational cleavage in which a conformational change required for activity occurs. This is a classical activation pathway, for example, for serine proteases in which a cleavage generates a new N-terminus that interacts with conserved regions of the catalytic machinery, such as catalytic residues, to induce conformational changes necessary for activation. activity. Activation can result in the production of multi-chain forms of the proteases. In some cases, the single chain forms of the protease may exhibit proteolytic activity. As used herein, "domain" refers to a portion of a molecule, such as encoding proteins or nucleic acids, that is structurally and / or functionally distinct from other portions of the molecule and is identifiable. An exemplary polypeptide domain is a part of the polypeptide that can form an independently folded structure within cboynn / i znz / B / v of a polypeptide composed of one or more structural motifs (for example, combinations of alpha helices and / or beta strands). connected by loop regions) and / or that is recognized by a particular functional activity, such as enzyme activity, dimerization, or substrate binding. A polypeptide can have one or more distinct domains, usually more than one. For example, the polypeptide can have one or more structural domains and one or more functional domains. An individual polypeptide domain can be distinguished based on structure and function. A domain can span a contiguous linear sequence of amino acids. Alternatively, a domain can span multiple non-contiguous amino acid portions, which are non-contiguous along the linear amino acid sequence of the polypeptide. Usually, a polypeptide contains a plurality of domains. For example, serine proteases can be characterized based on the sequence of protease domains. Those skilled in the art are familiar with polypeptide domains and can identify them by virtue of structural and / or functional homology to other such domains. For exemplification herein, definitions are provided, but it is understood that it is within the skill of the art to recognize particular domains by name. If necessary, appropriate software can be used to identify domains. As used herein, a polypeptide framework region is a region of the polypeptide that contains at least one framework domain. As used herein, a "protease domain" is the catalytically active portion of a protease. Reference to a protease domain of a protease includes single, double, and multichain forms of any of these proteins. A protease domain of a protein contains all the required properties of that protein required for its proteolytic activity, such as, for example, its catalytic center. As used herein, a "catalytically active portion" or "catalytically active domain" of a protease, for example, a u-PA polypeptide, refers to the protease domain, or any fragment or portion thereof that retains the protease activity. For example, a catalytically active portion of a u-PA polypeptide can be a u-PA protease domain including an isolated single chain form of the protease domain or an activated double chain form. Significantly, at least in vitro, single chain forms of proteases and catalytic domains or proteolytically active portions thereof (usually C-terminal truncations) exhibit protease activity. As used herein, a "nucleic acid encoding a protease domain or catalytically active portion of a protease" refers to a nucleic acid encoding only the aforementioned single-stranded protease domain or active portion thereof, and not the other contiguous portions of the protease as a continuous sequence. PbQynn / i znz / B / v As used herein, the statement that a polypeptide consists essentially of the protease domain means that the only portion of the polypeptide is a protease domain or a catalytically active portion thereof. The polypeptide can optionally, and generally includes additional sequences not derived from amino acid protease. As used herein, a "protease active site" refers to the substrate binding site where catalysis of the substrate occurs. The structure and chemical properties of the active site allow recognition and binding of the substrate and subsequent hydrolysis and cleavage of the cleaved bond in the substrate. The active site of a protease contains amino acids that contribute to the catalytic mechanism of peptide cleavage, such as the amino acids Gln His Ala Arg Ala Ser His Leu (C3 active site; residues 737-744 of SEQ ID NO:47), as well as amino acids that contribute to substrate sequence recognition, such as amino acids that contribute to extended substrate binding specificity. For example, cleavage at the C3 active site can inhibit its activity, such as: QHA R ( A S H L (residues 737-744 of SEQ ID NO:47) P4 P3 P2P1 ;ΡΓ P2'P3' P4'. As used herein, "substrate recognition site" or "cleavage sequence" refers to the sequence recognized by the active site of a protease that is cleaved by a protease. Typically, a cleavage sequence for a serine protease is six residues in length to match the extended substrate specificity of many proteases, but may be longer or shorter depending on the protease. Typically, for example, for a serine protease, a cleavage sequence is composed of amino acids P1-P4 and P1'-P4' in a substrate, where cleavage occurs after the P1 position. Typically, a cleavage sequence for a serine protease is six residues in length to match the extended substrate specificity of many proteases, but may be longer or shorter depending on the protease. As used herein, "target substrate" refers to a substrate that is cleaved by a protease. Usually, the target substrate is specifically cleaved at its substrate recognition site by a protease. At a minimum, a target substrate includes the amino acids that make up the cleavage sequence. Optionally, a target substrate includes a peptide containing the cleavage sequence and any other amino acids. A full-length protein, allelic variant, isoform, or any portion thereof, that contains a cleavage sequence recognized by a protease, is a target substrate for that protease. For example, for purposes herein where complement inactivation is intended, a target substrate is complement protein 03, or any portion or fragment thereof that contains a cleavage sequence recognized by a u-PA polypeptide. . These target substrates can be purified proteins, CbQjnn / l 7Π7 / Β / Υ or they may be present in a mixture, such as an in vitro mixture or an in vivo mixture. The mixtures can include, for example, blood or serum or other tissue fluids. Additionally, a target substrate includes a peptide or protein that contains an additional moiety that does not affect cleavage of the substrate by a protease. For example, a target substrate can include a four amino acid peptide or a full length protein chemically linked to a fluorogenic moiety. Proteases can be modified to exhibit increased substrate specificity for a target substrate. As used herein, "u-PA" or "uPA" or "u-PA polypeptide" refers to any u-PA polypeptide including, but not limited to, a recombinantly produced polypeptide, a synthetically produced polypeptide and a u-PA polypeptide extracted or isolated from cells or tissues including, but not limited to, liver and blood. Alternative names used interchangeably for u-PA include urokinase and urinary plasminogen activator and urokinase plasminogen activator and urinary type plasminogen activator and urokinase type plasminogen activator. u-PA includes related polypeptides from different species including, but not limited to, animals of human and non-human origin. Human u-PA includes u-PA, allelic variants, isoforms, synthetic nucleic acid molecules, protein isolated from human cells and tissues, and modified forms thereof. Exemplary unmodified human u-PA polypeptides include, but are not limited to, natural unmodified and wild-type mature u-PA polypeptides (SEQ ID NO:3), the unmodified precursor u-PA polypeptide and type wild type including a propeptide and / or signal peptides (such as the u-PA polypeptide set forth in SEQ ID NO:1) and the protease domain (such as the u-PA protease domain set forth in SEQ ID NO: 2). One skilled in the art would recognize that the referenced positions of the mature u-PA polypeptide (SEQ ID NO:3) differ by 20 amino acid residues when compared to the precursor u-PA polypeptide (SEQ ID NO:1), which is the u-PA polypeptide containing the signal peptide sequence. Thus, the first amino acid residue of SEQ ID NO:3 "corresponds to" the twenty-first (21st) amino acid residue of SEQ ID NO:1. Reference to "u-PA" encompasses the activated or two-chain form of the uPA polypeptide containing the N-terminal A chain (SEQ ID NO:3 amino acids 1-158) and the C-terminal B chain (amino acids 159 -411 of SEQ ID NO:3) linked by a disulfide bond between residues 148C and 279C (corresponding to the mature u-PA polypeptide set forth in SEQ ID NO:3). The two-chain form, or high molecular weight (HMW) u-PA, is formed from a mature u-PA polypeptide (eg, that set forth in SEQ ID NO:3) by proteolytic cleavage after the HMW residue. Lys158 amino acid before the Ile159 residue. Proteolytic cleavage can be carried out, for example, by plasmin, kallikrein, cathepsin B, cboynn / i znz / B / v matriptase and nerve growth factor gamma. The u-PA polypeptides provided herein may be further modified, such as by chemical modification or post-translational modification. These modifications include, but are not limited to, glycosylation, pegylation, albumination, farnisylation, carboxylation, hydroxylation, phosphorylation, and other polypeptide modifications known in the art. u-PA includes u-PA from any species, including human and non-human species, u-PA polypeptides of non-human origin include, but are not limited to, murine, canine, leporine, avian, bovine, sheep, pigs and other primates. Exemplary non-human origin u-PA polypeptides include, for example, mouse (Mus musculus, SEQ ID NO:52), rat (Rattus norvegicus, SEQ ID NO:53), cow (Bos taurus, SEQ ID NO : 54), pig (Sus scrofa, SEQ ID NO: 55), rabbit (Oryctolagus cuniculus, SEQ ID NO: 56), chicken (Gallus gallus, SEQ ID NO: 57), yellow baboon (Papio cynocephalus, SEQ ID NO: 58), Sumatran orangutan (Pongo abelii, SEQ ID NO: 59), dog (Canis lupus, SEQ ID NO: 60), sheep (Ovis aries, SEQ ID NO: 61), marmoset (Callithrix jacchus, SEQ ID NO: 62), rhesus macaque (Macaca mulatta, SEQ ID NO:63), northern white-cheeked gibbon (Nomascus leucogenys, SEQ ID NO:64) and chimpanzee (Pan trodyglotes, SEQ ID NO: 65). Reference to u-PA polypeptides also includes precursor polypeptides and mature u-PA polypeptides in single- or double-chain forms, truncated forms thereof that have activity, the protease domain isolated, and includes allelic variants and species variants. , variants encoded by splice variants, and other variants, including polypeptides that are at least or at least about 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90 %, 95%, 96%, 97%, 98%, 99% or more sequence identity with the precursor polypeptide set forth in SEQ ID NO:1 or the mature form thereof (SEQ ID NO:3) or the protease domain thereof (SEQ ID NO: 2). u-PA polypeptides include, but are not limited to, tissue-specific isoforms and allelic variants thereof, synthetic molecules prepared by translation of nucleic acids, proteins generated by chemical synthesis, such as synthesis involving ligation of polypeptides plus short, via recombinant methods, proteins isolated from human and non-human cells and tissue, chimeric u-PA polypeptides and modified forms thereof. u-PA polypeptides also include fragments or portions of u-PA that are of sufficient length or include appropriate regions to retain at least some activity (after activation if necessary) of a full length mature polypeptide. In one example, the u-PA portion is the protease domain, such as, for example, the protease domain set forth in SEQ ID NO: 2 which corresponds to amino acids 179431 of the u-PA sequence set forth in SEQ ID NO: 1. u-PA polypeptides also include those that contain chemical or post-translational modifications and those that CbQjnn / l 7Π7 / Β / Υ do not contain chemical or post-translational modifications. These modifications include, but are not limited to, pegylation, albumination, glycosylation, farnylation, carboxylation, hydroxylation, phosphorylation, HESylation (extension of half-life by coupling drug molecules to biodegradable hydroxyethyl starch (HES)), PASylation (conjugation by genetic fusion or chemical coupling of pharmacologically active compounds, such as low molecular weight proteins, peptides, and drugs, with natively disordered biosynthetic polymers made of the small L-amino acids Pro, Ala, and / or Ser) and other polypeptide modifications known in the art. technique. As used herein, "u-PA protease" or "u-PA protease domain" refers to any u-PA polypeptide including, but not limited to, a recombinantly produced polypeptide, a polypeptide produced synthetically and a uPA polypeptide extracted or isolated from cells or tissues including, but not limited to, liver and blood. The u-PA protease includes related polypeptides from different species including, but not limited to, animals of human and non-human origin. A human u-PA protease domain or u-PA protease includes u-PA, allelic variants, isoforms, synthetic nucleic acid molecules, protein isolated from human cells and tissues, and modified forms thereof. Exemplary reference human u-PA protease domains include, but are not limited to, wild-type, unmodified u-PA protease domain (SEQ ID NO:2) and an alternative protease domain (such as the u-PA protease domain set forth in SEQ ID NO: 5). One skilled in the art would recognize that the referenced positions of the u-PA protease domain (SEQ ID NO:2) differ by 178 amino acid residues when compared to the mature u-PA polypeptide (SEQ ID NO:1), which is the u-PA polypeptide containing the full length WT sequence. Thus, the first amino acid residue of SEQ ID NO:2 "corresponds to" the one hundred and seventy-nine (179) amino acid residue of SEQ ID NO:1. As used herein, a "modification" refers to a modification of an amino acid sequence of a polypeptide or a nucleotide sequence in a nucleic acid molecule and includes amino acid or nucleotide deletions, insertions, and replacements, respectively. Methods for modifying a polypeptide are routine to those skilled in the art, such as through the use of recombinant DNA methodologies. A distinction exists between modifications to the amino acid sequence of the polypeptide and modification of the polypeptide. The first refers to insertions, deletions and replacements or substitutions of amino acids; the second to modifications of the polypeptide, such as post-translational modifications, PEGylation and other modifications of proteins to alter properties and / or activities. As used herein, "substitution" or "replacement" refers to the replacement of cboynn / i znz / B / v one or more nucleotides or amino acids in a natural, target, wild-type, or other nucleic acid or polypeptide sequence with an alternative nucleotide or amino acid, without changing the length (as described by several residues) of the molecule. Thus, one or more substitutions in a molecule do not change the number of nucleotide or amino acid residues in the molecule. Amino acid replacements in comparison to a particular polypeptide can be expressed in terms of the amount of the amino acid residue along the length of the polypeptide sequence. For example, a modified polypeptide having an amino acid modification at the 35th position of the amino acid sequence that is a substitution / replacement of Arginine (Arg; R) with glutamine (Gln; Q) can be expressed as R35Q, Arg35Gln, or 35Q. Simply R35 can be used to indicate that the amino acid at the 35th modified position is an arginine. As used herein, a "modified u-PA" or "modified u-PA polypeptide" refers to a u-PA protease that exhibits altered activity, such as altered substrate specificity, compared to the normal form. not modified. These proteases include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more modifications (i.e., changes in amino acids) as compared with a wild-type u-PA, so that an activity, such as substrate specificity or selectivity, of the u-PA protease to cleave complement protein C3 is altered. A modified u-PA may be a full-length u-PA protease, or may be a portion thereof of a full-length protease, such as the u-PA protease domain, provided that the u-PA protease Modified PA contains modifications in regions that alter the activity or substrate specificity of the protease and the protease is proteolytically active. A modified u-PA protease, or a modified u-PA protease domain, can also include other modifications in regions that do not affect the substrate specificity of the protease. Therefore, a modified u-PA polypeptide usually has 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98 %, 99% or greater sequence identity to a corresponding amino acid sequence of a wild-type u-PA polypeptide. A modified full-length u-PA polypeptide or a catalytically active portion thereof or a protease domain thereof of a modified u-PA polypeptide can include polypeptides that are fusion proteins as long as the fusion protein possesses target specificity. As used herein, chymotrypsin numbering refers to the amino acid numbering of a mature chymotrypsin polypeptide of SEQ ID NO:76. Alignment of a protease domain of another protease, such as, for example, the protease domain of u-PA, can be performed with chymotrypsin. In that case, the amino acids of the u-PA polypeptide that correspond to chymotrypsin amino acids are given the cboynn / i znz / B / v chymotrypsin amino acid numbering. The corresponding positions can be determined for that alignment by one skilled in the art using manual alignments or using the many available alignment programs (eg, BLASTP). The corresponding positions can also be based on structural alignments, for example, through the use of computer simulated alignments of protein structure. Statement that amino acids in a polypeptide correspond to amino acids in a described sequence refers to amino acids identified upon alignment of the polypeptide with the described sequence to maximize identity or homology (where conserved amino acids align) using a standard alignment algorithm. , such as the GAP algorithm. The corresponding chymotrypsin numbers of amino acid positions 159-411 of the u-PA polypeptide set forth in SEQ ID NO:3 are provided in Table 1. The amino acid positions relative to the sequence set forth in SEQ ID NO:3 are in regular type, amino acid residues at those positions are in bold and the corresponding chymotrypsin numbers are in italics. For example, upon alignment of the serine protease domain of u-PA (SEQ ID NO:2) with mature chymotrypsin, isoleucine (I) at position 159 in u-PA is given the chymotrypsin numbering of 116. Consequently, subsequent amino acids are numbered. In one example, a phenylalanine (F) at amino acid position 173 of mature u-PA (SEQ ID NO:3) corresponds to amino acid position F30 based on chymotrypsin numbering. When a residue exists in a protease, but is not present in chymotrypsin, the amino acid residue is given a letter notation. For example, residues in chymotrypsin that are part of a loop with amino acid 60 based on chymotrypsin numbering, but are inserted into the sequence of u-PA as compared to chymotrypsin, are called, for example, D60a, Y60b or P60c. These residues correspond to D208, Y209 and P210, respectively, by numbering with respect to the mature u-PA sequence set forth in SEQ ID NO:3. Table 1. Chymotrypsin numbering of u-PA cboynn / i znz / B / v 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 1 74 175 176 177 178 179 180 181 182 183 184 185 186 187 188 A A I Y R R H R G G S V T Y V 31 32 33 34 35 36 37 37A 37B 37C 37D 38 39 40 41 189 190 191 192 193 194 195 196 197 198 201 202 203 44 45 46 47 48 49 50 51 52 53 54 55 56 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 A C F I D Y P K K E D Y I V Y 57 58 59 60 60A 60B 60 C 61 62 62A 63 64 65 6 6 67 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 L G R S R L N S N T Q G E M K 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 4 85 86 87 88 89 90 91 92 93 94 95 96 97 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 T L A H H N D I A L L K I R S 97A 97B 98 99 100 101 102 103 104 105 106 107 108 109 110 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 K E G R C A Q P S R T I Q T I 110 A 110 B 110 c 110 D 111 112 113 114 115 116 117 118 119 120 121 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 C L P S M Y N D P Q F G T S C 12 2 123 124 125 126 127 128 129 130 131 132 133 134 135 136 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 E I T G F G K E N S T D Y L Y 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 309 310 311 312 31 3 314 315 316 317 318 319 320 321 322 323 P E Q L K M T V V K L I S H R 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 E C Q Q P H Y Y G S E V T T K 167 168 169 170 170 A 170 B 171 172 173 174 175 176 177 178 179 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 M L C A A D P Q W K T D S C Q 180 181 182 183 184 185 185 A 185 B 186 187 188 189 190 191 192 cboynn / i ζηζ / Β / γ 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 G D S G G P L V C S L Q G R M 4 205 206 207 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 T L T G I V S W G R G C A L K 208 209 210 211 212 213 214 215 216 217 218 220 221 222 223 384 385 386 387 388 389 390 391 392 393 394 395 396 397 D W 223 A 224 225 226 227 228 229 230 231 232 233 234 235 236 237 399 400 401 402 403 404 405 406 407 408 409 410 411 I R S H T K E E N G L A L 238 239 240 241 242 243 244 245 246 247 248 249 250 CbQjnn / l 7Π7 / Β / Υ As used herein, kcat measures the catalytic activity of an enzyme; The units of kcat are seconds1. The reciprocal of kcat is the time required for an enzyme molecule to “convert a substrate molecule; kcat measures the number of substrate molecules converted by the enzyme molecule per second. kcat is sometimes called the refill number. In enzymology, kcat (also referred to as turnover number) is the maximum number of chemical conversions of substrate molecules per second that an individual catalytic site performs for a given enzyme. It is the maximum rate of reaction (Vmax) when all enzyme catalytic sites are saturated with substrate. As used herein, specificity for a target substrate refers to a preference for cleavage of a target substrate by a protease compared to another substrate, referred to as a non-target substrate. The specificity is reflected in the specificity constant (kcat / Km), which is a measure of the affinity of a protease for its substrate, and the efficiency of the enzyme, kcat / Km, a measure of the efficiency of the enzyme; a large value of kcat (fast turnover) or a small value of Km (high affinity for the substrate) makes kcat / Km large. As used herein, a specificity constant for cleavage is (kcat / Km), where Km is the Michaelis-Menton constant ([S] at mean Vmax) and kcat is the Vmax / [ET], where Et is the final concentration of the enzyme. The parameters kcat, Km and kcat / Kmse can be calculated by plotting the inverse of the substrate concentration versus the inverse of the substrate cleavage rate, and fitting the Lineweaver-Burk equation (1 / rate=(Km / Vmax)(1 / [S]) + 1 / Vmax; where VmáX=[ET]kcat). Any method to determine the rate of increase in cleavage over time in the presence of various concentrations of substrate can be used to calculate the specificity constant. For example, a substrate is bound to a fluorogenic moiety, which is released upon cleavage by a protease. By determining the rate of cleavage at different enzyme concentrations, kcat can be determined for a particular protease. The specificity constant can be used to determine the preference of a protease for a target substrate over another substrate. As used herein, substrate specificity refers to a protease's preference for one target substrate over another. Substrate specificity can be measured as a ratio of specificity constants. As used herein, a substrate specificity ratio is the ratio of specificity constants and can be used to compare specificities of two or more proteases or one protease to two or more substrates. For example, the substrate specificity of a protease for competing substrates or of competing proteases for a substrate can be compared by comparing kCat / Km. For example, a protease that has a specificity constant of 2 X 106M'1sec'1 for a target substrate and 2 X 104M_1sec_1 for a non-target substrate is more specific for the target substrate. Using the above specificity constants, the protease has a substrate specificity ratio of 100 for the target substrate. As used herein, substrate preference or specificity for a target substrate can be expressed as a substrate specificity ratio. The particular value of the ratio that reflects a preference is a function of the substrates and proteases in question. A substrate specificity ratio that is greater than 1 signifies a preference for a target substrate and a substrate specificity of less than 1 signifies a preference for a non-target substrate. In general, a ratio of at least or about 1 reflects a difference sufficient for a protease to be considered a candidate therapeutic. As used herein, altered specificity refers to a change in the substrate specificity of a modified protease compared to an initial wild-type protease. In general, the change in specificity is a reflection of the change in preference of a modified protease for a target substrate compared to a wild type protease substrate (referred to herein as a non-target substrate). Typically, the modified u-PA proteases provided herein exhibit a higher substrate specificity for complement C3 protein compared to the substrate specificity of the wild-type u-PA protease. For example, a modified protease that has a substrate specificity ratio of 100 for a target substrate versus a non-target substrate exhibits a 10-fold higher specificity compared to a scaffold cboynn / i znz / B / v protease with a ratio of 100. substrate specificity of 10. In another example, a modified protease having a substrate specificity ratio of 1 compared to a ratio of 0.1, exhibits a 10-fold increase in substrate specificity. To exhibit higher specificity compared to a scaffold protease, a modified protease has substrate specificity 1.5-fold, 2-fold, 5-fold, 10-fold, 50-fold, 100-fold, 200-fold, 300-fold, 400-fold, 500-fold or greater for any of the more than the complement proteins. As used herein, "selectivity" can be used interchangeably with specificity when referring to the ability of a protease to choose and cleave a target substrate among a mixture of competing substrates. The increased selectivity of a protease for a target substrate compared to any one or more other target substrates can be determined, for example, by comparing the specificity constants for cleavage of the target substrates by a protease. For example, if a protease has a cleavage specificity constant of 2 X 106M^sec-1 for a target substrate and 2 X 104M-1sec·1 for any of more substrates, the protease is more selective for the target substrate. As used herein, an "activity" or functional activity" of a polypeptide, such as a protease, refers to any activity exhibited by the polypeptide. These activities can be determined empirically. Example activities include, but are not limited to, the ability to interact with a biomolecule, eg, through substrate binding, DNA binding, or dimerization, enzyme activity, eg, kinase activity, or proteolytic activity. For a protease (including protease fragments), activities include, but are not limited to, the ability to specifically bind to a particular substrate, substrate binding affinity and / or specificity (e.g., high affinity and / or specificity). or low), effector functions, such as the ability to promote substrate (eg, protein, ie, C3) inhibition, neutralization, cleavage, or clearance, and in vivo activities, such as the ability to promote cleavage or clearance of proteins. Activity can be assessed in vitro or in vivo using recognized assays, such as ELISA, flow cytometry, surface plasmon resonance, or equivalent assays to measure histology and immunohistochemistry and immunofluorescence microscopy, cell-based assays, and binding assays. For example, for a protease, eg, a modified u-PA protease, activities can be assessed by measuring substrate protein cleavage, turnover, residual activity, stability, and / or in vitro and / or in vivo levels. Results of these in vitro assays indicating that a polypeptide exhibits activity can be correlated with the activity of the polypeptide in vivo, where the activity in vivo can be referred to as therapeutic activity or biological activity. The activity of a modified polypeptide can be any percentage level of activity of the unmodified polypeptide, including, but not limited to, at or about 1% activity, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , 100%, 200%, 300%, 400%, 500%, or more activity compared to the unmodified polypeptide. Assays to determine the functionality or activity of modified (or variant) proteases are well known in the art. Functional activities include, but are not limited to, biological activity, catalytic or enzymatic activity, antigenicity (ability to bind or compete with a polypeptide to bind to an anti-polypeptide antibody), immunogenicity, ability to form multimers, and ability to specifically bind. to a receptor or ligand for the polypeptide. As used herein, a functional activity in reference to a complement protein refers to a complement-mediated function including, but not limited to, anaphylaxis, opsonization, chemotaxis, or cell lysis. Examples of assays for evaluating complement activity activities include red blood cell hemolysis and detection of complement effector molecules such as by ELISA or SDS-PAGE. As used herein, catalytic activity or cleavage activity refers to the activity of a protease as assessed in in vitro proteolytic assays that detect proteolysis of a selected substrate. Cleavage activity can be measured by evaluating the catalytic efficiency of a protease. As used herein, activity towards a target substrate refers to cleavage activity and / or functional activity, or other measurement reflecting the activity of a protease on or towards a target substrate. A functional activity of a complement protein target substrate by a protease can be measured by evaluating an IC50 in a complement assay such as red blood cell lysis or other such assays known to a person skilled in the art or provided herein. to assess complement activity. Cleavage activity can be measured by evaluating the catalytic efficiency of a protease. For purposes herein, an activity is increased if a protease exhibits greater proteolysis or cleavage of a target substrate and / or modulates (ie, activates or inhibits) a functional activity of a complement protein as compared to the absence of the protease. As used herein, "increased activity" in reference to a modified u-PA polypeptide means that, when tested under the same conditions, the modified u-PA polypeptide exhibits increased activity as compared to a modified u-PA polypeptide. Unmodified PA that does not contain the amino acid replacements. For example, a modified u-PA polypeptide exhibits at least or about at least 110%, 120%, 130%, 140%, 150%, cboynn / i znz / B / v 160%, 170%, 180%, 190%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% or more of the activity of the polypeptide u- Unmodified or reference PA. As used herein, the term "same", when used in reference to antibody binding affinity, means that the EC50, association constant (Ka) or dissociation constant (Kd) is within about 1 to 100 times or 1 to 10 times that of the reference antibody (1-100 times higher affinity or 1-100 times lower affinity, or any numerical value or range or value within these ranges, than the reference antibody). As used herein, binding activity refers to characteristics of a molecule, eg, a polypeptide, related to whether and how it binds to one or more binding partners. Binding activities include the ability to bind to the binding partner, the affinity with which the binding partner binds (eg, high affinity), the strength of the binding to the binding partner, and / or the specificity for binding. with the union partner. As used herein, EC50, also called apparent Kd, is the concentration (eg, nM) of protease, where 50% of the maximum activity is observed at a fixed amount of substrate (eg, the concentration of polypeptide of modified u-PA needed to be cleaved through 50% of the available hC3). Typically, EC50 values ​​are determined from sigmoidal dose-response curves, where EC50 is the inflection point concentration. A high affinity of the protease for its substrate correlates with a low EC50 value and a low affinity corresponds to a high EC50 value. Affinity constants can be determined by standard kinetic methodology for protease reactions, eg, immunoassays, such as ELISA, followed by curve-fit analysis. As used herein, "affinity constant" refers to an association constant (Ka) used to measure the affinity or molecular binding strength between a protease and a substrate. The higher the affinity constant, the higher the affinity of the protease for the substrate. Affinity constants are expressed in units of reciprocal molarity (i.e., M-1> and can be calculated from the rate constant for the association and dissociation reaction as measured by standard kinetic methodology for protease substrate reactions. (eg, immunoassays, surface plasmon resonance, or other kinetic interaction assays known in the art.) The binding affinity of a protease can also be expressed as a dissociation constant, or Kd. The dissociation constant is the reciprocal of the association constant, Kd = 1 / Ka.Therefore, an affinity constant can also be represented by Kd.Affinity constants can be determined by standard kinetic methodology for association reactions. PbQynn / i znz / B / v protease, for example, immunoassays, surface plasmon resonance (SPR) (Rich and Myszka (2000) Curr. Opin. Biotechnol 11:54; Englebienne (1998) Analyst. 123:1599), isothermal titration calorimetry (ITC) or other kinetic interaction assays known in the art. technique (see, eg, Paul, ed., Fundamental Immunology, 2-ed., Raven Press, New York, pages 332-336 (1989)). Instruments and methods for real-time detection and monitoring of binding rates are known and commercially available (eg, BIAcore 2000, BIAcore AB, Uppsala, Sweden, and GE Healthcare Life Sciences; Malmqvist (2000) Biochem. Soc. Trans. 27:335). Methods for calculating affinity are well known, such as methods for determining EC50 values ​​or methods for determining association / dissociation constants, including those exemplified herein. For example, with respect to EC50, high binding affinity means that the protease specifically binds to a target protein with an EC50 that is less than about 10 ng / mL, 9 ng / mL, 8 ng / mL, 7 ng / mL, 6 ng / mL, 5 ng / mL, 3 ng / mL, 2 ng / mL, 1 ng / mL or less. High binding affinity can also be characterized by an equilibrium dissociation constant (Kd) of 10-6M or less, such as 10-7Μ, 10'8Μ, 10-10Μ, 10-11Μ or 10-12M or less. In terms of equilibrium association constant (Ka), high binding affinity is generally associated with Ka values ​​greater than or equal to approximately 106M1, greater than or equal to approximately 107M1, greater than or equal to approximately 108M-1, or greater than or equal to approximately 109Μ'1, 1010Μ'1, 1011Μ-1, or 1012M1. Affinity can be estimated empirically or affinities can be determined comparatively, eg, by comparing the affinity of two or more antibodies for a particular antigen, eg, by calculating even ratios of the affinities of the tested antibodies. For example, these affinities can be easily determined using standard techniques, such as by ELISA; balance dialysis; surface plasmon resonance; by radioimmunoassay using radiolabeled target antigen; or by another method known to the person skilled in the art. Affinity data can be analyzed, for example, by the method of Scatchard et al., Ann N.Y. Acad. Sel., 51:660 (1949) or by curve-fit analysis, for example, using a 4-parameter logistic nonlinear regression model using the equation: y = ((A-D) / (1+((x / C)AB))) + D, where A is the minimum asymptote, B is the slope factor, C is the inflection point (EC50), and D is the maximum asymptote. As used herein, ED50 is the dose (eg, mg / kg or nM) of a protease (eg, a modified u-PA) that produces a specific result (eg, cleavage of complement C3 protein) in 50% of the total population (eg, total amount of C3 present in the sample). As used herein, the term surface plasmon resonance is PbQynn / i znz / B / v refers to an optical phenomenon that allows analysis of interactions in real time by detecting alterations in protein concentrations within a biosensor array, for example, using the BIAcore system (GE Healthcare Life Sciences). As used herein, a human protein is one encoded by a nucleic acid molecule, such as DNA, present in the genome of a human, including all allelic variants and conservative variations thereof. A variant or modification of a protein is a human protein if the modification is based on the prominent sequence or wild type of a human protein. As used herein, naturally occurring α-amino acid residues are those 20 naturally occurring α-amino acid residues that are incorporated into protein by specific recognition of the tRNA molecule loaded with its cognate mRNA codon in humans. As used herein, non-naturally occurring amino acids refer to amino acids that are not genetically encoded. As used herein, "nucleic acid" refers to at least two linked nucleotides or nucleotide derivatives, including a deoxyribonucleic acid (DNA) and a ribonucleic acid (RNA) and analogs thereof, linked together, usually by bonds. of phosphodiester. Also included in the term "nucleic acid" are nucleic acid analogs such as peptide nucleic acid (PNA), phosphorothioate DNA and other analogs and derivatives or combinations thereof. Nucleic acids also include derivatives of DNA and RNA that contain, for example, a nucleotide analogue or a backbone that is not a phosphodiester bond, for example, a phosphotriester bond, a phosphoramidate bond, a phosphorothioate bond, a thioester bond or a peptide bond (peptide nucleic acid). The term also includes, as RNA or DNA equivalents, derivatives, variants, and analogs made from nucleotide analogs, single-stranded (sense or antisense), and double-stranded nucleic acids. Deoxyribonucleotides include deoxyadenosine, deoxycytidine, deoxyguanosine, and deoxythymidine. For RNA, the uracil base is uridine. Nucleic acids can be single or double stranded. When referring to probes or primers, which are optionally labeled, such as with a detectable label, such as a radioactive or fluorescent label, single stranded molecules are contemplated. These molecules are usually of a length such that their target is statistically unique or of low copy number (usually less than 5, generally less than 3) to probe or prime a library. In general, a probe or primer contains at least 14, 16, or 30 contiguous nucleotides of sequence complementary or identical to a gene of interest. Probes and primers can be 10, 20, 30, 50, 100 or more nucleotides long. cboynn / i znz / B / v As used herein, an isolated nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the natural source of the nucleic acid molecule. An isolated nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. Exemplary isolated nucleic acid molecules provided herein include isolated nucleic acid molecules encoding a provided u-PA protease. As used herein, "synthetic," referring to, for example, a synthetic nucleic acid molecule or synthetic gene or synthetic peptide refers to a nucleic acid molecule or polypeptide molecule that is produced by recombinant methods. and / or by chemical synthesis methods. As used herein, "polypeptide" refers to two or more covalently linked amino acids. The terms "polypeptide" and "protein" are used interchangeably herein. As used herein, a "peptide" refers to a polypeptide that is from 2 to about 40 amino acids in length. As used herein, amino acids occurring in the various amino acid sequences provided herein are identified according to their known three-letter or one-letter abbreviations (Table 2). The nucleotides that occur in the various nucleic acid fragments are designated by the standard single letter designations routinely used in the art. As used herein, an "amino acid" is an organic compound that contains an amino group and a carboxylic acid group. A polypeptide contains two or more amino acids. For purposes herein, amino acids include the twenty naturally occurring amino acids (Table 2), unnatural amino acids, and amino acid analogues (ie, amino acids where the α-carbon has a side chain). As used herein, amino acids, which occur in the various amino acid sequences of polypeptides appearing herein, are identified according to their familiar three-letter or one-letter abbreviations (see Table 2). Nucleotides, which occur in the various nucleic acid molecules and fragments, are designated by the standard single letter designations routinely used in the art. As used herein, "amino acid residue" refers to an amino acid formed upon chemical digestion (hydrolysis) of a polypeptide at its peptide bonds. The amino acid residues described herein are presumed to be in the "L" isomeric form. Residues in the "D" isomeric form, so designated, can be substituted for any L-amino acid residue as long as the polypeptide retains the PbQynn / i znz / B / v desired functional property. NH2 refers to the free amino group present at the amino terminus of a polypeptide. COOH refers to the free carboxyl group present at the carboxyl terminus of a polypeptide. In accordance with the standard polypeptide nomenclature described in J. Biol. Chem., 243: 3557-3559 (1968) and adopted at 37 C.F.R. §§ 1.821-1.822, the amino acid residue abbreviations are shown in Table 2: Table 2 - Correspondence table SYMBOL 1 Letters 3 Letters AMINO ACID Y Tyr Tyrosine G Gly Glycine F Phe Phenylalanine M Met Methionine A Ala Alanine S Ser Serine I lie Isoleucine L Leu Leucine T Thr Threonine V Val Valine P Pro Proline K Lys Lysine H His Histidine Q Gln Glutamine E Glu Glutamic acid Z Glx Glu and / or Gln W Trp Tryptophan R Arg Arginine D Asp Aspartic acid N Asn Asparagine B Asx Asn and / or Asp C Cys Cysteine ​​X Xaa Unknown or other All amino acid residue sequences represented herein by a formula have a left-to-right orientation in the conventional amino-to-carboxyl-terminus direction. The phrase "amino acid residue" includes the amino acids listed in the Correspondence Table (Table 2), modified, unnatural and unusual amino acids. Additionally, a hyphen at the beginning or end of a sequence of amino acid residues indicates a peptide bond to an additional sequence of one or more amino acid residues or to an amino-terminal group such as NH2 or a carboxyl-terminal group such as COOH. As used herein, "naturally occurring amino acids" refer to the 20 L-amino acids that occur in polypeptides. As used herein, naturally occurring α-amino acid residues are those 20 naturally occurring α-amino acid residues that are incorporated into protein by specific recognition of the tRNA molecule loaded with its codon. of cognate mRNA in humans. As used herein, "unnatural amino acid" refers to an organic compound that has a similar structure to a natural amino acid but has been structurally modified to mimic the structure and reactivity of a natural amino acid. Thus, non-naturally occurring amino acids include, for example, amino acids or amino acid analogs other than the 20 naturally occurring amino acids and include, but are not limited to, D-stereoisomers of amino acids. Exemplary unnatural amino acids are known to those skilled in the art, and include, but are not limited to, para-acetyl-phenylalanine, para-azido-phenylalanine, 2-Aminoadipic acid (Aad), 3-aminoadipic acid ( bAad), βalanine / p-amino-propionic acid (Bala), 2-aminobutyric acid (Abu), 4-aminobutyric / piperidinic acid (4Abu), 6-amino-caproic acid (Acp), 2-Aminoheptanoic acid ( Ahe), 2-Aminoisobutyric Acid (Aib), 3-Aminoisobutyric Acid (Baib), 2-Aminopimelic Acid (Apm), 2,4-Diaminobutyric Acid (Dbu), Desmosine (Des), 2,2'-diam¡ acid nopimelic (Dpm), 2,3-diaminopropionic acid (Dpr), N-ethylglycine (EtGly), N-ethylasparagine (EtAsn), hydroxylysine (Hyl), allo-hydroxylysine (Ahyl), 3-Hydroxyproline (3Hyp), 4-Hydroxyproline (4Hyp), Isodesmosine (Ide), Allo-Isoleucine (Aile), N-Methylglycine, Sarcosine (MeGly), N-Methylisoleucine (Melle), 6-NMethylisine (MeLys), N-Methylvaline (MeVal), Norvaline (Nva), Norleucine (Nle) and Ornithine (Orn). Exemplary unnatural amino acids that are known to those skilled in the art are described herein. As used herein, an isokinetic mixture is one in which the molar ratios of amino acids have been adjusted based on their reported reaction rates (see, eg, Ostresh et al. (1994) Biopolymers 34:1681). As used herein, a DNA construct is a single or double stranded, linear or circular DNA molecule that contains combined and juxtaposed DNA segments in a manner not found in nature. DNA constructs exist as a result of human manipulation and include clones and other copies of engineered molecules. As used herein, a DNA segment is a portion of a larger DNA molecule that has specified attributes. For example, a segment of DNA encoding a specific polypeptide is a portion of a larger DNA molecule, such as a plasmid or plasmid fragment, which, when read from the 5' to 3' direction, encodes the amino acid sequence of the specified polypeptide. As used herein, the term "ortholog" means a polypeptide or protein obtained from one species that is the functional counterpart of a polypeptide or protein from a different species. Sequence differences between orthologs are the result of speciation. cboynn / i znz / B / v As used herein, the term "polynucleotide" means a single or double stranded polymer of deoxyribonucleotides or ribonucleotide bases read from the 5' end to the 3' end. Polynucleotides include RNA and DNA, and can be isolated from natural sources, synthesized in vitro, or prepared from a combination of natural and synthetic molecules. The length of a polynucleotide molecule is given herein in terms of nucleotides (abbreviated nt") or base pairs (abbreviated "bp"). The term nucleotides is used for single and double stranded molecules where the context allows. When the term is applied to double-stranded molecules, it is used to denote the full length and is understood to be equivalent to the term base pairs. It is understood by those skilled in the art that the two strands of a double-stranded polynucleotide may differ slightly in length and that the ends thereof may stagger; therefore, all nucleotides within a double-stranded polynucleotide molecule cannot be paired. These unpaired ends generally do not exceed 20 nucleotides in length. As used herein, "sequence alignment" refers to the use of homology to align two or more nucleotide or amino acid sequences. Typically, two or more sequences that are related by 50% or more identity are aligned. An aligned set of sequences refers to 2 or more sequences that are aligned at corresponding positions and may include RNA-derived alignment sequences, such as ESTs and other cDNAs, aligned with genomic DNA sequences. Related or variant polypeptides or nucleic acid molecules can be aligned by any method known to those skilled in the art. Typically these methods maximize matches and include methods such as using manual alignments and using the many available alignment programs (eg, BLASTP) and others known to those skilled in the art. By aligning polypeptide or nucleic acid sequences, one skilled in the art can identify analogous portions or positions, using identical and conserved amino acid residues as guides. Additionally, one skilled in the art can also use conserved amino acid or nucleotide residues as guides to find corresponding amino acid or nucleotide residues between human and non-human sequences. The corresponding positions can also be based on structural alignments, for example, through the use of computer simulated alignments of protein structure. In other cases, corresponding regions can be identified. One skilled in the art can also use conserved amino acid residues as guides to find corresponding amino acid residues between human and non-human sequences. As used herein, "sequence identity" refers to the number of identical or similar amino acids or nucleotide bases in a comparison between a PbQynn / i znz / B / v test and a reference polypeptide or polynucleotide. Sequence identity can be determined by sequence alignment of the nucleic acid or protein sequences to identify regions of similarity or identity. For purposes herein, sequence identity is generally determined by alignment to identify identical residues. The alignment can be local or global. Matches, mismatches, and gaps can be identified between compared sequences. Gap are null amino acids or nucleotides inserted between residues of aligned sequences such that identical or similar characters align. In general, there may be internal and terminal separations. Sequence identity can be determined by considering gaps as the number of identical residues / length of the shortest sequence x 100. When gap penalties are used, sequence identity can be determined without penalty for trailing gaps (for example , terminal separations are not penalized). Alternatively, sequence identity can be determined without regard to gaps as the number of identical positions / length of the total aligned sequence x 100. As used herein, a position corresponding to or a statement that nucleotides or amino acid positions “correspond to nucleotides or amino acid positions in a described sequence, as set forth in the sequence listing, refers to nucleotides or amino acid positions identified by aligning to the disclosed sequence to maximize identity using a standard alignment algorithm, such as the GAP algorithm. For purposes herein, the alignment of a u-PA sequence is to the human u-PA protease domain amino acid sequence set forth in SEQ ID NO: 2 or 5, particularly a reference human u-PA of SEQ ID NO:5. By aligning the sequences, one skilled in the art can identify corresponding residues, eg, using identical, conserved amino acid residues as guides. In general, to identify corresponding positions, amino acid sequences are aligned such that the highest order match is obtained (see, for example: Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey , 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo et al. al (1988) SIAM J Applied Math 48:1073). Alternatively, one skilled in the art can number the residues by the chymotrypsin number, thereby identifying the corresponding residues. For closely related sequences, a computer algorithm is not needed; the alignment can be done visually cboynn / i znz / B / v. As used herein, a "global alignment" is an alignment that aligns two sequences from start to finish, aligning each letter in each sequence only once. An alignment occurs, regardless of whether there is similarity or identity between the sequences. For example, 50% sequence identity based on "overall alignment" means that in a complete sequence alignment of two compared sequences each 100 nucleotides in length, 50% of the residues are the same. It is understood that global alignment can also be used to determine sequence identity even when the lengths of the aligned sequences are not the same. Differences in the termini of the sequences are taken into account to determine sequence identity, unless "no penalty for end-offs" is selected. In general, a global alignment is used for sequences that share significant similarity over most of their length. Exemplary algorithms for performing global alignment include the Needleman-Wunsch algorithm (Needleman et al. (1970) J. Mol. Biol. 48:443). Example programs for performing global alignment are publicly available and include the Global Sequence Alignment Tool available on the National Center for Biotechnology Information (NCBI) website (ncbi.nlm.nih.gov / ), and the program available at deepc2.psi.iastate.edu / aat / align / align.html. As used herein, a "local alignment" is an alignment that aligns two sequences, but only aligns those parts of the sequences that share similarity or identity. Therefore, a local alignment determines whether subsegments of one sequence are present in another sequence. If there is no similarity, no alignment is returned. Local alignment algorithms include BLAST algorithm and Smith-Waterman {Adv. Appl. Math. 2:482 (1981)). For example, 50% sequence identity based on "local alignment" means that in a complete sequence alignment of two compared sequences of any length, a region of similarity or identity 100 nucleotides in length has 50% of the residues that they are the same in the region of similarity or identity. For purposes herein, sequence identity can be determined by standard alignment algorithm programs used with predetermined separation penalties set by each vendor. Default parameters for the GAP program may include: (1) a nail comparison matrix (containing a value of 1 for identities and 0 for non-identities) and the weighted comparison matrix of Gribskov et al. (1986) Nuci. Acids Res. 14: 6745, as described by Schwartz and Dayhoff, eds., Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, pp. 353358 (1979); (2) a 3.0 penalty for each gap and an additional 0.10 penalty for each symbol on each gap; and (3) no penalty for separations CbQjnn / l 7Π7 / Β / Υ late. If any two nucleic acid molecules have nucleotide sequences or any two polypeptides have amino acid sequences that are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical, or other similar variations citing percent identity, can be determined using known computer algorithms based on local or global alignment (see, for example, wikipedia.org / wiki / Sequence_alignment_software, providing links to dozens of databases and programs known and publicly available alignment patterns). In general, for purposes herein, sequence identity is determined using computational algorithms based on global alignment, such as the Needleman-Wunsch global sequence alignment tool available from NCBI / BLAST (blast.ncbi.nlm. n¡h.gov / Blast.cgi?CMD=Web&Page_TYPE=BlastHome); LAlign (William Pearson implementing the Huang and Miller algorithm (Adv. Appl. Math. (1991) 12:337-357)); and Xiaoqui Huang's program available at deepc2.psi.iastate.edu / aat / align / align.html. In general, when comparing nucleotide sequences herein, an alignment with penalty for end gaps is used. Local alignment can also be used when the sequences being compared are substantially the same length. As used herein, the term "identity" represents a comparison or alignment between a test and a reference polypeptide or polynucleotide. In one non-limiting example, "at least 90% identical to" refers to 90% to 100% percent identities to the reference polypeptide or polynucleotide. Identity at a level of 90% or more is indicative of the fact that, assuming for exemplification purposes a test and reference polypeptide or polynucleotide length of 100 amino acids or nucleotides are compared, they do not differ by more than 10% (i.e. , 10 out of 100) of amino acids or nucleotides in the test polypeptide or polynucleotide from that of the reference polypeptides. Similar comparisons can be made between a test and reference polynucleotide. These differences can be represented as randomly distributed point mutations throughout the length of an amino acid sequence, or they can be clustered at one or more locations of variable length up to the maximum allowable, e.g., 10 / 100 amino acid difference (approximately 90% of identity). Differences may also be due to deletions or truncations of amino acid residues. Differences are defined as nucleic acid or amino acid substitutions, insertions or deletions. Depending on the length of the sequences compared, at the level of homologies or identities above about 85-90%, the result may be independent of the program and set of separation parameters; these high levels of identity can be easily assessed, often without relying on software. As used herein, a disulfide bond (also called an S-S bond or cboynn / i znz / B / v a disulfide bridge) is a single covalent bond derived from the coupling of thiol groups. Disulfide bonds in proteins form between the thiol groups of cysteine ​​residues and stabilize interactions between polypeptide domains. As used herein, "coupled" or "conjugated" means linked through a covalent or non-covalent interaction. The conjugates provided herein contain a modified u-PA polypeptide protease domain (referred to as an SPD, see, for example, Figure 4), and all or part of the remaining u-PA polypeptide, bound directly or via a linker to another moiety, such as a polypeptide that confers a property, such as increased serum half-life (i.e., human serum albumin HSA), or facilitates expression or purification (i.e., SUMO, his-SUMO, TSG-6), or directs the protein to the receptor, such as an antibody that binds to a receptor. The polypeptide can be linked directly or via a polypeptide linker, generally few amino acids, about 4-20, such as combinations of Ser and Gly residues. Conjugates containing a polypeptide are generally fusion proteins. Conjugates also include modified u-PA polypeptides in which amino acid residues are attached to moieties, such as PEG moieties, glycosylation moieties, and the like. As used herein, "primer" refers to a nucleic acid molecule that can act as a starting point for template-directed DNA synthesis under appropriate conditions (for example, in the presence of four different nucleoside triphosphates and an agent). polymerization agents, such as DNA polymerase, RNA polymerase, or reverse transcriptase) in a suitable buffer and at a suitable temperature. It is understood by those skilled in the art that certain nucleic acid molecules can serve as a "probe" and as a "primer". However, one primer has a 3' hydroxyl group for extension. A primer can be used in a variety of methods, including, for example, polymerase chain reaction (PCR), reverse transcriptase (RT) POR, RNA PCR, LCR, multiplexed POR, narrow-band PCR, PCR capture, expression PCR, 3' and 5' RACE, in situ PCR, ligation-mediated PCR and other amplification protocols. As used herein, "primer" refers to an oligonucleotide containing two or more deoxyribonucleotides or ribonucleotides, usually more than three, from which synthesis of a primer extension product can be initiated. Experimental conditions conducive to synthesis include the presence of nucleoside triphosphates and an agent for polymerization and extension, such as DNA polymerase, and a suitable buffer, temperature, and pH. As used herein, "primer pair" refers to a set of primers that includes a 5' (upstream) primer that anneals to the 5' end of a sequence to be amplified (eg, by PCR ) and a 3' primer (downstream) that anneals to cboynn / i znz / B / v the complement of the 3' end of the sequence to be amplified. As used herein, "specifically hybridized" refers to attachment, by complementary base pairing, of a nucleic acid molecule (eg, an oligonucleotide) with a target nucleic acid molecule. Those skilled in the art are familiar with in vitro and in vivo parameters that affect specific hybridization, such as the length and composition of the particular molecule. Parameters particularly relevant to in vitro hybridization further include fixation and wash temperature, buffer composition, and salt concentration. Example wash conditions to remove non-specifically bound nucleic acid molecules at high stringency are 0.1 x SSPE, 0.1% SDS, 65°C and at medium stringency are 0.2 x SSPE, 0.1% SDS, 50°C. Equivalent stringency conditions are known in the art. One of skill in the art can easily adjust these parameters to achieve specific hybridization of a nucleic acid molecule with a suitable target nucleic acid molecule for a particular application. As used herein, substantially identical to a product means similar enough that the property of interest does not change enough that the substantially identical product can be used in place of the product. As used herein, the terms "substantially identical" or "similar" are also understood to vary with context as understood by those skilled in the relevant art. As used herein, the wild type form of a polypeptide or nucleic acid molecule is a form encoded by a gene or by a coding sequence encoded by the gene. Typically, a wild-type form of a gene, or the molecule encoded by it, does not contain mutations or other modifications that alter function or structure. The term wild type also encompasses forms with allelic variation such as occurs between two or more species. As used herein, a predominant form of a polypeptide or nucleic acid molecule refers to a form of the molecule that is the major form produced from a gene. A "predominant form" varies from source to source. For example, different types of cells or tissues can produce different forms of polypeptide, eg, through alternative splicing and / or alternative protein processing. In each type of cell or tissue, a different polypeptide may be a "predominant form." As used herein, an allelic variant or allelic variation refers to any of two or more alternative forms of a gene that occupy the same chromosomal locus. Allergic variation arises naturally through mutation and can result in phenotypic polymorphism within populations. Gene mutations may be subtle (no change in the encoded polypeptide) or may code for cboynn / i znz / B / v polypeptides having altered amino acid sequence. The term "allelic variant" is also used herein to denote a protein encoded by an allelic variant of a gene. Typically, the reference form of the gene codes for a wild type and / or predominant form of a polypeptide from a population or individual reference member of a species. Typically, allelic variants, including variants between two or more species, have at least 80%, 90%, or more amino acid identity with a wild-type and / or predominant form from the same species; the degree of identity depends on the gene and whether the comparison is between species or intraspecies. In general, intraspecies allelic variants have at least or at least approximately 80%, 85%, 90%, or 95% identity or more with a wild type and / or predominant form, including at least or at least approximately 96%, 97 %, 98%, 99% or more identity with a wild type and / or predominant form of a polypeptide. As used herein, "allele", used interchangeably herein with "allelic variant" refers to alternative forms of a gene or parts thereof. Alleles occupy the same locus or position on homologous chromosomes. When a subject has two identical alleles of a gene, the subject is said to be homozygous for that gene or allele. When a subject has two different alleles of a gene, the subject is said to be heterozygous for the gene. The alleles of a specific gene may differ from each other by a single nucleotide or by several nucleotides, and may include nucleotide substitutions, deletions, and insertions. An allele of a gene can also be a form of a gene that contains a mutation. As used herein, "species variants" refers to variants in polypeptides between different species, including different mammalian species, such as mouse and human. In general, species variants are approximately either 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99 % or more sequence identity. Corresponding residues between two or more species variants can be determined by sequence comparison and alignment to maximize the number of matching nucleotides or residues, for example, so that sequence identity is equal to or greater than 95%, equal or greater than 96%, equal to or greater than 97%, equal to or greater than 98%, or greater than 99%. The position of interest is then given the assigned number in the reference nucleic acid molecule. Alignment can be done manually or by eye, particularly when sequence identity is greater than 80%. As used herein, a splice variant refers to a variant produced by differential processing of a primary transcript of genomic DNA that results in more than one type of mRNA. As used herein, modification referring to modification of the CbQjnn / l 7Π7 / Β / Υ primary amino acid sequence of a polypeptide or a nucleotide sequence in a nucleic acid molecule and includes amino acid and nucleotide deletions, insertions, and replacements, respectively. This is in contrast to modifications of the polypeptide itself, which include post-translational modifications, such as glycosylation, farnisylation, pegylation, and fusions, such as fusions with other polypeptides to change a property, such as serum half-life, such as through albumin, albumin fusion. , such as human serum albumin, and other such modifications to the polypeptide. Thus, reference to amino acid sequence modifications refers to insertions, deletions, substitutions / replacements, and combinations thereof. Polypeptide modification refers to modifications that are added to the polypeptide that do not change the sequence thereof. For purposes herein, amino acid substitutions, deletions, and / or insertions may be made in any of the u-PA polypeptides or catalytically active fragments thereof as long as the resulting protein exhibits protease or other activity (or, if desired, these changes can be made to remove the activity). Modifications can be made by making conservative amino acid substitutions and also non-conservative amino acid substitutions. For example, amino acid substitutions may be made that desirably or advantageously alter the properties of proteins. In one embodiment, mutations can be made that prevent degradation of the polypeptide. Many proteases cleave after basic residues, such as R and K; to eliminate that cleavage, the basic residue is replaced with a nonbasic residue. The interaction of the protease with an inhibitor can be blocked while retaining catalytic activity by effecting a non-conservative change in the site of interaction of the inhibitor with the protease. Other activities can be altered as well. For example, receptor binding can be altered without altering catalytic activity. Contemplated amino acid substitutions include conservative substitutions, such as those set forth in Table 3, that do not abolish proteolytic activity. As described herein, substitutions that alter the properties of proteins, such as removal of cleavage sites and other similar sites, are also contemplated; these substitutions are generally not conservative, but can be easily made by those skilled in the art. As used herein, suitable conservative amino acid substitutions are known to those skilled in the art and can generally be made without altering the biological activity of the resulting molecule. Those skilled in the art generally recognize that individual amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, for example, Watson et al. Molecular Biology of the Gene, 4th Edition, 1987, The Benjamin / Cummings Pub. Co., p.224). These substitutions can be made according to those set forth in Table 3 as follows: PbQynn / i znz / B / v Table 3 Parent residue Example conservative substitution Ala (A) Gly; Ser Arg (R) Lys Asn (N) Gln; His Cys (C) Ser Gln (Q) Asn Glu (E) Asp Gly (G) Ala; Pro Hls(H) Asn; Gln He(I) Leu; Val Leu (L) He; Val Lys (K) Arg; glin; Glu Met (M) Leu; tyr; He Phe (F) Met; read; Tyr Ser(S) Thr Thr(T) Ser Trp(W) Tyr Tyr(Y) Trp; Phe Val(V)lie; read Other substitutions are also permissible and can be determined empirically or according to known conservative substitutions. As used herein, the term "promoter" means a portion of a gene that contains DNA sequences that provide for RNA polymerase binding and initiation of transcription. Promoter sequences are commonly, but not always, found in the 5' non-coding region of genes. As used herein, the isolated or purified polypeptide or protein or biologically active portion thereof is substantially free of contaminating cellular material or other proteins from the tissue cell from which the protein is derived, or substantially free of chemical precursors. or other chemicals when chemically synthesized. Preparations may be determined to be substantially free if they appear free of readily detectable impurities as determined by standard methods of analysis, such as thin layer chromatography (TLC), gel electrophoresis, and high performance liquid chromatography (HPLC), used by those skilled in the art to assess that purity, or pure enough that further purification does not detectably alter the physical and chemical properties, such as enzymatic and biological activities, of the substance. Methods for purification of the compounds to produce substantially chemically pure compounds are known to those skilled in the art. However, a substantially chemically pure compound may be a mixture of stereoisomers. In these cases, further purification can increase the specific activity of the compound. The term "substantially free of cellular material" includes protein preparations in which the protein is separated from the cellular components of the cells from which it is isolated or recombinantly produced. In one embodiment, the term "substantially free of cellular material" includes preparations of protease proteins that have less than about 30% (by dry weight) of non-protease proteins (also referred to herein as a contaminating protein), generally less than about 20% non-protease proteins or 10% non-protease proteins or less than about 5% non-protease proteins. When the protease protein or active portion thereof is recombinantly produced, it is also substantially free of culture medium, that is, the culture medium represents less than, about, or equal to 20%, 10%, or 5% of the volume of the protease protein preparation. As used herein, the term "substantially free of chemical precursors or other chemicals" includes protease protein preparations in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. The term includes preparations of protease proteins that have less than about 30% (by dry weight), 20%, 10%, 5% or less of chemical precursors or chemicals or non-protein components. As used herein, production by recombinant means using recombinant DNA methods refers to the use of well-known methods of molecular biology to express proteins encoded by cloned DNA. As used herein, "expression" refers to the process by which polypeptides are produced by transcription and translation of polynucleotides. The expression level of a polypeptide can be assessed using any method known in the art, including, for example, methods for determining the amount of the polypeptide produced from the host cell. These methods may include, but are not limited to, quantification of the polypeptide in the cell lysate by ELISA, Coomassie blue staining after gel electrophoresis, Lowry protein assay, and Bradford protein assay. As used herein, a "host cell" is a cell that is used to receive, maintain, reproduce, and / or amplify a vector. The host cells can also be used to express the polypeptide encoded by the vector. The nucleic acid contained in the vector is replicated when the host cell divides, thus amplifying the nucleic acids. As used herein, a "vector" or plasmid is a cboynn / i znz / B / v replicable nucleic acid from which one or more heterologous proteins can be expressed when the vector is transformed into a suitable host cell. Reference to a vector includes discrete elements that are used to introduce heterologous nucleic acid into cells for expression or replication thereof. Reference to a vector also includes those vectors into which a nucleic acid encoding a polypeptide or fragment thereof can be introduced, usually by restriction digest and ligation. Reference to a vector also includes those vectors that contain nucleic acid encoding a protease, such as a modified u-PA. The vector is used to introduce the nucleic acid encoding the polypeptide into the host cell for amplification of the nucleic acid or for expression / display of the polypeptide encoded by the nucleic acid. Vectors usually remain episomal, but can be designed to effect integration of a gene or part thereof into a chromosome of the genome. Vectors that are artificial chromosomes, such as yeast artificial chromosomes and mammalian artificial chromosomes, are also contemplated. The selection and use of these vehicles are well known to those skilled in the art. A vector also includes "virus vectors" or "viral vectors." Viral vectors are modified viruses that are operably linked to foreign genes to transfer (as vehicles or transporters) the foreign genes into cells. As used herein, an "expression vector" includes vectors capable of expressing DNA that is operatively linked to regulatory sequences, such as promoter regions, that are capable of effecting expression of these DNA fragments. These additional segments may include promoter and terminator sequences, and may optionally include one or more origins of replication, one or more selectable markers, an enhancer, a polyadenylation signal, and the like. Expression vectors are generally derived from plasmid or viral DNA, or may contain elements of both. Thus, an expression vector refers to a recombinant DNA or RNA construct, such as a plasmid, phage, recombinant virus, or other vector that, upon introduction into a suitable host cell, results in expression of the DNA. cloned. Appropriate expression vectors are well known to those skilled in the art and include those that are replicable in eukaryotic and / or protuberance cells and those that remain episomal or those that integrate into the host cell genome. As used herein, vector also includes "virus vectors" or "viral vectors." Viral vectors are modified viruses that are operably linked to foreign genes to transfer (as vehicles or transporters) the foreign genes into cells. As used herein, an adenovirus refers to any of a group of DNA-containing viruses that cause conjunctivitis and upper respiratory tract infections in humans. As used herein, naked DNA refers to DNA free of cboynn / i znz / B / v history that can be used for vaccines and gene therapy. Naked DNA is the genetic material that is passed from cell to cell during a processed gene transfer called transformation. In transformation, purified or naked DNA is taken up by the recipient cell which will give the recipient cell a new characteristic or phenotype. As used herein, "operably linked" with reference to nucleic acid sequences, regions, elements, or domains means that the nucleic acid regions are functionally related to one another. For example, the nucleic acid encoding a guide peptide can be operatively linked to the nucleic acid encoding a polypeptide, whereby the nucleic acids can be transcribed and translated to express a functional fusion protein, where the guide peptide effects the secretion of the fusion polypeptide. In some cases, nucleic acid encoding a first polypeptide (eg, a guide peptide) is operably linked to nucleic acid encoding a second polypeptide and the nucleic acids are transcribed as a single mRNA transcript, but translation of the mRNA transcription can result in the expression of one of two polypeptides. For example, an amber stop codon can be located between the nucleic acid encoding the first polypeptide and the nucleic acid encoding the second polypeptide, so that when introduced into a partial amber suppressor cell, the mRNA transcript The resulting individual protein can be translated to produce a fusion protein containing the first and second polypeptides, or can be translated to produce the first polypeptide alone. In another example, a promoter can be operably linked to nucleic acid encoding a polypeptide, whereby the promoter regulates or mediates transcription of the nucleic acid. As used herein, "primary sequence" refers to the sequence of amino acid residues in a polypeptide or the sequence of nucleotides in a nucleic acid molecule. As used herein, protein-binding sequence refers to a protein or peptide sequence that is capable of specifically binding to other protein or peptide sequences in general, to a set of protein or peptide sequences, or to a sequence of a particular protein or peptide. As used herein, a "tag" or an "epitope tag" refers to a sequence of amino acids, typically added to the C-terminus or N-terminus of a polypeptide, such as a u-PA provided in the present. Inclusion of tags fused to a polypeptide may facilitate purification and / or detection of polypeptides. Typically, a tag or tag polypeptide refers to a polypeptide that has enough residues to provide an epitope recognized by an antibody or can be used for detection or purification, but is short enough not to interfere. PbQynn / i znz / B / v with the activity of the polypeptide to which it is bound. The tag polypeptide is usually sufficiently unique that an antibody that specifically binds to it will not substantially cross-react with epitopes on the polypeptide to which it is bound. Epitope-tagged proteins can be affinity purified using highly specific antibodies raised against the tags. Suitable tag polypeptides are generally at least 5 or 6 amino acid residues and usually between about 8-50 amino acid residues, usually between 9-30 residues. The tags can be attached to one or more proteins and allow the detection of the protein or its recovery from a sample or mixture. These tags are well known and can be easily synthesized and designed. Exemplary tag polypeptides include those used for affinity purification and include Small Ubiquitin-like Modifier (SUMO) tags, FLAG tags, His tags, the influenza hemagglutinin (HA) tag polypeptide and its antibody 12CA5, (Field et al (1988) Mol Cell Biol 8:2159-2165); the c-myc tag and the 8F9, 3C7, 6E10, G4, B7 and 9E10 antibodies thereto (see, for example, Evan et al. (1985) Molecular and Cellular Biology 5:3610-3616); and the herpes simplex virus glycoprotein D (gD) tag and its antibody (Paborsky et al. (1990) Protein Engineering 3:547-553). An antibody used to detect an epitope-tagged antibody is commonly referred to herein as a secondary antibody. As used herein, metal binding sequence refers to a protein or peptide sequence that is capable of specifically binding to metal ions in general, to a set of metal ions, or to a particular metal ion. As used herein, the term "assessment" is intended to include quantitative and qualitative determination in the sense of obtaining an absolute value for the activity of a protease, or a domain thereof, present in the sample, and also of obtaining an index. , ratio, percentage, visual value or other value indicative of the level of activity. The evaluation can be direct or indirect and the chemical species actually detected need not, of course, be the proteolysis product itself, but can, for example, be a derivative thereof or some additional substance. For example, detection of a complement protein cleavage product, such as by SDS-PAGE and Coomassie blue staining of proteins. As used herein, "biological activity" refers to the in vivo activities of a compound or physiological responses that result from in vivo administration of a compound, composition, or other mixture. Biological activity, therefore, encompasses the therapeutic effects and pharmaceutical activity of these compounds, compositions, and mixtures. Biological activities can be observed in in vitro systems designed to test or use PbQynn / i znz / B / v these activities. Thus, for purposes herein, a biological activity of a protease is its catalytic activity in which a polypeptide is hydrolyzed. As used herein, equivalent, when referring to two nucleic acid sequences, means that the two sequences in question encode the same amino acid sequence or equivalent proteins. When equivalent is used in reference to two proteins or peptides, it means that the two proteins or peptides have substantially the same amino acid sequence with only amino acid substitutions (such as, but not limited to, conservative changes as set forth in Table 3, above) that do not substantially alter the activity or function of the protein or peptide. When "equivalent" refers to a property, the property need not be present to the same extent (for example, two peptides may exhibit different rates of the same type of enzyme activity), but the activities are usually substantially the same. Complementary, when referring to two nucleotide sequences, means that the two nucleotide sequences are capable of hybridizing, usually with less than 25%, 15%, or 5% mismatches between opposite nucleotides. If necessary, the percentage of complementarity shall be specified. Usually, the two molecules are selected to hybridize under high stringency conditions. As used herein, an agent that modulates the activity of a protein or the expression of a gene or nucleic acid decreases or increases or otherwise alters the activity of the protein or, in some way, up-regulates or down-regulates it. or otherwise alters the expression of the nucleic acid in a cell. As used herein, a "chimeric protein" or "fusion protein" protease refers to a polypeptide operatively linked to a different polypeptide. A chimeric or fusion protein provided herein may include one or more proteases or a portion thereof, such as single-chain protease domains thereof, and one or more additional polypeptides for any one or more control signals. transcriptional / translational, signal sequences, a localization tag, a purification tag, part of an immunoglobulin G domain and / or a targeting agent. These chimeric or fusion proteins include those produced by recombinant means as fusion proteins, those produced by chemical means, such as by chemical coupling, through, for example, coupling to sulfhydryl groups, and those produced by any other method through the which at least one protease, or a part thereof, is linked, directly or indirectly through linkers, to another polypeptide. As used herein, "operably linked" when referring to a fusion protein refers to a protease polypeptide and a non-protease polypeptide that are fused in-frame with each other. The non-protease polypeptide can be fused to the N-terminus or C-terminus of the protease polypeptide. As used herein, a targeting agent is any moiety, such as a protein or effective portion thereof, that provides specific binding of the conjugate to a cell surface receptor, which in some cases can internalize bound conjugates or moieties of the same. A targeting agent can also be one that promotes or facilitates, for example, the isolation or affinity purification of the conjugate; binding of the conjugate to a surface; or detection of the conjugate or complexes containing the conjugate. As used herein, "linker" refers to short amino acid sequences that bind two polypeptides (or nucleic acid encoding these polypeptides). "Peptide linker" refers to the short amino acid sequence that joins the two polypeptide sequences. Examples of polypeptide linkers are linkers that join two antibody chains into a synthetic antibody fragment such as an scFv fragment. Linkers are known and any known linker can be used in the provided methods. Examples of polypeptide linkers are (Gly-Ser)n amino acid sequences, with some Glu or Lys residues scattered to increase solubility. Other exemplary linkers are described herein; any of these and other known linkers can be used with the provided compositions and methods. As used herein, the derivative or analog of a molecule refers to a portion derived from or a modified version of the molecule. As used herein, disease or disorder refers to a pathological condition in an organism that results from a cause or condition including, but not limited to, infections, acquired conditions, genetic conditions, conditions related to environmental exposures, and human behaviors. , and conditions characterized by identifiable symptoms. Diseases or disorders include clinically diagnosed diseases, as well as disorders in the normal state of the body that have not been diagnosed as clinical disease. The diseases and disorders of interest herein are those that involve complement activation, including those mediated by complement activation and those in which complement activation plays a role in the etiology or pathology. Diseases and disorders of interest herein include those characterized by complement activation (eg, age-related macular degeneration and delayed kidney graft function). As used herein, macular degeneration occurs when the small central portion of the retina, known as the macula, deteriorates. There are two types of AMD: dry (atrophic) and wet (neovascular or exudative). Most AMD starts out as the PbQynn / i znz / B / v dry type and in 10-20% of individuals, it progresses to the wet type. Age-related macular degeneration is always bilateral (meaning it occurs in both eyes), but it doesn't necessarily progress at the same rate in both eyes. As used herein, age-related macular degeneration (AMD) is an inflammatory disease that causes visual impairment and blindness in older people. The proteins of the complement system are central to the development of this disease. Local and systemic inflammation in AMD was mediated by the dysregulated action of the alternative pathway of the complement system. As used herein, delayed graft function (DGF) is a manifestation of acute kidney injury (AKI) with attributes unique to the transplantation process. It occurs after transplant surgery. Delayed graft function (DGF) is a common complication frequently defined as the need for dialysis during the first week after transplantation. Intrinsic renal synthesis of the third complement component C3 (C3) contributes to acute rejection by priming a T cell-mediated response. For example, in brain-dead donors, local levels of renal C3 are higher in obtaining e inversely related to renal function 14 days after transplantation. As used herein, a complement-mediated disease or disorder is any disorder in which any one or more of the complement proteins play a role in the disease, either due to the absence or presence of a complement protein or protein. complement-related or activation or inactivation of a complement or complement-related protein. In some embodiments, a complement-mediated disorder is one that is due to a deficiency in one or more complement proteins. In other embodiments, as described herein, a complement-mediated disorder is one that is due to activation or overactivation of one or more complement proteins. A complement-mediated disorder is also one that is due to the presence of any one or more of the complement proteins and / or continued activation of the complement pathway. As used herein, "macular degeneration-related disorder" refers to any of a number of conditions in which the retinal macula degenerates or becomes dysfunctional (for example, as a consequence of decreased growth of macular cells). macula, increased death or rearrangement of cells in the macula (for example, RPE cells), loss of normal biological function, or a combination of these events). Macular degeneration results in the loss of integrity of the histoarchitecture of the cells and / or extracellular matrix of the normal macula and / or the loss of function of the cells of the macula. Examples of disorders related to macular degeneration include age-related macular degeneration (AMD), geographic atrophy (GA), North Carolina macular dystrophy, Sorsby fundus dystrophy, Stargardt disease, pattern dystrophy, Best's, dominant drusen and leventine malatia). Macular degeneration-related disorder also encompasses extramacular changes that occur before or after the dysfunction and / or degeneration of the macula. Therefore, the term "macular degeneration-related disorder" also broadly includes any condition that alters or impairs the integrity or function of the macula (for example, damage to the RPE or Bruch's membrane). For example, the term encompasses retinal detachments, chorioretinal degenerations, retinal degenerations, photoreceptor degenerations, RPE degenerations, mucopolysaccharidoses, cone-rod dystrophies, cone-rod dystrophies, and cone degenerations. A macular degeneration-related disorder described herein includes macular degeneration, such as, for example, AMD macular degeneration. A disorder related to macular degeneration includes disorders treated by anti-VEGF treatment, such as, for example, anti-VEGF antibodies, or treatment with laser or an implantable telescope. As used herein, treating a subject with a disease or condition means that the subject's symptoms are partially or fully alleviated, or remain static after treatment. Thus, treatment encompasses prophylaxis, therapy, and / or cure. Prophylaxis refers to the prevention of a potential disease and / or a prevention of the worsening of symptoms or the progression of a disease. Treatment also encompasses any pharmaceutical use of a modified u-PA polypeptide and compositions provided herein. As used herein, "prevention" or prophylaxis refers to methods in which the risk or likelihood of developing a disease or condition is reduced. As used herein, a "therapeutic agent," therapeutic, radioprotective, or chemotherapeutic regimen refers to conventional drugs and drug therapies, including vaccines, that are known to those skilled in the art. Radiotherapeutic agents are well known in the art. As used herein, "treatment" means any manner in which the symptoms of a condition, disorder, or disease are ameliorated or otherwise beneficially altered. Treatment also encompasses any pharmaceutical use of the compositions herein. As used herein, "improvement of symptoms" of a particular disease or disorder by treatment, such as by administration of a pharmaceutical composition or other therapeutic product, refers to any decrease, whether cboynn / i znz / B / v permanent or temporary, lasting or transient, of the symptoms that can be attributed to or associated with the administration of the composition or therapeutic product. As used herein, a "pharmaceutically effective agent" includes any therapeutic agent or bioactive agents, including, but not limited to, for example, anesthetics, vasoconstrictors, dispersing agents, and conventional therapeutic drugs, including small molecule drugs and proteins. therapeutic. As used herein, an "effective amount" of a compound or composition for treating a particular disease is an amount that is sufficient to ameliorate or in some way reduce the symptoms associated with the disease. This amount can be administered as a single dosage or can be administered according to a regimen, whereby it is effective. The amount can cure the disease, but it is usually administered for the purpose of ameliorating the symptoms of the disease. Repeated administration is usually required to achieve a desired improvement in symptoms. As used herein, a "therapeutically effective amount" or "therapeutically effective dose" refers to the amount of a compound-containing agent, compound, material, or composition that is at least sufficient to produce a therapeutic effect after administration. administration to a subject. Therefore, it is the amount needed to prevent, cure, ameliorate, stop, or partially stop a symptom of a disease or disorder. As used herein, a "therapeutic effect" means an effect resulting from treatment of a subject that alters, typically ameliorates or improves, the symptoms of a disease or condition or that cures a disease or condition. As used herein, a "prophylactically effective amount" or a "prophylactically effective dose" refers to the amount of a compound-containing agent, compound, material, or composition that, when administered to a subject, has the effect intended prophylactic, for example, to prevent or delay the onset or recurrence of disease or symptoms, reduce the likelihood of disease or symptom onset or recurrence, or reduce the incidence of viral infection. The full prophylactic effect does not necessarily occur after administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount can be administered in one or more administrations. As used herein, "administration of a non-complementary protease", such as a modified u-PA protease, refers to any method in which the non-complementary protease is brought into contact with its substrate. Administration can be effected in vivo or ex vivo or in vitro. For example, for ex vivo administration, a fluid is withdrawn CbQjnn / l 7Π7 / Β / Υ body, such as blood, of a subject and contacted outside the body with the modified non-complementary protease, such as a modified u-PA protease. For administration in vivo, the modified non-complementary protease, such as a modified uPA protease, can be introduced into the body, such as via local, topical, systemic and / or other routes of introduction. In vitro administration encompasses methods, such as cell culture methods. As used herein, "unit dosage form" refers to physically discrete units suitable for human and animal subjects and individually packaged as known in the art. As used herein, "patient or subject to be treated" includes humans and human or non-human animals. Mammals include; primates, such as humans, chimpanzees, gorillas, and monkeys; domesticated animals, such as dogs, horses, cats, pigs, goats, and cows; and rodents such as mice, rats, hamsters, and gerbils. As used herein, a "combination" refers to any association between two or more items. The association can be spatial or refer to the use of the two or more elements for a common purpose. The combination can be two or more separate items, such as two compositions or two collections, a mix thereof, such as a single mix of the two or more items, or any variation thereof. The elements of a combination are generally associated or functionally related. As used herein, a "composition" refers to any mixture of two or more products or compounds (eg, agents, modulators, regulators, etc.). It can be a solution, a suspension, a liquid, a powder, a paste, aqueous or non-aqueous formulations, or any combination thereof. As used herein, a stabilizing agent refers to a compound added to the formulation to protect the antibody or conjugate, such as under the conditions (eg, temperature) under which the formulations herein are stored or used. Thus, agents that prevent protein degradation of other components are included in the compositions. Examples of these agents are amino acids, amino acid derivatives, amines, sugars, polyols, salts and buffers, surfactants, inhibitors or substrates, and other agents as described herein. As used herein, fluid refers to any composition that can flow. Thus, fluids encompass compositions that are in the form of semi-solids, pastes, solutions, aqueous mixtures, gels, lotions, creams, and other similar compositions. As used herein, a “manufactured item” is a product that is manufactured and sold. As used throughout this application, the term is intended to encompass a cboynn / i znz / B / v therapeutic agent with a modified u-PA polypeptide or nucleic acid molecule contained therein or separate items of packaging. As used herein, a "kit" refers to a packaged combination, optionally including reagents and other products and / or components for practicing the methods using the elements of the combination. For example, kits containing a modified protease polypeptide, such as a modified u-PA protease provided herein, or nucleic acid molecule provided herein, and other item are provided for a purpose including, but not limited to a, administration, diagnosis and assessment of a biological activity or property. The kits optionally include instructions for use. As used herein, a "cell extract" refers to a preparation or fraction that is made from a lysed or disrupted cell. As used herein, "animal" includes any animal, such as, but not limited to; primates including humans, gorillas and monkeys; rodents, such as mice and rats; poultry, such as chickens; ruminants, such as goats, cattle, deer, sheep; porcine, such as pigs and other animals. Non-human animals exclude humans as the contemplated animal. The proteases provided herein are from any source, animal, plant, prokaryotic and fungal. Most proteases are of animal origin, including mammalian origin. As used herein, a "single dose" formulation refers to a formulation that contains a single dose of therapeutic agent for direct administration. Single-dose formulations generally do not contain any preservatives. As used herein, a multidose formulation refers to a formulation that contains multiple doses of a therapeutic agent and can be administered directly to provide multiple individual doses of the therapeutic agent. Doses can be administered over the course of minutes, hours, weeks, days, or months. Multi-dose formulations may allow for dose adjustment, dose mixing, and / or dose splitting. Because multi-dose formulations are used over time, they generally contain one or more preservatives to prevent microbial growth. As used herein, a "control" or "standard" refers to a sample that is substantially identical to the test sample, except that it is not treated with a test parameter or, if a plasma sample, may be from a normal unaffected volunteer with the condition of interest. A control can also be an internal control. For example, a control can be a sample, such as a virus, that has a known property or activity. As used herein, the singular forms "a", "a" and "the" include plural references unless the context clearly indicates otherwise. Thus, for example, reference to "an" agent includes one or more agents. cboynn / i znz / B / v As used herein, the term or is used to mean and / or unless explicitly stated that it refers only to alternatives or if the alternatives are mutually exclusive. As used herein, ranges and amounts may be expressed as "about" a particular value or range. Approximate also includes the exact amount. Therefore, "about 5 bases" means "about 5 bases" and also "5 bases." As used herein, "optional" or "optionally" means that the event or circumstance described below does or does not occur, and that the description includes instances where that event or circumstance occurs and instances where it does not. For example, "optionally substituted" means that the group is unsubstituted or substituted. As used herein, abbreviations for any protecting groups, amino acids and other compounds are, unless otherwise indicated, in accordance with their commonly used, recognized abbreviations or the IUPACIUB Biochemical Nomenclature Commission (see, (1972 ) Biochem. 11:1726). For clarity of description, and not by way of limitation, the detailed description is divided into the subsections that follow. B. STRUCTURE AND FUNCTION OF U-PA Urokinase-type plasminogen activator (u-PA, also called urinary plasminogen activator or urokinase) is a serine proteinase that catalyzes the hydrolysis of plasminogen to plasmin. u-PA is found in urine, blood, seminal fluids, and in many cancerous tissues. It is involved in a variety of biological processes, which are linked to its conversion of plasminogen to plasmin, which in turn is a serine protease. Plasmin has roles in a variety of normal and pathological processes including, for example, cell migration and tissue destruction through its cleavage of a variety of molecules including fibrin, fibronectin, proteoglycans, and laminin. u-PA is involved in tissue remodeling during wound healing, inflammatory cell migration, neovascularization, and tumor cell invasion. u-PA also cleaves and activates other substrates, including, but not limited to, hepatocyte growth / spread factor (HGF / SF), the latent form of membrane matrix metalloprotease type 1 (MTSP1), derived growth factors of platelets and others. Provided herein are modified urokinase-type plasminogen activator (u-PA) polypeptides that are modified so that they cleave inhibitory sequences in 03, such that activation of 03 in C3a and C3b fragments is inhibited. The activity / specificity of the modified u-PA polypeptides provided herein PbQynn / i znz / B / v is such that they cleave C3 with greater activity and / or specificity or kcat / km compared to unmodified u-PA polypeptide, particularly any of SEQ ID NO: 1-6. Modified u-PA polypeptides may also have reduced activity or specificity or both for a natural physiological substrate plasminogen of the unmodified u-PA polypeptide. Thus, the modified u-PA polypeptides provided herein inhibit complement activation in a complement pathway. Modified u-PA polypeptides also exhibit increased selectivity for cleaving C3 compared to other u-PA substrates, such as plasminogen. Therefore, the modified u-PA polypeptides provided herein do not exhibit unwanted cleavage activities against physiological natural u-PA substrates so that they do not exhibit unwanted side effects. In some embodiments, the modified u-PA polypeptide is a protease domain or single chain form; in those cases, the free cisterna (residue position 122 by chymotrypsin numbering) is replaced with a serine, to decrease or eliminate aggregation after protein preparation. In embodiments in which the modified u-PA polypeptide is full-length or another form in which it is activated by cleavage, the residue at position 122 (by chymotrypsin numbering) is generally not replaced with S so that the bond disulfide can be formed to produce the activated two-chain polypeptide. 1. Serine Proteases Serine proteases (SPs), which include secreted enzymes and enzymes sequestered in cytoplasmic storage organelles, have a variety of physiological roles, including in blood coagulation, wound healing, digestion, immune responses, and tumor invasion and metastasis. For example, chymotrypsin, trypsin, and elastase work in the digestive tract; Factor 10, Factor 11, Thrombin and Plasmin are involved in coagulation and wound healing; and C1r, C1s, C3-conversases play a role in complement activation. A class of cell surface proteins designated type II transmembrane serine proteases are proteases that are membrane-anchored proteins with extracellular domains. As cell surface proteins, they play a role in intracellular signal transduction and in mediating cell surface proteolytic events. Other serine proteases bind to the membrane and function in a similar way. Others are secreted. Many serine proteases exert their activity by binding to cell surface receptors and thus act on cell surfaces. Cell surface proteolysis is a mechanism for the generation of biologically active proteins that mediate a variety of cellular functions. Serine proteases, including secreted and transmembrane serine proteases, are cboynn / i 7f\7iw involved in processes including neoplastic development and progression. Although the precise role of these proteases has not been fully elaborated, serine proteases and their inhibitors are involved in the control of many intracellular and extracellular physiological processes, including degradative actions on cancer cell invasion and metastatic spread, and neovascularization of cancer cells. tumors that are involved in tumor progression. Proteases are involved in the degradation and remodeling of the extracellular matrix (ECM) and contribute to tissue remodeling, and are necessary for cancer invasion and metastasis. The activity and / or expression of some proteases has been shown to correlate with tumor progression and development. More than 20 families (denoted S1-S27) of serine proteases have been identified, and are grouped into 6 clans (SA, SB, SC, SE, SF, and SG) on the basis of structural similarity and other functional evidence (Rawlings ND et al. al (1994) Meth. Enzymol. 244: 19-61). There are similarities in the reaction mechanisms of various serine peptidases. The chymotrypsin, subtilisin, and carboxypeptidase C clans have a catalytic triad of serine, aspartate, and histidine in common: serine acts as a nucleophile, aspartate as an electrophile, and histidine as a base. The geometric orientations of the catalytic residues are similar between families, despite different protein folds. Linear arrangements of catalytic residues commonly reflect clan relationships. For example, the catalytic triad in the chymotrypsin (SA) clan is ordered HDS, but DHS is ordered in the subtilisin (SB) clan and SDH is ordered in the carboxypeptidase (SC) clan. Examples of serine proteases of the chymotrypsin superfamily include tissue-type plasminogen activator (tPA), trypsin, trypsin-like protease, chymotrypsin, plasmin, elastase, urokinase (or urinary-type plasminogen activator, u-PA), acrosin, protein Activated C, C1 esterase, cathepsin G, chymase, and proteases of the blood coagulation cascade including kallikrein, thrombin, and Factors Vlla, IXa, Xa, Xla, and Xlla (Barret, A.J., In: Proteinase Inhibitors, Ed. Barrett, A.J., et al., Elsevier, Amsterdam, Pages 322 (1986), Strassburger, W. et al., (1983) FEBS Lett., 157 :219-223, Dayhoff, M.O., Atlas of Protein Sequence and Structure, Vol 5, National Biomedical Research Foundation, Silver Spring, Md. (1972) and Rosenberg, R.D. et al. (1986) Hosp. Prac., 21:131-137). The activity of proteases in the family of serine proteases is dependent on a set of amino acid residues that form their active site. One of the residues is always a serine; hence their designation as serine proteases. For example, chymotrypsin, trypsin, and elastase share a similar structure, and their active serine residue is at the same position (Ser195) in all three. Despite their similarities, they have different substrate specificities; different peptide bonds are cleaved during protein digestion. For example, chymotrypsin prefers an aromatic side chain at the residue whose cboynn / i znz / B / v carbonyl carbon is part of the peptide bond to be cleaved. Trypsin prefers a positively charged Lys or Arg residue at this position. Serine proteases differ markedly in their substrate recognition properties: some are highly specific (ie, the proteases involved in blood coagulation and the immune complement system); some are only partially specific (ie, the mammalian digestive proteases trypsin and chymotrypsin); and others, such as subtilisin, a bacterial protease, are completely nonspecific. Despite these differences in specificity, the catalytic mechanism of serine proteases is well conserved. The mechanism of cleavage of a target protein by a serine protease is based on nucleophilic attack of the target peptide bond by a serine. Cysteine, threonine, or water molecules associated with aspartate or metals may also play this role. In many cases, the nucleophilic property of the group is enhanced by the presence of a histidine, held in a proton accepting state by an aspartate. Aligned side chains of serine, histidine, and aspartate construct the catalytic triad common to most serine proteases. For example, active site residues of chymotrypsin and serine proteases that are members of the same family as chymotrypsin, such as, for example, MTSP-1, are Asp102, His57, and Ser195. The catalytic domains of all serine proteases of the chymotrypsin superfamily have sequence homology and structural homology. Sequence homology includes conservation of: 1) characteristic active site residues (eg, Ser195, His57, and Asp102 in the case of trypsin); 2) the oxyanion hole (eg Gly193, Asp194 in the case of trypsin); and 3) cysteine ​​residues that form disulfide bridges in the scaffold (Hartley, B.S., (1974) Symp. Soc. Gen. Microbiol., 24: 152-182). Structural homology includes 1) a common fold characterized by two key Greek structures (Richardson, J. (1981) Adv. Prot. Chem., 34:167-339); 2) a common disposal of catalytic waste; and 3) detailed preservation of the structure within the core of the molecule (Stroud, R.M. (1974) Sci. Am., 231: 24-88). Throughout the chymotpsin family of serine proteases, the major enzyme-substrate interaction is fully conserved, but the side-chain interactions vary considerably. The identity of the amino acids contained in the S1-S4 cavities of the active site determines the substrate specificity of that particular cavity. The grafting of amino acids from one serine protease to another of the same fold modifies the specificity of one to the other. Usually, the amino acids of the protease that contain the S1-S4 cavities are those that have side chains within 4 to 5 angstroms of the substrate. The interactions that these amino acids have with the protease substrate are generally called "first layer" interactions because they come into direct cboynn / i znz / B / v contact with the substrate. However, there may be "second layer" and "third layer" interactions that ultimately position the first layer amino acids. The substrate binding effects of the first layer and the second layer are mainly determined by loops between beta-barrel domains. Because these loops are not core elements of the protein, fold integrity is maintained as long as loop variants with new substrate specificities can be selected during the course of evolution to fill the necessary metabolic or regulatory niches. molecular level. Typically for serine proteases, the following amino acids in the primary sequence are determinants of specificity: 195, 102, 57 (the catalytic triad); 189, 190, 191, 192 and 226 (S1); 57, the loop between 58 and 64, and 99 (S2); 192, 217, 218 (S3); the loop between Cys168 and Cys180, 215, and 97 to 100 (S4); and 41 and 151 (S2'), based on chymotrypsin numbering, where an amino acid at an S1 position affects P1 specificity, an amino acid at an S2 position affects P2 specificity, an amino acid at S3 position affects P3 specificity , and an amino acid at position S4 affects P4 specificity. Position 189 in a serine protease is a residue buried at the bottom of the cavity that determines S1 specificity. The structural determinants for u-PA are listed in Table 4, with protease domains for each of the designated proteases aligned with that of the chymotrypsin protease domain. The number under the Cys168-Cys182 and 60 loop column headings indicates the number of amino acids in the loop between the two amino acids and in the loop. The yes / no designation under the Cys191-Cys220 column headings indicates whether the disulfide bond is present in the protease. These regions are variable within the family of chymotrypsin-like serine proteases and represent structural determinants themselves. 2. Structure The u-PA cDNA has been cloned from numerous mammalian species. Exemplary u-PA precursor polypeptides, or prepro-urokinase polypeptides include, but are not limited to, u-PA polypeptides from human (SEQ ID NO:1 and encoded by SEQ ID NO:7), mouse (SEQ ID NO:7). NO:52), rat (SEQ ID NO:53), bovine (SEQ ID NO:54), pig (SEQ ID NO:55), rabbit (SEQ ID NO:56), chicken (SEQ ID NO:57), yellow baboon (SEQ ID NO:58), Sumatran orangutan (SEQ ID NO:59), dog (SEQ ID NO:60), sheep (SEQ ID NO: 61), marmoset (SEQ ID NO: 62), rhesus macaque (SEQ ID NO:63), northern white-cheeked gibbon (SEQ ID NO:64), and chimpanzee (SEQ ID NO:65). The mRNA transcript is usually translated to generate a precursor protein that contains a 20 amino acid signal sequence at the N-terminus. After transport to the ER, the signal peptide is removed to provide a prourokinase polypeptide. Examples of prourokinase polypeptides include, but are not limited to, u-PA polypeptides from human (SEQ ID NO:3), mouse (SEQ cboynn / i znz / B / v ID NO:66), rat (SEQ ID NO:67), bovine (SEQ ID NO:68), pig (SEQ ID NO:69), rabbit (SEQ ID NQ:70), chicken (SEQ ID NO:71), yellow baboon (SEQ ID NO:72), Sumatran orangutan (SEQ ID NO:73), dog (SEQ ID NO:74) and sheep (SEQ ID NO:75). For example, the human u-PA mRNA transcript is normally translated to form a 431 amino acid precursor protein (SEQ ID NO:1) containing a 20 amino acid signal sequence at the N-terminus, Met Arg Ala Leu Leu Ala Arg Leu Leu Cys Val Leu Val Ser Asp Ser Lys Gly (amino acid residues 1-20 of SEQ ID NO:1). Thus, upon transport to the ER and removal of the signal peptide, a 411 amino acid prourokinase polypeptide is produced with an amino acid sequence set forth in SEQ ID NO:3. As described in more detail below, prourokinase is a zymogen or proenzyme that is further processed by proteolytic cleavage to generate a mature two-chain u-PA polypeptide. Thus, for example, with reference to mature u-PA (SEQ ID NO:3), wild-type strand activated u-PA contains a first strand (A strand), residues 1-158 disulfide-linked to residues 159-411 (chain B) via a disulfide bond between Cys148 (chymotrypsin numbering C97a) and Cys279 (chymotrypsin numbering C122). Therefore, in the modified u-PA polypeptides provided herein, when the protease domain is produced, it contains the C122S replacement, but when an activated form that is a 2-chain form is produced, the residue at 122 (numbering of chymotpsin) is C so that it forms a disulfide bond with another C, generally in the activation sequence (see later discussion and Example 15). The human u-PA precursor has an amino acid sequence set forth in SEQ ID NO:1 and encoded by a nucleotide sequence set forth in SEQ ID NO:7. Human pro-u-PA, also called mature u-PA, lacking the signal sequence is set forth in SEQ ID NO:3. Two isoforms of human u-PA exist, produced by alternative splicing. Human u-PA isoform 1 is the previously described canonical form set forth in SEQ ID NO:1. In human u-PA isoform 2, amino acids 1-29 of SEQ ID NO: 1 are replaced with amino acids 1-12 of SEQ ID NO: 51, with the resulting protein containing 414 amino acids (set forth in SEQ ID NO:51). Allergic and other variants of human u-PA are known. For example, a uPA variant is known that contains the V15L amino acid modification in the amino acid sequence set forth in SEQ ID NO:1. In another example, a modified u-PA polypeptide is known to contain the amino acid modification C299S (C122S by chymotrypsin numbering) in the amino acid sequence set forth in SEQ ID NO:1 (corresponding to the amino acid sequence set forth in SEQ ID NO:1). NOT: 4). Additional variants include those containing amino acid modifications P121L, D130G, C131W, I194M, K211Q, G366c, and A410V in u cboynn / i znz / E / v Mature PA set forth in SEQ ID NO:3 (corresponding to amino acid modifications P141L, D150G, C151W, 1214M, K231Q, G386C and A430V in SEQ ID NO:1). u-PA polypeptides are synthesized and secreted as single-chain zymogen molecules (also called prourokinases or single-chain urokinases), which are converted to active double-chain u-PA by a variety of proteases including, for example, plasmin, kallikrein, cathepsin B, matriptase, and nerve growth factor gamma. Cleavage to generate the two-chain form occurs between residues 158 and 159 (SEQ ID NO:3) in the human prourokinase sequence (corresponding to amino acid residues 178 and 179 in SEQ ID NO:1). The two resulting chains are linked by a disulfide bond between Cys148 and Cys279, thus forming the two-chain form of u-PA. The two-chain form of u-PA is also called high molecular weight u-PA (HMW-u-PA). HMW-u-PA can be further processed into low molecular weight u-PA (LMW-u-PA) by cleavage of the A chain into a short A chain (A1, amino acids 136-157 of SEQ ID NO:3) and an amino-terminal fragment. Disulfide linked 21178 to 179-411 linked via Cys corresponding to Cys148 and Cys279 (SEQ ID NO:3). Urokinase-type plasminogen activator, u-PA, is a classical serine protease, containing a His-Asp-Ser catalytic triad, which cleaves a specific Arg-Val bond in plasminogen to form plasmin. Plasmin, in turn, can cleave u-PA at Lys158Ile159 of SEQ ID NO: 3 (corresponding to Lys15-lle16 by chymotrypsin numbering) forming the two-stranded form described above. The human u-PA catalytic triad includes the amino acids His204, Asp255 and Ser356 of SEQ ID NO: 3 (corresponding to His57, Asp102 and Ser195 by chymotrypsin numbering). Residues Ser138 and Ser303 of human uPA set forth in SEQ ID NO:3 are phosphorylated (Franco et al. (1997) J Cell Biol 137:779-791). Human u-PA contains O-linked glycosylation, eg, fucosylation, at amino acid residue Thr18 of SEQ ID NO: 3 (Buko et al. (1991) Proc Nati Acad Sci USA 88:3992-3996) and O-linked glycosylation N at amino acid residue Asn302 of SEQ ID NO:3. Mature human u-PA contains intrachain disulfide bonds between residues 011-C19, C13-C31, C33-C42, C50-C131, C71-C113, C102-C126, 01890205, C197-C268, 0293-0362, 0325- 0341 and 0352-0380 of SEQ ID N0:3 and an interchain disulfide bond between residues 0148-0279 of SEQ ID N0:3. The mature form of u-PA is a 411 residue protein (corresponding to amino acid residues 21 to 431 in the amino acid sequence set forth in SEQ ID N0:1 which is the precursor form containing a 20 amino acid signal peptide). u-PA contains three domains: the serine protease domain, the kringle domain, and the growth factor domain. In the mature form of human u-PA, amino acids 1-158 represent the N-terminal A chain that includes a growth factor domain (amino acids 1-49), a cboynn / i znz / B / v kringle domain (amino acids 50-131) and an interdomain linker region (amino acids 132158). Amino acids 159-411 represent the C-terminal or B-chain serine protease domain. u-PA is synthesized and secreted as a single-chain zymogen molecule, which is converted to an active double-chain u-PA by a variety of proteases including, for example, plasmin, kallikrein, cathepsin B, and nerve growth factor gamma. Cleavage in the two-stranded form occurs between residues 158 and 159 in a mature u-PA sequence (corresponding to amino acid residues 178 and 179 in SEQ ID NO:3). The two resulting chains are held together by a disulfide bond, thus forming the two-chain form of u-PA. Urokinase-type plasminogen activators contain three domains: a serine protease domain, a kringle domain, and a growth factor domain. In the zymogenic or proenzyme form of human u-PA, amino acids 1-158 of SEQ ID NO:3 represent the N-terminal A chain (or long A chain) which includes an epidermal growth factor domain (amino acids 1-49 ), a kringle domain (amino acids 50-131) and an interdomain linker region (amino acids 132-158) and amino acids 159-411 represent the catalytically active C-terminal serine protease domain or B chain. The epidermal growth factor domain it is responsible for the binding of u-PA to the cell surface anchored u-PA receptor (uPAR). In the extracellular matrix, u-PA binds to the cell membrane by binding to the u-PA receptor. LMW-u-PA is proteolytically active but does not bind to the u-PA receptor. The serine protease domain contains surface exposed loops around residues 37, 60, 96, 110, 170 and 185, by chymotrypsin numbering. Upon activation or cleavage, the amino terminus inserts into a hydrophobic binding cleft of the catalytic protease domain that forms hydrophobic interactions and a salt bridge to the Asp194 side pocket that stabilizes the substrate-binding pocket and oxyanion hole. in a catalytically productive conformation. Asp194, according to chymotrypsin numbering, participates in hydrogen bonding to the backbone amino group of Gly142 and the backbone carbonyl group of Lys143 (Bluuse et al. (2009) J Biol Chem 284:4647-4657). . The conformational changes after cleavage involve four disordered regions of the activation domain, including the activation loop (residues 16-21), the autolysis loop (residues 142-152), the oxyanion stabilizer loop (residues 184-194 ) and the S1 entry frame (residues 216-223), all numbered according to chymotrypsin numbering (see, Bluuse et al. (2009) J Biol Chem 284:4647-4657; Hedstrom (2002) Chem Rev 102: 4501-4524; Huber and Bode (1978) Acc Chem Res 11:114122; Madison et al. (1993) Science 262:419-421). The structural determinants for u-PA are set forth in Table 4 below with numbering based on the numbering of mature chymotrypsin. The number under the PbQynn / i znz / B / v loop column headers Cys168-Cys182 and 60 indicate the number of amino acids in the loop between the two amino acids and in the loop. The designation of yes under the Cys191-Cys220 column headings indicates that a disulfide bond is present. These regions are variable within the family of chymotrypsin-like serine proteases and represent structural determinants themselves. Modification of a u-PA polypeptide to alter any one or more of the amino acids in the S1-S4 pocket affects the specificity or selectivity of the u-PA polypeptide for a target substrate. Extended substrate specificity (P1-P4) reveals that u-PA has a high specificity for cleavage after P1 Arg, a preference for small amino acids at the P2 position, a preference for small polar amino acids (Thr and Ser) at the P3 position and no preference at the P4 position (Ke et al. (1997) J. Biol. Chem., 272:16603-16609; Harris et al. (2000) Proe Nati Acad Sel USA, 97:7754-7759). cboynn / i znz / B / v Table 4. Structural determinants for u-PA substrate cleavage (chymotrypsin numbering) Residues determining specificity S4 S3 S2 S1 171 17 4 180 215 Cys16 8 Cys18 2 192 21 8 99 57 Loop of 60 189 190 22 6 Cys19 1 Cys22 0 H S M W 15 Q R H H 11 D S G yes. 3. Function / activity Urokinase-type plasminogen activator is a serine protease that catalyzes the hydrolysis of plasminogen to plasmin. Plasmin acts directly on the degradation of extracellular matrix proteins (Andreasen et al. (2000) Cell. Mol. Life Sel. 57:25-40). u-PA plays an important role in cell adhesion, migration and invasion, tissue remodeling, and cancer (Blasi et al. (2002) RevMol Cell Biol3:932; Andreasen et al. (2000) Cell. Mol. Life Sci. 57:25 -40; Mondino and Blasi (2004) Trends Immunol 25:450; Ploug (2003) Curr Pharm Des 9:1499). Abnormal expression of u-PA has been associated with rheumatoid arthritis, allergic vasculitis, xeroderma pigmentosa, and the invasive capacity of malignant tumors. u-PA is regulated by binding to the high-affinity cell surface receptor uPAR. Binding of u-PA to uPAR increases the rate of plasminogen activation and enhances extracellular matrix degradation and cell invasion. The binary complex formed between uPAR and u-PA interacts with membrane-associated plasminogen to form higher-order activation complexes that lower the Km (ie, approximate affinity kinetic rate constant for a substrate) for plasminogen activation. (Bass et al. (2002) Biochem. Soc. Trans., 30: 189-194). Binding of u-PA to uPAR protects the protease from inhibition by the cognate inhibitor, ie, PAI-1. This is because single chain u-PA normally present in plasma is not susceptible to inhibition by PAI-1, and any active u-PA in plasma will be inhibited by PAI-1. The active u-PA that binds to the receptor is fully available for inhibition by PAI-1, however PAI-1 cannot access the bound active molecule (Bass et al. (2002) Biochem. Soc. Trans., 30: 189194). As a result, u-PA functions primarily on the cell surface and its functions correlate with the activation of plasmin-dependent pericellular proteolysis. u-PA also cleaves hepatocyte growth factor / dispersal factor (HGF / SF), the latent form of membrane matrix metalloprotease type 1 (MT-SP1; matriptase), platelet-derived growth factor C (PDGF- C), platelet-derived growth factor D (PDGF-D), platelet-derived growth factor DD (PDGFDD), and other proteins (see, for example, Hurst et al. (2012) Biochem J 441:909-918; Ustach and Kim (2005) Mol Cell Biol 5:6279-6288, Ehnman et al (2009) Oncogene 28(4):534-544). Plasmin degrades fibrin clots, cleaves fibrin, fibronectin, thrombospondin, laminin, and von Willebrand factor, proteolyzes complement system mediators, and activates collagenases. As such, plasmin is involved in thrombolysis or degradation of the extracellular matrix, binding plasmin to vascular disease and cancer. For example, components of the plasminogen activation system have been found to be highly expressed in malignant tumors. Hepatocyte growth / spread factor regulates cell growth, cell motility, and morphogenesis through the binding of activated HGF to the HGF receptor c-Met and its ability to stimulate mitogenesis, cell motility, and matrix invasion link it to the angiogenesis, tumorigenesis and tissue regeneration. Platelet-derived growth factors regulate cell growth and division and play a significant role in angiogenesis, which, when uncontrolled, is a hallmark of cancer. Once activated by proteolytic cleavage, PDGFs bind to PGDF receptor tyrosine kinases leading to phosphorylation and a number of downstream signaling pathways involved in cancer. Due to the role of u-PA and the aforementioned proteins in vascular diseases, the u-PA polypeptides provided herein are altered in ways that reduce selectivity towards these proteins. By virtue of changes in their specificity and activity, the modified u-PA polypeptides provided herein exhibit reduced activity or no substantial activity on natural substrates, and high activity, compared to unmodified u-PA on the C3 protein of complement. As a result, at therapeutic doses, the modified uPA polypeptides provided herein specifically inhibit complement activation but have little or no side effects from cleavage of natural u-PA targets. CbQjnn / l 7Π7 / Β / Υ C. COMPLEMENT INHIBITION BY TARGETING TO C3 The modified u-PA polypeptides provided herein exhibit increased specificity and / or activity for an inhibitory cleavage sequence in complement protein C3 n compared to u-PA that does not contain the amino acid modifications (eg, u-PA human wild-type (see SEQ ID NO:1 or 3)) or the catalytic domain or protease domain thereof (see SEQ ID NO:2) or corresponding unmodified u-PA polypeptides including the C122S replacement, by chymotrypsin numbering . Replacement with S at residue 122 does not alter the specificity or activity at C3, but reduces aggregation. Since C3 is involved in all 3 pathways of complement initiation (see, for example, FIGURE 1), targeting 03 by proteolytic inhibition provides a broad and general therapeutic target for inactivation of the complement cascade. Cleavage by inactivation of C3 blocks terminal complement activity as well as the amplification loop of the alternative pathway. The three pathways converge at 03 (see, for example, Figure 1). By virtue of the ability to inhibit complement activation, these modified u-PA polypeptides can be used to treat various diseases, conditions, and pathologies associated with complement activation, such as inflammatory responses and autoimmune diseases. Complement activation is associated with the development of diseases and conditions by promoting local inflammation and tissue damage caused in part by the generation of effector molecules and a membrane attack complex. In one example, such as in many autoimmune diseases, complement causes tissue damage because it is activated under inappropriate circumstances, such as by antibody to host tissues. In other situations, complement may be activated normally, such as by sepsis, but still contribute to disease progression, such as in respiratory distress syndrome. Pathologically, complement can cause substantial damage to blood vessels (vasculitis), renal basement membrane and fused endothelial and epithelial cells (nephritis), joint synovium (arthritis), and red blood cells (hemolysis) if not adequately controlled. The role of 03 in complement activation is discussed in more detail below. 1. Complement protein C3 and its role in complement initiation The complement system involves more than 30 soluble and cell membrane-bound proteins that function not only in the antibody-mediated immune response, but also in the innate immune response to recognize and destroy pathogens such as bacteria, virus-infected cells, and parasites. . Complement activation is initiated at pathogenic surfaces via three distinct pathways: the classical pathway, the alternative pathway, and the lectin pathway. These pathways are distinct in that the components required for their initiation are different, but the pathways ultimately generate the same set of effector molecules (eg, C3-convertases) that cleave complement protein C3 to trigger the formation of the complement complex. membrane attack (MAC) (see, for example, Figure 1). Thus, the complement C3 protein is an attractive target for a therapeutic agent since modulation of C3 results in modulation of various opsonins, anaphylatoxins, and the MAC. Additionally, naturally occurring complement inhibitory proteins including factor H (FH), CR1, complement receptor Ig (CR1g), DAF, and MCP inhibit at the C3 level. There are three (3) pathways of complement activation (see Figure 1, which illustrates these pathways). The complement pathways are distinct; each depends on different molecules and mechanisms for initiation. The pathways are similar in that they converge to generate the same set of effector molecules, ie C3-convertases. In the classical and lectin pathways C4b2b acts as a C3-convertase; in the alternative pathway, C3bBb is a C3-convertase (see Table 5). Cleavage of C3 generates C3b, which acts as an opsonin and as the main effector molecule of the complement system for subsequent complement reactions, and C3a, which is a peptide mediator of inflammation. The addition of C3b to each C3-convertase forms a C5-convertase that generates C5a and C5b. C5a, like C3a, is a peptide mediator of inflammation. C5b mediates the "late" events of complement activation by initiating the sequence of reactions that culminate in the generation of the membrane attack complex (MAC). Although all three pathways produce different C3 and C5-conversases, all pathways produce the split products of 03 and 05 and form MACs. Alternatively, C3 can be cleaved and activated by extrinsic proteases, such as lysosomal enzymes and elastase (Markiewski and Lambris (2007) Am J Pathology 171:715-727; Ricklin and Lambris (2007) Nat Biotechnol25:1265-1275). cboynn / i znz / B / v Table 5. Complement cascades Alternative pathway Classic pathway Lectin pathway Activators Surface molecules of pathogen LPS, teichoic acid, zymosan IgM and IgG bound to antigen; nonimmune molecules Pathogens by surface carbohydrate recognition C3 convertase C3bBb C4b2b C4b2b C5 convertase C3bBb3b C4b2b3b C4b2b3b MAC C5678poly9 C5678poly9 C5678poly9 anaphylatoxins C3a, C5a C3a, C4a, C5a C3a, C4a, C 5a to. classical route C1q is the first component of the classical complement pathway. C1q is a calcium-dependent binding protein associated with the collector protein family due to a general shared structural homology (Malhotra et al., (1994) Clin Exp Immunol. 97(2):4-9; Holmskov et al. (1994). ) Immunol Today 15(2):67-74). The collections, often called pattern recognition molecules, generally function as opsonins to target pathogens for phagocytosis by immune cells. Unlike conventional collections such as MBL, the carboxy-terminal globular recognition domain of C1q has no lectin activity but can serve as a "charged" pattern recognition molecule due to marked differences in the electrostatic surface potential of its domains. globular (Gaboriaud et al. (2003) J. Biol. Chem. 278(47):46974-46982). C1 q initiates the classical complement pathway in two different ways. First, the classical pathway is activated by interaction of C1 q with immune complexes (ie, antigen-antibody complexes or aggregated IgG or IgM antibody), thus linking the antibody-mediated humoral immune response with complement activation. When the Fab part (the variable region) of IgM or IgG binds to antigen, the conformation of the Fe (constant) region is altered, allowing 01q to bind. 01q must bind to at least 2 Fe regions to be activated. 01 q, however, is also capable of activating complement in the absence of antibody, thus functioning in the innate or immediate immune response to infection. In addition to initiation by an antibody, complement activation is also achieved through the interaction of 01q with non-immune molecules such as polyanions (bacterial lipopolysaccharides, DNA and RNA), certain small polysaccharides, viral membranes, 0-reactive protein (CRP) , serum amyloid P component (SAP) and bacterial, fungal and viral membrane components. q is part of the 01 complex that contains a single 01 q molecule bound to two molecules each of the 01 r and 01 zymogens. Binding of more than one of the 01q globular domains to a target surface (such as aggregated antibody or a pathogen), causes a conformational change in the (C1r:C1s)2 complex that results in activation of C1r protease to cleave C1 s to generate an active serine protease. Active C1 s cleaves downstream complement components 04 and 02 to generate C4b and C2b, which together form the C3-convertase of the classical pathway. C3-convertase cleaves 03 into C3b, which binds covalently to the surface of the pathogen and acts as an opsonin, and C3a, which stimulates inflammation. Some C3b molecules associate with C4b2b complexes producing C4b2b3b which is the classical C5-convertase cascade. Table 6 summarizes the proteins involved in the classical complement pathway. cboynn / i znz / B / v Table 6. Proteins of the Classical Pathway Native Component Active Form Function of Active Form C1 (C1q:(C1r:C1s)2) C1q Binds directly to pathogenic surfaces or indirectly to pathogen-bound antibody 01 r Cleaves 01 s on an active protease C1s Cleaves 04 and 02 Table 6. Proteins of the Classical Pathway Native component Active form Function of the Active Form C4 C4b Binds to the pathogen and acts as an opsonin; binds C2 for cleavage by C1 s C4a Peptide mediator of inflammation C2 C2b Active enzyme of the classical C3 / C5-convertase pathway; cleaves C3 and C5 C2a Precursor to quinine C2 vasoactive C3 C3b Binds to pathogen surfaces and acts as an opsonin; initiates amplification via the alternative pathway; binds C5 for cleavage by C2b C3a Peptide mediator of inflammation CbQjnn / l 7Π7 / Β / Υ b. alternative pathway The alternative pathway is initiated by foreign pathogens in the absence of antibody. Complement initiation via the alternative pathway occurs through the spontaneous hydrolysis of C3 into C3b. A small amount of C3b is always present in body fluids, due to protease activity in serum and tissue. Host autocells normally contain high levels of membrane sialic acid that inactivate C3b if bound, but bacteria contain low levels of external sialic acid and therefore bind C3b without inactivating it. C3b on pathogenic surfaces is recognized by the Factor B zymogen protease. Factor B is cleaved by Factor D. Factor D is the only complement system activating protease that circulates as an active enzyme rather than a zymogen, but since Factor B is the only substrate for Factor D, the presence of low levels of an active protease in normal serum is generally safe for the host. Cleavage of Factor B by Factor D produces the active product Bb which can associate with C3b to form C3bBb, the alternative pathway C3-convertase. Similar to the classical pathway, C3-convertase produces more C3b and C3a from 03. C3b binds covalently to the pathogen surface and acts as an opsonin and further initiates the alternative pathway, while C3a stimulates inflammation. Some of C3b binds to the complex to form C3bBb3b, the alternative pathway C5-convertase. C3bBb3b is stabilized by the plasma protein properdin or Factor P which binds to microbial surfaces and stabilizes convertase. Table 7 summarizes the proteins involved in the alternative complement pathway. Table 7. Alternative Pathway Proteins Native Component Active Form Function of Active Form C3 C3b Binds to pathogen surface, binds Factor B for cleavage by Factor D Ba Small fragment of Factor B, function unknown Table 7. Proteins of the Alternative Pathway Native component Active form Function of the Active Form Factor B Bb Active enzyme of C3-convertase and C5-convertase Factor D D Plasma serine protease, cleaves Factor B when it binds C3b to Ba and Bb Factor P (properdin) P Plasma proteins with affinity for C3bBbconvertase in bacterial cells; stabilizes convertase CbQjnn / l 7Π7 / Β / Υ c. Lectin pathway The lectin pathway (also called the MBL pathway) is initiated after recognition and binding of pathogen-associated molecular patterns (PAMPs; ie, carbohydrate moieties) by lectin proteins. Examples of lectin proteins that activate the complement lectin pathway include mannose-binding lectin (MBL) and ficolins (ie, Lficolin, M-ficolin, and H-ficolin). MBL is a member of the collectin family of proteins and therefore exists as an oligomer of subunits composed of identical polypeptide chains, each containing a collagen-like lectin, or carbohydrate recognition, domain. rich in cysteine. MBL acts as a pattern recognition molecule to recognize portions of carbohydrates, particularly neutral sugars such as mannose or N-acetylglucosamine (GIcNAc) on the surface of pathogens via its globular lectin domain in a calcium-dependent manner. MBL also acts as an opsonin to facilitate the phagocytosis of bacterial, viral, and fungal pathogens by phagocytic cells. Additional initiators of the lectin pathway include ficolins including L-ficolin, M-ficolin and H-ficolin (see, for example, Liu et al. (2005) J Immunol. 175:3150-3156). Similar to MBL, ficolins recognize carbohydrate moieties such as, for example, N-acetyl glucosamine and mannose structures. Activation of the alternative pathway by MBL or ficolins is analogous to activation of the classical pathway by C1q whereby a single lectin molecule interacts with two protease zymogens. In the case of the lectin proteins, the zymogens are MBL-associated serine proteases, MASP-1 and MASP-2, which are closely homologous to the C1r and C1s zymogens of the classical pathway. Upon recognition of a PAMP by a lectin protein, such as, for example, by binding to a pathogen surface, MASP-1 and MASP-2 are activated to cleave C4 and C2 to form the C3-convertase cascade. from MBL. C3b then binds to the complex to form the C5-convertase of the MBL cascade. MASP activation is implicated not only in responses to microorganisms, but in any response that involves exposing neutral sugars, including but not limited to tissue injury, such as seen in organ transplants. Like the alternative cascade, the MBL cascade is activated independently of the antibody; Like the classical cascade, the MBL cascade uses C4 and C2 to form C3-convertase. Table 8 summarizes the proteins involved in the complement lectin pathway. cboynn / i znz / B / v Table 8. Lectin Pathway Proteins Native Component Active Form Function of Active Form MBL MBL Recognizes PAMP, such as on pathogenic surfaces (eg, through carbohydrate recognition) Ficolins L-Ficolin; MFicoline, or HFicholine Recognizes PAMP, such as on pathogenic surfaces (eg, through carbohydrate recognition) MASP-1 MASP-1 Cleaves 04 and C2 MASP-2 MASP-2 Cleaves 04 and 02 d. Complement-mediated effector functions Regardless of the route of initiation used, the end result is the formation of activated fragments of complement proteins (for example, C3a, C4a, and C5a anaphylatoxins and C5-9 membrane attack complexes), which act as effector molecules. to mediate various effector functions. Recognition of complement effector molecules by cells for the initiation of effector functions (eg, chemotaxis and opsonization) was mediated by a diverse group of complement receptors. Complement receptors are distributed in a wide variety of cell types including erythrocytes, macrophages, B cells, neutrophils, and mast cells. Upon binding of a complement component to the receptor, the receptors initiate an intracellular signaling cascade that results in cellular responses such as stimulating phagocytosis of bacteria and secreting inflammatory molecules from the cell. For example, the complement receptors CR1 and CR2 that recognize C3b, C4b, and their products are important in stimulating chemotaxis. CR3 (CD11b / CD18) and CR4 (CD11c / CD18) are integrans that are equally important in phagocytic responses, but also play a role in leukocyte adhesion and migration in response to ¡C3b. C5a and C3a receptors are G protein-coupled receptors that play a role in many of the proinflammatory functions mediated by C5a and C3a anaphylatoxins. For example, the receptors for C3a, C3aR, exist on mast cells, eosinophils, neutrophils, basophils, and monocytes and are directly involved in the proinflammatory effects of C3a. Thus, through complement receptors, these complement effector molecule fragments mediated several functions including leukocyte chemotaxis, macrophage activation, vascular permeability, and cell lysis (Frank, M. and Fries, L. Complement. In Paul, W. (ed.) Fundamental Immunology, Raven Press, 1989). A summary of some effector functions of complement products is listed in Table 9. Table 9: Molecules and Functions of the Complement Effector Product Activity C2b (prokinin) accumulation of body fluid C3a (anaphylatoxin) Degranulation of basophils and mast cells; improved vascular permeability; smooth muscle contraction; induction of C3b suppressor T cells and their opsonization products; activation of phagocytes C4a (anaphylatoxin) activation of basophils and mast cells; smooth muscle contraction; enhanced vascular permeability C4b opsonization C5a (anaphylatoxin; chemotactic factor) activation of basophils and mast cells; improved vascular permeability; smooth muscle contraction; chemotaxis; neutrophil aggregation; stimulation of oxidative metabolism; stimulation of leukotriene release; induction of helper T cells C5b67 chemotaxis; binding to other cell membranes and lysis of transient cells C5b6789 (C5b-9) lysis of target cells cboynn / i znz / B / v Yo. Complement-mediated lysis: Membrane Attack Complex The final step of the complement cascade through the three pathways is the formation of the membrane attack complex (MAC) (Figure 1). C5 can be cleaved by any C5-convertase into C5a and C5b. C5b combines with C6 and C7 in solution, and the C5b67 complex associates with the pathogenic lipid membrane via hydrophobic sites on 07. C8 and several C9 molecules, which also have hydrophobic sites, bind to form the membrane attack complex , also called C5b6789 or C5b-9. C5b-9 forms a pore in the membrane through which water and solutes can pass, resulting in osmotic lysis and cell death. If complement is activated on an antigen without a lipid membrane to which C5b67 can bind, the C5b67 complex can bind to nearby cells and initiate lysis of bystanders. A single MAC can lyse an erythrocyte, but nucleated cells can endocytose MACs and repair the damage unless multiple MACs are present. Gram-negative bacteria, with their exposed outer membrane and enveloped virus, are generally susceptible to complement-mediated lysis. Less susceptible are Gram-positive bacteria, whose plasma membrane is protected by its thick peptidoglycan layer, bacteria with a capsule or slime layer around their cell wall, or viruses that do not have a lipid envelope. Also, the MAC can be disrupted by proteins that bind to the complex prior to membrane insertion, such as streptococcal inhibitor of complement (SIC) and clusterin. MAC commonly helps to destroy gram-negative bacteria as well as human cells displaying foreign antigens (virus-infected cells, tumor cells, etc.) by causing their lysis, and can also damage the envelope of enveloped viruses. ii. Inflammation Inflammation is a process in which blood vessels dilate and become more permeable, thus allowing the body's defense cells and defense chemicals to leave the blood and enter the tissues. Activation of complement results in the formation of various proinflammatory mediators such as C3a, C4a, and C5a. Intact anaphylatoxins in serum or plasma are rapidly converted to the more stable and less active C3adesArg, C4a-desArg, or C5a-desArg forms by carboxypeptidase N. C3a, C4a, and C5a, and to a lesser extent their desArg derivatives, are bioactive polypeptides potent, called anaphylatoxins because of their inflammatory activity. Anaphylatoxins bind to receptors on various cell types to stimulate smooth muscle contraction, increase vascular permeability, and activate mast cells to release inflammatory mediators. C5a, the most potent anaphylatoxin, acts primarily on white blood cells, particularly neutrophils. C5a stimulates the adherence of leukocytes to blood vessel walls at the site of infection by stimulating increased expression of adhesion molecules so that leukocytes can be squeezed out of blood vessels and into tissues, a process called diapedesis. C5a also stimulates neutrophils to produce reactive oxygen species for extracellular destruction, proteolytic enzymes, and leukotrienes. C5a can also further amplify the inflammatory process indirectly by inducing the production of chemokines, cytokines, and other proinflammatory mediators. C5a also interacts with mast cells to release vasodilators such as histamine so that blood vessels become more permeable. C3a also interacts with white blood cells, with important effects on eosinophils suggesting a role for C3a in allergic inflammation. C3a induces smooth muscle contraction, improves vascular permeability, and causes basophil degranulation and release of histamine and other vasoactive substances. C2a can be converted to C2 quinine, which regulates blood pressure by causing blood vessels to dilate. Although not technically considered an anaphylatoxin, ¡C3b, an inactive derivative of C3b, functions to induce leukocyte adhesion to vascular endothelium and induce production of the proinflammatory cytokine IL-1 through binding to its surface integrin receptors. cell phone. C5b-9 also indirectly stimulates leukocyte adhesion, activation, and chemotaxis by inducing the expression of cell adhesion molecules such as Eselectin and inducing interleukin-8 secretion (Bhole et al. (2003) Crit Care Med 31 (1): 97104). C5b-9 also stimulates the release of secondary mediators that contribute to inflammation, such as, for example, prostaglandin E2, leukotriene B4, and thromboxane. Conversion of human complement components C3 and C5 to produce their respective anaphylatoxin products has been implicated in certain naturally occurring disease states including: autoimmune disorders such as systemic lupus erythematosus, cboynn / i znz / B / v rheumatoid arthritis, malignancy , myocardial infarction, Purtscher retinopathy, sepsis and respiratory distress syndrome in adults. Increased circulating levels of C3a and C5a have been detected in certain conditions associated with iatrogenic complement activation, such as: cardiopulmonary bypass surgery, renal dialysis, and nylon fiber leukemia. iii. chemotaxis Chemotaxis is a process by which cells are directed to migrate in response to chemicals in their environment. In the immune response, a variety of chemokines direct the movement of cells, such as phagocytic cells, to sites of infection. For example, C5a is the major chemotactic factor for circulating neutrophils, but it can also induce monocyte chemotaxis. Phagocytes move toward increasing concentrations of C5a and subsequently bind, through their CR1 receptors, to antigen-bound C3b molecules. The chemotactic effect of C5a, observed with basophils, eosinophils, neutrophils, and mononuclear phagocytes, is active at concentrations as low as 10-10M. iv. opsonization An important action of complement is to facilitate the uptake and destruction of pathogens by phagocytic cells. This occurs through a process called opsonization whereby complement components bound to target bacteria interact with complement receptors on the surface of phagocytic cells such as neutrophils or macrophages. In this case, the complement effector molecules are called opsonins. Pathogen opsonization is a major function of C3b and C4b. C3b also functions as an opsonin. C3a and C5a increase the expression of C3b receptors on phagocytes and increase their metabolic activity. C3b and, to a lesser extent, C4b help clear harmful immune complexes from the body. C3b and C4b bind immune complexes to CR1 receptors on red blood cells. The erythrocytes then deliver the complexes to fixed macrophages within the spleen and liver for destruction. Immune complexes can lead to harmful Type III hypersensitivity. v. Activation of the Humoral Immune Response B cell activation requires ligation of the B cell receptor (BCR) by antigen. However, complement has been shown to play a role in lowering the threshold of B cell responses to antigen by up to 1000-fold. This occurs by the binding of C3d or C3dg, complementary products generated from the C3 breakdown fragments, to CR2 receptors on B cells that can co-ligate with the BCR. Co-ligation coregulation occurs when antigenic particles, such as cboynn / i znz / B / v as, for example, immune complexes, opsonized with C3d bind to the CR2 receptor through C3d, as well as to the BCR through antigen. . Co-ligation of antigen complexes can also occur when C3d binds to antigens enhancing its uptake by antigen presenting cells, such as dendritic cells, which can then present the antigen to B cells to enhance the antibody response. CR2-deficient mice show defects in B-cell function that result in reduced levels of natural antibody and impaired humoral immune responses. 2. Structure and Function of C3 The variant u-PA polypeptides provided herein cleave complement protein C3 or its proteolytic fragments thereby inhibiting complement. Human complement protein C3 (Uniprot accession number P01024) is a 1663 amino acid single chain preproprotein having an amino acid sequence set forth in SEQ ID NO:47. The protein is encoded by a 41 kb gene located on chromosome 19 (nucleotide sequence set forth in SEQ ID NO:46). The pre-proprotein contains a 22 amino acid signal peptide (SEQ ID NO:47 amino acids 1-22) and a tetra-arginine sequence (SEQ ID NO:47 amino acids 678-681) that is removed by an enzyme similar to function that results in the formation of a mature two-chain protein containing a beta chain (SEQ ID NO:47 amino acids 23-667) and an alpha chain (SEQ ID NO:47 amino acids 672-1663), which are linked by an interchain disulfide bond between amino acid residues Cys559 and Cys816. The mature 2-chain protein has an amino acid sequence set forth in SEQ ID NO:77. During the complement cascade, the complement C3 protein is further processed by proteolytic cleavage to form various C3 proteolytic fragments. As described above, all three complement initiation pathways converge in C3 convertases C4b2b and C3bBb. C3-convertases cleave C3 between residues 748 and 749 of SEQ ID NO:47 (see Table 10 below) generating anaphylatoxin C3a (amino acids 672-748 of SEQ ID NO:47) and opsonin C3b (C3b alpha' chain ; amino acids 749-1663 of SEQ ID NO:47). C3a is involved in inflammation and C3b forms the C5 convertases that ultimately lead to C5a anaphylatoxin and MAC. The variant uPA polypeptides provided herein inhibit complement and, as such, do not cleave C3 at this GLAR cleavage site. C3b has binding sites for several complement components including C5, properdin (P), factors Η, B, and I, complement receptor 1 (CR1), and membrane cofactor protein (MCP) (see Sahu and Lambris (2001). ) Immunological Reviews 180:35-48). Binding of Factor I, a plasma protease, in the presence of cofactors H, CR1, and MCP results in inactivation of C3b, while binding of factors B and P in the presence of cboynn / i znz / B / v factor D results in amplification of convertase 03 and initiation of MAC. Factor I cleaves C3b in the presence of cofactors between residues 1303-1304, 1320-1321 and 954-955 of SEQ ID NO:47 (see Table 10 below) generating ¡C3b fragments (amino acids 749-1303 of SEQ ID NO:47) and C3f (amino acids 1304-1320 of SEQ ID NO:47). Subsequently, Factor I cleaves ¡C3b generating C3c (C3c alpha' chain Fragment 1; amino acids 749-954 of SEQ ID NO:47) and C3dg (amino acids 955-1303 of SEQ ID NO:47). The end result is that C3b is permanently inactivated (see Sahu and Lambris (2001) Immunological Reviews 180:35-48). Since Factor I inactivates C3b, Factor I cleavage sites are candidates for cleavage by the variant u-PA polypeptides provided herein. Additional C3b proteolytic fragments include C3g (SEQ ID NO:47 amino acids 955-1001), C3d (SEQ ID NO:47 amino acids 1002-1303) and C3c alpha' chain Fragment 2 (SEQ ID NO:1321-1663 amino acids NO:47). The cleavage sequences in the complement protein C3 are set forth in Table 10 below, which lists the P4-P1 residues, the cleavage site amino acid residues (P1-P1' site) and the protease responsible for cleavage. The modified u-PA polypeptides provided herein do not cleave at these sites. PbQynn / i znz / B / v TABLE 10: Complement C3 Protein Cleavage Sequences P4-P1 Residues Cleavage Site (in SEQ ID NO:47) Between Residues Protease SEQ ID NO. GLAR 748-749 C3 convertase 78 RLGR 954-955 Factor I 79 LPSR 1303-1304 Factor I 80 SLLR 1320-1321 Factor I 81 to. C3a C3a (amino acids 672-748 of SEQ ID NO:47) is an anaphylatoxin that is involved in inflammation, basophil and mast cell degranulation, enhanced vascular permeability, smooth muscle contraction, and induction of suppressor T cells. b. C3b C3b (SEQ ID NO:47 amino acids 749-1663) has several functions in the complement cascade. C3b is an opsonin that facilitates the uptake and destruction of pathogens by phagocytic cells. Additionally, C3b combines with C3-convertases to generate C5-convertases that activate complement protein C5, thus generating C5a and C5b anaphylatoxin, which combines with C6, C7, C8, and C9 to form the C5-convertase complex. membrane attack. Additionally, as described in section 1b above, C3b is involved in the alternative complement initiation pathway. C3b is regulated by the complement regulatory protein Factor I, a plasma protease that degrades C3b into several fragments, including ¡C3b, C3c, C3d, C3f, and C3dg, thereby permanently inactivating C3b. C3b plays a critical role in complement-mediated effector functions by virtue of its ability to bind to the C3-convertases C4b2b and C3bBb, thereby generating the C5-convertases C4b2b3b and C3bBb3b. C5-convertases cleave the C5 zymogen into its active fragments, namely C5a and C5b anaphylatoxin. C5a is involved in chemotaxis and inflammation and C5b is involved in MAC formation. c. C3b inhibitors C3b has binding sites for several complement components including C5, properdin (P), factors Η, B, and I, complement receptor 1 (CR1), and membrane cofactor protein (MCP) (see Sahu and Lambris (2001). ) Immunological Reviews 180:35-48). Binding of Factor I, a plasma protease, in the presence of cofactors H, CR1, and MCP results in inactivation of C3b, while binding of factors B and P in the presence of factor D results in amplification of the convertase. C3 and the start of MAC. Factor I cleaves C3b in the presence of cofactors between residues 1303-1304, 1320-1321 and 954-955 of SEQ ID NO:47 generating C3b (amino acids 749-1303 of SEQ ID NO:47) and C3f ( amino acids 1304-1320 of SEQ ID NO:47). Although not technically considered an anaphylatoxin, ¡C3b, an inactive derivative of C3b, functions to induce leukocyte adhesion to vascular endothelium and induce production of the proinflammatory cytokine IL-1 through binding to its surface integrin receptors. cell phone. The ¡C3b protein functions as an opsonin. Subsequently, Factor I cleaves ¡C3b, C3c generating fragments (C3c alpha' chain Fragment 1: amino acids 749-954 of SEQ ID NO:47 and C3c alpha' chain Fragment 2: amino acids 1321-1663 of SEQ ID NO :47) and C3dg (amino acids 955-1303 of SEQ ID NO:47). The end result is that C3b is permanently inactivated (see Sahu and Lambris (2001) Immunological Reviews 180:35-48). C3dg can be further cleaved to generate C3g (SEQ ID NO: 47 amino acids 955-1001) and C3d (SEQ ID NO: 47 amino acids 1002-1303) fragments. D. MODIFIED U-PA POLYPEPTIDES THAT CLEVE C3 Modified or variant urokinase-like plasminogen activator (u-PA) polypeptides are provided herein. Conjugates, such as fusion proteins, are also provided that contain modified u-PA polypeptides so that the resulting activated forms of these cleave C3. The modified u-PA polypeptides provided herein exhibit altered activities or properties as compared to a natural, wild-type or reference u-PA polypeptide. For example, the u-PA polypeptides provided herein contain modifications compared to a natural, wild-type, or reference u-PA polypeptide set forth in any of SEQ ID cboynn / i znz / B / v NO:1-6, or in a polypeptide having at least 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, particularly at least 95% sequence identity to any of SEQ ID NO:1-6, such as the protease domain of u -Reference PA set forth in SEQ ID NO:5. Modified u-PA polypeptides provided herein include u-PA polypeptides that alter (inhibit) complement activation by effecting inhibitory cleavage of complement protein C3. Among the modified u-PA polypeptides provided herein are those that effect inhibitory cleavage of complement C3 protein. Included are those that effect inhibitory cleavage of C3 with higher activity or specificity, Kcat / Km, compared to a corresponding form of u-PA that does not contain the modification (the replacement, deletion, and / or insertion) or compared to the u-PA form. corresponding unmodified u-PA whose sequences are set forth in any of SEQ ID NO:1-6. Modified u-PA polypeptides may also have decreased specificity and / or selectivity for substrates and targets cleaved or recognized by unmodified u-PA, including plasminogen cleavage and / or uPAR binding, compared to u-PA polypeptide. Corresponding PA that does not contain the amino acid modifications. The modified u-PA polypeptides provided herein inhibit or inactivate complement through inhibitory or inactivating cleavage of complement C3 protein. The modified u-PA polypeptides provided herein inhibit or inactivate complement by cleaving complement protein C3 at a cleavage site that results in inhibition or inactivation of C3. The inactivation or cleavage by inhibition of complement protein C3 can be in any sequence in C3 as long as the resulting cleavage of C3 results in the inactivation or inhibition of complement activation. Since the modified u-PA polypeptides provided herein inhibit complement activation, the modified u-PA polypeptides do not effect cleavage of the C3 zymogen to generate the C3a and C3b activated fragments. Thus, the modified u-PA polypeptides provided herein do not cleave C3 between residues 748-749 of SEQ ID NO: 47, which would result in the generation of C3a and C3b. Cleavage sites of complement protein 03 inhibition or inactivation can be determined or identified empirically. If necessary, a modified u-PA polypeptide can be tested for its ability to inhibit complement as described in section E below and as exemplified in the Examples. The modified u-PA polypeptides provided herein catalyze the inhibitory or inactivating cleavage of complement C3 protein. The modified u-PA polypeptides provided herein cleave complement C3 protein at cboynn / i znz / B / v any cleavage sequence as long as the resulting C3 fragments are inactive or unable to activate a complement-mediated effector function. The modified u-PA polypeptides provided herein have altered (ie, decreased) specificity and / or selectivity for natural u-PA targets, including plasminogen and uPAR. In one example, the modified u-PA polypeptides provided herein have reduced specificity for plasminogen cleavage. In another example, the modified u-PA polypeptides provided herein have reduced selectivity for binding to uPAR. In some examples, the modified u-PA polypeptides provided herein have reduced specificity for plasminogen cleavage and reduced selectivity for uPAR binding. In other examples, the modified u-PA polypeptides provided herein have higher specificity for cleavage of complement protein C3 and lower specificity for cleavage of plasminogen. In other examples, the modified u-PA polypeptides provided herein have higher selectivity for complement protein C3 and lower selectivity for plasminogen and / or uPAR. The modified u-PA polypeptides provided herein and described in the examples are, for example, isolated protease domains from u-PA. Smaller portions of these that retain protease activity are also contemplated. The modified u-PA polypeptides provided herein are mutants of the uPA protease domain, particularly modified u-PA polypeptides in which the Cys residue in the protease domain is free (i.e., does not form disulfide bonds). with no other Cys residues in the protein) is substituted with another amino acid substitution, preferably with a conservative amino acid substitution or a substitution that does not abolish activity, such as, for example, Serine substitution, and modified u-PA polypeptides wherein one or more glycosylation sites are removed. Modified u-PA polypeptides are also contemplated in which other conservative amino acid substitutions in which catalytic activity is retained are also contemplated (see, eg, Table 3, for examples of amino acid substitutions). The modified u-PA polypeptides provided herein contain one or more amino acid modifications such that they cleave complement protein 03 in a manner that results in complement inactivation or inhibition. The modifications can be a single amino acid modification, such as single amino acid replacements (substitutions), insertions or deletions, or multiple amino acid modifications, such as multiple amino acid replacements, insertions or deletions. Examples of modifications are amino acid replacements, including single or multiple amino acid replacements. The amino acid replacement can be a conservative substitution, as set forth in Table 3, or a non-conservative substitution, such as any cboynn / i znz / B / v described herein. The modified u-PA polypeptides provided herein may contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20 or more positions modified compared to the u-PA polypeptide that does not contain the modification. The modifications described herein can be made in any u-PA polypeptide. For example, the modifications are made in a human u-PA polypeptide having an amino acid sequence that includes or is set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4. , SEQ ID NO: 5 or SEQ ID NO: 6, or allelic variants thereof; a mouse u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:52 or 66; a rat u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:53 or 67; a cow u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:54 or 68; a porcine u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:55 or 69; a rabbit u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:56 or 70; a chicken u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:57 or 71; a yellow baboon u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:58 or 72; a Sumatran orangutan u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:59 or 73; a dog u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NQ:60 or 74; an ovine u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:61 or 75; a marmoset u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:62; a rhesus macaque u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:63; a northern white-cheeked gibbon u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:64; and a chimpanzee u-PA polypeptide having an amino acid sequence that is included or set forth in SEQ ID NO:65; or in sequence variants or catalytically active fragments exhibiting at least 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93 %, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of SEQ ID NO:1 -6 and 52-75. Modified u-PA polypeptides provided herein may be modified in any region or domain of a u-PA polypeptide provided herein, as long as the modified u-PA polypeptide retains its ability to effect inactivation or inhibitory cleavage. of the complement protein C3. The modified u-PA polypeptides provided herein may be u-PA polypeptides CbQjnn / l 7Π7 / Β / Υ single chain or two chains, species variants, allelic variants, isoforms or catalytically active fragments thereof, such as, for example, the protease domain thereof. The u-PA polypeptides provided herein can be full-length or truncated u-PA polypeptides. The modified u-PA polypeptides provided herein can be the u-PA protease domain or a modified form of the u-PA protease domain. Also contemplated for use herein are zymogenic, precursor, or mature forms of modified u-PA polypeptides, provided that the u-PA polypeptides retain their ability to effect inhibitory cleavage or inactivation of complement protein 03. Modifications to a u-PA polypeptide can also be made to a u-PA polypeptide that also contains other modifications, including primary sequence modifications and modifications that are not in the primary sequence of the polypeptide. For example, a modification described herein can be in a u-PA polypeptide that is a fusion polypeptide or chimeric polypeptide. Modified u-PA polypeptides provided herein also include polypeptides that are conjugated to a polymer, such as a PEG reagent. For purposes herein, reference to positions and amino acids for modification, including amino acid replacements or substitutions, herein are with reference to the u-PA polypeptide set forth in any of SEQ ID NO:1-6. It is within the level of one of ordinary skill in the art to make any of the modifications provided herein in another u-PA polypeptide by identifying the corresponding amino acid residue in another u-PA polypeptide, such as the u-PA polypeptide. set forth in any of SEQ ID NO:1-6 or a variant thereof exhibiting at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to a u-PA polypeptide set forth in any of SEQ ID NO:1-6. Corresponding positions in another u-PA polypeptide can be identified by aligning the u-PA polypeptide with the reference to a u-PA polypeptide set forth in any of SEQ ID NO:1-6. For modification purposes (eg, amino acid replacement), the corresponding amino acid residue can be any amino acid residue, and need not be identical to the residue set forth in any of SEQ ID NO:1-6. Usually, the corresponding amino acid residue identified by alignment with, for example, residues in SEQ ID NO:5 is an amino acid residue that is identical to SEQ ID NO:5, or is an amino acid residue conservative or semi-conservative thereto. It is also understood that exemplary replacements provided herein can be made at the corresponding residue in a u-PA polypeptide, such as the u-PA protease domain, as long as the replacement is different from what exists in the u-PA polypeptide. unmodified form of the u-PA polypeptide, such as the u-PA protease domain. Based on PbQynn / i znz / B / v In this description and in the description elsewhere herein, it is well within the level of one skilled in the art to generate a modified u-PA polypeptide containing any of the mutations described, and to analyze each one for detecting a property or activity as described herein. The modified u-PA polypeptides provided herein alter complement activity through proteolysis-mediated inhibition or inactivation of complement protein C3. Additionally, the modified u-PA polypeptides provided herein have decreased specificity for plasminogen cleavage and / or uPAR binding. For example, the modified u-PA polypeptides provided herein exhibit less than 100% of the wild-type activity of a u-PA polypeptide for plasminogen cleavage, such as less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or less of the plasminogen-cleaving activity of a wild-type or reference u-PA polypeptide, such as the corresponding polypeptide not containing the modification of amino acids. In another example, the modified u-PA polypeptides provided herein exhibit less than 100% of the wild-type binding activity of a u-PA polypeptide for uPAR, such as less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or less of the uPAR-binding activity of a wild-type or reference u-PA polypeptide, such as the corresponding polypeptide not containing the modification of amino acids. Nucleic acid molecules encoding any of the modified u-PA polypeptides provided herein are also provided herein. In some examples, the encoding nucleic acid molecules can also be modified to contain a heterologous signal sequence to alter (eg, increase) the expression and secretion of the polypeptide. The modified u-PA polypeptides and encoding nucleic acid molecules provided herein can be produced or isolated by any method known in the art including isolation from natural sources, isolation of recombinantly produced proteins in cells, tissues, and organisms, and by recombinant methods and by methods including in silico steps, synthetic methods and any method known to those skilled in the art. The modified polypeptides and encoding nucleic acid molecules provided herein can be produced by standard recombinant DNA techniques known to one of skill in the art. Any method known in the art can be used to effect mutation of one or more amino acids in a target protein. Methods include standard random or site-directed mutagenesis of encoding nucleic acid molecules or methods of solid phase polypeptide synthesis. For example, nucleic acid molecules encoding a u-PA polypeptide can be subjected to mutagenesis, such as random mutagenesis of PbQynn / i znz / B / v coding nucleic acid, error-prone POR, site-directed mutagenesis, overlapping POR, gene shuffling, or other recombinant methods. Nucleic acid encoding the polypeptides can then be introduced into a host cell for heterologous expression. Therefore, nucleic acid molecules encoding any of the modified polypeptides provided herein are also provided herein. In some examples, the modified u-PA polypeptides are produced synthetically, such as using solution phase or solid phase peptide synthesis. The nucleic acid molecules can be provided in gene therapy vectors, such as AAV or adenovirus vectors, for expression of the encoded modified u-PA polypeptide in vivo, such as in the eye or for systemic administration. The encoded u-PA polypeptide can be a full-length polypeptide or a protease domain or other form that is active or capable of being activated. The u-PA polypeptides provided herein have been modified to have greater specificity and / or selectivity for cleavage inhibitory cleavage sequence or inactivation of complement protein 03 polypeptides. u-PA can be modified using any method known in the art for protein modification. These methods include random and site-directed mutagenesis. Assays such as assays for complement activation biological function provided herein and known in the art can be used to assess the biological function of a modified u-PA polypeptide to determine whether the modified u-PA polypeptide is targets complement protein 03 for cleavage and inactivation. Example methods for identifying a u-PA polypeptide and modified u-PA polypeptides are provided herein. 1. Example Modified u-PA Polypeptides Provided herein are modified u-PA polypeptides that contain one or more, including 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and more modifications of amino acids in a u-PA polypeptide and that cleave complement protein 03 such that complement is either inhibited or inactivated. Modifications are in the primary amino acid sequence and include amino acid residue replacements, deletions, and insertions. The modification alters the specificity / activity of the uPA polypeptide, when in an active form. The herein modified u-PA polypeptides are selected to recognize and cleave a target site on a complement protein, particularly 03, to inactivate it. They can also be further modified and tested for reduced specificity / activity on in vivo substrates, such as plasminogen. They can be selected and identified by any suitable protease detection method. The herein modified u-PA polypeptides were initially identified using the screening method described in US Pat. cboynn / i znz / B / v 8,211,428, wherein a library of modified proteases is reacted with a cognate serpin or other inhibitory serpin that is modified to include a target sequence in the reactive site loop to capture modified proteases that would cleave said target. The modified u-PA polypeptides provided herein show increased activity or specificity or Kcat / Km for complement C3 protein at a site that inactivates C3, and may also have decreased activity or specificity for plasminogen and / or display increased selectivity, specificity and / or activity for a target site complement 03 protein, thus the modified u-PA polypeptide inactivates 03. The modified u-PA polypeptides exhibit increased activity to cleave and inactivate 03 compared to the corresponding wild-type form or wild type with the C122S replacement (by chymotrypsin numbering). In particular, the protease domain of the modified u-PA polypeptide exhibits increased C3-inactivation cleavage activity compared to the u-PA protease domain of SEQ ID NO: 5 (u-PA protease domain with C122S ). The increase in activity can be 10%, 20%, 50%, 100%, 1 time, 2 times, 3 times, 4, 5, 6, 7, 8, 9, 10 times and more compared to the u- PA not modified. The modified u-PA polypeptide may have reduced activity for a natural substrate, such as plasminogen. For example, modified u-PA polypeptides can exhibit 0 to 99% of the u-PA activity of a wild-type or reference u-PA polypeptide, such as the u-PA polypeptide set forth in SEQ ID NO. :5, for plasminogen and at least 0.5 times, 1 time, 2 times, 3 times or more to cleave C3 to inactivate it. For example, the modified u-PA polypeptides provided herein exhibit less than or less than about 99%, 95%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20% , 10% or less of the u-PA activity of a wild-type or reference u-PA polypeptide, such as the corresponding polypeptide that does not contain the amino acid modification (eg, amino acid replacement), eg, a domain of u-PA protease set forth in SEQ ID NO:5. For example, exemplary positions that can be modified, for example by amino acid replacement or substitution, include, but are not limited to, any of the positions that correspond to position 173,178, 179, 180, 181, 185, 186, 187,188, 208, 209, 249, 250, 252, 306, 314 or 353 with reference to the amino acid sequence set forth in SEQ ID NO: 3 (corresponding to positions 30, 35, 36, 37, 37a, 38, 39, 40, 41, 60a, 60b, 97a, 97b, 99, 149, 157 or 192 according to chymotrypsin numbering). For example, the amino acid positions can be replacements at positions that correspond to the replacement of phenylalanine (F) at one or more of positions 173, R178, R179, H180, R181, V185, T186, Y187, V188, D208, Y209, T249. , L250, H252, Y306, M314 or Q353 with reference to amino acid positions set forth in SEQ ID NO:3 (corresponding to cboynn / i znz / B / v F30, R35, R36, Η37, R37a, V38, T39, Y40, V41, D60a, Y60b, T97a, L97b, H99, Y149, M157 and Q192, respectively according to chymotrypsin numbering). Example amino acid replacements at any of the above positions are set forth in Table 11. Reference to the corresponding position in Table 11 refers to the positions set forth in SEQ ID NO:3. (See also the Examples below). It is understood that replacements can be made at the corresponding position in another u-PA polypeptide by alignment with the sequence set forth in SEQ ID NO:3, whereby the corresponding position is the aligned position. For example, the replacement can be made in the u-PA protease domain with the sequence set forth in SEQ ID NO: 2 or a reference u-PA protease domain with the sequence set forth in SEQ ID NO: 5. In For some examples, the amino acid replacement(s) may be at the corresponding position in a u-PA polypeptide as set forth in SEQ ID NO: 5 or a variant thereof that is at least or at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 86%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% , 98%, 99%, particularly 95%, or greater sequence identity thereto, provided that the resulting modified uPA polypeptide exhibits altered (i.e., enhanced) specificity toward complement protein C3 as compared to u-PA activity towards plasminogen and / or an altered selectivity for complement protein C3. In one example, any one or more of the replacements are found in any of SEQ ID NOs:1-6, so long as the resulting modified u-PA polypeptide exhibits altered (i.e., enhanced) specificity for complement protein C3 in comparison with an activity of u-PA towards plasminogen and / or an altered selectivity for the complement protein 03. PbQynn / i znz / B / v Table 11. Example mutations resulting in increased activity for C3 cleavage Corresponding Position (in SEQ ID NO:3) Corresponding Position (chymotrypsin numbering) Replacement 173 30 Y 178 35 W YQ 179 36 H 180 37 E P D N G KY 181 37a Q P E N S 185 38 D E 186 39 W Y F 187 40 H FQ 188 41 R L 208 60a PT 209 60b QHSATL 249 97a E I 250 97b AG 252 99 Q 279 122 S 306 149 K R 314 157 K 353 192 H > ί Ν C ι\ C C -X ο ί σ Exemplary amino acid modifications in the modified u-PA polypeptides provided herein include, but are not limited to, tyrosine (Y) replacement at a position corresponding to position 173 (30 by chymotrypsin numbering); W at a position corresponding to position 178 (35 by chymotrypsin numbering); And in a position that corresponds to position 178; Q at a position corresponding to position 178; H at a position corresponding to position 179 (36 by chymotrypsin numbering); E at a position corresponding to position 180 (37 by chymotrypsin numbering); P at a position corresponding to position 180; D at a position corresponding to position 180; N at a position corresponding to position 180; G at a position corresponding to position 180; K at a position corresponding to position 180; And in a position that corresponds to position 180; Q at a position corresponding to position 181 (37a by chymotrypsin numbering); P at a position corresponding to position 181; E at a position corresponding to position 181; N at a position corresponding to position 181; S at a position corresponding to position 181; D at a position corresponding to position 185 (38 by chymotrypsin numbering); E at a position corresponding to position 185; W at a position corresponding to position 186 (39 by chymotrypsin numbering); And in a position that corresponds to position 186; F at a position corresponding to position 186; H at a position corresponding to position 187 (40 by chymotrypsin numbering); F at a position corresponding to position 187; Q at a position corresponding to position 187; R at a position corresponding to position 188 (41 by chymotrypsin numbering); L at a position corresponding to position 188; P at a position corresponding to position 208; T at a position corresponding to position 208 (60a by chymotrypsin numbering); Q at a position corresponding to position 209 (60b by chymotrypsin numbering); H at a position corresponding to position 209; S at a position corresponding to position 209; A at a position corresponding to position 209; T at a position corresponding to position 209; L at a position corresponding to position 209; E at a position corresponding to position 249 (97a by chymotrypsin numbering); I at a position corresponding to position 249; A at a position corresponding to position 250 (97b by chymotrypsin numbering); G at a position corresponding to position 250; Q at a position corresponding to position 252 (99 by chymotrypsin numbering); K at a position corresponding to position 306 (149 by chymotrypsin numbering); R at a position corresponding to position 306; K at a position corresponding to position 314 (157 by chymotrypsin numbering); or H at a position corresponding to position 353 (192 by chymotrypsin numbering); each with reference to the amino acid positions set forth in SEQ ID NO:3. S at a position corresponding to position 279 (122S) by chymotrypsin numbering) replaces a free Cys to thereby reduce a tendency for aggregation. Exemplary modified u-PA polypeptides containing 2 or more amino acid modifications are set forth in Table 12 below, and their activity to cleave C3 described in Table 14. Sequence ID NO. refers to an exemplary u-PA protease domain containing the aforementioned replacements, including the replacement at C122S to reduce or eliminate aggregation. C122 is a free cysteine, which can result in crosslinking between protease polypeptides. It is understood that the protease domain is exemplary, and full-length and precursor molecules, as well as other catalytically active portions of the protease domain, full-length polypeptide, and precursor may include such replacements, to form u-PA polypeptides. full-length modified activated and other forms. CbQjnn / l 7Π7 / Β / Υ | Table 12. Modified u-PA polypeptides Mature u-PA numbering Chymotrypsin numbering Example SEQ ID NO F173ΥΛ / 185D / Y187ΗΛ / 188R / L250A / H 252Q / C279S / M314K F30Y / V38D / Y40HA / 41 R / L97bA / H 99Q / C122S / M157K 8 F17 3Y / R178W / R179H / H180E / V185E / T186W / Y187H / V188R / Y209Q / T249E / L 250A / H252Q / C279S / Y306K / M314K F30Y / R35W / R36H / H37EA / 38E / T3 9W / Y40H / V41 R / Y60bQ / T97aE / L9 7bA / H99Q / C122S / Y149K / M157K 9 F173Y / R178W / R179H / H180DA / 185E / Τ186Y / Y187F / V188R / T249I / L250A / H2 52Q / C279S / Y306R / M314K F30Y / R 35W / R36H / H37D / V38E / T3 9Y / Y40F / V41 R / T97al / L97bA / H99 Q / C122S / Y149R / M157K 10 R178W / R179H / H180N / V185E / T186F / Y187FA / 188R / T249I / L250A / H252Q / C2 79S / Y306R / M314K / Q353H R35W / R36H / H37N / V38E / T39F / Y4 0F / V41 R / T97al / L97bA / H99Q / C12 2S / Y149R / M157K / Q192H 11 F173Y / R178Y / R179H / H180KA / 185E / T 186F / Y187FA / 188R / T 249I / L250A / H25 2Q / C279S / Y306R / M314K F30Y / R35Y / R36H / H37KA / 38E / T3 9F / Y40F / V41 R / T97al / L97bA / H99 Q / C122S / Y149R / M157K 12 F173Y / R178W / R179H / H180 ΝΛ / 185E / Τ186Y / Y187F / V188R / Y209S / T249E / L2 50A / H252Q / O279S / Y306K / M314K F30Y / R35W / R36H / H37N / V38E / T3 9Y / Y40F / V41 R / Y60bS / T97aE / L97 bA / H99Q / C12 2S / Y149K / M157K 13 F173Y / R178W / R179H / H180P / V185E / Τ186Y / Y187F / V188R / Y209S / T249E / L2 50A / H252Q / C279S / Y306K / M314K F30Y / R35W / R36H / H37PA / 38 E / T3 9Y / Y40F / V41 R / Y60bS / T97aE / L97 bA / H99Q / C122S / Y149K / M157K 14 V185E / Y187Q / V188L / Y209L / L250A / H 252Q / C279S V38E / Y40Q / V41 L / Y60bL / L97bA / H 99Q / C122 S 15 F173Y / R178Q / R179H / H180G / R181E / F30Y / R35Q / R36H / H37G / R37aE / V 16 100 | Table 12. Modified u-PA polypeptides NUMERATION OF MATURE NUMBERPSINE NUMBER SEQ NO OF EXAMPLE V185E / T186F / Y187F / V188R / D208P / Y 209S / T249I / L250A / H252Q / C279S / Y30 6R / M314K 38E / T39F / Y40F / V41 R / D60AP / D60AP Y60 bS / T97al / L97bA / H99Q / C122S / Y1 49R / M157K F173Y / R178Y / R179H / H180P / R181Q / V 185E / T186Y / Y187F / V188R / Y209H / T2 49I / L250A / H252Q / C2 79S / Y306R / M31 4K F30Y / R35Y / R36H / H37P / R37aQ / V 38E / T39Y / Y40F / V41 R / Y60bH / T97al / L97bA / H99Q / C122S / Y149R / M1 57K 17 R178Q / H180Y / R181E / V185E / T186Y / V 188R / D208T / Y209T / T249I / L250A / H25 2Q / C279S / Y306R R35Q / H37Y / R37aE / V38E / T39Y / V 41 R / D60aT / Y60bT / T97al / L97bA / H 99Q / C122S / Y149R 18 R178W / H180P / R181N / V185E / T186Y / V188R / D208P / Y209L / T249I / L250A / H2 52Q / C279S / Y306R R35W / H37P / R37aN / V38E / T39Y / V 41 R / D60aP / Y60bL / T97al / L97bA / H 99Q / C122S / Y149R 19 R178W / H180D / R181P / V185Ε / Γ186W / V188R / Y209A / T249I / L250A / H252Q / C2 79S / Y306R R35W / H37D / R37aP / V38E / T39W / V41 R / Y60bA / T97al / L97bA / H99Q / C122S / Y 149R 20 R178Q / H180Y / R181E / V185Ε / Γ186Y / V 188R / D208P / Y209Q / T249I / L250A / H25 2Q / C279S / Y306R R35Q / H37Y / R37aE / V38E / T39Y / V 41 R / D60aP / Y60bQ / T97al / L97bA / H99 Q / C122S / Y149R 21 H180Y / R181E / V185E / T186Y / V188R / D 208P / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R H37Y / R37aE / V38E / T39Y / V41 R / D 60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R 22 R178Q / R181E / V185E / T186Y / V188R / D 208P / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R R35Q / R37aE / V38E / T39YA / 41 R / D 60aP / Y60bQ / T97al / L9 7bA / H99Q / C122S / Y149R 23 R178Q / H180Y / V185E / T186Y / V188R / D 208P / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R R35Q / H37Y / V38E / T39Y / V41R / D6 0aP / Y60bQ / T97 AL / L97bA / H99Q / C 122S / Y149R 24 R178Q / H180Y / R181E / T186Y / V188R / D 208P / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R R35Q / H37Y / R37aE / T39Y / V41 R / D 60aP / Y60bQ / T 97al / L97bA / H99Q / C122S / Y149R 25 R178Q / H180Y / R181E / V185E / V188R / D208P / Y209Q / T249I / L250A / H252Q / C 279S / Y306R R35Q / H37Y / R37aE / V38E / V41 R / D 60aP / Y60 bQ / T97al / L97bA / H99Q / C122S / Y149R 26 R178Q / H180Y / R181E / V185Ε / Γ186Y / D 208P / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R R35Q / H37Y / R37aE / V38E / T39Y / D 60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R 27 R178Q / H180Y / R181E / V185E / T186Y / V 188R / Y209Q / T249I / L250A / H252Q / C27 9S / Y306R R35Q / H37Y / R37aE / V38E / T39Y / V 41 R / Y60 bQ / T97al / L97bA / H99Q / C 122S / Y149R 28 R178Q / H180Y / R181E / V185E / T186Y / V 188R / D208P / T249I / L250A / H252Q / C27 9S / Y306R R35Q / H37Y / R37aEA / 38E / T39Y / V 41 R / D6 0aP / T97al / L97bA / H99Q / C 122S / Y149R 29 R178Q / H180Y / R181E / V185Ε / Γ186Y / V 188R / D208P / Y209Q / L250A / H252Q / C2 79S / Y306R R35Q / H37Y / R37aE / V38E / T39Y / V 41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / Y149R 30 R178Q / H180Y / R181E / V185E / T186Y / V 188R / D208P / Y209Q / T249I / H252Q / C2 79S / Y306R R35Q / H37Y / R37aE / V38E / T3 9Y / V 41 R / D60aP / Y60bQ / T97al / H99Q / C 122S / Y149R 31 ctzQynn / i ζπζ / β / υιλι 101 | Table 12. Modified u-PA polypeptides Mature u-PA numbering Chymotrypsin numbering Example SEQ ID NO R178Q / H180Y / R181E / V185Ε / Γ186Y / V 188R / D208P / Y209Q / T249I / L250A / C27 9S / Y306R R35Q / H37Y / R37aE / V38E / T39 Y / V 41 R / D60aP / Y60bQ / T97al / L97bA / C122S / Y149R 32 R178Q / H180Y / R181E / V185E / T186Y / V 188R / D208P / Y209Q / T249I / L250A / H25 2Q / C279S R35Q / H3 7Y / R37aE / V38E / T39Y / V 41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S 33 Y187Q / V188L / Y209L / L250A / H252Q / C279S Y40Q / V41 L / Y60bL / L97bA / H99Q / C 122S 34 V185E / Y18 7Q / Y209L / L250A / H252Q / C279S V38E / Y40Q / Y60bL / L97bA / H99Q / C122S 35 V185E / Y187Q / V188L / L250A / H252Q / C279S V38E / Y40Q / V41 L / L97bA / H99Q / C 122S 36 V185E / Y187Q / V188L / Y209L / H252Q / C279S V38E / Y40Q / V41 L / Y60bL / H99Q / C 122S 37 V185E / Y187Q / V188L / Y209L / L250A / C279S V38E / Y40Q / V41 L / Y60bL / L97bA / C 122S 38 Y187Q / V188L / L250 A / H252Q / C279S Y40Q / V41 L / L97bA / H99Q / C122S 39 Y187Q / V188L / L250A / C279S Y40Q / V41 L / L97bA / C122S 40 R181S / V188R / L250G / H252Q / C279S R37aS / V41 R / L97bG / H99Q / C122S 41 Τ186Y / V188R / L250A / H252Q / C279S T39Y / V41 R / L97bA / H99Q / C122S 42 T186Y / V188R / Y209Q / L250A / H252Q / C 279S T39Y / V41 R / Y60bQ / L97bA / H99Q / C122S 43 T186Y / V188 R / D208P / L250A / H252Q / C 279S T39Y / V41 R / D60aP / L97bA / H99Q / C122S 44 cboynn / i znz / B / v 2. Additional Modifications Any of the modified u-PA polypeptides provided herein may contain one or more additional modifications. Additional modifications can include, for example, any amino acid substitution, deletion, or insertion known in the art, typically any that increases the specificity toward complement protein C3 compared to the activity of u-PA toward plasminogen and / or alters the selectivity for complement protein C3. In addition, modifications that alter any other activity of interest are contemplated. It has long been known in the art that primary sequence amino acid modifications are additive (see, eg, Wells (1990) Biochem 29:8509-8517). Any modified u-PA polypeptide provided herein may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20 or more additional amino acid modifications to provide additional activities or alter activities. Examples of additional modifications that can be included in the modified u-PA polypeptides provided herein include, but are not limited to, those described in US Pat. 4,997,766; 5,126,134; 5,129,569; 5,275,946; 102 5,571,708; 5,580,559; 5,648,253; 5,728,564; 5,759,542; 5,811,252; 5,891,664; 5,932,213; 5,980,886; 6,248,712; 6,423,685; 7,070,925; 7,074,401; 7,807,457; 7,811,771; 8,211,428; United States Patent Publication Nos. 2002 / 0106775; 2004 / 0265298; 2004 / 0146938; 2009 / 0010916; 2011 / 0055940; 2008 / 0020416; 2006 / 0142195; International Patent Publication Nos. WO1988 / 008451; WO1989 / 010401; WO1990 / 004635; WO1996 / 013160; WO 2002 / 40503; Petersen et al. (2001) Eur J Biochem 268:4430-4439; Skeldal et al. (2006) FEBS J 273:5143-5149; Sun et al. (1997) J Biol Chem 272:23818-23823; Blouse et al. (2009) J Biol Chem 284:4647-4657; Nelles et al. (1987) JBC 262:5682-5689; Crowley et al. (1993) Proc. nati. Acad. Sel. USA 90:5021-5025; Zeslawska et al. (2000) J Mol Biol 301:465-475; Zeslawska et al. (2003) J Mol Biol 328:109-118; Quax et al. (1998) Arterioscler Thromb Vaso Biol 18:693-701; Homandberg and Wai (1990) Thrombin Res 58:403412; Zaitsev et al. (2010) Blood 115:5241-5248; Yang et al. (1994) Biochemistry 33:606-612; Davidow et al. (1991) Protein Eng 4:923-928; Boutad and Castellino (1993) Arch Biochem Biophys 303:222-230; Tsujikawa et al. (1996) Yeast 12:541-553; Garriere et al. (2002) Biol Chem 383:107-113; Stopelli et al. (1985) Proc. nati. Acad. Sel. USA 82:4939-4943; Stoppelli et al. (1987) J Biol Chem 262:4437-4440; Franco et al. (1998) J Biol Chem 273:27734-27740; Franco et al. (1997) J Cell Biol 137:779-791; Li et al. (1995) J Biol Chem 270:30282-30285; Botkjaer et al. (2009) Biochemistry 48:9606-9617; Bdeir et al. (2003) Blood 102:3600-3608; Eguchi et al. (1990) J Biochem 108:72-79; Miyake et al. (1988) J Biochem 104:643-647; Bergstrom et al. (2003) Biochem 42:5395-5402; Sun and Liu (2005) Proteins 61:870-877; Sun et al. (1998) Biochemistry 37:2935-2940; Anderson et al. (2008) Biochem J 412:447-457; Li et al. (1992) Biochim Biophys Acta 1159:37-43; Lijnen et al. (1988) Eur J Biochem 177:575-582; Lijnen et al. (1988) Eur J Biochem 172:185-188; Lijnen et al. (1992) Eur J Biochem 205:701709; Lijnen et al. (1994) Eur J Biochem 224:567-574; Lijnen et al. (1990) J Biol Chem 265:5232-5236; Yoshimoto et al. (1996) Biochim Biophys Acta 1293:83-89; Magdolen et al. (1996) Eur J Biochem 237:743-751; Nienaber et al. (2000) J Biol Chem 275:7239-7248; Gurewich et al. (1988) J Clin Invest 1956-1962; Liu et al. (1996) Biochemistry 35:14070-14076; Liu et al. (2002) Circ Res 90:757-763; Mukhina et al. (2000) J Biol Chem 275:16450-16458; Peng et al. (1997) Biochem Mol Biol Int 41:887-894; Turkmen et al. (1997) Electrophoresis 18:686-689; Peng et al. (1999) Biotechnol Lett 21:979-985; Ueshima et al. (1994) Thromb Haemost 71:134-140; and Melnick et al. (1990) J Biol Chem 265:801-807. Non-limiting examples of amino acid modifications described in the art include any one or more of S9A, C13A, T18A, C19A, V20A, S21A, N22Y, N22A, N22Q, N22R, K23A, K23H, K23Q, K23E, Y24A, F25A , S26A, S26F, N27A, N27R, I28A, H29A, H29R, W30A, W30R, W30F, N32S, K35A, G38R, E43A, I44A, D45A, K46A, S47A, S47G, K48A, K48P, T49A, Y51A, N54A, L80H , Q81R, Q82P, T83R, H99Y, P105A, D106A, N107A, R108A, R108D, R109A, R110A, 103 G118N, L119R, K120R, Κ120Α, P121L, L122T, L122R, V123Y, V123W, Q124A, Ε125Α, Η129Α, D130G, C131W, K135G, K135S, Κ135Υ, K135Q, Κ136Ρ, S138E, C148S, C148A, Κ151Ε, Τ152Α, R154G, R154P, R154A, P155R, P155L, Ρ155Α, Ρ155Ν, P155S, P155G, P155Q, R156P, R156A, R156H, R156S, R156Y, R156E, R156G, R156L, F157L, F157T, F157G, F157Q, F157D, F157E, K158R, Κ158Ε, Κ158Α, Κ158Η, K158S, Κ158Υ, K158G, K158W, K158V, Κ158Μ, I159R, Ι159Α, Ι159Ρ , I159G, Ι160Α, Ι160Κ, G162R, Ε163Α, F164V, F164A, F164V, I167L, Ρ171L, F173I, F173V, F173L, F173T, F173G, F173M, A175S, Υ177Α, R178A, R179A, Η180Α, R181A, S184A, Τ186Α, Τ186Ε, T186D, Υ187Α, Υ187Η, V188A, S192N, Ι194Μ, S195A, Η204Α, H204Q, F206A, D208A, Υ209Α, Ρ210Α, Κ211Α, Κ211Q, K212A, E213A, D214A, Y215A, 1216A, Y218A, R221A, S222L, R223G, R223A, L224A, L2 24P, N225A, S226P, N227A, Q229A, E231G, K233E, K233A, F234A, E235K, E235A, E237A, I240V, K243E, K243A, D244A, Y245A, D255A, R262A, K264A, E265A, R267A, C268Y, C279S, C279A, F289L, G290D, E2 94G, I295T, G297D, F298A, G299A, G299H, K300A, K300H, K300W, E301D, E301A, E301H, N302A, N302Q, N302V, N302L, N302I, N302S, N302T, S303E, S303A, S303E, T304A, T304V, T304M, D3 05A, Y306A, Y306G, Y306V, Y306H, L307A, Y308A, P309A, P309S, P309T, P309V, P309G, P309N, P309L, P309D, P309R, P309H, P309F, P309W, E310A, Q311A, L312P, L312V, L312M, K313Y, K313T, K313A, K313H, T315A, T315I, V316A, V317A, Y330H, A343T, D344A, Q346A, W3 47A, K348A, K348E, T349I, D350A, S351A, Q353A, G354R, D355A, S356A, G357E, G366C, R378C, R378A, K383A, K385A, R400A, H402A, K404A, E405A, E406A G408A, or A410V, according to the amino acid sequence set forth in SEQ ID NO:3. Additional modifications include amino acid replacements that introduce a glycosylation site. Modified u-PA polypeptides include those that contain chemical or post-translational modifications. In some examples, the modified u-PA polypeptides provided herein do not contain chemical or post-translational modifications. Chemical and post-translational modifications include, but are not limited to, pegylation, sialation, albumination, glycosylation, farnisylation, carboxylation, hydroxylation, PASylation, HESylation, phosphorylation, binding to one or more multimerization domains, such as Fe, and other modifications of polypeptides known in the art. In addition to one or more amino acid modifications, such as amino acid replacements, insertions, deletions, and combinations thereof, provided herein, the modified u-PA polypeptides provided herein may be conjugated or fused to any moiety using any method known in the art, including chemical and recombinant methods, providing the resulting polypeptide, when in active form, retains the ability to effect inhibitory or inactivating cleavage of complement protein 03. PbQynn / i znz / B / v 104 For example, in addition to one or more amino acid modifications, such as amino acid replacements, provided herein, the modified u-PA polypeptides provided herein may also contain other modifications that are or are not in the primary sequence of the polypeptide, including, but not limited to, modification with a carbohydrate moiety, a polyethylene glycol (PEG) moiety, a silation moiety, an immunoglobulin G Fc domain, or any other domain or moiety. For example, such additional modifications can be made to increase the stability or serum half-life of the protein. to. decreased immunogenicity The modified u-PA polypeptides provided herein can be modified to have decreased immunogenicity. Decreased immunogenicity can be effected by sequence changes that remove antigenic epitopes from the polypeptide or by altering post-translational modifications. One of skill in the art is familiar with methods for identifying antigenic epitopes on a polypeptide (see, for example, Liang et al. (2009) BMC Bioinformatics, 10:302; Yang et al. (2009) Rev. Med. Virol., 19:77-96). In some examples, one or more amino acids may be modified to remove or alter an antigenic epitope. In another example, altering the glycosylation of a protein can also affect immunogenicity. For example, it is contemplated to alter peptide glycosylation, as long as the polypeptides retain the ability to effect inhibitory or inactivating cleavage of complement C3 protein. Glycosylation sites can be removed by simple mutations. Glycosylation sites can be added by introduction of a canonical sequence, such as by insertion or single or multiple mutations, such as NXS(T), where X is not a proline. Glycosylation sites may also increase serum half-life. b. Faith Domain Modified u-PA polypeptides can be linked to the Fe region of an immunoglobulin polypeptide. Typically, this fusion retains at least one functionally active hinge, Ch2 and Ch3 domains of the constant region of an immunoglobulin heavy chain. For example, a full length Fe sequence of lgG1 includes amino acids 99-330 of the sequence set forth in SEQ ID NO: 45 below. Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 15 1015 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 2530 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 4045 cboynn / i znz / B / v 105 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 5560 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 70 7580 Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 9095 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120125 Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 130 135140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200205 Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 210 215220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250255 Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys. 325330 106 An exemplary Fe sequence for hlgG1 is set forth in SEQ ID NO: 50: Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 15 1015 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 2530 Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 4045 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 5560 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 70 7580 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 9095 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105110 Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gln Pro Arg 115 120125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155160 lie Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215220 Leu Ser Leu Ser Pro Gly Lys 225230 It contains almost all of the hinge sequence corresponding to amino acids 100-110 of SEQ ID NO:45; the complete sequence for the Ch2 and Ch3 domain as set forth in SEQ ID NO:45. ctzQynn / i ζπζ / β / υιλι 107 Another exemplary Fc polypeptide is disclosed in PCT Application Publication No. WO 93 / 10151, and is a single chain polypeptide that extends from the N-terminal hinge region to the native C-terminus of the Fc region of an antibody. human IgG1 (SEQ ID NQ:50). The precise site at which binding is made is not critical: particular sites are well known and can be selected in order to optimize the biological activity, secretion, or binding characteristics of the HABP polypeptide. For example, other exemplary Fc polypeptide sequences begin at amino acid C109 or P113 of the sequence set forth in SEQ ID NO: 45 (see, eg, US Publication No. 2006 / 0024298 ). In addition to hlgG1 Fc, other Fc regions and other multimerization domains can also be used. For example, where effector functions mediated by Fc / FcyR interactions are to be minimized, fusion with IgG isotypes that poorly recruit effector cells or complement is contemplated, such as, for example, IgG2 or IgG4 Fc. In addition, Fc fusions may contain immunoglobulin sequences that are substantially encoded by immunoglobulin genes belonging to any of the antibody classes, including, but not limited to, IgG (including human subclasses IgG1, IgG2, IgG3, or IgG4), IgA (including human lgA1 and lgA2 subclasses), IgD, IgE and IgM classes of antibodies. Linkers can be used to covalently link Fc to another polypeptide to generate an Fc chimera. Modified Fc domains are also well known. In some examples, the Fc region is modified such that it exhibits impaired binding to an FcR to result in an altered effector function (ie, more or less) than the effector function of an Fc region of a wild-type immunoglobulin heavy chain. . Thus, a modified Fc domain may have altered affinity, including but not limited to, increased or low or no affinity for the Fc receptor. For example, different IgG subclasses have different affinities for FcRs, with IgG1 and IgG3 usually binding substantially better to receptors than IgG2 and IgG4. Different FcR mediate different effector functions. FcyR1, FcyRIIa / c, and FcyRIIIa are positive regulators of immune complex-activated activation, characterized by having an intracellular domain that has a tyrosine-based immunoreceptor activation motif (ITAM). FcyRIIb, however, has a tyrosine-based immunoreceptor inhibition motif (ITIM) and is therefore inhibitory. Altering the affinity of an Fc region for a receptor can modulate effector functions and / or pharmacokinetic properties associated with the Fc domain. Modified Fc domains are known to one skilled in the art and are described in the literature, see, for example, US Patent No. 5,457,035; United States Patent Publication No. US 2006 / 0024298; and International Patent Publication No. WO 2005 / 063816 for cboynn / i znz / B / v 108 example mods. The resulting chimeric polypeptides containing Fe moieties, and multimers formed therefrom, can be readily purified by affinity chromatography on protein A or protein G columns. In another example, the modified u-PA polypeptide can be linked to human serum albumin (HSA), such as HSA residues 25-608, or the full length, or portion thereof: 20 30 4050 MKWVTFISLL FLFSSAYSRG VFRRDAHKSE VAHRFKDLGE ENFKALVLIA 70 80 90100 FAQYLQQCPF EDHVKLVNEV TEFAKTCVAD ESAENCDKSL HTLFGDKLCT 110 120 130 140150 VATLRETYGE MADCCAKQEP ERNECFLQHK DDNPNLPRLV RPEVDVMCTA 160 170 180 190200 FHDNEETFLKKYLYEIARRH PYFYAPELLF FAKRYKAAFT ECCQAADKAA 210 220 230 240250 CLLPKLDELR DEGKASSAKQ GLKCASLQKF GERAFKAWAV ARLSQRFPKA 260 270 280 290300 EFAEVSKLVT DLTKVHTECC HGDLLECADD RADLAKYICE NQDSISSKLK 310 320 330 340350 ECCEKPLLEK SHCIAEVEND EMPADLPSLA ADFVGSKDVC KNYAEAKDVF 360 370 380 390400 LGMFLYEYAR RHPDYSVVLL LRLAKTYETT LEKCCAAADP HECYAKVFDE 410 420 430 440450 FKPLVEEPQN LIKQNCELFE QLGEYKFQNA LLVRYTKKVP QVSTPTLVEV 460 470 480 490500 SRNLGKVGSKCCKHPEAKRM PCAEDCLSVF LNQLCVLHEK TPVSDRVTKC 510 520 530 540550 CTESLVNGRP CFSALEVDET YVPKEFNAET FTFHADICTL SEKERQIKKQ 560 570 580 590600 TALVELVKHK PKATKEQLKA VMDDFAAFVE KCCKADDKET CFAEEGKKLV AASQAALGL c. polymer conjugation In some examples, the modified u-PA polypeptides provided herein are conjugated to other polymers. Polymers can increase the size of the CbQjnn / l 7Π7 / Β / Υ 109 polypeptide to reduce renal clearance and therefore increase half-life or to modify the structure of the polypeptide to increase half-life or reduce immunogenicity. Exemplary polymers that can be conjugated to u-PA polypeptides include natural and synthetic homopolymers, such as polyols (i.e., poly-OH), polyamines (i.e., poly-NHs), and polycarboxylic acids (i.e., poly -COOH), and other heteropolymers, that is, polymers comprising one or more different coupling groups, for example, a hydroxyl group and amine groups. Examples of suitable polymeric molecules include polymeric molecules selected from polyalkylene oxides (PAO), such as polyalkylene glycols (PAG), including polyethylene glycols (PEG), methoxypolyethylene glycols (mPEG) and polypropylene glycols, PEG-glycidyl ethers (Epox-PEG) , PEG-oxycarbonylimidazole (CDI-PEG), branched polyethylene glycols (PEG), polyvinyl alcohol (PVA), polycarboxylates, polyvinylpyrrolidone, poly- D,L-amino acids, polyethylene-co-maleic acid anhydride, polystyrene-co-acid anhydride maleic, dextrans including carboxymethyl dextrans, heparin, homologous albumin, celluloses including methyl cellulose, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, carboxyethyl cellulose and hydroxypropyl cellulose, chitosan hydrolysates, starches such as hydroxyethyl starches and hydroxypropyl starches, glycogen, agaroses and derivatives thereof, guar gum, pullulan, inulin, xanthan gum, carrageenan, pectin, alginic acid hydrolysates and biopolymers. Usually, the polymers are polyalkylene oxides (PAO), such as polyethylene oxides, such as PEG, usually mPEG, which have few reactive groups capable of crosslinking. Typically, the polymers are non-toxic polymeric molecules such as (methoxy¡)polyethylene glycol (mPEG) that can be covalently conjugated to u-PA polypeptides (for example, to binding groups on the protein surface) using a relatively simple chemistry. Suitable polymeric molecules for binding to u-PA polypeptides include, but are not limited to, polyethylene glycol (PEG) and PEG derivatives such as methoxy polyethylene glycols (mPEG), PEG-glycidyl ethers (Epox-PEG), PEG-oxycarbonylimidazole (CDIPEG), branched PEG, and polyethylene oxide (PEO) (see, for example, Roberts et al., Advanced Drug Delivery Review 2002, 54: 459-476; Harris and Zalipsky (eds.) "Poly(ethylene glycol), Chemistry and Biological Applications" ACS Symposium Series 680, 1997; Mehvar et al., J. Pharm. Pharmaceut. Sci., 3(1):125-136, 2000; Harris and Chess (2003) Nat Rev Drug Discov. 2( 3):214-21 and Tsubery, J Biol. Chem 279(37):38118-24, 2004). The polymer molecule can have a molecular weight usually ranging from about 3 kDa to about 60 kDa. In some embodiments, the polymer molecule that is conjugated to a u-PA polypeptide provided herein has a molecular weight of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 or more than 60 kDa. cboynn / i znz / B / v 110 Methods for modifying polypeptides by covalent attachment (conjugation) of a PEGF or PEG derivative (ie, "PEGylation") are well known in the art (see, for example, U.S. 2006 / 0104968; U.S. 5,672,662; U.S. 6,737,505; and U.S. 2004 / 0235734). Techniques for PEGylation include, but are not limited to, specialized linkers and coupling chemistries (see, for example, Harris, Adv. Drug Deliv. Rev. 54:459-476, 2002), attachment of multiple PEG moieties to a single conjugation site (such as through the use of branched PEGs; see, for example, Veronese et al., Bioorg. Med. Chem. Lett. 12:177-180, 2002), site-specific PEGylation and / or mono-PEGylation (see, eg, Chapman et al. Nature Biotech. 17:780-783, 1999) and site-directed enzymatic PEGylation (see, eg, Sato, Adv. Drug Deliv. Rev., 54:487-504, 2002). (see also, for example, Lu and Felix (1994) Int. J. Peptide Protein Res. 43:127-138; Lu and Felix (1993) Peptide Res. 6:142-6, 1993; Felix et al. ( 1995) Int J Peptide Res 46:253-64 Benhar et al (1994) J Biol Chem 269:13398-404 Brumeanu et al (1995) J Immunol 154:3088-95; see also Caliceti et al (2003) Adv Drug Deliv Rev 55(10):1261-77 and Molineux (2003) Pharmacotherapy 23 (8 Pt 2):3S-8S). Methods and techniques described in the art can produce proteins that have 1,2,3, 4, 5, 6, 7, 8, 9,10 or more than 10 PEG or PEG derivatives attached to a single protein molecule (see eg, U.S. 2006 / 0104968). Numerous reagents for PEGylation have been described in the art. These reagents include, but are not limited to, N-hydroxysuccinimidyl (NHS) activated PEG, mPEG succinimidyl, mPEG2-N-hydroxysuccinimide, mPEG succinimidyl alpha-methylbutanoate, mPEG succinimidyl propionate, mPEG succinimidyl butanoate , mPEG Carboxymethyl 3-Hydroxybutanoic Acid Succinimidyl Ester, Homobifunctional PEG Succinimidyl Propionate, PEG Homobifunctional Propionaldehyde, PEG Homobifunctional Butyraldehyde, PEG Maleimide, PEG Hydrazide, PEG p-Nitrophenylcarbonate, mPEG-Benzotriazole Carbonate, PEG Propionaldehyde, mPEG Butyral dehyde, mPEG2 branched butyraldehyde, mPEG acetyl, mPEG piperidone, mPEG methyl ketone, mPEG unbonded maleimide, mPEG vinyl sulfone, mPEG thiol, mPEG orthopyridylthioester, mPEG orthopyridyl disulfide, Fmoc-PEG-NHS, BocPEG-NHS, vinylsulfone PEG-NHS, PEG-NHS acrylate, PEG-NHS fluorescein and PEGNHS biotin (see, for example, Monfardini et al., Bioconjugate Chem. 6:62-69, 1995; Veronese et al., J. Bioactive Compatible Polymers 12:197-207, 1997; U.S. 5,672,662; U.S. 5,932,462; U.S. 6,495,659; U.S. 6,737,505; U.S. 4,002,531; U.S. 4,179,337; U.S. 5,122,614; U.S. 5,183,550; U.S. 5,324, 844; U.S. 5,446,090; U.S. 5,612,460; U.S. 5,643,575; U.S. 5,766,581; U.S. 5,795, 569; U.S. 5,808,096; U.S. 5,900,461; U.S. 5,919,455; U.S. 5,985,263; U.S. 5,990, 237; U.S. 6,113,906; U.S. 6,214,966; U.S. 6,258,351; U.S. 6,340,742; U.S. 6,413,507; U.S. 6,420,339; U.S. 6,437,025; U.S. 6,448,369; U.S. 6,461,802; U.S. 6,828,401; U.S. 6,858,736; U.S. 2001 / 0021763; U.S. 2001 / 0044526; U.S. 2001 / 0046481; U.S. 2002 / 0052430; U.S. cboynn / i znz / E / v 111 2002 / 0072573; U.S. 2002 / 0156047; U.S. 2003 / 0114647; U.S. 2003 / 0143596; U.S. 2003 / 0158333; U.S. 2003 / 0220447; U.S. 2004 / 0013637; US 2004 / 0235734; U.S. 2005 / 000360; U.S. 2005 / 0114037; U.S. 2005 / 0171328; U.S. 2005 / 0209416; EP 01064951; EP 0822199; WO 00176640; WO 0002017; WO 0249673; WO 9428024; and WO 0187925). d. Protein transduction domain The modified u-PA polypeptides provided herein can be linked, such as a fusion protein containing an antibody, or antigen-binding fragment thereof, conjugated to a protein transduction domain (PTD) that enhances retention. of the antibody at a target site for therapy, such as a mucosal site, such as the eye. Any PTD can be employed as long as the PTD promotes binding to target cell surfaces at the therapeutic site (eg, mucosal site) and / or uptake of the modified u-PA polypeptide by target cells at the therapeutic site (eg, mucosal site). , mucosal site, such as the eye). In general, PTDs include short cationic peptides that can bind to the cell surface through electrostatic attachment to the cell membrane and can be taken up into the cell by membrane translocation (Kabouridis (2003) TRENDS Biotech 21(11) 498 -503). The provided PTDs generally interact with a target cell by binding to glycosaminoglycans (GAGs), such as, for example, hyaluronic acid, heparin, heparan sulfate, dermatan sulfate, keratin sulfate, or chondroitin sulfate and their derivatives. The protein transduction domain can be of any length. In general, the length of the PTD ranges from 5 or about 5 to 100 or about 100 amino acids in length. For example, the length of the PTD can range from 5 or about 5 to 25 or about 25 amino acids in length. In some examples, the PTD is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids. of length. A single or multiple PTDs can be conjugated to a modified u-PA polypeptide. These are advantageously used for the treatment of ocular or ophthalmic disorders, such as diabetic retinopathies or macular degeneration, including AMD. For example, multiple copies of the same PTD (eg, dimer, trimer, tetramer, pentamer, hexamer, heptamer, octamer, nonamer, decamer, or larger multimer) or different PTDs can be conjugated to the modified u-PA polypeptide. Several proteins and their peptide derivatives possess cellular internalization properties. Exemplary PTDs are known in the art and include, but are not limited to, PTDs listed in the Table below, including, for example, PTDs derived from human immunodeficiency virus 1 (HIV-1) TAT (SEQ ID NOS:125 -135; Rubén et al. (1989) J. Virol. 63:1-8), the herpesvirus VP22 tegument protein (SEQ ID NO: 140; Elliott and O'Hare cboynn / i znz / B / v 112 (1997) Cell 88:223-233), the Drosophila melanogaster Antennapedia protein (Antp) homeotic protein (Penetratin PTD; SEQ ID NO: 112; Derossi et al. (1996) J. Biol. Chem. 271: 18188-18193), the antimicrobial protegrin 1 (PG-1) peptide SynB (eg, SynB1 (SEQ ID NO: 121), SynB3 (SEQ ID NO: 122), and SynB4 (SEQ ID NO: 123); Kokryakov et al (1993) FEBS Lett. 327:231-236) and Kaposi's fibroblast growth factor (SEQ ID NO: 105; Lin et al., (1995) J. Biol. Chem. 270-14255-14258) . Other proteins and their peptide derivatives have been found to possess similar cellular internalization properties. The carrier peptides that have been derived from these proteins show little sequence homology to each other, but are all highly cationic and rich in arginine or lysine. In fact, synthetic polyarginine peptides have been shown to be internalized with a high level of efficiency and can be selected for conjugation with the provided antibody (Futaki et al. (2003) J. Mol. Recognit. 16:260-264 Suzuki et al (2001) J Biol Chem 276:5836-5840). The PTD may also be selected from one or more synthetic PTDs, including but not limited to, transport (SEQ ID NO: 136; Pooga et al. (1988) FASEB J. 12:67-77; Pooga et al. (2001 ) FASEB J. 15:1451-1453), MAP (SEQ ID NO: 103; Oehlke et al. (1998) Biochim. Biophys. Acta. 1414:127-139), KALA (SEQ ID NO: 101; Wyman et al (1997) Biochemistry 36:3008-3017) and other cationic peptides, such as, for example, various β-cationic peptides (Akkarawongsa et al. (2008) Antimicrob. Agents and Chemother. 52(6):2120-2129) . Additional PTD peptides and variant PTDs are also provided, for example, in US Patent Publication Nos. US 2005 / 0260756, US 2006 / 0178297, US 2006 / 0100134, US 2006 / 0222657, US 2007 / 0161595, US 2007 / 0129305, European Patent Publication No. EP 1867661, PCT Publication Nos. WO 2000 / 062067, WO 2003 / 035892, WO 2007 / 097561, WO 2007 / 053512 and Table 13 herein (below). Any of the PTDs provided herein or known in the art can be conjugated to a provided therapeutic antibody. PbQynn / i znz / B / v Table 13: Known Protein Transduction Domains Protein Transduction Domain (PTD) Protein Source SEQ ID NO TRSSRAGLQFPVGRVHRLLRK Buforin II 82 RKKRRRESRKKRRRES DPV3 83 GRPRESGKKRKRKRLKP DPV6 84 GKRKKKGKLGKKRDP DPV7 85 GKRK KKGKLGKKRPRSR DPV7b 86 RKKRRRESRRARRSPRHL DPV3 / 10 87 SRRARRSPRESGKKRKRKR DPV10 / 6 88 VKRGLKLRHVRPRVTRMDV DPV1047 89 VKRGLKLRHVRPRVTRDV DPV1048 90 SRRARRSPRHLGSG DPV10 91 LRRERQSRLRRERQSR DPV15 92 113 Table 13: Known Protein Transduction Domains Protein Transduction Domain (PTD) Protein Source SEQ ID NO GAYDLRRRERQSRLRRRERQSR DPV15b 93 WEAALAEALAEALAEHLAEALAEALEALAA GALA 94 KGSWYSMRKMSMKIRPFFPQQ Fibrinogen beta chain 95 KTRYYSMKKTTMKI IPFNRL Fibrinogen gamma chain precursor 96 RGADYSLRAVRMKIRPLVTQ Fibrinogen alpha chain 97 LGTYTQDFNKFHTFPQTAIGVGAP hCT(9-32) 98 TSPLNIHNGQKL HN-1 99 NSAAFEDLRVLS Influenza Virus Nucleoprotein (NLS) 100 WEAKLAKALAKALAKHLAKALAKALKACEA KALA 101 VPMLKPMLKE Ku70 102 KLALKLALKALKAALKLA MAP 103 GALFLGFLGAAGSTM GAWSQPKKKRKV MPG 104 AAVALLPAVLLALLAP Human fibroblast growth factor 4 (Kaposi fibroblast growth factor ) 105 VQRKRQKLM N50 (NLS of NF-kB P50) 106 KETWWETWWTEWSQPKKKRKV Pep-1 107 SDLWEMMMVSLACQY Pep-7 108 RQIKIWFQNRRMKWKK Penetratin 109 GRQIKIWFQNRRMKWKK Penetratin Variant 110 RRMKWKK Penetratin Short 111 E RQIKIWFQNRRMKWKK Penetratin 42-58 112 RRRRRRR Poly arginine - R7 113 RRRRRRRRR Poly arginine-R9 114 RVIRVWFQNKRCKDKK pISL 115 MANLGYWLLALFVTMWTDVGLCKKRPKP Mouse PrPc1-28 Prion 116 LLIILRRRIRKQAHAHSK pVEC 117 LLIILRRRIRKQAHAH pVEC Variant 118 VRLPPPVRLPPPVRLPPP SAP 1 19 PKKKRKV SV-40 (NLS) 120 RGGRLSYSRRRFSTSTGR SynB1 121 RRLSYSRRRF SynB3 122 AWSFRVSYRGISYRRSR SynB4 123 YGRKKRRQRRRPPQ Tat 47-60 124 YGRKKRRQRRR Tat 47-57 125 YGRKKRRQRR Tat 47-56 126 GRKKRRQRR Tat 48-56 127 GRKKRRQRRR Tat 48-57 128 RKKRRQRRR Tat 49-57 129 RKKRRQRR Tat 49-56 130 GRKKRRQRRRPPQ Tat 4 8-60 131 GRKKR Tat 48-52 132 PbQ>nn / l 7f\7IW 114 Table 13: Known Protein Transduction Domains Protein Transduction Domain (PTD) Protein Source SEQ ID NO CFITKALGISYGRKKRRQRRRPPQFSQTHQVSLSKQ Tat 37-72 133 FITKALGISYGRKKRRQRRRPQFSQTHQVSLSKQ Tat 38-72 134 YGRK KRRQRRRRPP Tat 47-59 135 GWTLNSAGYLLGKINLKALAALAKKIL Carry 136 AGYLLGKINLKALAALAKKIL Carry 10 137 GWTLNSAGYLLG Derived from transport 138 INLKALAALAKKIL Derivative from transport 139 DAATATRGRSAASRPTERPRAPARSASRPRRPVD VP22 140 DPKGDPKGVTVTVTVTVTGKGDPKPD VT5 141 GALFLGWLGAAGSTMGAWSQPKKKRKV Based on signal peptide sequence 142 KLALKLALKALKAALKLA An phyphilic model peptide 143 KFFKFFKFFK Bacterial cell wall permeability 144 LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES LL-37 145 SWLSKTAKKLENSAKKRISEGIAIAIQGGPR Cecropin P1 146 ACYC RIP ACIAG E R R YGTCIYQG R L WAFCC alpha defensin 147 DHYNCVSSGGQCLYSACPIFTKIQGTCYRGKAKCCK beta defensin 148 RKCRIWIRVCR Bactenecin 149 RRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPR FPGKR PR- 39 150 ILPWKWPWWPWRR Indolicidin 151 GALFLGWLGAAGS TMGAWSQPKKKRKV MPS 152 PVIRRVWFQNKRCKDKK pls1 153 cboynn / i znz / B / v In some examples, PTDs can be modified by replacing a lysine or arginine with another basic amino acid, such as by replacing a lysine with an arginine or by replacing an arginine with a lysine. E. ASSAYS TO ASSESS OR MONITOR U-PA ACTIVITY IN COMPLEMENT MEDIATED FUNCTIONS The modified u-PA polypeptides provided herein exhibit altered specificity and / or selectivity for complement protein C3. Exemplary modified u-PA polypeptides specifically cleave complement protein C3 and thus alter complement activation. In addition, the exemplary modified u-PA polypeptides provided herein have altered, or reduced, specificity and / or selectivity for cleavage of natural u-PA substrates, such as plasminogen, and binding to uPAR. Various in vitro and in vivo assays can be used to monitor or analyze u-PA polypeptides for their ability to cleave complement protein C3 and for their effects on complement activation and complement mediated diseases and disorders. 115 complement. These assays are known to those skilled in the art. One of skill in the art can test a particular u-PA polypeptide for cleavage of the complement C3 protein and / or test for any change in the effects of a u-PA on complement-mediated activity compared to the absence. of a protease. Some of these tests are exemplified herein. Exemplary in vitro and in vivo assays are provided herein to compare an activity of a modified u-PA polypeptide on complement C3 protein function. As discussed below, numerous assays, such as assays to measure complement activation, are known to one skilled in the art. Also provided herein are exemplary assays for determining the activity of modified u-PA polypeptides for wild-type u-PA activities, such as plasminogen cleavage or uPAR binding. Assays to determine the specificity of the modified u-PA polypeptides for complement protein C3 are also provided. Example tests are described below. 1. Methods to assess u-PA activity on complement protein C3 function A modified u-PA protease may exhibit alterations in specificity and / or selectivity to any one or more complement proteins and thus inactivate any one or more complement proteins, such as, for example, C3, compared to the corresponding full-length, scaffold, or wild-type form of the modified u-PA protease. Modified u-PA proteases retain their protease activity, but may exhibit increased specificity and / or selectivity to any one or more complement proteins. Exemplary modified u-PA proteases specifically cleave any one or more complement proteins, such as, for example, C3, and thereby alter the activity of a complement protein. All of these modified u-PA proteases with increased specificity and / or selectivity to any one or more complement proteins are candidate therapeutic agents. When the modified u-PA protease exhibits increased specificity and / or selectivity to any of one or more complement proteins, in vitro and in vivo assays can be used to monitor or analyze proteases for effects on complement-mediated functions. These assays are known to those skilled in the art. One skilled in the art can test a modified u-PA protease for cleavage of any one or more complement proteins, such as, for example, C3, and / or test to assess any changes in the effects of a complement proteinase. modified u-PA in a complement-mediated activity compared to the absence of a modified u-PA protease. Some of these tests are exemplified herein. cboynn / i znz / B / v 116 Exemplary in vitro and in vivo assays are provided herein to compare an activity of a modified u-PA protease against the function of one or more targeting complement proteins. Many of the assays are applicable to other proteases and modified proteases. As discussed above, assays for complement activities include, but are not limited to, assays that measure activation products of complement activation, such as, for example, the MAC C5b-9 complex, and the generation of one. or more complement cleavage products such as C4a, C5a, C3b and C3d. Assays to measure complement activation also include functional assays that measure the functional activity of specific components of the complement pathways, such as, for example, hemolytic assays used to measure activation of any of the classical, lectin, or alternative pathways. Assays to assess the effects of proteases and modified proteases on complement proteins and / or complement-mediated functions include, but are not limited to, SDS analysis followed by Western Blot or Coomassie Brilliant Blue staining, enzyme immunoassays, and hemolytic assays. In one example, in vitro assays can be performed using purified complement proteins. In another example, in vivo assays can be performed by testing the serum of a species, including human or mammalian species, for functional complement activation. Example tests are described below. In one example, in vitro assays can be performed using purified complement protein 03, as exemplified in Example 2-4. In another example, in vitro assays can be carried out in physiologically relevant solutions (i.e., vitreous humor), as exemplified in Example 5. In another example, in vitro assays can be carried out using peptide libraries to assess the specificity of cleavage. In another example, assays can be performed to assess the normal functions of the modified u-PA polypeptides, ie, activity towards normal substrates. Various disease models known to one of skill in the art can be used to assess the efficacy of u-PA polypeptides provided herein in various complement-mediated diseases and disorders. a.Protein detection Protein detection is a means of measuring individual complement components in a sample. Complement proteins can be detected to directly assess the effects of a u-PA polypeptide on complement protein 03 cleavage, or alternatively, complement proteins can be measured as a means of assessing complement activation. Complement protein 03, treated in the presence or absence of a u-PA polypeptide, can be analyzed by one or more assays including SDS-PAGE followed by Coomassie staining or Western blot, cboynn / i znz / B / v 117 enzyme immunoassay, immunohistochemistry, flow cytometry, nephelometry, agar gel diffusion, or radial immunodiffusion. Example assays for protein detection are described below. Yo. SDS-PAGE analysis Analysis of complement proteins in the presence or absence of increasing concentrations of a u-PA polypeptide can be performed by analysis of proteins on SDS-PAGE followed by detection of those proteins. In these examples, complement proteins can be detected by staining for total protein, such as by Coomassie Brilliant Blue staining, Silver staining, or by any other method known to one of skill in the art, or by Western blotting using polyclonal antibodies. or monoclonal specific for a specific protein. Typically, a purified complement protein, such as, for example, complement protein C3, can be incubated in the presence or absence of a u-PA polypeptide. The treated complement protein can be redissolved on an SDS-PAGE gel followed by a method of detecting protein in the gel, eg, by Coomasie Brilliant Blue staining. The treated protein can be compared to its cognate full-length protein and the degradation products formed by protease cleavage of the protein can be determined. In another embodiment, a sample, such as, for example, human serum or plasma, may be treated in the presence or absence of a u-PA polypeptide or may be collected after treatment of an animal or human with or without a u-PA polypeptide. The u-PA-treated sample can be analyzed on SDS-PAGE and a specific complement protein, such as, for example, 03, 05 or Factor B, can be detected by Western Blotting using monoclonal or polyclonal antibodies against the protein. Complement protein cleavage can be compared to a sample that was not treated with a u-PA polypeptide. Additionally, the sample can be stimulated to initiate complement activation such as by incubation with IgG which stimulates activation of the classical pathway or by LPS which stimulates activation of the alternative pathway. The sample can be redissolved using SDS-PAGE for the detection of any one or more of the native complement proteins to determine the presence or absence of cleavage products of a specific protein compared to a sample of the untreated protein with a u-PA polypeptide. In these examples, native complement protein cleavage effector molecules can also be analyzed by Western Blot using monoclonal and polyclonal antibodies to assess activation of one or more of the complement pathways. Examples of complement effector molecules may include, but are not limited to, C3a, C3d, IC3b, Bb, and C5-b9. For example, decreased expression in a Bb sample may indicate that a u-PA polypeptide inhibited activation of the pathway. PbQynn / i znz / B / v...

Claims

1. A modified urokinase-like plasminogen activator (u-PA) polypeptide comprising one or more amino acid modifications selected from modifications corresponding to R35Q, H37Y, V41R, Y40Q, D60aP, L97bA, T97a1, and H99Q, and amino acid modifications conserving the same, wherein the modified u-PA polypeptide has increased activity / specificity for a complement protein compared to the unmodified active form of the u-PA polypeptide, wherein: the amino acid modifications are selected from replacements, insertions, and deletions in the primary sequence of the modified u-PA polypeptide; the modified u-PA polypeptide cleaves a complement protein to inhibit or thereby reduce complement activation compared to the unmodified u-PA polypeptide that does not contain the amino acid modifications; the complement protein is O3; the residues are numbered by chymotrypsin numbering;The unmodified u-PA polypeptide comprises the sequence exposed in any of SEQ ID NO: 1-6, which exposes wild-type (WT) full-length u-PA, WT protease domain u-PA, WT mature u-PA, full-length u-PA with C122S, by chymotrypsin numbering, C122S protease domain u-PA, C122S mature u-PA, or a catalytically active fragment thereof including the amino acid modification position(s); the modified u-PA polypeptide has at least 85% sequence identity with a polypeptide from any of SEQ ID NO: 1-6; and the conservative modifications are selected from R35Y, W, F or N; H37R, Q, E, W or F; V41K; D60aS; T97aD, L or V; L97bG or S; and H99N.

2. The modified u-PA polypeptide of claim 1, comprising one or more amino acid modifications selected from modifications corresponding to R35Q, H37Y, V41 R, Y40Q, D60aP, L97bA, T97al and H99Q.

3. The modified u-PA polypeptide of claim 1 or claim 2, wherein the modified u-PA polypeptide has reduced activity or specificity for cleavage of a substrate sequence in plasminogen.

4. The modified u-PA polypeptide of any of claims 1-3 having increased activity for O3 cleavage that is at least 3 times greater than the unmodified u-PA polypeptide comprising the protease domain of SEQ ID NO: 5, or a corresponding form of u-PA exposed in any of SEQ ID NO: 1-4 and 6.

5. The modified u-PA polypeptide of any of claims 1-4, wherein the unmodified u-PA polypeptide consists of the amino acid sequence set out in any of SEQ ID NO: 1-6.

6. The modified u-PA polypeptide of any of claims 1-5, wherein the unmodified u-PA polypeptide consists of the amino acid sequence set out in SEQ ID NO: 2 or SEQ ID NO:

5.

7. The modified u-PA polypeptide of any of claims 1-4 and 6, wherein the modified u-PA polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the polypeptides of any of SEQ ID NO: 1-6 (full length u-PA WT, protease domain u-PA WT, mature u-PA WT, full length u-PA with C122S, protease domain u-PA with C122S, mature u-PA with C122S) or a catalytically active fragment thereof.

8. The modified u-PA polypeptide of any of claims 1-4 and 6, wherein the modified u-PA polypeptide has 1 or up to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid replacements, insertions or deletions, compared to the unmodified u-PA polypeptide of any of claims 1-6 or a catalytically active portion thereof.

9. The modified u-PA polypeptide of any of claims 1-8, comprising the V41R replacement.

10. The modified u-PA polypeptide of any of claims 1-8, comprising the V41L replacement.

11. The modified u-PA polypeptide of claim 9 or claim 10, further comprising the V38E replacement.

12. The modified u-PA polypeptide of any of claims 1-11, comprising the H37Y replacement.

13. The modified u-PA polypeptide of any of claims 1-11, comprising the H37Y / V38E replacements.

14. The modified u-PA polypeptide of any of claims 1-11, comprising replacements R35Y / H37K or R35Q / H37K.

15. The modified u-PA polypeptide of any of claims 1-11, comprising replacements R35Y / H37K / V38E or R35Q / H37K / V38E.

16. The modified u-PA polypeptide of any of claims 1-11, comprising the L97bA replacement.

17. The modified u-PA polypeptide of any of claims 1-11, comprising R35Q. PbQynn / i znz / B / v 286 18. The modified u-PA polypeptide of any of claims 1-11, comprising H99Q.

19. The modified u-PA polypeptide of any of claims 1-11, comprising D60aP.

20. The modified u-PA polypeptide of any of claims 1-11, comprising T97al.

21. The modified u-PA polypeptide of any of claims 1-20, further comprising the amino acid replacement corresponding to T39Y, T39W, T39F or selected preservative replacements of T39M or T39L.

22. The modified u-PA polypeptide of any of claims 1-21, further comprising the replacement of amino acids T39Y.

23. The modified u-PA polypeptide of any of claims 1-22, further comprising the amino acid replacements R35Q / H37Y or V38E / V41R / Y149R or R35Q / H37Y / R37aEA / 38E / T39Y / V41R / D60aT / Y60bT / T97a1 / L97bA / H99Q / C122S / Y149R 24. The modified u-PA polypeptide of any of claims 1-23, wherein the modified u-PA polypeptide has an ED50 for C3 inactivation cleavage of less than either 100 nM, 50 nM, 30 nM, or 25 nM in an in vitro assay.

25. The modified u-PA polypeptide of claim 24, wherein: the polypeptide comprises the modifications in the protease domain set out in Table 14; and has an ED50 of 100 nM or less, has an ED50 of less than 50 nM or less, or has an ED50 of less than 30 nM, or has an ED50 of less than 25 nM.

26. The modified u-PA polypeptide of claim 24 or claim 25, wherein the assay comprises incubating human C3 substrate complement protein with different concentrations of each modified protease for 1 hour at 37°C to determine the ED50.

27. The modified u-PA polypeptide of any of claims 1-26 cleaving C3 with an ED50 of 50 nM or less.

28. The modified u-PA polypeptide of claim 27, comprising the V41R modification.

29. The modified u-PA polypeptide of claim 27, comprising modifications V38E / V41R.

30. The modified u-PA polypeptide of claim 28, further comprising a replacement in one or more of positions R35, H37 and V38.

31. The modified u-PA polypeptide of claim 30, wherein the replacement at V38 is E. cboynn / i 7f\7iw 287 32. The modified u-PA polypeptide of any of claims 1-31, comprising R35Y / H37S / V38E / V41R.

33. The modified u-PA polypeptide of any of claims 1-32, comprising the modifications selected from: R35Y / H37S / R37aP / V38E / T39Y / V41R / D60aP / Y60bD / T97a1 / L97bA / H99Q / C122S / Y151L; R35W / R36Q / H37S / V38P / T39Y / V41R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151L; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / K82R / T97al / L97bA / H99Q / K110aR / C122S / Y1 49R / M157K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K / K1 79R; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / K92R / T97al / L97bA / H99Q / C122S / Y149R / M1 57K; F30Y / R35V / R36H / H37G / V38E / T39W / Y40H / V41 R / Y60bW / T97al / L97bA / H99Q / C122S / Y149E / M157K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / K92S / T97al / L97bA / H99Q / O122S / Y149R / M15 7K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41R / K61 R / K62R / T97al / L97bA / H99Q / C122S / Y149 R / M157K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K / K1 79S; R35W / H37P / R37aN / V38E / T39Y / V41 R / D60aP / Y60bL / T97al / L97bA / H99Q / C122S;F30Y / R35W / R36T / H37SA / 38S / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y 151L / M157R / Q192Y; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / M157K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41R / K61 S / K62S / T97al / L97bA / H99Q / C122S / Y149 R / M157K; R35A / H37E / R37aG / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L; R35W / R36Q / H37S / V38T / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151 P / M157R; F30Y / R35W / H37YA / 38E / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R; V38E / T39WA / 41 R / D60aW / Y60bP / L97bG / H99L / C122S; R35W / R36K / H37SA / 38E / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151 L / M157S / Q192H; R35Q / H37Y / R37aPA / 38E / T39Y / V41 R / D60aQ / Y60bP / T97al / L97bA / H99Q / C122S / Y149R; 117V / F30Y / R35Q / R36H / H37W / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / M157K / T158A; R35Y / H37S / R37aP / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 cboynn / i znz / B / v 288 Η; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157Κ;R35W / R36N / H37S / V38E / T39Y / Y40M / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157S; R35Y / H37D / V38E / T39W / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / K82S / T97al / L97bA / H99Q / K110aS / C122S / Y1 49R / M157K; R35W / H37P / R37aN / V38E / T39Y / V41R / D60aP / Y60bL / D97T / T97aE / L97bG / A98S / H99L / C122S; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41R / T97al / L97bA / H99Q / Y149R / M157K; R35Y / H37S / R37aP / V38E / T39Y / V41R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S; F30Y / R35W / H37SA / 38E / T39Y / Y40H / V41 R / Y60bN / T97al / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35W / H37SA / 38E / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / M157K; R35HA / 38E / T39YA / 41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / T158A; R35Q / R36H / H37Y / V38E / T39Y / Y40L / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K; R35W / H37P / R37aG / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; V38D / V41 Q / D60aH / Y60bS / T97aW / L97bR / H99E / C122S / Y151 L / E175D / R217E / K224R; F30Y / R35W / R36H / H37P / R37aQ / V38E / T39Y / Y40FA / 41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y149R / M157K;F30Y / R35W / R36Q / H37S / V38P / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157R; F30H / R35W / R36T / H37S / V38P / T39YA / 41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151 L / M157S; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40HA / 41 R / Y60bD / T97al / L97bA / H99Q / C122S / M157K; F30Y / R35Y / R36H / H37N / V38E / T39F / Y40F / V41R / K61 E / R72H / T97al / L97bA / H99Q / C122S / Y149 R / M157K / Q169K; R35W / R36Q / H37S / V38S / T39Y / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151L / M157 S / Q192H; R35W / H37G / R37aE / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 T; R35W / H37P / R37aN / V38E / T39Y / V41 R / D60aP / Y60bL / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35W / H37SA / 38E / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S; F30Y / R35V / R36H / H37G / V38E / T39W / Y40H / V41 R / Y60bA / T97al / L97bA / H99Q / C122S / Y149R / M 157K; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40HA / 41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R; F30Y / R35W / H37SA / 38E / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40HA / 41 R / Y60bN / T97aE / L97bA / H99Q / C122S / M157K;CtzQjnn / l 7Π7 / Ε / ΥΙΛΙ 289 F30Y / R35W / R36H / H37S / V38E / T39Y / Y40H / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S; F30Y / R35W / R36S / H37S / V38Q / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151L / M157S / Q192N; F30Y / R35W / R36H / H37P / R37aD / V38E / T39Y / Y40F / V41R / D60aE / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R / M157K; R35Q / H37Y / R37aS / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; R37aS / V38E / Y40V / V41 R / H99L / C122S / Y151L / R217V; V38D / V41 R / L97bG / H99Q / C122S / Y151L / R217E; R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S; F30Y / R35V / R36H / H37D / V38E / T39W / Y40H / V41 R / Y60bP / T97al / L97bA / H99Q / C122S / Y149R / M 157K; 117V / F30Y / R35Q / R36H / H37W / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / M157K; F30Y / R35V / R36H / H37S / V38E / T39F / Y40H / V41 R / Y60bS / T97aM / L97bA / H99Q / C122S / Y149W / M157K; R35Y / H37S / R37aP / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; N26D / F30Y / R35Y / R36H / H37E / V38E / T39F / Y40F / V41 R / K61 E / T97al / L97bA / H99Q / R110dS / P11 4S / C122S / Y149R / M157K;F30Y / R35W / R36H / H37P / R37aE / V38E / T39Y / Y40F / V41R / Y60bA / T97aE / L97bA / H99Q / C122S / Y 149R / M157K; R35L / H37D / R37aN / V38E / T39Y / V41R / D60aP / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35W / R36H / H37P / V38E / T39Y / Y40F / V41R / Y60bS / T97aE / L97bA / H99Q / C122S / Y149K / M157K; R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / T97al / L97bA / H99Q / C122S / Y149R; R35H / V38E / T39YA / 41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / T158S / E167K; R35Q / H37YA / 38E / T39YA / 41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35Y / R36H / H37D / V38E / T39W / Y40H / V41R / Y60bE / T97al / L97bA / H99Q / C122S / Y149R / M 157K; F30Y / R35Y / R36H / H37K / V38E / T39F / Y40F / V41R / T97al / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35Y / R36H / H37E / V38E / T39F / Y40F / V41R / K61E / T97al / L97bA / H99Q / C122S / Y149R / M157K / T242A; F30Y / R35L / V38D / Y40H / V41 R / L97bA / H99Q / C122S / M157K / T158A; F30Y / R35Y / R36H / H37P / R37aQ / V38E / T39Y / Y40FA / 41R / Y60bH / T97aE / L97bA / H99Q / C122S / Y 149R / M157K; V38D / V41 R / L97bR / H99E / C122S / Y151L / R217E; H37G / R37aD / G37bD / V38F / T39H / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151 L / E175D / R21 7E / K224R;cboynn / i znz / B / v 290 R35Y / H37V / R37aW / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y151L / Q192 T; F30Y / R35M / R36H / H37G / V38E / T39F / Y40H / V41 R / Y60bP / T97aF / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35W / R36Q / H37S / V38T / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y 151L / M157K / Q192T; R35W / H37D / R37aP / V38E / T39W / V41 R / D60aR / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; 117V / F30Y / R35W / R36H / H37EA / 38E / T39W / Y40H / V41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y 149K / M157K; 117V / F30Y / R35Q / H37WA / 38D / Y40HA / 41 R / Y60bN / L97bA / H99Q / C122S / Y149H / M157K / T158A; R35HA / 38E / T39YA / 41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / I138V / E167K; F30Y / R35W / R36Q / H37S / V38E / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151L / M157T / Q192H; R35H / G37bD / V38E / T39Y / V41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / T158S; R35H / H37P / R37aG / V38E / T39FA / 41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40HA / 41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157K; V38D / T39Y / Y40L / V41 R / L97bl / H99E / C122S / R217E;F30Y / R36H / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / M157K / T158A; F30H / R35W / R36H / H37S / V38E / T39Y / Y40M / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157K; R35V / R36H / H37D / V38E / T39W / Y40MA / 41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K; R35W / R36K / H37SA / 38A / T39Y / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151L / M157 R / Q192T; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40H / V41 R / Y60bA / T97al / L97bA / H99Q / C122S / Y149R / M 157K; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40F / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / N145S / S146V / T147M / D148G / Y149Q / L150F / M157K; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / Y149R / M157K; F30Y / R35I / R36H / H37D / V38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K; R35V / V38E / Y40Q / V41 L / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R; R35Q / H37Y / R37aPA / 38E / T39Y / V41 R / D60aN / Y60bN / T97al / L97bA / H99Q / C122S / Y149R; V38E / Y40Q / V41 L / L97bG / H99Q / C122S / R217T; R35HA / 38E / T39YA / 41 R / T56S / D60aP / Y60bQ / L97bA / H99Q / C122S / T158S; F30H / R35Q / H37W / V38D / V41 R / L97bA / H99Q / C122S / Y151 L / M157K;R35Q / H37YA / 38E / T39YA / 41 R / D60aP / T97al / L97bA / H99Q / C122S / Y149R; CbQjnn / l 7Π7 / Β / Υ 291 R35W / H37P / R37aN / V38E / T39Y / V41K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 A; V38D / V41 R / Y60bR / T97aW / L97bR / H99E / C122S / E175D / R217E / K224R; F30Y / R35W / R36H / H37S / V38E / Y40H / Y60bN / T97al / L97bA / H99Q / C122S / Y149R; V38D / V41L / Y60bP / T97aM / L97bR / H99E / C122S / Y151L / E175D / R217E / K224R; F30Y / R35W / R36H / H37D / V38E / T39F / Y40H / V41 R / Y60bE / T97al / L97bA / H99Q / C122S / Y149R / M 157K; F30Y / R35W / R36H / H37N / V38E / T39Y / Y40F / V41 R / Y60bS / T97aE / L97bA / H99Q / C122S / Y149K / M157K; F30Y / R35W / R36K / H37S / V38D / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y 151L / M157R / Q192T; V41R / H99Q / C122S / Y151L / R217V; V38E / Y40P / V41L / L97bG / H99L / C122S / Y151Q / R217E; V38E / Y40L / V41R / H99L / C122S / Y151L / R217S; V38E / Y40Q / V41L / L97bG / H99Q / C122S / Y151P / R217T; V38E / T39Y / V41 R / D60aW / Y60bP / L97bR / H99l / C122S; R35W / H37D / R37aP / V38E / T39W / V41R / Y60bA / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R36H / H37F / V38E / T39Y / Y40H / V41 R / Y60bD / T97aV / L97bA / H99Q / C122S / Y149L / M157K;F30Y / R35W / R36H / H37E / S37dP / V38E / T39W / Y40H / V41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y149K / M157K; R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aA / Y60bP / T97al / L97bA / H99Q / C122S / Y149R; R36H / V38D / V41 R / A96D / D97E / A98G / T97adel / H99L / L97bdel / O122S / T178S / R217D; V38D / V41 R / L97bG / H99Q / C122S / Y151L / R217A; F30H / R35Q / R36H / H37Y / V38E / T39Y / Y40L / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35W / R36K / H37E / V38E / T39W / Y40H / V41 R / Y60bQ / K61 E / l65T / T97aE / L97bA / H99Q / C12 2S / Y149K / M157K; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40H / V41 R / Y60bS / T97aL / L97bA / H99Q / C122S / Y149L / M157K; F30Y / R35Q / R36H / H37Y / R37aEA / 38E / T39Y / Y40F / V41R / D60aS / Y60bP / T97aE / L97bA / H99Q / C 122S / Y149R / M157K; R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aG / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; I24N / F30Y / R35W / R36H / H37E / V38E / T39W / Y40L / V41 R / Y60bQ / N87D / T97aE / L97bA / H99Q / C1 22S / Y149K / M157K; V38E / Y40Q / V41 L / L97bA / H99Q / C122S; F30Y / R35Q / H37W / V38D / Y40H / V41 R / Y60bN / L97bA / H99Q / C122S / Y149H / M157K / T158A; F30Y / R35W / R36A / H37E / V38E / T39W / Y40H / V41 R / Y60bQ / K61 D / l65R / T97aE / L97bA / H99Q / C1 292 22S / Y149K / M157K;F30Y / R35Q / R36H / H37W / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / M157K; F30Y / R35Y / R36H / H37K / V38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K / K1 87R / K223R / K224R; R35W / R36Q / H37S / V38E / T39Y / Y40L / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151 L / M157S / Q192T; R35HA / 38E / T39YA / 41 R / D60aP / Y60bQ / P60cS / L97bA / H99Q / C122S / I138V / E167K; R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aT / Y60bT / T97al / L97bA / H99Q / C122S / Y149R; 117V / F30Y / R36HA / 38E / Y40HA / 41 R / T97al / L97bA / H99Q / C122S / M157K; R35Y / R36H / H37K / V38E / T39F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K; R35HA / 38E / T39YA / 41 R / T56A / D60aP / Y60bQ / L97bA / H99Q / C122S; F30Y / V38D / Y40F / V41 L / L97bA / H99Q / C122S / Y151L / M157R; V38E / Y40A / V41 L / L97bG / H99Q / C122S / R217T; I24T / F30Y / R35W / R36H / H37EA / 38E / T39W / Y40H / V41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y1 49K / M157K; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40F / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157K; 117V / F30Y / V38D / Y40H / V41 R / L97bA / H99Q / C122S / M157K / T158A; R35W / H37P / R37aN / V38E / T39Y / V41 K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 T;F30H / R35L / H37DA / 38D / V41 R / L97bA / H99Q / C122S / Y151L / M157K / R217E; F30Y / R35W / R36H / H37D / R37aEA / 38E / T39Y / Y40F / V41R / D60aE / Y60bF / T97aE / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35L / R36H / H37GA / 38E / T39Y / Y40HA / 41 R / Y60bP / T97aE / L97bA / H99Q / C122S / Y149M / M157K; 117V / F30H / V38D / V41R / L97bA / H99Q / C122S / Y151L / M157K; F30Y / R35V / R36H / H37K / V38E / T39F / Y40H / V41 R / Y60bN / T97al / L97bA / H99Q / C122S / Y149R / M 157K; R35W / R36H / H37N / V38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K / Q192H; R35V / Y40Q / V41 L / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R; F30Y / R35M / R36H / H37G / R37aE / V38E / T39Y / Y40F / V41R / D60aP / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R / M157K; R35QA / 38DA / 41 R / L97bG / H99Q / C122S / Y151 L; R37aSA / 38E / Y40P / V41 L / L97bG / H99Q / C122S / Y151Q / R217T; R35V / R37aE / V38E / Y40Q / V41 L / T97aE / L97bA / H99Q / C122S / Y149R; F30H / V38D / V41 R / A96G / L97bA / H99Q / C122S / Y151L / M157K; cboynn / i 7f\7iw 293 T39L / Y40L / V41 R / T97al / L97bA / H99Q / C122S; F30Y / R35W / R36H / H37E / V38E / T39Y / Y40F / V41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y149R / M157K Y40Q / V41 L / Y60bL / L97bA / H99Q / C122S;F30Y / R36H / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / S146F / M157K / Q192H / K243Q Y40Q / V41 L / L97bA / H99Q / C122S / Y149R; F30Y / R35W / R36Q / H37E / V38E / T39W / Y40H / V41 R / Y60bQ / K61 L / l65V / T97aE / L97bA / H99Q / C12 2S / Y149K / M157K; R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; R35Q / V38D / V41 R / T97aS / L97bA / H99Q / C122S / Y151 L; V41 R / L97bR / H99Q / C122S / Y151L / R217V; R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / Y149R; F30Y / R35V / R36H / H37SA / 38E / T39Y / Y40H / V41 R / Y60bP / T97aE / L97bA / H99Q / C122S / Y149E / M 157K; R35A / H37T / R37aD / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192S; R35S / V38D / V41 R / L97bA / H99Q / C122S / Y151L; R35S / V38D / V41 L / L97bG / H99Q / C122S / Y151L / R217Q; F30Y / R35H / V38D / Y40H / V41 R / L97bA / H99Q / C122S / M157K / T158S; R35Q / H37S / R37aE / V38E / T39Y / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; H37G / R37aD / V38F / T39H / V41 R / Y60bK / L97bR / H99E / C122S / Y151L / E175D / Q192T / R217E / K22 4R; H37G / R37aD / V38F / T39H / V41 R / Y60bK / L97bR / H99E / C122S / Y151L / E175D / Q192T / R217E;R35W / H37D / R37aS / V38E / T39Y / V41 R / D60aE / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; R35Q / H37G / R37aD / V38E / T39Y / V41 R / D60aP / Y60bA / T97al / L97bA / H99Q / C122S / Y149R; R35Q / H37D / R37aK / V38E / T39F / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; R35Y / R36H / H37S / V38D / T39Y / V41 R / Y60bN / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R36H / V38E / Y40H / V41 R / K61 E / T97al / L97bA / H99Q / C122S / M157K; R37aSA / 38D / V41 Q / L97bG / H99Q / C122S / Y151L / R217T; F30H / V38D / V41 R / L97bA / H99Q / C122S / Y151 L / M157K; F30Y / R35K / R36H / H37E / R37aK / V38E / T39F / Y40F / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C1 22S / Y149R / M157K; F30Y / R35Q / R36H / H37G / R37aE / V38E / T39Y / Y40F / V41R / D60aP / Y60bG / T97al / L97bA / H99Q / C 122S / Y149R / M157K; F30Y / R35W / R36Q / H37E / V38A / T39W / Y40H / V41 R / Y60bQ / K61 D / l65V / T97aE / L97bA / H99Q / C1 22S / Y149K / M157K; cboynn / i znz / B / v 294 F30Y / R35H / V38D / Y40H / V41 R / T56A / L97bA / H99Q / C122S / M157K; R35N / H37T / R37aY / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; R37aH / V38E / T39Y / V41 R / T56A / D60aP / Y60bQ / L97bA / H99Q / C122S / T158A; F30H / R35Q / H37T / V38D / V41 R / L97bA / H99Q / C122S / Y151 L / M157K;F30H / R36L / V38E / V41 R / K82R / L97bA / H99Q / C122S / Y151 L / M157K; V38D / V41R / H99Q / C122S / Y151L / R217V; R35Q / H37G / R37aP / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35Q / R36H / H37Y / R37aEA / 38E / T39Y / Y40FA / 41R / D60aE / Y60bA / T97aE / L97bA / H99Q / C 122S / Y149R / M157K; R35Q / H37Y / R37aD / V38E / T39L / V41 R / D60aE / Y60bT / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35L / R36H / H37E / V38E / T39N / Y40FA / 41R / T97al / L97bA / H99Q / C122S / Y149R / M157K; R36S / V38E / Y40L / V41 N / L97bG / H99Q / C122S / Y151L / R217T; T39W / V41 R / L97bG / H99Q / C122S; F30Y / R35W / R36H / H37EA / 38E / T39W / Y40H / V41 R / Y60bQ / T97aE / L97bA / H99Q / C122S / Y149K / M157K; R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; R35S / R37aAA / 38E / Y40Q / V41 L / L97bA / H99Q / C122S / Y149V; F30Y / R35W / R36H / H37Q / V38E / T39H / Y40H / V41 R / T97aE / L97bA / H99Q / C122S / Y149L / M157K; F30Y / R35Q / R36H / H37Y / R37aD / V38E / T39Y / Y40F / V41R / Y60bV / T97aE / L97bA / H99Q / C122S / Y149R / M157K; F30H / V38D / V41 R / L97bA / H99Q / Y151 L / M157K; F30H / R35H / H37I / V38D / V41 R / L97bA / H99Q / C122S / Y149W / Y151L / M157K / R217S;V38D / T39Y / Y40HA / 41 R / T97al / L97bA / H99Q / C122S; R35F / H37D / R37aN / V38E / T39Y / V41 R / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; T39Y / V41 R / Y60bQ / L97bG / H99Q / C122S; T39Y / V41 R / D60aP / Y60bQ / L97bA / H99Q / C122S; V38D / V41 R / L97bR / H99Q / C122S / Y151L / R217E; R36S / V38D / T39L / Y40L / V41 R / L97bl / H99E / C122S / R217T; R35S / R37aD / V38E / Y40Q / V41 L / Y60bV / T97aL / L97bA / H99Q / C122S / Y149L; Y40Q / V41 L / Y60bT / T97aE / L97bA / H99Q / C122S / Y149R; F30Y / V38E / Y40H / V41 R / T56A / L97bA / H99Q / C122S / M157K / K243M; F30Y / R36H / R37aH / V38E / Y40H / V41 R / K61 E / T97al / L97bA / H99Q / C122S / M157K; F30H / R35Q / V38D / V41 R / L97bA / H99Q / C122S / Y151L / M157K; V38D / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151 L / E175D / Q192T / R217E / K224R; H37G / G37bD / V38F / T39H / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151 L / E175D / Q192T / R21 7E / K224R; 295 R35S / R37aD / V38E / Y40Q / V41 L / T97aE / L97bA / H99Q / C122S / Y149R; R35V / R37aE / V38E / Y40Q / V41 L / Y60bS / T97aE / L97bA / H99Q / C122S; Y40QA / 41 L / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R; F30Y / R35H / V38D / Y40HA / 41 R / L97bA / H99Q / C122S / I138V / M157K; T39Y / V41 R / Y60bQ / L97bA / H99Q / C122S;F30Y / R35H / R36H / H37D / R37aE / V38E / T39Y / Y40FA / 41R / D60aP / Y60bD / T97al / L97bA / H99Q / C 122S / Y149R / M157K; F30Y / V38D / Y40HA / 41 R / L97bA / H99Q / C122S / M157K / T158A; V38E / T39WA / 41 R / D60aP / Y60bD / L97bA / H99L / C122S; F30Y / R36H / V38E / Y40H / V41 R / l65T / T97al / L97bA / H99Q / C122S / M157K; V38D / V41 R / L97bR / H99Q / C122S / Y151L / R217V; R35Q / H37S / R37aPA / 38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; R35W / R36H / H37S / V38E / T39Y / V41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157K; R36S / V38E / Y40Q / V41 R / L97bG / H99L / C122S / Y151P / R217E; V38E / Y40Q / V41 L / Y60bL / L97bA / H99Q / C122S; H37G / R37aD / G37bD / V38F / T39H / V41 R / Y60bK / L97bR / H99E / C122S / Y151L / E175D / Q192T / R2 17E; H37G / R37aD / V38F / T39H / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151L / E175D / Q192T / R21 7E; F30Y / R35Y / R36H / H37KA / 38E / T39F / Y40F / V41 R / T97al / L97bA / H99Q / C122S / Y149R / M157K / K1 87S / K223S / K224Y; Y40Q / V41 L / L97bA / H99Q / C122S; F30H / R35H / V38D / V41R / K61 E / L97bA / H99Q / C122S / Y151L / M157K / R206H; F30Y / V38D / Y40HA / 41 R / L97bA / H99Q / C122S / M157K; F30Y / R36H / V38E / Y40H / V41 R / T97aE / L97bA / H99Q / C122S / Y149R / M157K;R35A / R37aE / V38E / Y40Q / V41 L / L97bA / H99Q / C122S / Y149R; V38D / V41 L / L97bG / H99Q / C122S / Y151 L / R217Q; F30H / R35Q / H37W / V38D / V41 R / D60aE / L97bA / H99Q / C122S / Y149L / Y151 L / M157K / R217D; F30Y / R35F / R36H / H37G / V38E / T39Y / Y40H / V41 R / Y60bS / T97aD / L97bA / H99Q / C122S / Y149R / M157K; T39Y / V41 R / L97bG / H99Q / C122S; F30Y / R35I / R36H / H37EA / 38E / T39Y / Y40H / V41 R / Y60bS / T97aV / L97bA / H99Q / C122S / Y149L / M1 57K; R35S / R37aDA / 38E / Y40Q / V41 L / L97bA / H99Q / C122S / Y149R; Y40H / V41 Q / L97bG / H99Q / C122S / R217T; R35W / H37D / V38D / T39YA / 41 R / Y60bS / L97bA / H99Q / C122S / Y149R; cboynn / i znz / B / v 296 V38D / T39F / Y40L / V41 R / T97aW / L97bA / H99Q / C122S; V38D / T39Y / Y40L / V41 R / T97aE / L97bA / H99Q / C122S; F30Y / R35Q / R36H / H37G / R37aE / V38E / T39F / Y40F / V41R / D60aP / Y60bS / T97aE / L97bA / H99Q / C 122S / Y149R / M157K; V38D / T39L / Y40L / V41 R / T97al / L97bA / H99Q / C122S; V38D / T39Y / Y40L / V41 R / T97aW / L97bA / H99Q / C122S; F30Y / R36H / V38D / Y40HA / 41R / L97bA / H99L / C122S / F141L / M157K / T158A; F30Y / R35Q / R36H / H37G / R37aE / V38E / T39Y / Y40F / V41R / D60aA / Y60bS / T97aE / L97bA / H99Q / C 122S / Y149R / M157K;F30Y / R35Q / R36H / H37G / R37aE / V38E / T39Y / Y40F / V41R / D60aP / Y60bS / T97aE / L97bA / H99Q / C 122S / Y149R / M157K; T39Y / V41R / Y60bP / L97bG / H99Q / C122S; F30H / R36H / V38D / V41 R / T56A / L97bA / H99Q / O122S / Y151 L / M157K; F30Y / R35E / R36H / H37D / R37aN / V38E / T39Y / Y40FA / 41R / Y60bN / T97aE / L97bA / H99Q / C122S / Y 149R / M157K; V38E / Y40Q / V41 L / D60aP / Y60bL / L97bA / H99Q / C122S / Y149W; F30Y / R36H / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / M157K; F30H / R35QZH37W / V38DA / 41 R / D60aE / Y60bS / L97bA / H99Q / C122S / Y149L / Y151L / M157K; R35Q / H37G / R37aE / V38E / T39Y / V41 R / D60aP / Y60bT / T97al / L97bA / H99Q / C122S / Y149R; F30Y / R35W / R36H / H37SA / 38E / T39Y / Y40HA / 41 R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / Y151P / M157K / Q192H; F30Y / R35M / R36H / H37D / R37aD / V38E / T39Y / Y40F / V41R / D60aP / Y60bS / T97aE / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40H / V41R / Y60bT / T97aD / L97bA / H99Q / C122S / Y149R / M157K; V38D / T39L / Y40L / V41R / T97aV / L97bA / H99Q / C122S; V38D / V41 R / Y60bS / T97al / L97bR / H99E / C122S / Y151 L / E175D / Q192F / R217E / K224R; T39Y / V41 R / Y60bP / L97bA / H99Q / C122S;R36H / V38D / Y40F / V41 R / D97E / A98G / T97adel / H99L / L97bdel / O122S / Y151 L / Q192E / R217D; R35IWH37G / R37aD / V38E / T39W / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; F30Y / V38D / Y40L / V41 R / L97bA / H99Q / C122S / Y151L / M157K / Q192H; F30H / V38D / Y40F / V41 R / L97bA / H99Q / C122S / Y151L / M157F; H37M / R37aD / V38E / T39A / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; F30H / V38D / V41 R / L97bA / H99Q / Y151 L / M157K; T22I / F30Y / R35SA / 38D / Y40H / V41 R / L97bA / H99Q / C122S / I138V / M157K; R35L / H37D / R37aS / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; Cboynn / i znz / B / v 297 F30Y / R35L / V38D / Y40H / V41 R / N76S / L97bA / H99Q / C122S / M157K / K187E; F30H / V38D / V41 R / L97bA / H99Q / C122S / Y151 L / M157S; R35W / H37D / V38D / T39Y / V41 R / Y60bH / L97bA / H99Q / C122S / Y149R; F30Y / R36H / H37G / V38E / T39W / Y40H / V41 R / Y60bA / T97aE / L97bA / H99Q / C122S / Y149Q / M157K; R35Q / H37G / R37aE / V38W / T39Y / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151L / E175D / Q19 2T / R217E / K224R; H37G / R37aD / G37bD / V38F / T39H / V41 R / Y60bK / L97bR / H99E / C122S / Y151L / E175D / Q192T / R2 17E / K224R; V38D / T39Y / Y40M / V41 R / T97aE / L97bA / H99Q / C122S;R35Q / H37N / V38D / T39Y / V41 R / Y60bP / L97bA / H99Q / C122S; F30Y / R35W / R36H / H37D / V38E / T39Y / Y40F / V41 R / Y60bS / T97aE / L97bA / H99Q / C122S / Y149K / M157K; R35Q / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; V38E / T39L / V41 R / D60aN / Y60bP / L97bG / H99Q / C122S; F30Y / R36H / H37A / V38E / T39Y / Y40H / V41 R / Y60bQ / T97aV / L97bA / H99Q / C122S / Y149R / M157K; F30Y / R35W / R36H / H37E / R37aP / V38E / T39Y / Y40F / V41R / Y60bN / T97aE / L97bA / H99Q / C122S / Y149Q / M157K; H37T / R37aL / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151 L / Q192R; H37G / R37aD / V38F / T39H / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151L / E175D / Q192T / R21 7E / K224R; F30Y / R35W / R36H / H37S / V38E / Y40H / Y60bN / T97aE / L97bA / H99Q / C122S / Y149R / M157K; V38D / T39W / Y40L / V41 R / T97aL / L97bA / H99Q / C122S; H37G / R37aD / G37bD / T39H / V41 R / Y60bK / T97aS / L97bR / H99E / C122S / Y151L / E175D / Q192T / R 217E / K224R; T39Y / V41 R / L97bA / H99Q / C122S; V38D / T39L / Y40L / V41 R / T97aW / L97bA / H99Q / C122S; F30Y / R36H / V38E / Y40H / V41 R / T97al / L97bA / H99Q / C122S / Y149N / L150V / M157K; R35S / V38D / L97bA / H99Q / C122S / Y151L / M157Y;R37aS / V38D / T39Y / Y40F / V41 R / H99L / C122S / R217T; Y40Q / V41 L / Y60bE / L97bA / H99Q / C122S / Y149R; Y40H / V41 T / L97bG / H99Q / C122S / R217T; and any of these polypeptides in which C122S is C122C, by chymotrypsin numbering.

34. The modified u-PA polypeptide of any of claims 1-33, further comprising one or more of the amino acid modifications R35Q, Y60bQ and Y149R.

35. The modified u-PA polypeptide of any of claims 1-34, further comprising the modification of amino acids R37aE or R37aS.

36. The modified u-PA polypeptide of any of claims 1-35, comprising replacements R35Q / H37Y / T39Y / V41R or R35Q / H37Y / T39Y / V41R / C122S.

37. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications R35Q / H37Y / T39YA / 41 R / L97bA / H99Q / C122S or R35Q / H37Y / T39Y / V41 R / L97bA / H99Q.

38. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications T39Y / V41 R / Y60bQ / L97bA / H99Q or T39Y / V41 R / Y60bQ / L97bA / H99Q / C122S.

39. The modified u-PA polypeptide of any of claims 1-35, comprising replacements T39Y / V41R / D60aP / L97bA / H99Q / C122S or T39Y / V41R / D60aP / L97bA / H99Q.

40. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications corresponding to Y40Q / V41 L / L97bA / C122S or Y40QA / 41 R / L97bA / C122S or Y40Q / V41 L / L97bA or Y40Q / V41 R / L97bA.

41. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications corresponding to R37aSA / 41 R / L97bG / H99Q or R37aS / V41 R / L97bG / H99Q / C122S.

42. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications corresponding to T39Y / V41 L / L97bA / H99Q / C122S or T39Y / V41 R / L97bA / H99Q / C122S or T39Y / V41 L / L97bA / H99Q or T39Y / V41 R / L97bA / H99Q.

43. The modified u-PA polypeptide of claim 40, further comprising the replacement corresponding to H99Q.

44. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications corresponding to R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aT / Y60bT / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aT / Y60bT / T97al / L97bA / H99Q / Y149R.

45. The modified u-PA polypeptide of claim 44, wherein the unmodified polypeptide consists of the polypeptide of SEQ ID NO: 2 or SEQ ID NO: 5, the protease domain.

46. ​​The modified u-PA polypeptide of claim 44, wherein the unmodified polypeptide consists of the mature u-PA of SEQ ID NO: 3 or SEQ ID NO:

6.

47. The modified u-PA polypeptide of claim 44, wherein the unmodified cboynn / i znz / B / v 299 polypeptide consists of the pro-peptide of SEQ ID NO: 1 or SEQ ID NO:

4.

48. The modified u-PA polypeptide of any of claims 1-35, comprising the amino acid modifications: H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Y / H37S / R37aP / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151 L; or R35O / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / V38E / T39YA / 41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aE / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R; or R35O / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / T97al / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / L97bA / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / H99Q / C122S / Y149R; or R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / C122S / Y149R;o R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S; o R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aA / Y60bP / T97al / L97bA / H99Q / C122S / Y149R; o R35L / H37D / R37aS / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; o R35IWH37G / R37aD / V38E / T39W / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; o R35Q / H37G / R37aP / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y149R; o R35A / H37G / R37aE / V38E / T39FA / 41 R / D60aE / Y60bP / T97al / L97bA / H99Q / C122S / Y149R; o R35Q / H37S / R37aE / V38E / T39Y / V41 R / D60aP / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; o R35Q / H37T / R37aP / V38E / T39Y / V41 R / D60aE / Y60bD / T97al / L97bA / H99Q / C122S / Y149R; o R35Q / H37G / R37aE / V38E / T39H / V41 R / D60aP / Y60bA / T97al / L97bA / H99Q / C122S / Y149R; o R35W / H37D / R37aS / V38E / T39Y / V41 R / D60aE / Y60bS / T97al / L97bA / H99Q / C122S / Y149R; o R35Q / H37G / R37aE / V38E / T39Y / V41 R / D60aP / Y60bT / T97al / L97bA / H99Q / C122S / Y149R; o R35W / H37P / R37aN / V38E / T39Y / V41 R / D60aP / Y60bL / D97T / T97aE / L97bG / A98S / H99L / C122S;cboynn / i 7f\7iw 300 R35W / H37P / R37aN / V38E / T39Y / V41 K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 A; or R35Y / H37V / R37aW / V38E / T39Y / V41 R / D60aP / Y60bE / T97al / L97bA / H99Q / C122S / Y151L / Q192 T; or R35Y / H37S / R37aP / V38E / T39Y / V41 R / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151 L; or R35W / H37P / R37aN / V38E / T39Y / V41 K / D60aP / Y60bD / T97al / L97bA / H99Q / C122S / Y151L / Q192 T; or each without replacement in C122.; 49. The modified u-PA polypeptide of any of claims 1-35 comprising the amino acid modifications corresponding to R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R, wherein the unmodified u-PA polypeptide comprises the protease domain exposed in SEQ ID NO: 2 or SEQ ID NO:

5.

50. The modified u-PA polypeptide of any of claims 1-49, wherein the modified u-PA polypeptide comprises the amino acid residue sequence set out in any of SEQ ID NO: 8-44.

51. The modified u-PA polypeptide of any of claims 1-50, wherein the modified u-PA polypeptide comprises the amino acid residue sequence set forth in SEQ ID NO: 21 or 987.

52. The modified u-PA polypeptide of any of claims 1-50, wherein the modified u-PA polypeptide comprises the amino acid residue sequence set out in SEQ ID NO:

18.

53. The modified u-PA polypeptide of any of claims 1-52, wherein the unmodified u-PA polypeptide consists of the amino acid residue sequence set out in SEQ ID NO:

5.

54. The modified u-PA polypeptide of claim 48, wherein the unmodified u-PA polypeptide consists of the amino acid residue sequence set out in SEQ ID NO: 2 or SEQ ID NO:

5.

55. The modified u-PA polypeptide of claim 49, wherein the unmodified u-PA polypeptide consists of the amino acid residue sequence set out in SEQ ID NO:

5.

56. The modified u-PA polypeptide of any of claims 1-55 that is cleaved within the QHARASHLG residues (residues 737-745) of human C3 (SEQ ID NO: 47).

57. The modified u-PA polypeptide of claim 56, wherein R1-RΓ is RA.

58. The modified u-PA polypeptide of any of claims 1-57 having a stability of more than 50% after incubation in PBS, or a body fluid for 7 days.

59. The modified u-PA polypeptide of any of claims 1-58 having stability of more than 80% after incubation in PBS, or a body fluid for 7 days.

60. The modified u-PA polypeptide of claim 58 or claim 59, wherein the incubation is carried out in a body fluid.

61. The modified u-PA polypeptide of claim 60, wherein the body fluid is aqueous humor.

62. The modified u-PA polypeptide of any of claims 1-61, wherein the modified u-PA polypeptide, when in its active form, has at least 100-fold decreased activity in plasmin compared to a corresponding unmodified u-PA polypeptide form.

63. The modified u-PA polypeptide of any of claims 1-62 conjugated to another portion or polymer, either directly or by means of a ligand.

64. The modified u-PA polypeptide of claim 63 which is a fusion protein.

65. The modified u-PA polypeptide of claim 63 or claim 64 that is PEGylated.

66. The modified u-PA polypeptide of any of claims 1-65 comprising a polymer that increases serum half-life and / or reduces immunogenicity or both.

67. The modified u-PA polypeptide of claim 66, wherein the polymer comprises a polypeptide.

68. The modified u-PA polypeptide of any of claims 63-67 that binds directly or indirectly to serum albumin.

69. The modified u-PA polypeptide of claim 68, wherein the serum albumin is human serum albumin (HSA).

70. The modified u-PA polypeptide of claim 68 or claim 69, wherein the HSA comprises the sequence set forth in SEQ ID NO: 991, or a form having at least 90% or at least 95% sequence identity thereto.

71. The modified u-PA polypeptide of claim 63, wherein the polymer is an Fc domain.

72. The modified u-PA polypeptide of claim 71, wherein the Fc domain comprises the sequence disclosed in SEQ ID NO: 50 or SEQ ID NO: 992 or a form having at least 90% or at least 95% sequence identity thereto. 302 73. The modified u-PA polypeptide of any of claims 63-72, wherein the polymer is directly linked to the modified u-PA polypeptide.

74. The modified u-PA polypeptide of any of claims 63-73, wherein the polypeptide or polymer is linked by a peptide linker to the modified u-PA polypeptide.

75. The modified u-PA polypeptide of claim 74, wherein the linker comprises Gly and / or Ser.

76. The modified u-PA polypeptide of claim 74 or 75, wherein the linker contains up to 20, 30, 40 or 50 amino acid residues.

77. The modified u-PA polypeptide of claim 74, wherein the linker comprises (GS)n(G)n(S)n(GGS)n(SGG)n(GGSSGG)n(AGS)n, wherein n is 1 to 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 or a linker containing up to 25 residues containing S, G, E and K.

78. The modified u-PA of claim 77, comprising the amino acid residue sequence exposed in any of SEQ ID NO: 1001-1003, 1024-1029 and multimers thereof and sequences having at least 99% sequence identity to the same.

79. A fusion protein, comprising a modified u-PA polypeptide or a catalytically active portion of a modified u-PA polypeptide of any of claims 1-78 fused to a non-protease polypeptide or a portion thereof.

80. The fusion protein of claim 79, wherein the non-protease polypeptide is a multimerization domain or a protein transduction domain (PTD).

81. The fusion protein of claim 79, wherein the non-protease polypeptide is a multimerization domain that is an Fe domain.

82. The fusion protein of claim 79, comprising a furin activation site.

83. The fusion protein of claim 82, wherein the furin activation site comprises QSGQKTLRRRKR (SEQ ID NO: 996) or QCGQKTLRRRKR (SEQ ID NO: 995) or QSGQKTLRRKR (SEQ ID NO: 1044) or a furin activation site having at least 98% sequence identity to the same.

84. A fusion protein, comprising a modified u-PA polypeptide or catalytically active portion thereof of any of claims 1-78, wherein the fusion protein comprises a signal sequence and the modified u-PA polypeptide or catalytically active portion thereof.

85. The fusion protein of claim 84, wherein the signal sequence is II-2, u-PA, or IgGK. PbQynn / i znz / B / v 303 86. A fusion protein of any of claims 79-85, comprising a fusion partner.

87. The fusion protein of claim 86, wherein the fusion partner is a multimerization domain.

88. The fusion protein of claim 86, wherein the fusion partner is albumin, or an Fe domain, or a single-chain antibody or other antigen-binding fragment of an antibody, or a hyaluronic acid-binding domain (HABD).

89. The fusion protein of claim 88, wherein the fusion partner is a HABD that is tumor necrosis factor-stimulated gene 6 (TSG-6).

90. The fusion protein of claim 88, wherein the fusion partner is HSA.

91. The fusion protein of claim 88, wherein the fusion partner is Fe from IgG.

92. The fusion protein of claim 88, wherein the fusion partner is an anti-type II collagen antibody scFv fragment or an anti-VEGFR antibody or fragment thereof.

93. The fusion protein of any of claims 79-92, comprising an activation sequence.

94. The fusion protein of claim 93, wherein the activation sequence comprises a cysteine, and the modified u-PA polypeptide comprises a free cysteine, whereby, upon activation, the resulting activated polypeptide comprises two chains.

95. The fusion protein of claim 93, wherein the activation sequence is a u-PA activation sequence or a furin activation sequence.

96. The fusion protein of claim 95, wherein the activation sequence has the sequence exposed in any of SEQ ID NO: 995-998, 1041 and 1044 or a sequence having at least 98% or 99% sequence identity to the same.

97. The fusion protein of any of claims 79-96, comprising an activation sequence, a modified u-PA polypeptide, and HSA.

98. A fusion protein of claim 79, comprising the amino acid sequence set forth in any of SEQ ID NO: 1006, 1007, 1009 and 1010.

99. A fusion protein of claim 79, comprising the amino acid sequence set forth in any of SEQ ID NO: 1004-1019 and 1034-1040.

100. The fusion protein of claim 79, comprising the amino acid sequence set forth in SEQ ID NO: 1015 or 1019.

101. The fusion protein of any of claims 79-100 having no signal sequence or having the signal sequence removed after cboynn / i znz / B / v 304 expression.

102. A fusion of any of claims 79-101 that is an activated two-chain form containing a chain A and a chain B.

103. The fusion protein of claim 102, wherein chain B begins at residues IIGG of the modified u-PA polypeptide and terminates at the C-terminal end of the fusion protein.

104. A fusion protein of claim 102, comprising a modified u-PA polypeptide and HSA.

105. The fusion protein of claim 102 or claim 103, comprising the amino acid sequence set forth in SEQ ID NO: 1005, 1011, 1014, 1015, 1016, 1019 and 1036, but lacking the signal sequence.

106. The fusion protein of claim 105, comprising a chain A of residues 21-178 and a chain B of residues 179- to the C-terminal end of the protein with a disulfide bond between residues 168-299.

107. The fusion protein of any of claims 79-106, which is a two-chain activated fusion protein, comprising chain A and chain B, wherein chain A consists of residues 21-178 of SEQ ID NO: 1015 and chain B consists of residues 179-1022; and chains A and B are linked by a disulfide bridge between C168 and C299 of SEQ ID NO: 1015.

108. The fusion protein of claim 87, which is a dimer through interaction of complementary multimerization domains.

109. The fusion protein of claim 108, wherein the multimerization domain is an Fc domain.

110. A nucleic acid molecule, comprising a nucleotide sequence encoding for a modified u-PA polypeptide or fusion protein of any of claims 1-109.

111. A vector, comprising the nucleic acid molecule of claim 110.

112. The vector of claim 111, which is a prokaryotic vector.

113. The vector of claim 111 which is a eukanthotic vector.

114. The vector of claim 111 which is a viral vector.

115. The vector of claim 111 which is a yeast vector.

116. The vector of any of claims 111-115, which is a herpes simplex virus vector, or a vaccinia virus vector, or an adenoviral vector, or an adeno-associated viral vector, or a retroviral vector, or an insect vector.

117. The vector of any of claims 111-116, which is an expression vector. cboynn / i znz / B / v 305 118. The vector of any of claims 111-116 which is a gene therapy vector.

119. Use of the nucleic acid of claim 110 or the vector of any of claims 111-118 for gene therapy to treat a disease or condition mediated by or involving complement activation, wherein inhibition of complement activation effects the treatment or improvement of the disease or condition.

120. The vector of any of claims 111-118 for use in treating a complement-mediated disease by inhibiting complement activation.

121. A method for treating a disease or condition mediated by or involving complement activation, comprising administering the nucleic acid molecule of claim 110 or the vector of any of claims 111-118.

122. The method of claim 121, wherein the complement-mediated disease or condition is selected from inflammatory diseases and conditions, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), sepsis, rheumatoid arthritis (RA), a cardiovascular disease, membranoproliferative diseases and conditions, ophthalmic or ocular diseases or disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), organ transplant rejection, and ischemia-reperfusion injury.

123. The method of claim 121, wherein the disease or condition is an eye or ophthalmic disease or is rejection or inflammation due to a transplanted organ.

124. The method of claim 121, wherein the disease or condition is diabetic retinopathy or macular degeneration.

125. The method of claim 121, wherein the disease or condition is macular degeneration.

126. The method of claim 124, wherein macular degeneration is age-related macular degeneration (AMD).

127. The method of claim 121, wherein the disease or condition is delayed renal graft function (DGF).

128. The method of claim 121, wherein the disease or condition is atypical hemolytic uremic syndrome (aHUS).

129. The method of claim 121, wherein the disease or condition is complement 3 glomerulopathy (C3G).

130. The use of claim 119, wherein the complement-mediated disease or condition is selected from inflammatory diseases and conditions, 306 complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), sepsis, rheumatoid arthritis (RA), an eye disease, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

131. The use of claim 119, wherein the disease or condition is an ophthalmic or ocular disease or is rejection or inflammation due to a transplanted organ.

132. The use of claim 119, wherein the disease or condition is diabetic retinopathy or macular degeneration.

133. The use of claim 119, wherein the disease or condition is macular degeneration.

134. The use of claim 132, wherein macular degeneration is age-related macular degeneration (AMD).

135. The use of claim 119, wherein the disease or condition is delayed renal graft function (DGF).

136. The use of claim 119, wherein the disease or condition is atypical hemolytic uremic syndrome (aHUS).

137. The use of claim 119, wherein the disease or condition is complement 3 glomerulopathy (C3G).

138. An isolated cell or a cell culture, comprising the nucleic acid molecule of claim 110 or the vector of any of claims 111-118.

139. A non-human cell, comprising the nucleic acid molecule of claim 110 or the vector of any of claims 111-118.

140. An isolated cell, comprising the nucleic acid molecule of claim 110 or the vector of any of claims 111-118, wherein the isolated cell is not a human zygote.

141. A method for inhibiting complement activation, comprising contacting a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109, in active form, with complement protein C3, whereby the complement protein C3 is cleaved so that complement activation is reduced or inhibited.

142. The method of claim 141, wherein the contact of the modified u-PA polypeptide with the complement protein O3 is carried out in vitro.

143. The method of claim 11, wherein the contact of the modified u-PA polypeptide with the complement protein O3 is carried out in vivo.

144. The method of any of claims 141-143, wherein the inhibition of complement activation leads to a reduction of inflammatory symptoms associated with a complement-mediated disease or disorder selected from an inflammatory disorder, a neurodegenerative disorder, and a cardiovascular disorder.

145. The method of claim 144, wherein the complement-mediated disease or disorder is selected from inflammatory diseases and conditions, sepsis, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), rheumatoid arthritis (RA), eye disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

146. The method of claim 145, wherein the ischemia-reperfusion injury is caused by an event or treatment selected from myocardial infarction (MI), stroke, angioplasty, coronary artery bypass graft, cardiopulmonary bypass (CPB), and hemodialysis.

147. The method of any of claims 141-146, wherein the complement-mediated disease or disorder results from the treatment of a subject.

148. The method of any of claims 141-147, wherein the treatment with the modified u-PA polypeptide is performed prior to the treatment of a subject.

149. A method for treating a disease or condition mediated by or involving complement activation, comprising administering a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109, in active form, wherein inhibition of complement activation effects the treatment or improvement of the disease or condition.

150. The method of claim 149, wherein the complement-mediated disease or condition is selected from inflammatory diseases and conditions, sepsis, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), rheumatoid arthritis (RA), an eye disease, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

151. The method of claim 149, wherein the disease or condition is an eye disease or is rejection or inflammation due to a transplanted organ.

152. The method of claim 149, wherein the disease or condition is diabetic retinopathy or macular degeneration.

153. The method of claim 149, wherein the disease or condition is macular degeneration. PbQynn / i znz / B / v 308 154. The method of claim 153, wherein macular degeneration is age-related macular degeneration (AMD).

155. The method of claim 149, wherein the disease or condition is delayed renal graft function (DGF).

156. The method of claim 149, wherein the disease or condition is atypical hemolytic uremic syndrome (aHUS).

157. The method of claim 149, wherein the disease or condition is complement 3 glomerulopathy (C3G).

158. The method or use of any of claims 119-137 and 141-157, wherein the modified u-PA polypeptide or vector or nucleic acid encoding molecule is administered intravenously.

159. The method or use of any of claims 119-137 and 141-157, wherein the modified u-PA polypeptide or vector or nucleic acid encoding molecule is administered subcutaneously.

160. The method or use of any of claims 119-137 and 141-157, wherein the modified u-PA polypeptide or vector or nucleic acid encoding molecule is delivered to the eye.

161. The method or use of claim 160, wherein administration to the eye is effected by intravitreal or subretinal or intraretinal injection.

162. The method or use of claim 160 or claim 161, wherein the modified u-PA polypeptide is linked to a transduction domain to facilitate transduction in the vitreous humor.

163. The method or use of any of claims 119-137 and 141-162, wherein the modified u-PA polypeptide is PEGylated.

164. The method or use of any of claims 119-137 and 141-162, wherein the modified u-PA polypeptide is modified by albumination, glycosylation, farnisylation, carboxylation, hydroxylation, phosphorylation, HESylation and / or PASylation.

165. The method or use of any of claims 119-137 and 141-164, wherein an individual dose is 0.1 to 10 mg depending on the route of administration.

166. The method or use of any of claims 119-137 and 141-165, wherein the treatment is repeated a plurality of times.

167. The method or use of any of claims 119-137 and 141-166, wherein the treatment is repeated every 2 days, 3 days, 4 days, 5 days, 6 days, weekly, bimonthly or monthly.

168. The method of any of claims 119-137 and 141-167, wherein the treatment is repeated every two months, every three months, or every four months. PbQynn / i znz / B / v 309 169. A combination comprising: (a) a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109; and (b) a second agent or agents for treating a complement-mediated disease or disorder.

170. The combination of claim 169, wherein the second agent or agents are one or more anti-inflammatory or anticoagulant agents.

171. The combination of claim 169 or claim 170, wherein the second agent is one or more anti-inflammatory agents selected from one or more of a non-steroidal anti-inflammatory drug (NSAID), antimetabolite, corticosteroid, analgesic, cytotoxic agent, pro-inflammatory cytokine inhibitor, anti-inflammatory cytokine, B-cell targeting agent, T-cell targeting compound antigens, adhesion molecule blocker, chemokine receptor antagonist, kinase inhibitor, PPAR-γ (gamma) ligand, complement inhibitor, heparin, warfarin, acenocoumarol, phenindione, EDTA, citrate, oxalate, argatroban, lepirudin, bivalirudin, and ximelagatran.

172. The method, use or combination of any of claims 119-137 and 141-168, wherein the modified u-PA polypeptide comprises the V41R or V41L modification.

173. The method, use or combination of any of claims 119-137 and 141-168, wherein the modified u-PA polypeptide comprises the V41R modification.

174. The method, use or combination of any of claims 119-137 and 141-168, wherein the modified u-PA polypeptide comprises modifications V38E / V41R.

175. The method, use or combination of any of claims 119-137 and 141-168, wherein the modified u-PA polypeptide comprises the modifications Y40QA / 41 R / L97bA or Y40Q / V41 L / L97bA or R37aS / V41 R / L97bG / H99Q.

176. The method, use or combination of any of claims 119-137 and 141-168, wherein the modified u-PA polypeptide comprises the modifications: R35Q / H37Y / R37aEA / 38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R.

177. The method, use, or combination of any of claims 172-176, wherein the unmodified u-PA polypeptide comprises the amino acid residue sequence set forth in SEQ ID NO: 2 or SEQ ID NO: 5 178. The modified u-PA polypeptide of any of claims 1-78 or the fusion protein of any of claims 79-109, method, use or combination of any of claims 119-137 and 141-177, wherein the modified u-PA polypeptide or fusion protein comprises the amino acid residue sequence set forth in SEQ ID NO: 21 or 987 or in any of SEQ ID NO: 40-44 or 40-44 without the CbQjnn / l 7P7 / B / Y 310 modification at C122, by chymotrypsin numbering.

179. A method for treating DGF, comprising intravenously administering a modified u-PA polypeptide of any of claims 1-78 or the fusion protein of any of claims 79-109.

180. A method for treating atypical hemolytic uremic syndrome (aHUS), comprising administering a modified u-PA polypeptide of any of claims 1-78 or the fusion protein of any of claims 79-109.

181. A method for treating complement 3 glomerulopathy (C3G), comprising administering a modified u-PA polypeptide of any of claims 1-78 or the fusion protein of any of claims 79-109.

182. The method of any of claims 179-181, wherein an individual dose is 0.1 to 3 mg, or 0.1 to 2 mg, or 1 to 3 mg, or 1 to 10 mg.

183. The method of any of claims 179-181, wherein the modified u-PA polypeptide or fusion protein comprises the amino acid residue sequence set out in SEQ ID NO: 21 or 987 or in any of SEQ ID NO: 40-44 or 40-44 without the modification at C122, by chymotrypsin numbering.

184. The method of any of claims 179-183, wherein the treatment is repeated a plurality of times.

185. The method of any of claims 179-184, wherein the treatment is repeated every 2 days, 3 days, 4 days, 5 days, 6 days, weekly, bimonthly or monthly.

186. The method of any of claims 179-185, wherein the treatment is repeated every two months, every three months, or every four months.

187. The method of any of claims 101-105, wherein the modified u-PA polypeptide comprises the replacements R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R.

188. The method of any of claims 179-187, wherein the modified u-PA polypeptide or fusion protein comprises the protease domain exposed in SEQ ID NO: 21 or 987 or a catalytically active portion thereof.

189. A method for treating an ophthalmic disorder or eye disorder, comprising administering a modified u-PA polypeptide of any of claims 1-71 or the fusion protein of any of claims 79-109 to the eye.

190. The method of claim 189, wherein the disorder is macular degeneration or diabetic retinopathy.

191. The method of claim 190, wherein the disorder is AMD.

192. The method of any of claims 189-191, wherein an individual dose is from 0.1 mg to 2 mg / eye or 3 mg / eye. cboynn / i 7f\7iw 311 193. The method of any of claims 189-191, wherein an individual dose is 1 mg to 2 mg / eye.

194. The method of any of claims 189-191, wherein an individual dose is 0.1 to 1 mg.

195. The method of any of claims 179-194, wherein the modified u-PA polypeptide comprises the amino acid residue sequence set out in any of SEQ ID NO: 21 or 987 or any of SEQ ID NO: 40-44 or 40-44 without the modification at C122, by chymotrypsin numbering.

196. The method of any of claims 179-195, wherein the treatment is repeated a plurality of times.

197. The method of any of claims 179-196, wherein the treatment is repeated every 2 days, 3 days, 4 days, 5 days, 6 days, weekly, bimonthly or monthly, every two months, every three months or every four months.

198. The method of any of claims 179-197, wherein the modified u-PA polypeptide comprises the replacements R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R.

199. The method of any of claims 179-198, wherein the modified u-PA polypeptide comprises the replacements Y40Q / V41L / L97bA / C122S or Y40Q / V41 R / L97bA / C122S or Y40Q / V41 L / L97bA or Y40QA / 41 R / L97bA.

200. The method of any of claims 179-199, wherein the modified u-PA polypeptide comprises the protease domain exposed in SEQ ID NO: 21 or 987 or a catalytically active portion thereof.

201. Use of a modified u-PA polypeptide to treat AMD or DGF, wherein the modified u-PA polypeptide comprises the replacements R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / C122S / Y149R or R35Q / H37Y / R37aE / V38E / T39Y / V41 R / D60aP / Y60bQ / T97al / L97bA / H99Q / Y149R or Y40Q / V41 L / L97bA / C122S or Y40Q / V41 R / L97bA / C122S or Y40QA / 41 L / L97bA or Y40Q / V41 R / L97bA.

202. The use of a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109, in active form, to inhibit complement activation.

203. The use of claim 202, wherein: the modified u-PA polypeptide is contacted with complement protein C3, whereby the complement protein C3 is cleaved so that complement activation is reduced or inhibited; and the contact of the modified u-PA polypeptide with the complement protein C3 is carried out in vitro or in a non-human animal.

204. The use of claim 202, wherein the contact of the modified u-PA polypeptide with the complement protein C3 is carried out in vivo.

205. The use of any of claims 202-204, wherein inhibition of complement activation leads to a reduction of inflammatory symptoms associated with a complement-mediated disease or disorder selected from an inflammatory disorder, a neurodegenerative disorder, and a cardiovascular disorder.

206. The use of claim 205, wherein the complement-mediated disease or disorder is selected from inflammatory diseases and conditions, sepsis, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), rheumatoid arthritis (RA), eye disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

207. The use of claim 206, wherein the ischemia-reperfusion injury is caused by an event or treatment selected from myocardial infarction (MI), stroke, angioplasty, coronary artery bypass graft, cardiopulmonary bypass (CPB), and hemodialysis.

208. The use of any of claims 202-207, wherein the complement-mediated disease or disorder results from the treatment of a subject.

209. The use of any of claims 202-208, wherein treatment with the modified u-PA polypeptide is performed prior to the treatment of a subject.

210. The use of a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109, in active form, for treating a disease or condition mediated by or involving complement activation, wherein inhibition of complement activation effects the treatment or improvement of the disease or condition.

211. The use of claim 210, wherein the complement-mediated disease or condition is selected from inflammatory diseases and conditions, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), sepsis, rheumatoid arthritis (RA), an eye disease, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

212. The use of a nucleic acid encoding a modified u-PA polypeptide CbQjnn / l 7P7 / B / Y 313 of any of claims 1-41 or a vector comprising the nucleic acid for gene therapy.

213. The use of any of claims 210-212, wherein the disease or condition is an eye disease or is rejection or inflammation due to a transplanted organ.

214. The use of claim 210 or claim 212, wherein the disease or condition is diabetic retinopathy or macular degeneration.

215. The use of claim 210 or claim 212, wherein the disease or condition is macular degeneration.

216. The use of claim 215, wherein macular degeneration is age-related macular degeneration (AMD).

217. The use of claim 210 or claim 212, wherein the disease or condition is delayed renal graft function (DGF).

218. The use of claim 2210 or claim 212, wherein the disease or condition is atypical hemolytic uremic syndrome (aHUS).

219. The use of claim 210 or 212, wherein the disease or condition is complement 3 glomerulopathy (C3G).

220. The use of any of claims 212-217 for intravitreal or intraretinal or subretinal injection for the treatment of an ophthalmic disorder.

221. The use of claim 220, wherein an individual dose is 0.1 to 1 mg.

222. The use of claim 220, wherein an individual dose is from 0.1 mg to 2 mg / eye or 3 mg / eye.

223. The use of claim 220, wherein an individual dose is 1 mg to 2 mg / eye.

224. The use of any of claims 202-223, wherein the modified u-PA polypeptide comprises the amino acid residue sequence set out in SEQ ID NO: 21 or 987 or in any of SEQ ID NO: 40-44 or 40-44 without the modification at C122, by chymotrypsin numbering.

225. The use of any of claims 202-224, wherein the treatment is repeated a plurality of times.

226. The use of any of claims 202-225, wherein the treatment is repeated every 2 days, 3 days, 4 days, 5 days, 6 days, weekly, bimonthly or monthly, every two months, every three months or every four months.

227. A method for preparing a modified u-PA polypeptide, comprising: introducing a nucleic acid or vector encoding a polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109 into a cell; culturing the cell under conditions whereby the polypeptide is expressed; and optionally, isolating the expressed polypeptide.

228. The method of claim 227, wherein the cell is a eukaryotic cell.

229. The method of claim 228, wherein the cell is a mammalian cell or a yeast cell.

230. A pharmaceutical composition, comprising a modified u-PA polypeptide of any of claims 1-78 or a fusion protein of any of claims 79-109 in a pharmaceutically acceptable carrier.

231. The pharmaceutical composition of claim 230, wherein the modified u-PA polypeptide or fusion protein is in an activated two-chain form.

232. The pharmaceutical composition of claim 230 or claim 231, wherein the modified u-PA polypeptide or fusion protein comprises a protease domain of SEQ ID NO: 21 or SEQ ID NO:

987.

233. The pharmaceutical composition of any of claims 230-232, wherein the modified u-PA polypeptide or fusion protein has the sequence set out in SEQ ID NO: 1015 or 1019.

234. The pharmaceutical composition of any of claims 230-233 for use in the treatment of a complement-mediated disease, disorder, or condition.

235. The pharmaceutical composition of claim 234, wherein the complement-mediated disease or disorder is selected from an inflammatory disorder, a neurodegenerative disorder, and a cardiovascular disorder.

236. The pharmaceutical composition of claim 234, wherein the complement-mediated disease or disorder is selected from inflammatory diseases and conditions, complement 3 glomerulopathy (C3G), atypical hemolytic uremic syndrome (aHUS), sepsis, rheumatoid arthritis (RA), eye disorders, membranoproliferative glomerulonephritis (MPGN), multiple sclerosis (MS), myasthenia gravis (MG), asthma, inflammatory bowel disease, immune complex-mediated acute inflammatory tissue injury (IC), Alzheimer's disease (AD), and ischemia-perfusion injury.

237. The pharmaceutical composition of claim 234, wherein the disease or disorder is an eye disease or is rejection or inflammation due to a transplanted organ.

238. The pharmaceutical composition of claim 237, wherein the disease or condition is an eye disease that is diabetic retinopathy or macular degeneration.

239. The pharmaceutical composition of claim 238, wherein the disease, disorder, or condition is macular degeneration. cboynn / i znz / B / v 315 240. The pharmaceutical competition of claim 239, wherein macular degeneration is age-related macular degeneration (AMD).

241. The pharmaceutical composition of claim 234, wherein the disease or condition is delayed renal graft function (DGF).

242. The pharmaceutical composition of claim 234, wherein the disease or condition is atypical hemolytic uremic syndrome (aHUS).

243. The pharmaceutical composition of claim 234, wherein the disease or condition is complement 3 glomerulopathy (C3G).