AU2005265319A1 - Therapeutic peptides - Google Patents

Description

WO 2006/010057 PCT/US2005/024373 THERAPEUTIC PEPTIDES [00011 This application claims priority of U.S. provisional patent application Serial No. 60/586,419 filed July 8, 2004, the disclosure of which is incorporated by reference in its entirety. FIELD OF THE INVENTION [00021 Generally, the invention relates to compounds that exhibit improved bioefficacy in multidose administration. More specifically, the invention relates to polypeptides or peptides modified to include an antibody Fc region and one or more water soluble polymers. BACKGROUND OF THE INVENTION [0003] Recombinant proteins are an emerging class of therapeutic agents. Such recombinant therapeutics have engendered advances in protein formulation and chemical modification. Modifications have been identified that can protect therapeutic proteins, primarily by blocking their exposure to proteolytic enzymes. Protein modifications may also increase the therapeutic protein's stability, circulation time, and biological activity. A review article describing protein modification and fusion proteins is Francis (1992), Focus on Growth Factors 3:4-10 (Mediscript, London), which is hereby incorporated by reference. [0004] One useful modification is combination of a polypeptide with an "Fe" domain of an antibody. Antibodies comprise two functionally independent parts, a variable domain known as "Fab," which binds antigen, and a constant domain known as "Fc," which links to such effector functions as complement activation and attack by phagocytic cells. An Fc has a long serum half-life, whereas an Fab is short-lived. Capon et al. (1989), Nature 337: 525-3 1. When constructed together with a therapeutic protein, an Fe domain can provide longer half life or incorporate such functions as Fc receptor binding, protein A binding, complement fixation and perhaps even placental transfer. Id. Table 1 summarizes use of Fc fusions known in the art.

WO 2006/010057 PCT/US2005/024373 2 Table 1-Fc fusion with therapeutic proteins Form of Fc Fusion Therapeutic partner implications Reference IgG1 N-terminus of Hodgkin's disease; U.S. Patent No. 5,480,981 CD30-L anaplastic lymphoma; T-cell leukemia Murine Fcy2a IL-10 anti-inflammatory; Zheng eta]. (1995), J. transplant rejection Immunol. 154: 5590-600 IgGI TNF receptor septic shock Fisher et al. (1996), N. Engl. J. Med. 334: 1697 1702; Van Zee, K. et al. (1996), 3. Immunol. 156: 2221-30 IgG, IgA, IgM, TNF receptor inflammation, autoimmune U.S. Pat. No. 5,808,029, or IgE disorders issued September 15, 1998 (excluding the first domain) IgG1 CD4 receptor AIDS Capon et al. (1989), Nature 337: 525-31 IgG1, N-terminus anti-cancer, antiviral Harvill et al. (1995), IgG3 of IL-2 Immunotech. 1: 95-105 IgGI C-terminus of osteoarthritis; WO 97/23614, published OPG bone density July 3, 1997 IgG1 N-terminus of anti-obesity PCT/US 97/23183, filed leptin December 11, 1997 Human Ig CTLA-4 autoimmune disorders Linsley (1991), J. Exp. C1 1 Med. 174:561-9 [0005] Polyethylene glycol ("PEG") fusion proteins and peptides have also been studied for use in pharmaceuticals, on artificial implants, and other applications where biocompatibility is of importance. Various derivatives of PEG have been proposed that have an active moiety for permitting PEG to be attached to pharmaceuticals and implants and to molecules and surfaces generally. For example, PEG derivatives have been proposed for coupling PEG to surfaces to control wetting, static buildup, and attachment of other types of molecules to the surface, including proteins or protein residues. [00061 In other studies, coupling of PEG ("PEGylation") has been shown to be desirable in overcoming obstacles encountered in clinical use of biologically active molecules. Published PCT Publication No. WO 92/16221 states, for example, that many potentially therapeutic proteins have been found to have a short half life in blood serum. [0007] PEGylation decreases the rate of clearance from the bloodstream by increasing the apparent molecular weight of the molecule. Up to a certain size, the rate of glomerular filtration of proteins is inversely proportional to the size of the protein. The ability of PEGylation to decrease clearance, therefore, is generally not a function of how many PEG WO 2006/010057 PCT/US2005/024373 3 groups are attached to the protein, but the overall molecular weight of the altered protein. Decreased clearance can lead to increased efficacy over the non-PEGylated material. See, for example, Conforti et al., Pharm. Research Commun. vol. 19, pg. 287 (1987) and Katre et., Proc. Natl. Acad. Sci. U.S.A. vol. 84, pg. 1487 (1987). [0008] In addition, PEGylation can decrease protein aggregation, (Suzuki et al., Biochem. Biophys. Acta vol. 788, pg. 248 (1984)), alter protein immunogenicity (Abuchowski et al., J. Biol. Chem. vol. 252 pg. 3582 (1977)), and increase protein solubility as described, for example, in PCT Publication No. WO 92/16221. [0009] PEGylation of proteins illustrates some of the problems that have been encountered in attaching PEG to surfaces and molecules. The vast majority of PEGylating reagents react with free primary amino groups of the polypeptide. Most of these free amines are the epsilon amino group of lysine amino acid residues. Typical proteins possess a large number of lysines. Consequently, random attachment of multiple PEG molecules often occurs leading to loss of protein activity. [00101 In addition, if the PEGylated protein is intended for therapeutic use, the multiple species mixture that results from the use of non-specific PEGylation leads to difficulties in the preparation of a product with reproducible and characterizable properties. This non-specific PEGylation makes it difficult to evaluate therapeutics and to establish efficacy and dosing information. Site selective PEGylation of such proteins, however, leads to reproducibly-modified materials that gain the desirable attributes of PEGylation without the loss of activity. [0011] A variety of means have been used to attach the polyethylene glycol molecules to the protein. Amino groups, such as those on lysine residues or at the N-terminus, are convenient for such attachment. For example, Royer (U.S. Pat. No. 4,002,531, incorporated herein by reference in its entirety) states that reductive alkylation was used for attachment of polyethylene glycol molecules to an enzyme. EP 0 539 167 (incorporated herein by reference in its entirety), published Apr. 28, 1993, Wright, "Peg Imidates and Protein Derivates Thereof" states that peptides and organic compounds with free amino group(s) are modified with a derivative of PEG or related water-soluble organic polymers. U.S. Pat. No. 4,904,584 (incorporated herein by reference in its entirety), Shaw, issued Feb. 27, 1990, relates to the modification of lysine residues in proteins for the attachment of polyethylene glycol molecules via reactive amine groups. An increasing number of therapeutic proteins modified WO 2006/010057 PCT/US2005/024373 4 in the manner described above to include PEG have been previously described. See also, for example, European patent application publications EP 0 401 384; EP 0473 268; EP 0 335 423; EP 0 442 724; EP 0 154 316, each of which is incorporated by reference in its entirety. [0012] Other methods for attaching a polymer to a protein involve using a moiety to act as a linking group. Such moieties may, however, be antigenic. A tresyl chloride method involving no linking group is available, but this method may be difficult to use to produce therapeutic products as the use of tresyl chloride may produce toxic by-products. See Francis et al., In: Stability of protein pharmaceuticals: in vivo pathways of degradation and strategies for protein stabilization (Eds. Ahern., T. and Manning, M. C.) Plenum, N. Y., 1991) and Delgado et al., "Coupling of PEG to Protein By Activation With Tresyl Chloride, Applications In Immunoaffinity Cell Preparation", In: Fisher et al., eds., Separations Using Aqueous Phase Systems, Applications In Cell Biology and Biotechnology, Plenum Press, N.Y.N.Y., 1989 pp. 211-213, each of which is incorporated by reference in its entirety. [0013] In general, the interaction of a protein ligand with its receptor often takes place at a relatively large interface. However, as demonstrated in the case of human growth hormone bound to its receptor, only a few key residues at the interface actually contribute to most of the binding energy. Clackson, T. et al., Science 267:383-386 (1995). This observation and the fact that the bulk of the remaining protein ligand serves only to display the binding epitopes in the right topology makes it possible to find active ligands of much smaller size. Thus, molecules of only "peptide" length (2 to 40 amino acids) can bind to the receptor protein of a given large protein ligand. Such peptides may mimic the bioactivity of the large protein ligand ("peptide agonists") or, through competitive binding, inhibit the bioactivity of the large protein ligand ("peptide antagonists"). [0014] Phage display peptide libraries have emerged as a powerful method in identifying such peptide agonists and antagonists. See, for example, Scott et al. (1990), Science 249: 386; Devlin et al. (1990), Science 249: 404; U.S. Pat. No. 5,223,409, issued Jun. 29, 1993; U.S. Pat. No. 5,733,731, issued Mar. 31, 1998; U.S. Pat. No. 5,498,530, issued Mar. 12, 1996; U.S. Pat. No. 5,432,018, issued Jul. 11, 1995; U.S. Pat. No. 5,338,665, issued Aug. 16, 1994; U.S. Pat. No. 5,922,545, issued Jul. 13, 1999; WO 96/40987, published Dec. 19, 1996; and WO 98/15833, published Apr. 16, 1998, each of which is incorporated by reference. In such libraries, random peptide sequences are displayed by fusion with coat proteins of filamentous phage. Typically, the displayed peptides are affinity-eluted against an antibody-immobilized extracellular domain of a receptor. The retained phages may be WO 2006/010057 PCT/US2005/024373 5 enriched by successive rounds of affinity purification and repropagation, and the best binding peptides are sequenced to identify key residues within one or more structurally related families of peptides. See, e.g., Cwirla et al. (1997), Science 276: 1696-9, in which two distinct families were identified. The peptide sequences may also suggest which residues may be safely replaced by alanine scanning or by mutagenesis at the DNA level. Mutagenesis libraries may be created and screened to further optimize the sequence of the best binders. Lowman (1997), Ann. Rev. Biophys. Biomol. Struct. 26: 401-24. [0015] Other methods compete with phage display in peptide research. A peptide library can be fused to the carboxyl terminus of the lac repressor and expressed in E. coli. Another E. coli-based method allows display on the cell's outer membrane by fusion with a peptidoglycan-associated lipoprotein (PAL). These and related methods are collectively referred to as "E. coli display." Another biological approach to screening soluble peptide mixtures uses yeast for expression and secretion. See Smith et al. (1993), Mol. Pharmacol. 43: 741-8. The method of Smith et al. and related methods are referred to as "yeast-based screening." In another method, translation of random RNA is halted prior to ribosome release, resulting in a library of polypeptides with their associated RNA still attached. This and related methods are collectively referred to as "ribosome display." Other methods employ chemical linkage of peptides to RNA; see, for example, Roberts & Szostak (1997), Proc. Natl. Acad. Sci. USA, 94: 12297-303. This and related methods are collectively referred to as "RNA-peptide screening." Chemically derived peptide libraries have been developed in which peptides are immobilized on stable, non-biological materials, such as polyethylene rods or solvent-permeable resins. Another chemically derived peptide library uses photolithography to scan peptides immobilized on glass slides. These and related methods are collectively referred to as "chemical-peptide screening." Chemical-peptide screening may be advantageous in that it allows use of D-amino acids and other unnatural analogues, as well as non-peptide elements. Both biological and chemical methods are reviewed in Wells & Lowman (1992), Curr. Opin. Biotechnol. 3: 355-62. [0016] In the case of known bioactive peptides, rational design of peptide ligands with favorable therapeutic properties can be carried out. In such an approach, stepwise changes are made to a peptide sequence and the effect of the substitution upon bioactivity or a predictive biophysical property of the peptide (e.g., solution structure) is determined. These techniques are collectively referred to as "rational design." In one such technique, a series of peptides is made in which a single residue at a time is replaced with alanine. This technique is WO 2006/010057 PCT/US2005/024373 6 commonly referred to as an "alanine walk" or-an "alanine scan." When two residues (contiguous or spaced apart) are replaced, it is referred to as a "double alanine walk." The resultant amino acid substitutions can be used alone or in combination to result in a new peptide entity with favorable therapeutic properties. [0017] Structural analysis of protein-protein interaction may also be used to suggest peptides that mimic the binding activity of large protein ligands. In such an analysis, the crystal structure may suggest the identity and relative orientation of critical residues of the large protein ligand, from which a peptide may be designed. See, e.g., Takasaki et al. (1997), Nature Biotech. 15: 1266-70. These and related methods are referred to as "protein structural analysis." These analytical methods may also be used to investigate the interaction between a receptor protein and peptides selected by phage display, which may suggest further modification of the peptides to increase binding affinity. [0018] Conceptually, peptide mimetics of any protein can be identified using phage display and the other methods mentioned above. These methods have also been used for epitope mapping, for identification of critical amino acids in protein-protein interactions, and as leads for the discovery of new therapeutic agents. E.g., Cortese et al. (1996), Curr. Opin. Biotech. 7: 616-21. Peptide libraries are now being used most often in immunological studies, such as epitope mapping. Kreeger (1996), The Scientist 10(13): 19-20. [0019] Of particular interest is use of peptide libraries and other techniques in the discovery of pharmacologically active peptides. A number of such peptides identified in the art are summarized in Table 2. The peptides are described in the listed publications, each of which is hereby incorporated by reference. The pharmacologic activity of the peptides is described, and in many instances is followed by a shorthand term therefor in parentheses. Some of these peptides have been modified (e.g., to form C-terminally cross-linked dimers). Typically, peptide libraries were screened for binding to a receptor for a pharmacologically active protein (e.g., EPO receptor). In at least one instance (CTLA4), the peptide library was screened for binding to a monoclonal antibody.

WO 2006/010057 PCT/US2005/024373 7 Table 2-Pharmacologically active peptides Binding Form of partner/ Pharmacologic Reference peptide protein of activity interest intrapeptide EPO receptor EPO-mimetic Wrighton eal. (1996), disulfide- Science 273: 458-63; U.S. bonded Pat. No. 5,773,569, issued June 30, 1998 to Wrighton et al. C-terminally EPO receptor EPO-mimetic Livnah etal. (1996), cross-linked Science 273: 464-71; dimer Wrighton et al. (1997), Nature Biotechnology 15: 1261-5; International patent application WO 96/40772, published Dec. 19, 1996 linear EPO receptor EPO-mimetic Naranda etal. (1999), Pr. Natl. Acad. Sci. USA, 96: 7569-74; WO 99/47151, published September 23, 1999 linear c-Mpl TPO-mimetic Cwirla et al.(1997) Science 276: 1696-9; U.S. Pat. No. 5,869,451, issued Feb. 9, 1999; U.S. Pat. No. 5,932,946, issued Aug. 3, 1999 C-terminally c-Mpl TPO-mimetic Cwirla et al. (1997), cross-linked Science 276: 1696-9 dimer disulfide- stimulation of hematopoiesis Paukovits et al. (1984), linked dimer ("G-CSF-mimetic") Hoppe-Seylers Z. Physiol. Chem. 365: 303-11; Laerum et al. (1988), EU. Hemat. 16: 274-80 alkylene- G-CSF-mimetic Bhatnagar et al. (1996), L linked dimer Med. Chem. 39: 3814-9; Cuthbertson at al. (1997), J Med. Chem. 40: 2876-82; King et al. (1991), EV. Hematol. 19:481; King et al. (1995), Blood 86 (Suppl. 1): 309a linear IL-I receptor inflammatory and U.S. Pat. No. 5,608,035; autoimmune diseases U.S. Pat. No. 5,786,331; ("IL-1 antagonist" or U.S. Pat. No. 5,880,096; "IL-ira-mimetic") Yanofsky etal. (1996), Proc. Nat]. Acad. Sci. 93: 7381-6; Akeson et al. (1996), 1. Biol. Chem. 271: 30517-23; Wiekzorek et al. (1997), Pol. J. Pharmacol. The protein listed in this column may be bound by the associated peptide (e.g., EPO receptor, IL-I receptor) or mimicked by the associated peptide. The references listed for each clarify whether the molecule is bound by or mimicked by the peptides.

WO 2006/010057 PCT/US2005/024373 8 49: 107-17; Yanofsky (1996), PNAs, 93:7381 7386. linear Facteur thymique stimulation of lymphocytes Inagaki-Ohara et al. (1996), serique (FTS) ("FTS-mimetic") Cellular Immuno1. 171: 30 40; Yoshida (1984), Int. J. Immunopharmacol, 6:141 6. intrapeptide CTLA4 MAb CTLA4-mimetic Fukumoto etal. (1998), disulfide Nature Biotech. 16: 267-70 bonded exocyclic TNF-a receptor TNF-a antagonist Takasaki ea. (1997), Nature Biotech. 15:1266 70; WO 98/53842, published December 3, 1998 linear TNF-a receptor TNF-a antagonist Chirinos-Rojas L Imm., 562 1-5626. intrapeptide C3b inhibition of complement Sahu e (1996) J disulfide activation; autoimmune Immunol. 157:884-91; bonded diseases Morikis et al. (1998), --- ("CUb-antagonist") protein Sci. 7: 619-27 linear vinculin cell adhesion processes- Adey et al. (1997), cell growth, differentiation, Biochem. J. 324: 523-8 wound healing, tumor metastasis ("vinculin binding") linear C4 binding anti-thrombotic Linse et al. (1997), J. Biol. protein (C4BP) Chem. 272: 14658-65 linear urokinase receptor processes associated with Goodson et al. (1994), urokinase interaction with its Proc. Nati. Acad. Sci. 91: receptor (e.g., angiogenesis, 7129-33; International tumor cell invasion and application WO 97135969, metastasis); ("UKR published October 2, 1997 antagonist") linear Mdm2, Hdm2 Inhibition of inactivation of Picksley et al. (1994), p53 mediated by Mdm2 or Oncogene 9: 2523-9; hdm2; anti-tumor Bottger etal (1997) J.Mol. ("Mdmhdm antagonist) Biol. 269: 744-56; Bottger et al. (1996), Oncogene 13: 2141-7 linear p21 WA 1 anti-tumor by mimicking the Ball metal. (1997), Curr. activity of p21 wAF Biol. 7: 71-80 linear fa4esyl anti-cancer by preventing Gibbs et al. (1994), Cell transferase activation of ras oncogene 77:175-176 linear Ras effector anti-cancer by inhibiting Moodie et al. (1994), domain biological function of the ras Trends Genet 10: 44-48 oncogene RodFriguez et al. (1994), Nature 370:527-532 linear SIJ2iSH3 anti-cancer by inhibiting Pawson et al (1993), Curr domains tumor growth with activated Biol. 3:434-432 tyrosine kinases; treatment Yu et al. (1994), Cell of SH3-mediated disease 76:933-945; Rickles et al. states ("b3 antagonist") (1994), EMBO . 13: 5598 5604; Sparks et al.* (1994), . Biol. Chem. 269:e23853 6; Sparks metal. (1996), Proc. Nat. Acad. Sci. 93: 1540-4; US Pat. No.

WO 2006/010057 PCT/US2005/024373 9 5,886,150, issued March 23, 1999; US Pat. No. 5,888,763, issued March 30, 1999 linear p 1 6 1NK4 anti-cancer by mimicking Fahraeus et al. (1996), activity of p16; e.g., Curr. Biol. 6:84-91 inhibiting cyclin D-Cdk complex ("p 16-mimetic") linear Src, Lyn inhibition of Mast cell Stauffer et al. (1997), activation, IgE-related Biochem. 36: 9388-94 conditions, type I hypersensitivity ("Mast cell antagonist") linear Mast cell protease treatment of inflammatory International application disorders mediated by WO 98/33812, published release of tryptase-6 August 6, 1998 ("Mast cell protease inhibitors") linear HBV core antigen treatment of HBV viral Dyson & Muray (1995), (HBcAg) infections ("anti-HBV") Proc. Natl. Acad. Sci. 92: 2194-8 linear selectins neutrophil adhesion; Martens et al. (1995), L inflammatory diseases Biol. Chem. 270: 21129 ("selectin antagonist") 36; European patent application EP 0 714 912, published June 5, 1996 linear, cyclized calmodulin calmodulin antagonist Pierce et al. (1995), Molec. Diversity 1: 259-65; Dedman etal. ( 1 993), L Biol. Chem. 268: 23025 30; Adey & Kay (1996), Gene 169: 133-4 linear, integrins tumor-homing; treatment for International applications cyclized- conditions related to WO 95/14714, published integrin-mediated cellular June 1, 1995; WO events, including platelet 97/08203, published March aggregation, thrombosis, 6,1997; WO 98/10795. wound healing, published March 19, 1998; osteoporosis, tissue repair, WO 99/24462, published angiogenesis (e.g., for May 20, 1999; Kraft et al. treatment of cancer), and (1999), J. Biol. Chem. 274: tumor invasion 1979-1985 ("integrin-binding") cyclic, linear fibronectin and treatment of inflammatory WO 98/09985, published extracellular and autoimmune conditions March 12,1998 matrix components of T cells and macrophages linear somatostatin and treatment or prevention of European patent application cortistatin hormone-producing tumors, 0 911393, published April acromegaly, giantism, 28,1999 dementia, gastric ulcer, tumor growth, inhibition of hormone secretion, modulation of sleep or neural activity linear bacterial antibiotic; septic shock; U.S. Pat. No. 5,877,151, lipopolysac- disorders modulatable by issued March 2, 1999 WO 2006/010057 PCT/US2005/024373 10 charide CAP37 linear or pardaxin, mellitin antipathogenic WO 97/31019, published cyclic, 28 August 1997 including D amino acids linear, cyclic VIP impotence, WO 97/40070, published neurodegenerative disorders October 30, 1997 linear CTLs cancer EP 0 770 624, published May 2, 1997 linear THF-gamma2 Burnstein (1988), Biochem., 27:4066-71. linear Amylin Cooper (1987), Proc. Natl. Acad. Sci., 84:8628-32. linear Adrenomedullin Kitamura (1993), BBRC, 192:553-60. cyclic, linear VEGF anti-angiogenic; cancer, Fairbrother (1998), rheumatoid arthritis, diabetic Biochem., 37:17754-17764. retinopathy, psoriasis ("VEGF antagonist") cyclic MMP inflammation and Koivunen (1999), Nature autoimmune disorders; Biotech., 17:768-774. tumor growth (MMIP inhibitor") HGH fragment treatment of obesity U.S. Pat. No. 5,869,452 Echistatin inhibition of platelet Gan (1988), 1. Biol. Chem., aggregation 263:19827-32. linear SLE autoantibody SLE WO 96/30057, published October 3, 1996 GDlalpha suppression of tumor Ishikawa etal. (1998), metastasis FEBS Lett. 441 (1): 20-4 antiphospholipid endothelial cell activation, Blank eal. (1999), Proc. beta-2- antiphospholipid syndrome Natl. Acad. Sci. USA 96: glycoprotein-I (APS), thromboembolic 5164-8 (P2GPI) phenomena, antibodies thrombocytopenia, and recurrent fetal loss linear T Cell Receptor diabetes WO 96/11214, published beta chain April 18, 1996. Antiproliferative, antiviral WO 00/01402, published January 13, 2000. anti-ischemic, growth WO 99/62539, published hormone-liberating December 9, 1999. anti-angiogenic WO 99/61476, published December 2, 1999. linear Apoptosis agonist; treatment WO 99/38526, published of T cell-associated Aug. 5, 1999. disorders (e.g., autoimmune diseases, viral infection, T cell leukemia, T cell lymphoma) linear MHC class II treatment of autoimmune US Pat. No. 5,880,103, diseases issued March 9, 1999. linear androgen R, p75, proapoptotic, useful in WO 99/45944, published MJD, DCC, treating cancer September 16, 1999. huntingtin linear von Willebrand inhibition of Factor VII WO 97/41220, published Factor; Factor interaction; anticoagulants April 29, 1997. VII linear lentivirus LLPI antimicrobial US Pat. No. 5,945,507, WO 2006/010057 PCT/US2005/024373 11 issued Aug. 31, 1999. linear Delta-Sleep sleep disorders Graf (1986), Peptides Inducing Peptide 7:1165. linear C-Reactive inflammation and cancer Barna (1994), Cancer Protein (CRP) Immunol. Immunother. 38:38 (1994). linear Sperm-Activating infertility Suzuki (1992), Comp. Peptides Biochem. Physiol. 102B:679. linear angiotensins hematopoietic factors for Lundergan (1999), . hematocytopenic conditions Periodontal Res. 34(4):223 from cancer, AIDS, etc. 228. linear HIV-1 gp4l anti-AIDS Chan (1998), Cell 93:681 684. linear PKC inhibition of bone resorption Moonga (1998), E. Physiol. 83:717-725. linear defensins (HNP- antimicrobial Harvig (1994), Methods 1, -2, -3, -4) Enz. 236:160-172. linear p1 851 ""/neu, C- AHNP-mimetic:anti-tumor Park (2000), Nat. erbB-2 Biotechnol. 18:194-198. linear gpl30 IL-6 antagonist WO 99/60013, published Nov. 25, 1999. linear collagen, other autoimmune diseases WO 99/50282, published joint, cartilage, Oct. 7, 1999. arthritis-related proteins linear HIV- 1 envelope treatment of neurological WO 99/51254, published protein degenerative diseases Oct. 14, 1999. linear IL-2 autoimmune disorders (e.g., WO 00/04048, published graft rejection, rheumatoid Jan. 27, 2000; WO arthritis) 00/11028, published March 2,2000. [0020] Peptides identified by peptide library screening have been regarded as "leads" in development of therapeutic agents rather than being used as therapeutic agents themselves. Like other proteins and peptides, they would be rapidly removed in vivo either by renal filtration, cellular clearance mechanisms in the reticuloendothelial system, or proteolytic degradation. Francis (1992), Focus on Growth Factors 3: 4-11. As a result, the art presently uses the identified peptides to validate drug targets or as scaffolds for design of organic compounds that might not have been as easily or as quickly identified through chemical library screening. Lowman (1997), Ann. Rev. Biophys. Biomol. Struct. 26: 401-24; Kay et al. (1998), Drug Disc. Today 3: 370-8. [0021] In spite of the availability of recombinant therapeutic proteins, previously identified peptide mimetic, and modifications thereof, there thus remains a need in the art to provide additional compounds that have improved bioefficacy, particularly when administered in a multidose regimen.

WO 2006/010057 PCT/US2005/024373 12 SUMMARY OF THE INVENTION [0022] The present invention relates to polypeptides or peptides modified to include an antibody Fe region and one or more water soluble polymers. [00231 In one aspect, a substantially homogenous compound is provided comprising a structure set out in Formula I, Formula I: [(X)aF'-(X 2 )b-(L')c-WSPd wherein: F1 is a vehicle;

X

1 is selected from

P-(L

2 )e_

P

2

-(L

3 )f -P'-(L 2 )e

P

3

-(L

4 )gP 2

-(L

3 )f p(L 2 )e- and

P

4 -(L)h-P 3

(

4 )gp 2

L

3

)PL

2 e _

X

2 is selected from: -2 1 -(L2 )Pi),-P

_{L

2 )e~pl(L)f _p2

-(L

2 )ep1(L 3 )rP 2

-(L

4

)-P

3 , and -(L2 )p1(L 3 )rP 2

-(L

4 )g -P 3

-(L

5 )h-P 4 wherein P', P2, P', and P4 are each independently sequences of pharmacologically active peptides; L', L 2 , L, L, and L are each independently linkers; a, b, c, e, f, g, and h are each independently 0 or 1, provided that at least one of a and b is 1; d is at least 1; and WSP is a water soluble polymer, the attachment of which is effected at any reactive moiety in Fl; WO 2006/010057 PCT/US2005/024373 13 said compound having a property of improved bioefficacy when administered in a multidose regimen. In one aspect, the compound a multimer, and in another aspect, the compound is a dimer. [0024] In one embodiment, the invention provides a compound of Formula I comprising a structure set out in Formula II Formula II: [X'-F]-(L')-WSPd wherein F' is an Fe domain and is attached at the C-terminus of X1, and one or more WSP is attached to the Fc domain, optionally through linker L'. Compounds having this structure are provided as a multimer in one aspect and a dimer in another aspect. [0025] In another embodiment, the invention provides a compound of Formula I comprising a structure set out in Formula III Formula III: [F 1

-X

2

]-(L

1 )c-WSPd wherein F' is an Fc domain and is attached at the N-terminus of X 2 , and one or more WSP is attached to the Fc domain, optionally through linker L'. Multimers and dimers of a compound having this structure are also provided. [0026] The invention also provides a compound of Formula I comprising a structure set out in Formula IV Formula IV: [F' (L)e-Pl]-(L')c-WSPd wherein F1 is an Fe domain and is attached at the N-terminus of -(L )c-Pl and one or more WSP is attached to the Fe domain, optionally through linker L'. Multimers and dimers of a compound having this structure are also provided. [00271 The invention further contemplates a compound of Formula I comprising a structure set out in Formula V Formula V: [F(L')e-P(L 2 )rP 2 ]-(Ll)c-WSPd wherein F' is an Fe domain and is attached at the N-terminus of -L'-P'-L 2 _p 2 and one or more WSP is attached to the Fe domain, optionally through linker L'. Multimers and dimers of a compound having this structure are also provided.

WO 2006/010057 PCT/US2005/024373 14 [0028] In one aspect, a compound of the invention is provided as described above wherein P1 and/or P 2 are independently selected from a peptide set out in any one of Tables 4 through 20. In one aspect, P and/or P 2 have the same amino acid sequence. [0029] In another aspect, a compound of the invention is provided as described above wherein F1 is an Fc domain. In another aspect, a compound of the invention is provided wherein WSP is PEG. In yet another aspect, a compound as described above is provided wherein F1 is an Fe domain and WSP is PEG. [0030] In another embodiment, a substantially homogenous compound is provided comprising the structure: WSP-Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ ID NO: 1) wherein WSP is a water soluble polymer, said compound having a property of binding to c Mpl and stimulating platelet production, said compound having a property of improved bioefficacy in multidose administration. Multimers and dimers of a compound having this structure are also contemplated. [0031] In another embodiment, a substantially homogenous compound is provided comprising the structure: PEG-Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ ID NO: 2) said compound having a property of binding to c-Mpl and stimulating platelet production, said compound having a property of improved bioefficacy in multidose administration. Multimers and dimers of a compound having this structure are contemplated. [0032] In one aspect, the PEG component of a compound of the invention has a molecular weight of between about 2 kDa and 100 kDa. In another aspect, the PEG component of a compound of the invention has a molecular weight of between about 6 kDa and 25 kDa. [0033] The invention further provides a composition comprising a compound of the invention wherein the composition comprises at least 50% PEGylated compound. In another aspect, the composition of the invention comprises at least 75% PEGylated compound, at least 85% PEGylated compound, at least 90% PEGylated compound, at least 95% PEGylated compound, and at least 99% PEGylated compound.

WO 2006/010057 PCT/US2005/024373 15 [0034] In one embodiment, the invention also provides a substantially homogenous compound having the structure: PEG-Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ ID NO: 2) wherein PEG has a molecule weight of about 20 kD, said compound of SEQ ID NO: 2 being a dimer and having a property of binding to c-Mpl and stimulating platelet production, and said compound having a property of improved bioefficacy in multidose administration. [0035] The invention further provides a pharmaceutical composition comprising: (a) a substantially homogenous compound having the structure: PEG-Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ ID NO: 2) wherein PEG has a molecule weight of about 20 kD, (b) at least 95% diPEGylated compound; and (c) a pharmaceutically acceptable diluent, adjuvant or carrier, said composition having a property of improved bioefficacy in multidose administration. [0036] In one embodiment, a pharmaceutical composition is provided comprising: (a) a substantially homogenous compound having the structure: PEG-Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ ID NO: 2) wherein PEG has a molecule weight of about 20 kD and the compound of SEQ ID NO: 2 is a dimer, (b) at least 95% diPEGylated compound; and (c) a pharmaceutically acceptable diluent, adjuvant or carrier, said composition having a property of improved bioefficacy in multidose administration. [0037] The invention also provide a method of treating a hematopoietic disorder comprising administering a compound or composition of the invention in a regimen effective to treat said disorder. BRIEF DESCRIPTION OF THE FIGURE [0038] Numerous other aspects and advantages of the present invention will therefore be apparent upon consideration of the following detailed description thereof, reference being made to the drawing wherein: WO 2006/010057 PCT/US2005/024373 16 [0039] FIG. 1 shows increase in platelet production in vivo in a multidose administration regimen with a PEG-Fc-TMP of the invention, wherein results using a PEG Fc-TMP are shown with triangles (A) and results using an Fc-TMP are shown with circles (e). Squares (a) represent a control. DETAILED DESCRIPTION OF THE INVENTION [00401 DEFINITION OF TERMS [0041] The term "comprising" means that a compound may include additional amino acids on either or both of the N- or C- termini of the given sequence. Of course, these additional amino acids should not significantly interfere with the activity of the compound. [0042] "Substantially homogenous" as used herein with reference to a preparation of the invention means that the preparation includes a single species of a therapeutic compound detectable in the preparation of total therapeutic molecules in the preparation, unless otherwise stated at a specific percentage of total therapeutic molecules. In general, a substantially homogenous preparation is homogenous enough to display the advantages of a homogenous preparation, e.g., ease in clinical application in predictability of lot to lot pharmacokinetics. [0043] "Bioefficacy" refers to the capacity to produce a desired biological effect. Bioefficacy of different compounds, or different dosages of the same compound, or different administrations of the same compound are generally normalized to the amount of compound(s) to permit appropriate comparison. [0044] "Multidose administration" refers to a therapeutic or prophylactic treatment regimen which includes administration of more than one amount of a compound over a period of time. [0045] The term "vehicle" refers to a molecule that prevents degradation and/or increases half-life, reduces toxicity, reduces immunogenicity, or increases biological activity of a therapeutic protein. Exemplary vehicles include an Fc domain as well as a linear polymer; a branched-chain polymer (see, for example, U.S. Patent No. 4,289,872 to Denkenwalter et al., issued September 15, 1981; 5,229,490 to Tam, issued July 20, 1993; WO 93/21259 by Frechet et al., published 28 October 1993); a lipid; a cholesterol group; a carbohydrate or oligosaccharide; or any natural or synthetic protein, polypeptide or peptide that binds to a salvage receptor. Vehicles are further described hereinafter.

WO 2006/010057 PCT/US2005/024373 17 [0046] The term "native Fc" refers to molecule or sequence comprising the sequence of a non-antigen-binding fragment resulting from digestion of whole antibody, whether in monomeric or multimeric form. The original immunoglobulin source of the native Fc is in one aspect of human origin and may be any of the immunoglobulins. A native Fc is a monomeric polypeptide that may be linked into dimeric or multimeric forms by covalent association (i.e., disulfide bonds), non-covalent association or a combination of both. The number of intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from one to four depending on class (e.g., IgG, IgA, IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgA1, IgGA2). One example of a native Fc is a disulfide-bonded dimer resulting from papain digestion of an IgG. Ellison et al. (1982), Nucleic Acids Res. 10: 4071 9. The term "native Fc" as used herein is generic to the monomeric, dimeric, and multimeric forms. [0047] The term "Fc variant" refers to a molecule or sequence that is modified from a native Fc, but still comprises a binding site for the salvage receptor, FcRn. International applications WO 97/34631 (published 25 September 1997) and WO 96/32478 describe exemplary Fe variants, as well as interaction with the salvage receptor, and are hereby incorporated by reference. In one aspect, the term "Fc variant" comprises a molecule or sequence that is humanized from a non-human native Fc. In another aspect, a native Fc comprises sites that may be removed because they provide structural features or biological activity that are not required for the fusion molecules of the present invention. Thus, the term "Fc variant" comprises a molecule or sequence that lacks one or more native Fc sites or residues that affect or are involved in (1) disulfide bond formation, (2) incompatibility with a selected host cell (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC). Fc variants are described in further detail hereinafter. [0048] The term "Fc domain" encompasses native Fc and Fc variant molecules and sequences as defined above. As with Fc variants and native Fcs, the term "Fc domain" includes molecules in monomeric or multimeric form, whether digested from whole antibody or produced by other means. In one embodiment, for example, the Fc region can be: DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE

KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN

WO 2006/010057 PCT/US2005/024373 18 YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K (SEQ ID NO:1696). [00491 The term "multimer" as applied to Fc domains or molecules comprising Fe domains refers to molecules having two or more polypeptide chains associated covalently, noncovalently, or by both covalent and non-covalent interactions. IgG molecules typically form dimers; IgM, pentamers; IgD, dimers; and IgA, monomers, dimers, trimers, or tetramers. Multimers may be formed by exploiting the sequence and resulting activity of the native Ig source of the Fe or by derivatizing (as defined below) such a native Fc. [0050] The terms "derivatizing," "derivative" or "derivatized" comprise processes and resulting compounds in which, for example and without limitation, (1) the compound has a cyclic portion; for example, cross-linking between cysteinyl residues within the compound; (2) the compound is cross-linked or has a cross-linking site; for example, the compound has a cysteinyl residue and thus forms cross-linked dimers in culture or in vivo; (3) one or more peptidyl linkage is replaced by a non-peptidyl linkage; (4) the N-terminus is replaced by NRR 1 , NRC(O)R 1 , -NRC(O)OR 1 , -NRS(O) 2

R

1 , -NHC(O)NHR, a succinimide group, or substituted or unsubstituted benzyloxycarbonyl-NH-, wherein R and R 1 and the ring substituents are as defined hereinafter; (5) the C-terminus is replaced by -C(O)R 2 or -NR 3

R

4 wherein R 2 , R 3 and R 4 are as defined hereinafter; and (6) compounds in which individual amino acid moieties are modified through treatment with agents capable of reacting with selected side chains or terminal residues. Derivatives are further described hereinafter. [0051] The term "peptide" refers to molecules of 2 to 40 amino acids, molecules of 3 to 20 amino acids, and those of 6 to 15 amino acids. For example, peptides having a size selected from no greater than 35, no greater than 30, no greater than 25, no greater than 20 amino acids and/or no greater than 15 amino acids, are contemplated herein. Exemplary peptides may be randomly generated by any of the methods cited described herein, carried in a peptide library (e.g., a phage display library), derived by digestion of proteins, or chemically synthesized. Peptides include D and L form, either purified or in a mixture of the two forms. [0052] The term "randomized" as used to refer to peptide sequences refers to fully random sequences (e.g., selected by phage display methods) and sequences in which one or more residues of a naturally occurring molecule is replaced by an amino acid residue not appearing in that position in the naturally occurring molecule. Exemplary methods for WO 2006/010057 PCT/US2005/024373 19 identifying peptide sequences include phage display, E. coli display, ribosome display, yeast based screening, RNA-peptide screening, chemical screening, rational design, protein structural analysis, and the like. [0053] The term "pharmacologically active" means that a substance so described is determined to have activity that affects a medical parameter (e.g., blood pressure, blood cell count, cholesterol level) or disease state (e.g., cancer, autoimmune disorders). Thus, pharmacologically active peptides comprise agonistic or mimetic and antagonistic peptides as defined below. [0054] The terms "-mimetic peptide" and "-agonist peptide" refer to a peptide having biological activity comparable to a protein (e.g., EPO, TPO, G-CSF) that interacts with a protein of interest. These terms further include peptides that indirectly mimic the activity of a protein of interest, such as by potentiating the effects of the natural ligand of the protein of interest; see, for example, the G-CSF-mimetic peptides listed in Tables 2 and 7. As an example, the term "EPO-mimetic peptide" comprises any peptides that can be identified or derived as described in Wrighton et al. (1996), Science 273: 458-63, Naranda et al. (1999), Proc. Nati. Acad. Sci. USA 96: 7569-74, or any other reference in Table 2 identified as having EPO-mimetic subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [0055] As another example, the term "TPO-mimetic peptide" or "TMP" comprises peptides that can be identified or derived as described in Cwirla et al. (1997), Science 276: 1696-9 , U.S. Pat. Nos. 5,869,451 and 5,932,946 and any other reference in Table 2 identified as having TPO-mimetic subject matter, as well as International application WO 00/24770 published May 4, 2000, hereby incorporated by reference. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [0056] As another example, the term "G-CSF-mimetic peptide" comprises any peptides that can be identified or described in Paukovits et al. (1984), Hoppe-Seylers Z. Physiol. Chem. 365: 303-11 or any of the references in Table 2 identified as having G-CSF mimetic subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries.

WO 2006/010057 PCT/US2005/024373 20 [0057] The term "CTLA4-mimetic peptide" comprises any peptides that can be identified or derived as described in Fukumoto et al. (1998), Nature Biotech. 16: 267-70. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [0058] The term "-antagonist peptide" or "inhibitor peptide" refers to a peptide that blocks or in some way interferes with the biological activity of the associated protein of interest, or has biological activity comparable to a known antagonist or inhibitor of the associated protein of interest. Thus, the term "TNF-antagonist peptide" comprises peptides that can be identified or derived as described in Takasaki et al. (1997), Nature Biotech. 15: 1266-70 or any of the references in Table 2 identified as having TNF-antagonistic subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [0059] The terms "IL-I antagonist" and "IL- Ira-mimetic peptide" comprises peptides that inhibit or down-regulate activation of the IL-I receptor by IL-1. IL-1 receptor activation results from formation of a complex among IL-i, IL-1 receptor, and IL-i receptor accessory protein. IL-I antagonist or IL- lra-mimetic peptides bind to IL-1, IL-1 receptor, or IL-I receptor accessory protein and obstruct complex formation among any two or three components of the complex. Exemplary IL-1 antagonist or IL- Ira-mimetic peptides can be identified or derived as described in U.S. Pat. Nos. 5,608,035, 5,786,331, 5,880,096, or any of the references in Table 2 identified as having IL-ira-mimetic or IL-1 antagonistic subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [0060] The term "VEGF-antagonist peptide" comprises peptides that can be identified or derived as described in Fairbrother (1998), Biochem. 37: 17754-64, and in any of the references in Table 2 identified as having VEGF-antagonistic subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries.

WO 2006/010057 PCT/US2005/024373 21 [00611 The term "MMP inhibitor peptide" comprises peptides that can be identified or derived as described in Koivunen (1999), Nature Biotech. 17: 768-74 and in any of the references in Table 2 identified as having MMP inhibitory subject matter. Those of ordinary skill in the art appreciate that each of these references enables one to select different peptides than actually disclosed therein by following the disclosed procedures with different peptide libraries. [00621 The term "myostatin inhibitor peptide" comprisies peptides that can be identitifed by their ability to reduce or block myostatin activity or signaling as demonstrated in in vitro assays such as, for example the pMARE C2C12 cell-based myostatin activity assay or by in vivo animal testing as described in U.S. patent application Publication No US20040181033A1 and PCT application publication No. W02004/058988. Exemplary myostatin inhibitor pepetides are set out in Tables 21-24. [0063] The term "integrin/adhesion antagonist" comprises peptides that inhibit or down-regulate the activity of integrins, selectins, cell adhesion molecules, integrin receptors, selectin receptors, or cell adhesion molecule receptors. Exemplary integrin/adhesion antagonists comprise laminin, echistatin, the peptides described in Tables 25-28. [00641 The term "bone resorption inhibitor" refers to such molecules as determined by the assays of Examples 4 and 11 of WO 97/23614:, which is hereby incorporated by reference in its entirety. Exemplary bone resorption inhibitors include OPG and OPG-L antibody, which are described in WO 97/23614 and W098/46751, respectively, which are hereby incorporated by reference in their entirety. [0065] The term "nerve growth factor inhibitor" or "nerve growth factor agonist" comprises a peptide that binds to and inhibits nerve growth factor (NGF) activity or signaling. Exemplary peptides of this type are set out in Table 29. [0066] The term "TALL-1 modulating domain" refers to any amino acid sequence that binds to the TALL-1 and comprises naturally occurring sequences or randomized sequences. Exemplary TALL-I modulating domains can be identified or derived by phage display or other methods mentioned herein. Exemplary pepetides of this type are set out in Tables 30 and 31. [0067] The term " TALL-1 antagonist" refers to a molecule that binds to the TALL-1 and increases or decreases one or more assay parameters opposite from the effect on those parameters by full length native TALL-1. Such activity can be determined, for example, by WO 2006/010057 PCT/US2005/024373 22 such assays as described in the subsection entitled "Biological activity of AGP-3" in the Materials & Methods section of the patent application entitled, "TNF-RELATED PROTEINS", WO 00/47740, published August 17, 2000. [0068] The term "Ang 2-antagonist peptide" comprises peptides that can be identified or derived as having Ang-2-antagonistic characteristics. Exemplary peptides of this type are set out in Tables 32-38 . [0069] Additionally, physiologically acceptable salts of the compounds of this invention are also contemplated. By "physiologically acceptable salts" is meant any salts that are known or later discovered to be pharmaceutically acceptable. Some specific examples are: acetate; trifluoroacetate; hydrohalides, such as hydrochloride and hydrobromide; sulfate; citrate; tartrate; glycolate; and oxalate. [0070] The term "WSP" refers to a water soluble polymer which prevents a peptide, protein or other compound to which it is attached from precipitating in an aqueous environment, such as, by way of example, a physiological environment. A more detailed description of various WSP embodiments contemplated by the invention follows. [0071] STRUCTURE OF COMPOUNDS [0072] In General. In preparations in accordance with the invention, a peptide is attached to a vehicle through the N-terminus or C-terminus of the peptide, and the resulting structure further modified with a covalently attached WSP which is attached to the vehicle moiety in the vehicle-peptide product. Thus, the WSP-vehicle-peptide molecules of this invention may be described by the following formula I: I

[(X

1 )a-F-(X 2 )b(L')c-WSPd wherein:

F

1 is a vehicle; X1 is selected from Pl-(L2).

P

2 -(L) -P -(L 2 )e_

P

3

(L

4 )g-P 2 -(L)f -P(L 2 )e- and

P

4 -(L)hP g-P 2

_L

3 )pi 2 WO 2006/010057 PCT/US2005/024373 23

X

2 is selected from: -( 2 1 3 2 -(L2)eP1-(L)c -P2

-(L

2 eP'-(L 3

)P

2

-(L

4 )gP 3 , and

(L

2 )_Pi-(L 3

)P

2

-(L

4

-P

3

-(L

5 )h-P 4 wherein P , P2 ,3 , and P4 are each independently sequences of pharmacologically active peptides; L', L2, L3, , and L are each independently linkers; a, b, c, e, f, g, and h are each independently 0 or 1, provided that at least one of a and b is 1; d is at least 1; and WSP is a water soluble polymer, the attachment of which is effected at any reactive moiety in Fl. [0073] Thus, compound I comprises compounds of the formulae II [X'-F]-(L)c-WSPd including multimers thereof, wherein F1 is an Fe domain and is attached at the C-terminus of X1, and one or more WSP is attached to the Fe domain, optionally through linker L'; III

[F

1

-X

2 ]-(L)c-WSPd including multimers thereof, wherein F' is an Fe domain and is attached at the N-terminus of

X

2 , and one or more WSP is attached to the Fe domain, optionally through linker L'; IV [F 1

-(L

1 )e-P t ]-(L)c-WSPd including multimers thereof, wherein F1 is an Fc domain and is attached at the N-terminus of -(L)c-P 1 and one or more WSP is attached to the Fe domain, optionally through linker LI; and V [F'(L)e-P(L 2 )Pp2]-(L')c-WSPd WO 2006/010057 PCT/US2005/024373 24 including multimers thereof, wherein F1 is an Fc domain and is attached at the N-terminus of

-L'-P-L

2 _p 2 and one or more WSP is attached to the Fc domain, optionally through linker L'. [0074] Peptides. Any number of peptides may be used in conjunction with the present invention. Of particular interest are peptides that mimic the activity of EPO, TPO, growth hormone, G-CSF, GM-CSF, IL-1ra, leptin, CTLA4, TRAIL, TGF-a, and TGF-s. Peptide antagonists are also of interest, particularly those antagonistic to the activity of TNF, leptin, any of the interleukins (IL- 1, 2, 3, ...), and proteins involved in complement activation (e.g., C3b). Targeting peptides are also of interest, including tumor-homing peptides, membrane-transporting peptides, and the like. All of these classes of peptides may be discovered by methods described in the references cited in this specification and other references. [0075] Phage display, in particular, is useful in generating peptides for use in the present invention. It has been stated that affinity selection from libraries of random peptides can be used to identify peptide ligands for any site of any gene product. Dedman et al. (1993), J. Biol. Chem. 268: 23025-30. Phage display is particularly well suited for identifying peptides that bind to such proteins of interest as cell surface receptors or any proteins having linear epitopes. Wilson et al. (1998), Can. J. Microbiol. 44: 313-29; Kay et al. (1998), Drug Disc. Today 3: 370-8. Such proteins are extensively reviewed in Herz et al. (1997), J. Receptor & Signal Transduction Res. 17(5): 671-776, which is hereby incorporated by reference. Such proteins of interest are preferred for use in this invention. [0076] By way of example and without limitation, a group of peptides that bind to cytokine receptors are provided. Cytokines have recently been classified according to their receptor code. See Inglot (1997), Archivum Immunologiae et Therapiae Experimentalis 45: 353-7, which is hereby incorporated by reference. Among these receptors are the CKRs (family I in Table 3). The receptor classification appears in Table 3.

WO 2006/010057 PCT/US2005/024373 25 Table 3-Cytokine Receptors Classified by Receptor Code Cytokines (ligands) Receptor Type family subfamily family subfamily 1. Hematopoietic 1. IL-2, IL-4, IL-7, I. Cytokine R I. shared Wr, IL cytokines 11L-9, 11-13, IL- (CKR) 9R, IL-4R 15 2. IL-3, 1L-5, GM- 2. shared GP 140 CSF 3. IL-6, IL-11, IL- 13 12, LIF, OSM, 11-6 R, Leptin R CNTF, Leptin (OB) 4. "single chain" R, 4. G-CSF, EPO, GCSP-R, TPO TPO, PRL, GH R, GH-R 5. otherR 5. IL-17, HVS-IL II. IL-1ligands IL-10,CBCRF-1, R. a-r R HSV-EL10 III. Interferons 1. IFN-cd, a2, (x4, III. Interferon R 1 . IPNAR m, t, IFN-I 2. I 2. IFNGR IV. 1L- I and EL-1 1. IL-lIca, EL-13,IL- IV. IL-IR 1. IL-IR, IL like ligands IRa 1RAcP 2. IL- 18, IL-1I8BP 2. IL-1 8R, 11 18RAcP V. TNF family 1. TNF-t, TNF-3 3. NGF/TNFR 4 TNF-RI AGP-3R, (LT), FASL, DR4, DR5, 0X40, CD40 L, OPG, TACI, CD4O, CD3OL, CD27 FAS, ODR L, OX4RL, OPGL, TRAIL, APRIL, AGP-3, BLys, TL5, Ntn 2, KAY, Neutrokine-oa VI. Chemokines 1. c5 chemokines: 4. Chemokine R 1. CXCR S -8, GRO I-,1 , IF- , PF-4, SDF- 1 2. F chemokines: 2. CCR MIP1 a, MI IL-1 MCP-1,2,3,4, RATES, eotaxin 3. 7 chemokines: 3. CR lymphotactin 4. DAR 5 IL-17R - belongs to CKR family but is unassigned to 4 indicated subjamilies. 2 Other IFN type I subtypes remain unassigned. Hematopoietic cytokines, JIL-10 ligands and interferons do not possess functional intrinsic protein kinases. The signaling molecules for the cytokines are JAK's, STATs and related non-receptor molecules. 11-14, 1-16 and 11L-18 have been cloned but according to the receptor code they remain unassigned. 3TNF receptors use multiple, distinct intracellular molecules for signal transduction including "death domain" 'of FAS R and 55 kDa TNF-LOR that participates in their cytotoxic effects. NGF/TNF R can bind both NGF and related factors as well as TNF ligands. Chemokine receptors are seven transmembrane (7TM, serpentine) domain receptors. They are G protein coupled. 4The Duffy blood group antigen (DARC) is an erythrocyte receptor that can bind several diffcrcnt chemokines. 11-IR belongs to the iminunoglobulin superfamily but their signal transduction events characteristics remain unclear.

WO 2006/010057 PCT/US2005/024373 26 VII. Growth factors 1.1 SCF, M-CSF, VII. RKF 1. TK sub-family PDGF-AA, AB, 1.1 IgTK III R, BB, KDR, FLT- VEGF-RI, 1, FLT-3L, VEGF-RII VEGF, SSV PDGF, HGF, SF 1.2 FGFa, FGFP 1.2 IgTK IV R 1.3 EGF, TGF-a, 1.3 Cysteine-rich VV-F19 (EGF- TK-I like) 1.4 IGF-I, IGF-II, 1.4 Cysteine rich Insulin TK-II, IGF-RI 1.5 NGF, BDNF, 1.5 Cysteine knot NT-3, NT-4 6 TK V 2. TGF-01,32,03 2. Serine-threonine kinase subfamily

(STKS)

7 [00771 Other proteins of interest as targets for peptide generation in the present invention include the following: aV 1 Ang-2 B7 B7RPl CRP1 Calcitonin CD28 CETP cMet Complement factor B C4b CTLA4 Glucagon Glucagon Receptor LIPG MPL splice variants of molecules preferentially expressed on tumor cells; e.g., CD44, CD30 unglycosylated variants of mucin and Lewis Y surface glycoproteins CD19, CD20, CD33, CD45 prostate specific membrane antigen and prostate specific cell antigen matrix metalloproteinases (MMPs), both secreted and membrane-bound (e.g., MMP-9) Cathepsins 5 The neurotrophic cytokines can associate with NGF/TNF receptors also. 6 STKS may encompass many other TGF-@-related factors that remain unassigned. The protein kinases are intrinsic part of the intracellular domain of receptor kinase family (RKF). The enzymes participate in the signals transmission via the receptors.

WO 2006/010057 PCT/US2005/024373 27 TIE-2 receptor heparanase urokinase plasminogen activator (UPA), UPA receptor parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP), PTH-RI, PTH-RII Her2 Her3 Insulin Myostatin TALL-1 Nerve growth factor Integrins and receptors Selectins and receptors thereof Cell adhesion molecules and receptors thereof [0078] Exemplary peptides appear in Tables 4 through 38 below. These peptides may be prepared by any methods disclosed in the art, many of which are discussed herein. In most tables that follow, single letter amino acid abbreviations are used. The X in these sequences (and throughout this specification, unless specified otherwise in a particular instance) means that any of the 20 naturally occurring amino acid residues may be present. Any of these peptides may be linked in tandem (i.e., sequentially), with or without linkers, and a few tandem-linked examples are provided in the table. Linkers are listed as "A" and may be any of the linkers described herein. Tandem repeats and linkers are shown separated by dashes for clarity. Any peptide containing a cysteinyl residue may be cross-linked with another Cys-containing peptide, either or both of which may be linked to a vehicle. A few cross-linked examples are provided in the table. Any peptide having more than one Cys residue may form an intrapeptide disulfide bond, as well; see, for example, EPO-mimetic peptides in Table 5. A few examples of intrapeptide disulfide-bonded peptides are specified in the table. Any of these peptides may be derivatized as described herein, and a few derivatized examples are provided in the table. Derivatized peptides in the tables are exemplary rather than limiting, as the associated underivatized peptides may be employed in this invention, as well. For derivatives in which the carboxyl terminus may be capped with an amino group, the capping amino group is shown as -NH 2 . For derivatives in which amino acid residues are substituted by moieties other than amino acid residues, the substitutions are denoted by a, which signifies any of the moieties described in Bhatnagar etal. (1996), J. Med. Chem. 39: 3814-9 and Cuthbertson etal. (1997), J. Med. Chem. 40: 2876-82, which are incorporated by reference. The J substituent and the Z substituents (Z 5 , Z 6 , ...Z 40 ) are as defined in U.S. Pat. Nos. 5,608,035 ,5,786,331, and 5,880,096, which are incorporated by reference. For the EPO-mimetic sequences (Table 5), the substituents X 2 through X 1 and the WO 2006/010057 PCT/US2005/024373 28 integer "n" are as defined in WO 96/40772, which is incorporated by reference. Also for the EPO-mimetic sequences, the substituents Xna, Xia, X2a, X3a, X4a, X5a and Xca follow the definitions of XI, X 1 , X 2 , X 3 , X 4 , X 5 , and X,, respectively, of WO 99/47151, which is also incorporated by reference. The substituents "T," "e," and "+" are as defined in Sparks et al. (1996), Proc. Natl. Acad. Sci. 93: 1540-4, which is hereby incorporated by reference. X 4 , X 5 ,

X

6 , and X 7 are as defined in U.S. Pat. No. 5,773,569, which is hereby incorporated by reference, except that: for integrin-binding peptides, X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , and X8 are as defined in International applications WO 95/14714, published June 1, 1995 and WO 97/08203, published March 6, 1997, which are also incorporated by reference; and for VIP mimetic peptides, X 1 , X 1 ', X 1 ", X 2 , X 3 , X 4 , X 5 , X 6 and Z and the integers m and n are as defined in WO 97/40070, published October 30, 1997, which is also incorporated by reference. Xaa and Yaa below are as defined in WO 98/09985, published March 12, 1998, which is incorporated by reference. AA 1 , AA 2 , AB 1 , AB 2 , and AC are as defined in International application WO 98/53 842, published December 3, 1998, which is incorporated by reference. X 1 , X 2 , X 3 , and X 4 in Table 17 only are as defined in European application EP 0 911 393, published April 28, 1999. Residues appearing in boldface are D-amino acids. All peptides are linked through peptide bonds unless otherwise noted. Abbreviations are listed at the end of this specification. In the "SEQ ID NO." column, "NR" means that no sequence listing is required for the given sequence. Table 4-IL-1 antagonist peptide sequences Sequence/structure SEQ ID NO: ZuIZ7Z 8

QZ

5

YZ

6

Z

9 Zio 3

XXQZ

5

YZ

6 XX 4

Z

7

XQZ

5

YZ

6 XX 5

Z

7

Z

8

QZ

5 YZZ9ZiO 6 ZulZ 7 ZsQZ 5

YZ

6

Z

9 ZIO 7

Z

12

Z

13 Zi 4

Z

15

Z

16

Z

17

Z

18

Z

19

Z

2 0

Z

2 1

Z

22 ZuLZ 7 ZSQZsYZ 6 Z9ZiOL 8

Z

23 NZ2 4

Z

39 Z2 5

Z

26

Z

27 Z28Z 29

Z

3 0

Z

4 0 9 TANVSSFEWTPYYWQPYALPL 10 SWTDYGYWQPYALPISGL II ETPFTWEESNAYYWQPYALPL 12 ENTYSPNWADSMYWQPYALPL 13 SVGEDHNFWTSEYWQPYALPL 14 DGYDRWRQSGERYWQPYALPL 15 FEWTPGYWQPY 16 FEWTPGYWQHY 17 FEWTPGWYQJY 18 AcFEWTPGWYQJY 19 FEWTPGWpYQJY 20 FAWTPGYWQJY 21 FEWAPGYWQJY 22 WO 2006/010057 PCT/US2005/024373 29 FEWVPGYWQJY 23 FEWTPGYWQJY 24 AcFEWTPGYWQJY 25 FEWTPaWYQJY 26 FEWTPSarWYQJY 27 FEWTPGYYQPY 28 FEWTPGWWQPY 29 FEWTPNYWQPY 30 FEWTPvYWQJY 31 FEWTPecGYWQJY 32 FEWTPAibYWQJY 33 FEWTSarGYWQJY 34 FEWTPGYWQPY 35 FEWTPGYWQHY 36 FEWTPGWYQJY 37 AcFEWTPGWYQJY 38 FEWTPGW-pY-QJY 39 FAWTPGYWQJY 40 FEWAPGYWQJY 41 FEWVPGYWQJY 42 FEWTPGYWQJY 43 AcFEWTPGYWQJY 44 FEWTPAWYQJY 45 FEWTPSarWYQJY 46 FEWTPGYYQPY 47 FEWTPGWWQPY 48 FEWTPNYWQPY 49 FEWTPVYWQJY 50 FEWTPecGYWQJY 51 FEWTPAibYWQJY 52 FEWTSarGYWQJY 53 FEWTPGYWQPYALPL 54 1NapEWTPGYYQJY 55 YEWTPGYYQJY 56 FEWVPGYYQJY 57 FEWTPSYYQJY 58 FEWTPNYYQJY 59 TKPR 60 RKSSK 61 RKQDK 62 NRKQDK 63 RKQDKR 64 ENRKQDKRF 65 VTKFYF 66 VTKFY 67 VTDFY 68 SBLYWQPYSVQ 69 TLVYWQPYSLQT 70 RGDYWQPYSVQS 71 VHVYWQPYSVQT 72 RLVYWQPYSVQT 73 SRVWFQPYSLQS 74 NMVYWQPYSIQT 75 SVVFWQPYSVQT 76 TFVYWQPYALPL 77 TLVYWQPYSIQR 78 WO 2006/010057 PCT/US2005/024373 30 RLVYWQPYSVQR 79 SPVFWQPYSIQI 80 WIEWWQPYSVQS 81 SLIYWQPYSLQM 82 TRLYWQPYSVQR 83 RCDYWQPYSVQT 84 MRVFWQPYSVQN 85 KIVYWQPYSVQT 86 RHLYWQPYSVQR 87 ALVWWQPYSEQI 88 SRVWFQPYSLQS 89 WEQPYALPLE 90 QLVWWQPYSVQR 91 DLRYWQPYSVQV 92 ELVWWQPYSLQL 93 DLVWWQPYSVQW 94 NGNYWQPYSFQV 95 ELVYWQPYSIQR 96 ELMYWQPYSVQE 97 NLLYWQPYSMQD 98 GYEWYQPYSVQR 99 SRVWYQPYSVQR 100 LSEQYQPYSVQR 101 GGGWWQPYSVQR 102 VGRWYQPYSVQR 103 VHIVYWQPYSVQR 104 QARWYQPYSVQR 105 VHVYWQPYSVQT 106 RSVYWQPYSVQR 107 TRVWFQPYSVQR 108 GRIWFQPYSVQR 109 GRVWFQPYSVQR 110 ARTWYQPYSVQR 111 ARVWWQPYSVQM 112 RLMFYQPYSVQR 113 ESMWYQPYSVQR 114 BFGWWQPYSVHM 115 ARFWWQPYSVQR 116 RLVYWQ PYAPIY 117 RLVYWQ PYSYQT 118 RLVYWQ PYSLPI 119 RLVYWQ PYSVQA 120 SRVWYQ PYAKGL 121 SRVWYQ PYAQGL 122 SRVWYQ PYAMPL 123 SRVWYQ PYSVQA 124 SRVWYQ PYSLGL 125 SRVWYQ PYAREL 126 SRVWYQ PYSRQP 127 SRVWYQ PYFVQP 128 EYEWYQ PYALPL 129 IPEYWQ PYALPL 130 SRIWWQ PYALPL 131 DPLFWQ PYALPL 132 SRQWVQ PYALPL 133 IRSWWQ PYALPL 134 WO 2006/010057 PCT/US2005/024373 31 RGYWQ PYALPL 135 RLLWVQ PYALPL 136 EYRWFQ PYALPL 137 DAYWVQ PYAIPL 138 WSGYFQ PYALPL 139 NIEFWQ PYALPL 140 TRDWVQ PYALPL 141 DSSWQ PYLPL142 IGNWQ PYLPL143 NLRWQ PYLPL144 LPE____________________________ 145 DSYWQ PYLPL146 RSQYYQ PYALP ___________________ 147 ARFWQ PYLPL148 NSYFQ PYLPL149 RFMYWQYSVQR150 -ABLWQPYVQR151 WWQPALPL152 YYQPALPL153 YFQPYAL154 SNMQPQRLS160 ___ __ ___ __ ___ __ ___ __ ___ __ ___ __ ___ __ __ 167 TFVYQPYSVQ~ITGVTM168 EVWQPYL WARXPGF 6 TFWWQPYRGPWIVR 6 LFVYQPYALLELEAR 6 GVWQPYRGLAG PLQV16 AWYQPYATPLDE 167'__ MWYQPYSSQPAE 6 GWTQPYSQQQE17 FQPYSIQSDE17 PWEQPYARGG 177 RLWQPYSVQVD 178 T1YWQPYSQIA 179 RDWQPYSQT 180 DWIFQPYALPLD 186 EWDS YWQPSQTLL 182 WEQN YWQPYSVQ A 18 SDV YWQPYSVQSE 184 YYDG YWQPYSQVP 185 SWQVPYALPL 186 QRWVWQ PYLPL LL 187 SRJWWQ PYALPL 188 RSLYWQ PYALPL 189 TIIWEQ PYALPL 190 WO 2006/010057 PCT/US2005/024373 32 WETWYQ PYALPL 191 SYDWEQ PYALPL 192 SRIWCQ PYALPL 193 EIvFWQ PYALPL 194 DYVWQQ PYALPL 195 MDLLVQ WYQPYALPL 196 GSKVIL WYQPYALPL 197 RQGANI WYQPYALPL 198 GGGDEP WYQPYALPL 199 SQLERT WYQPYALPL 200 ETWVRE WYQPYALPL 201 KKGSTQ WYQPYALPL 202 LQARMN WYQPYALPL 203 EPRSQK WYQPYALPL 204 VKQKWR WYQPYALPL 205 LRRHDV WYQPYALPL 206 RSTASI WYQPYALPL 207 ESKEDQ WYQPYALPL 208 EGLTMK WYQPYALPL 209 EGSREG WYQPYALPL 210 VIEWWQ PYALPL 211 VWYWEQ PYALPL 212 ASEWWQ PYALPL 213 FYEWWQ PYALPL 214 EGWWVQ PYALPL 215 WGEWLQ PYALPL 216 DYVWEQ PYALPL 217 AHTWWQ PYALPL 218 FIEWFQ PYALPL 219 WLAWEQ PYALPL 220 VMEWWQ PYALPL 221 ERMWQ PYALPL 222 NXXWXX PYALPL 223 WGNWYQ PYALPL 224 TLYWEQ PYALPL 225 VWRWEQ PYALPL 226 LLWTQ PYALPL 227 SRIWXX PYALPL 228 SDIWYQ PYALPL 229 WGYYXX PYALPL 230 TSGWYQ PYALPL 231 VHPYXX PYALPL 232 EHSYFQ PYALPL 233 XXIWYQ PYALPL 234 AQLHSQ PYALPL 235 WANWFQ PYALPL 236 SRLYSQ PYALPL 237 GVTFSQ PYALPL 238 SIVWSQ PYALPL 239 SRDLVQ PYALPL 240 HWGH VYWQPYSVQ DDLG 241 SWHS VYWQPYSVQ SVPE 242 WRDS VYWQPYSVQ PESA 243 TWDA VYWQPYSVQ KWLD 244 TPPW VYWQPYSVQ SLDP 245 YWSS VYWQPYSVQ SVHS 246 WO 2006/010057 PCT/US2005/024373 33 YWY QPY ALGL 247 YWY QPY ALPL 248 EWI QPY ATGL 249 NWE QPY AKPL 250 AFY QPY ALPL 251 FLY QPY ALPL 252 VCK QPY LEWC 253 ETPFTWEESNAYYWQPYALPL 254 QGWLTWQDSVDMYWQPYALPL 255 FSEAGYTWPENTYWQPYALPL 256 TESPGGLDWAKIYWQPYALPL 257 DGYDRWRQSGERYWQPYALPL 258 TANVSSFEWTPGYWQPYALPL 259 SVGEDHNFWTSE YWQPYALPL 260 MNDQTSEVSTFP YWQPYALPL 261 SWSEAFEQPRNL YWQPYALPL 262 QYAEPSALNDWG YWQPYALPL 263 NGDWATADWSNY YWQPYALPL 264 THDEHI YWQPYALPL 265 MLEKTYTTWTPG YWQPYALPL 266 WSDPLTRDADL YWQPYALPL 267 SDAFTTQDSQAM YWQPYALPL 268 GDDAAWRTDSLT YWQPYALPL 269 AIIRQLYRWSEM YWQPYALPL 270 ENTYSPNWADSM YWQPYALPL 271 MNDQTSEVSTFP YWQPYALPL 272 SVGEDHNFWTSE YWQPYALPL 273 QTPFTWEESNAY YWQPYALPL 274 ENPFTWQESNAY YWQPYALPL 275 VTPFTWEDSNVF YWQPYALPL 276 QIPFTWEQSNAY YWQPYALPL 277 QAPLTWQESAAY YWQPYALPL 278 EPTFTWEESKAT YWQPYALPL 279 TTTLTWEESNAY YWQPYALPL 280 ESPLTWEESSAL YWQPYALPL 281 ETPLTWEESNAY YWQPYALPL 282 EATFTWAESNAY YWQPYALPL 283 EALFTWKESTAY YWQPYALPL 284 STP-TWEESNAY YWQPYALPL 285 ETPFTWEESNAY YWQPYALPL 286 KAPFTWEESQAY YWQPYALPL 287 STSFTWEESNAY YWQPYALPL 288 DSTFTWEESNAY YWQPYALPL 289 YIPFTWEESNAY YWQPYALPL 290 QTAFTWEESNAY YWQPYALPL 291 ETLFTWEESNAT YWQPYALPL 292 VSSFTWEESNAY YWQPYALPL 293 QPYALPL 294 Py-1-NapPYQJYALPL 295 TANVSSFEWTPG YWQPYALPL 296 FEWTPGYWQPYALPL 297 FEWTPGYWQJYALPL 298 FEWTPGYYQJYALPL 299 ETPFrWEESNAYYWQPYALPL 300 FTWEESNAYYWQJYALPL 301 ADVL YWQPYA PVTLWV 302 WO 2006/010057 PCT/US2005/024373 34 GDVAE YWQPYA LPLTSL 303 SWTDYG YWQPYA LPISGL 304 FEWTPGYWQPYALPL 305 FEWTPGYWQJYALPL 306 FEWTPGWYQPYALPL 307 FEWTPGWYQJYALPL 308 FEWTPGYYQPYALPL 309 FEWTPGYYQJYALPL 310 TANVSSFEWTPGYWQPYALPL 311 SWTDYGYWQPYALPISGL 312 ETPFTWEESNAYYWQPYALPL 313 ENTYSPNWADSMYWQPYALPL 314 SVGEDHNFWTSEYWQPYALPL 315 DGYDRWRQSGERYWQPYALPL 316 FEWTPGYWQPYALPL 317 FEWTPGYWQPY 318 FEWTPGYWQJY 319 EWTPGYWQPY 320 FEWTPGWYQJY 321 AEWTPGYWQJY 322 FAWTPGYWQJY 323 FEATPGYWQJY 324 FEWAPGYWQJY 325 FEWTAGYWQJY 326 FEWTPAYWQJY 327 FEWTPGAWQJY 328 FEWTPGYAQJY 329 FEWTPGYWQJA 330 FEWTGGYWQJY 331 FEWTPGYWQJY 332 FEWTJGYWQJY 333 FEWTPecGYWQJY 334 FEWTPAibYWQJY 335 FEWTPSarWYQJY 336 FEWTSarGYWQJY 337 FEWTPNYWQJY 338 FEWTPVYWQJY 339 FEWTVPYWQJY 340 AcFEWTPGWYQJY 341 AcFEWTPGYWQJY 342 INap-EWTPGYYQJY 343 YEWTPGYYQJY 344 FEWVPGYYQJY 345 FEWTPGYYQJY 346 FEWTPsYYQJY 347 FEWTPnYYQJY 348 SHLY-Nap-QPYSVQM 349 TLVY-Nap-QPYSLQT 350 RGDY-Nap-QPYSVQS 351 NMVY-Nap-QPYSIQT 352 VYWQPYSVQ 353 VY-Nap-QPYSVQ 354 TFVYWQJ-YALPL 355 FEWTPGYYQJ-Bpa 356 XaaFEWTPGYYQJ-Bpa 357 FEWTPGY-Bpa-QJY 358 WO 2006/010057 PCT/US2005/024373 35 AcFEWTPGY-Bpa-QJY 359 FEWTPG-Bpa-YQJY 360 AcFEWTPG-Bpa-YQJY 361 AcFE-Bpa-TPGYYQJY 362 AcFE-Bpa-TPGYYQJY 363 Bpa-EWTPGYYQJY 364 AcBpa-EWTPGYYQJy ~365 VYWQPYSVQ 366 RLVYWQPYSVQR 367 RLVY-Nap-QPYSVQR 368 RLDYWQPYSVQR 369 RLVWFQPYSVQR 370 RLVYWQPYSIQR 371 DNSSWYDSFLL 372 DNTAWYESFLA 373 DNTAWYENFLL 374 PARE DNTAWYDSFLI WC 375 TSEY DNTTWYEKFLA SQ 376 SQIP DNTAWYQSFLL HG 377 SPFI DNTAWYENFLL TY 378 EQIY DNTAWYDHFLL SY 379 TPFI DNTAWYENFLL TY 380 TYTY DNTAWYERFLM SY 381 TMTQ DNTAWYENFLL SY 382 TI DNTAWYANLVQ TYPQ 383 TI DNTAWYERFLA QYPD 384 HI DNTAWYENFLL TYTP 385 SQ DNTAWYENFLL SYKA 386 QI DNTAWYERFLL QYNA 387 NQ DNTAWYESFLL QYNT 388 TI DNTAWYENFLL NHNL 389 HY DNTAWYERFLQ QGWH 390 ETPFTWEESNAYYWQPYALPL 391 YIPFTWEESNAYYWQPYALPL 392 DGYDRWRQSGERYWQPYALPL 393 pY-lNap-pY-QJYALPL 394 TANVSSFEWTPGYWQPYALPL 395 FEWTPGYWQJYALPL 396 FEWTPGYWQPYALPLSD 397 FEWTPGYYQJYALPL 398 FEWTPGYWQJY 399 AcFEWTPGYWQJY 400 AcFEWTPGWYQJY 401 AcFEWTPGYYQJY 402 AcFEWTPaYWQJY 403 AcFEWTPaWYQJY 404 AcFEWTPaYYQJY 405 FEWTPGYYQJYALPL 406 FEWTPGYWQJYALPL 407 FEWTPGWYQJYALPL 408 TANVSSFEWTPGYWQPYALPL 409 AcFEWTPGYWQJY 410 AcFEWTPGWYQJY 411 AcFEWTPGYYQJY 412 AcFEWTPAYWQJY 413 AcFEWTPAWYQJY 414 WO 2006/010057 PCT/US2005/024373 36 AcFEWTPAYYQJY 415_ Table 5-EPO-mimetic peptide sequences Sequence/structure SEQ ID NO: YXCXXGPXTWXCXP 416 YXCXXGPXTWXCXP-YXCXXGPXTWXCXP 417 YXCXXGPXTWXCXP-A-YXCXXGPXTWXCNP 418 YXGXXGPXTWXCXP-A- faminee) 419 K PIA YXCXXG PXTWXCXP-A- (ct-anmine) GGTYSCHFGPLTWVCKPQGG 420 GGDYHCRMGPLTWVCKPLGG 421 GGVYACRMGPITWVCSPLGG 422 VGNYMCIIFGPITWVCRPGGG 423 GGLYLCRFGPVTWDcGyKaa 424 GGTYSCLIPGPLTWVcKpQGG- 425 GGTYSCHFGPLTWVCKPQGG GGTYSCHFGPLTWVCKPQGG -A- 426 GGTYSCHFGPLTWVCKPQGG_____ GGTYSCHFGPLTWVCKPQGGSSK 427 GGTYSCHFGPLTWVCKPQGGSSK- 428 GGTYSCHFGPLTWVCKPQGGSSK GGTYSCHFGPLTWVCKPQGGSSK-A- 429 GGTYSCBFGPLTWVCKPQGGSSK GGTYSCHFGPLTWVCKPQGGSS amn)430 /K P1A GGTYSCHFGPLTWVCKPQGGSS/ (OC-amine) GGTYSCHFGPLTWVCKPQGGSSK(A -biotin) 431

CX

4

X

5

GPX

6 TWX-,C 432 GGTYSCHGPLTWVCKPQGG 433 VGNYMAH]MGPITWVCRPGG 434 GGPHHVYACRMGPLTWIC 435 GGTYSCIIFGPLTWVCKPQ 436 GGLYACHMGPMTWVCQPLRG 437 TIAQYICYMGPETWECRPSPKA 438 YSCHIFGPLTWVCK 439 YCBFGPLTWVC 440

X

3

X

4

X

5

GPX

6

TWX

7 X, 441

YX

2

X

3

X

4

X

5

GPX

6 TWXJXS 442

X

1

YX

2

X

3

X

4

X

5

GPX

6

TWX

7

X

8

X

9

X

0

X

1 443

X

1

YX

9

CX

4

X

5

GPX

6

TWX

7

CXX

0

X

1 444 GGLYLCRFGPVTWDCGYKGG 445 GGTYSCBFGPLTWVCKPQGG 446 GGDYHCRMGPLTWVCKPLGG 447 WO 2006/010057 PCT/US2005/024373 37 VGNYMCHFGPITWYVCRPGGG 448 GGVYACRMGPITWVCSPLGG 449 VGNYMAHMGPITWVCRPGG 450 GGTYSCHFGPLTWVCKPQ 451 GGLYACHMGPMTWVCQPLRO 452 TIAQYICYMGPETWErFCRPSPKA 453 YSCHFGPLTWVCK 454 YCHFGPLTWVC 455 SCHFGPLTWVCK 456

(AX

2

).X

3

X

4 XsGPX 6

TWX

7

X

8 457 X~CXiX 2

GWYGX

3

CX

4

X

5 WXC 458 Table 6-TPO-minietic peptide sequences Sequence/structure SEQ ____ ____ ___ ____ ____ __ ED NO: TEGPTLRQWLAARA 459 IEGPTLRQWLAAKA 460 IEGPTrLREWLAARA 461 IEGPTLRQWLAARA-A4IEGPTLRQWLAARA 462 IEGPTLRQWLAAKA-A-IEGPTLRQWLAAKA 463 IEGPTLRQCLAARA-A-IEGP-fLRQCLAARA 464 1 PLQLAR--(EC-A-EPLQLAI 46 IEGPTLRQWLAARA-A-K(PEG)-A-IEGPTLRQWLAARA 466 1EGPTLRQWLAARA-A-K( AIEPQ L1 LAA 467 JEGPTLRQWLAARA-A-IEGPTLRQcLAARA 468 47 -TLREWL474 GRVRDVAGW475 GRVKDIAQL476 GVRDQILWAL 479 GVRQISL 470

GRVRDQI(X)

3 L 48 CTLRQWLQGC 482 CTLQEFLEGC 483 CTRTEWLHGC 484 CTLREWvLHGGFC 485 CTLREWVFAGLC 486 CTLRQWLTILLGMC 487 CTLAEFLASGVEQC 488 CSLQEELSHGGYVG 489 CTLRFFLDPTTAVC 490 CTLKEWLVSHEVWC 491

CTLREW'L(X)

2

.

6 C 492 WO 2006/010057 PCT/US2005/024373 38 REGPTLRQWM 493 EGPTLRQWLA 494 ERGPFWAKAC 495 REGPRCVMWM 496 CGTEGPTLSTWLDC 497 CEQDGPTLLEWLKC 498 CELVGPSLMSWLTC 499 CLTGPFVTQWLYEC 500 CRAGPTLLEWLTLC 501 CADGPTLREWISFC 502 C(X)i- 2

EGPTLREWL(X)

1

-

2 C 503 GGCTLREWLHGGFCGG 504 GGCADGPTLREWISFCGG 505 GNADGPTLRQWLEGRRPKN 506 LAIEGPTLRQWLHGNGRDT 507 HGRVGPTLREWKTQVATKK 508 TIKGPTLRQWLKSREHTS 509 ISDGPTLKEWLSVTRGAS 510 SIEGPTLREWLTSRTPHS 511 Table 7-G-CSF-mimetic peptide sequences Sequence/structure SEQ ID NO: EEDCK 512 EEDCK 513 EEDCK EEDoK 514 EEDaK 515 EEDaK pGluEDaK 516 pGluEDaK 517 pGluEDaK PicSDaK 518 PicSDaK 519 PicSDaK EEDCK-A-EEDCK 520 EEDXK-A-EEDXK 521 WO 2006/010057 PCT/US2005/024373 39 Table 8-TNF-antagonist peptide sequences Sequence/structure SEQ ID NO: YCFTASENHCY 522 YCFTNSENHCY 523 YCFTRSENHCY 524 FCASENHCY 525 YCASENHCY 526 FCNSENHCY 527 FCNSENRCY 528 FCNSVENRCY 529 YCSQSVSNDCF 530 FCVSNDRCY 531 YCRKELGQVCY 532 YCKEPGQCY 533 YCRKEMGCY 534 FCRKEMGCY 535 YCWSQNLCY 536 YCELSQYLCY 537 YCWSQNYCY 538 YCWSQYLCY 539 DFLPHYKNTSLGHRP 540 AAi-ABi NR AC

AA

2

-AB

2 Table 9-Integrin-binding peptide sequences Sequence/structure SEQ ID NO:

RXIETX

2

WX

3 541 RXiETX 2

WX

3 542 RGDGX 543 CRGDGXC 544

CXIX

2

RLDX

3

X

4 C 545 CARRLDAPC 546 CPSRLDSPC 547 XiX 2

X

3

RGDX

4

X

5

X

6 548

CX

2

CRGDCX

5 C 549 CDCRGDCFC 550 CDCRGDCLC 551 CLCRGDCIC 552 XiX 2

DDX

4

X

5

X

7 XB 553

X

1

X

2

X

3

DDX

4

X

5

X

6

X

7

X

8 554 CWDDGWLC 555 CWDDLWWLC 556 CWDDGLMC 557 CWDDGWMC 558 CSWDDGWLC 559 CPDDLWWLC 560 NGR NR GSL NR RGD

NR

WO 2006/010057 PCT/US2005/024373 40 CGRECPRLCQSSC 561 CNGRCVSGCAGRC 562 CLSGSLSC 563 RGD NR NGR NR GSL NR -NGRAHA564 CNGRC565 CDCRDCFC566 CGSLVRC567 -DLXXL568 RTD______________________________ 569 -RTDLSLRTY570 RTD__________________________ 571 RTD___________________________ 572 -GDLLLKLLTL573 GDLHSLQLLSR574 -RDDHMLRQLW575 SSDLHAKKRYG576 RGDLKQSELTW577 RGDLAALSAPPV 578 Table lO-Selectin antagonist peptide sequences Sequence/structure SEQ ID NO: DITWDQLWDLMK 579 DITWDELWKLMN 580 DYTWFELWDMMQ 581 QITWAQLWNMMK 582 DMTWHDLWTLMS 583 DYSWHDLWVEMMS 584 EITWDQLWEVMN 585 HVSWEQLWDIM4N 586 HITWDQLWRIMT 587 RNMSWLELWEHMK 588 AEWTWDQLWHVMNPAESQ 589 HBRAEWLALWEQMSP 590 KKEDWvLALWRIMSV 591 ITWDQLWDLMK 592 DITWDQLWDLMK 593 DITWDQLWDLMK 594 DITWDQLWDLMK 595 CQNRYTDLVAIQNKNE 596 AENWADNEPNNKRNNED 597 RKNNYTWTWGTKKLTNE598 KKALNEAEWAD599 CQXRYDLVAINKXE600 -RKXNXWTWGTXKLTEE601 ____________________________________ 602 CXXXYXLVAINKXE603 RKXXXXWXWVGTXKXLTXE 604 AXNWXXXEPNNXXXED 605 XKXKTXEAXNWXX 606 WO 2006/010057 PCT/US2005/024373 41 Table 11-Antipathogenic peptide sequences Sequence/structure SEQ ID NO: GFFALIPKIISSPLFKTLLSAVGSALSSSGGQQ 607 GFFALIPKIISSPLFKTLLSAVGSALSSSGGQE 608 GFFALIPKIISSPLFKTLLSAV 609 GFFALIPKIISSPLFKTLLSAV 610 KGFFALIPKIISSPLFKTLLSAV 611 KKGFFALIPKIISSPLFKTLLSAV 612 KKGFFALIPKIISSPLFKTLLSAV 613 GFFALIPKIIS 614 GIGAVLKVLT TGLPALISWIKRKRQQ 615 GIGAVLKVLTTGLPALISWIKRKRQQ 616 GIGAVLKVLTTGLPALISWIKRKRQQ 617 GIGAVLKVLTTGLPALISWIKR 618 AVLKVLTTGLPALISWIKR 619 KLLLLLKLLLLK 620 KLLLKLLLKLLK 621 KLLLKLKLKLLK 622 KKLLKLKLKLKK 623 KLLLKLLLKLLK 624 KLLLKLKLKLLK 625 KLLLLK 626 KLLLKLLK 627 KLLLKLKLKLLK 628 KLLLKLKLKLLK 629 KLLLKLKLKLLK 630 KAAAKAAAKAAK 631 KVVVKVVVKVVK 632 KVVVKVKVKVVK 633 KVVVKVKVKVK 634 KVVVKVKVKVVK 635 KLILKL 636 KVLHLL 637 LKLRLL 638 KPLHLL 639 KLILKLVR 640 KVFHLLHL 641 HKFRILKL 642 KPFHILHL 643 KIIIKIKIKTIK 644 KIIIKIKIKIIK 645 KIIKIKIK 646 KIPIKIKIKIPK 647 KIPIKIKIKIVK 648 RIIIRIRIRIIR 649 RIIIRIRIRIIR 650 RIIIRIRIRIIR 651 RIVIRIRIRLIR 652 RIIVRIRLRIIR 653 RIGIRLRVRJIR 654 KIVIRIRIRLIR 655 RIAVKWRLRFIK 656 KIGWKLRVRIIR 657 KKIGWLIIRVRR 658 WO 2006/010057 PCT/US2005/024373 42 RIrHRRUR 659 RIIVRIIRLRflRVR 660 RIGIRLRVRMRRV 661 11VRIRRIPRIR 662 KIGIKARVRI[IRVKII 664 RIIVHRLRIHURL665 I-HGKAHVI[IRHIII666 RIYVH-IRYIKIRL667 -KIGHARVIMYKU668 _____________________________________ 669 KPGEF-, _____________________________ 670 KIVIRRIRLZIREYIV671 RIIVIRLRIKKILIKK672 KIGWKRVRIIRVKGRLR673 KIVIIRIRERIRUGVKKRIR674 REAVKIRLRIIKKIRLIKIKRVIK 675 KAGWKLRVRURVKIGRLRKIGWKKRVRIK 676 RIYVPI-IRYIKIRL677 I<PGKARBIIRKII678 KIVHHUFHURIKIV679 RIIVIRLREKKILIKK680 RIYVSISIYIKIRL681 KIVITRERTSUZRSIV682 -KPEKARPIIRYIM683 CYC~ICCKGFFALIPKJJSSPLFKTLLSAVC 684 CKKGnFFALLEPKIISSPLFKTLLsAvc 685 CKKKGFFALEPKHSSPLFKTLLSAVC 686 CyclicCRIVIRIRIRLIRURC 687 CyclicCKPGHIKARPHIIYKIIC 688 CyclicCRFAVKIRLPfKIMRLIKKIRKRv1KC 689 KLLLKLLL KLLKC 690 KLLLKLLLLK 691 KLLKKLLLC 692 KLLLKLLLKL 693 Table 12-VIP-mimetic peptide sequences Sequnce/trucureSEQ ____ ___ ____ ___ ____ ___ ID NO: HSDAVFYDNYTR LRKQMAVKKYLN SILN 694 NMe HSDAVFYDNYTR LRKQMAVKKYLN SILN 695 xf X, Xi Xz696 X3_______________X4_______________ 697 NH CH COKKYX5 NHCH COX6 698 (CH2)mZ -(CH2)n KKYL 699 NSULN 700 KKYL 701 K-KYA 702 AVKKYL703 NSILN704 YKYV705 SI____________________________ 706 WO 2006/010057 PCT/US2005/024373 43 KKYLNle 707 NSYLN 708 NSIYN 709 KKYLPPNSILN 710 LauKKYL 711 CapKKYL 712 KYL 713 KKYNIe 714 VKKYL 715 LNSILN 716 YLNSILN 717 KKYLN 718 KKYLNS 719 KKYLNSI 720 KKYLNSIL 721 KKYL 722 KKYDA 723 AVKKYL 724 NSILN 725 KKYV 726 SILauN 727 NSYLN 728 NSIYN 729 KKYLNIe 730 KKYLPPNSILN 731 KKYL 732 KKYDA 733 AVKKYL 734 NSILN 735 KKYV 736 SILauN 737 LauKKYL 738 CapKKYL 739 KYL 740 KYL 741 KKYNle 742 VKKYL 743 LNSILN 744 YLNSILN 745 KKYLNIe 746 KKYLN 747 KKYLNS 748 KKYLNSI 749 KKYLNSIL 750 KKKYLD 751 cyclicCKKYLC 752 CKKYLK 753 1 1 S-CHrCO KKYA 754 WWTDTGLW 755 WWTDDGLW 756 WWDTRGLWVWTI 757 FWGNDGIWLESG 758 DWDQFGLWRGAA 759 RWDDNGLWVVVL 760 WO 2006/010057 PCT/US2005/024373 44 SGMWSHYGIWMG 761 GGRWDQAGLWVA 762 KLWSEQGIWMGE 763 CWSMHGLWLC 764 GCWDNTGIWVPC 765 DWDTRGLWVY 766 SLWDENGAWI 767 KWDDRGLWMH 768 QAWNERGLWT 769 QWDTRGLWVA 770 WNVHGIWQE 771 SWDTRGLWVE 772 DWDTRGLWVA 773 SWGRDGLWIE 774 EWTDNGLWAL 775 SWDEKGLWSA 776 SWDSSGLWMD 777 Table 13-Mdni/hdm antagonist peptide sequences Sequence/structure SEQ ID NO: TFSDLW 778 QETFSDLWKLLP 779 QPTFSDLWKLLP 780 QETFSDYWKLLP 781 QPTFSDYWKLLP 782 MPRFMDYWEGLN 783 VQNFIDYWTQQF 784 TGPAFTHYWATF 785 IDRAPTFRDHWFALV 786 PRPALVFADYWETLY 787 PAFSRFWSDLSAGAH 788 PAFSRFWSKLSAGAH 789 PXFXDYWXXL 790 QETFSDLWKLLP 791 QPTFSDLWKLLP 792 QETFSDYWKLLP 793 QPTFSDYWKLLP 794 Table 14-Calmodulin antagonist peptide sequences Sequence/structure SEQ ID NO: SCVKWGKKEFCGS 795 SCWKYWGKECGS 796 SCYEWGKLRWCGS 797 SCLRWGKWSNCGS 798 SCWRWGKYQICGS 799 SCVSWGALKLCGS 800 SCIRWGQNTFCGS 801 SCWQWGNLKICGS 802 SCVRWGQLSICGS 803 LKKFNARRKLKGAILTTMLAK 804 RRWKKNFIAVSAANRFKK 805 WO 2006/010057 PCT/US2005/024373 45 RKWQKTGHAVRAIGRLSS 806 INLKALAALAKKIL 807 KIWSILAPLGTTLVKLVA 808 LKKLLKLLKKLLKL 809 LKWKKLLKLLKKLLKKLL 810 AEWPSLTEIKTLSHFSV 811 AEWPSPTRVISTTYFGS 812 AELAHWPPVKTVLRSFT 813 AEGSWLQLLNLMKQMNN 814 AEWPSLTEIK 815 Table 15-Mast cell antagonists/Mast cell protease inhibitor peptide sequences Sequence/structure SEQ ID NO: SGSGVLKRPLPILPVTR 816 RWLSSRPLPPLPLPPRT 817 GSGSYDTLALPSLPLHPMSS 818 GSGSYDTRALPSLPLHPMSS 819 GSGSSGVTMYPKLPPHWSMA 820 GSGSSGVRMYPKLPPHWSMA 821 GSGSSSMRMVPTIPGSAKHG 822 RNR NR QT NR RQK NR NRQ NR RQK NR RNRQKT 823 RNRQ 824 RNRQK 825 NRQKT 826 RQKT 827 Table 16-SH3 antagonist peptide sequences Sequence/structure SEQ ID NO: RPLPPLP 828 RELPPLP 829 SPLPPLP 830 GPLPPLP 831 RPLPIPP -832 RPLPIPP 833 RRLPPTP 834 RQLPPTP 835 RPLPSRP 836 RPLPTRP 837 SRLPPLP 838 RALPSPP 839 RRLPRTP 840 RPVPPIT 841 ILAPPVP 842 RPLPMLP 843 RPLPILP 844 RPLPSLP 845 WO 2006/010057 PCT/US2005/024373 46 RPLPSLP 846 RPLPMIP 847 RPLPLIP 848 RPLPPTP 849 RSLPPLP 850 RPQPPPP 851 RQLPIPP 852 XXXRPLPPLPXP 853 XXXRPLPPIPXX 854 XXXRPLPPLPXX 855 RXXRPLPPLPXP 856 RXXRPLPPLPPP 857 PPPYPPPPIPXX 858 PPPYPPPPVPXX 859 LXXRPLPXTP 860 TXXRPLPXLP 861 PPXOXPPPTP 862 +PP'PPXKPXWL 863 RPXTPR+SXP 864 PPVPPRPXXTL 865 IPTLPVK 866 +ODXPLPXLP 867 Table 17-Somatostatin or cortistatin mimetic peptide sequences Sequence/structure SEQ ID NO: X -X2-Asn-Phe-Phe-Trp-Lys-Thr-Phe-X3-Ser-X4 868 Asp Arg Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 869 Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 870 Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 871 Asp Arg Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 872 Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 873 Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 874 Asp Arg Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 875 Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 876 Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 877 Asp Arg Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 878 Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 879 Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 880 Asp Arg Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 881 Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 882 Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 883 Asp Arg Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 884 Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 885 Cys Arg Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 886 Asp Arg Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 887 Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 888 Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys Lys 889 Asp Arg Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 890 Met Pro Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 891 Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 892 WO 2006/010057 PCT/US2005/024373 47 Table 18-UKR antagonist peptide sequences Sequnce/trucureSEQ AEPWHLNFSOLWYTID NO: 893 AEHTSSLDTYSLAF894 AELDWNHUIYPLFSNR895 AESSWTRY~vTSPSY896 AEWHPGLSFGSYLWSKT 897 AEPALLNWSFFFNPGLH 898 AEWSFYNLHLPEPQTIW 899 AEPLDLWSLYSLPPLAM 900 AEPTLWQLYQFPLRLSG 901 AEISFSELMWLRSTPAF 902 AELSEADLWTTWFGMGS 903 AESSLWRIFSPSALMMvS 904 AESLPThTSILWGKESV 905 AETLFMDLWHDKHIILLT 906 AEJILNFPLWHEPLWSTE 907 AESQTGTLNTLFWNTLR 908 AEPVYQYELDSYLRSYY 909 AELDLSTFYDIQYLLRT 910 AEEFKLGPNGYVYLHSA 911 FK-LXXXGYVYL 912 AESTYBIRLSLGYMIYTLN 913 YHXLXXGYMYfT 914 Table 19-Macrophage and/or T-ceII inhibiting peptide sequences Sequence/structure SEQ ED NO: Xaa-Yaa-Arg NR. Arg-Yaa-Xaa NR. Xaa-Arg-Yaa NR. Yaa-Arg-Xaa NR Ala-Arg NR. Arg-Arg NR. Asn-Arg NR Asp-Arg NR. Cys-Arg NR. Gln-Arg NR. Glu-Arg NR. Gly-Arg NR. His-arg NR. fle-Arg NR Leu-Arg NR Lys-Arg NR____ Met-Arg NR. Phe-Arg NR. Ser-Arg NR. Thr-Arg NR. Trp-Arg NR. Tyr-ArgNP Val-Arg

NR.

WO 2006/010057 PCT/US2005/024373 48 Ala-Gin-Arg NR Arg-Glu-Arg NR Asni-Glu-Arg N Asp-Glu-Arg NR____ Cys-Glu-Arg NqR Gln-Glu-Arg NR Glu-Glu-Arg NR Gly-Glu-Arg N His-Glu-Arg NR Ile-Glu-Arg N Leu-Glu-Arg NR Lys-Glu-Arg NR Met-Glu-Arg NR Phe-Glu-Arg NR Pro-Glu-Arg N Ser-Glu-Arg NR Thr-Glu-Arg NR Trp-Glu-Arg N Tyr-Glu-Arg NR Val-Glu-Arg NR Ag-Ala NR Arg-Asp N Arg-Cys NR____ Arg-Gln NR_____ Arg-Glu NR_____ Arg-Gly NR Arg-His NR Arg-le NR Arg-Leu NR Arg-Lys NR Arg-Met NR Arg-Phe N Arg-Pro NR Arg-Ser NR Arg-GlnLy NR Arg-GluMe NR Arg-GhiPh NR Mrg-GinPr NR Arg-Glu-Ser -NR Arg-Glu-Thr NR Arg-Glu-Asp NR Arg-Glu-Tyr

NR

WO 2006/010057 PCT/US2005/024373 49 Arg-Glu-Val NR Ala-Arg-Glu NR Arg-Arg-Glu NR Asn-Arg-Glu NR Asp-Arg-Glu NR Cys-Arg-Glu NR Gln-Arg-Glu NR Glu-Arg-Glu NR Gly-Arg-Glu NR His-Arg-Glu NR Ile-Arg-Glu NR Leu-Arg-Glu NR Lys-Arg-Glu NR Met-Arg-Glu NR Phe-Arg-Glu NR Pro-Arg-Glu NR Ser-Arg-Glu NR Thr-Arg-Glu NR Trp-Arg-Glu NR Tyr-Arg-Glu NR VaI-Arg-Glu NR Glu-Arg-Ala, NR Glu-Arg-Arg NR Glu-Arg-Asn NR Glu-Arg-Asp NR Glu-Arg-Cys NR Glu-Arg-Gln NR Glu-Arg-Gly NR Glu-Arg-His NR Glu-Arg-Ile NR Glu-Arg-Leu NR Glu-Arg-Lys NR Glu-Arg-Met NR Glu-Arg-Phe NR Glu-Arg-Pro NR____ Glu-Arg-Ser NR Glu-Arg-Thr NR Glu-Arg-Trp NR Glu-Arg-Tyr NR Glu-Arg-Val

NR

WO 2006/010057 PCT/US2005/024373 50 Table 20-Additional Exemplary Pharmacologically Active Peptides Sequencelstructure SEQ Activity ID NO: VEPNCDIHVMWEWECFERL 915 VEGF-antagonist GERWCFDGPLTWVCGEES 916 VEGF-antagonist RGWVEICVADDNGMCVTEAQ 917 VEGF-antagonist GWDECDVARMWEWECFAGV 918 VEGF- antagonist GERWCFDGPRAWVCGWEI 919 VEGF- antagonist EELWCFDGPRAWVCGYVK 920 VEGF- antagonist RGWVEICAADDYGRCLTEAQ 921 VEGF- antagonist RGWVEICESDVWGRCL 922 VEGF- antagonist RGWVEICESDVWGRCL 923 VEGF- antagonist GGNECDLARMWEWECFERL 924 VEGF- antagonist RGWVEICAADDYGRCL 925 VEGF-antagonist CTTHWGFTLC 926 MMP inhibitor CLRSGXGC 927 MMP inhibitor CXXHWGFXXC 928 MMP inhibitor CXPXC 929 MMP inhibitor CRRHWGFEFC 930 MMP inhibitor STTHWGFTLS 931 MMP inhibitor CSLHWGFWWC 932 CTLA4-mimetic GFVCSGIFAVGVGRC 933 CTLA4-mimetic APGVRLGCAVLGRYC 934 CTLA4-mimetic LLGRMK 935 Antiviral (HBV) ICVVQDWGHHRCTAGHMANLTSHASAI 936 C3b antagonist ICVVQDWGHHRCT 937 C3b antagonist CVVQDWGHHAC 938 C3b antagonist STGGFDDVYDWARGVSSALTTTLVATR 939 Vinculin-binding STGGFDDVYDWARRVSSALTTTLVATR 940 Vinculin-binding SRGVNFSEWLYDMSAAMKEASNVFPSRRSR 941 Vinculin-binding SSQNWDMEAGVEDLTAAMLGLLSTIHSSSR 942 Vinculin-binding SSPSLYTQFLVNYESAATRIQDLLIASRPSR 943 Vinculin-binding SSTGWVDLLGALQRAADATRTSIPPSLQNSR 944 Vinculin-binding DVYTKKELIECARRVSEK 945 Vinculin-binding EKGSYYPGSGIAQFHIDYNNVS 946 C4BP-binding SGIAQFHIDYNNVSSAEGWHVN 947 C4BP-binding LVTVEKGSYYPGSGIAQFHIDYNNVSSAEGWHVN 948 C4BP-binding SGIAQFIDYNNVS 949 C4BP-binding LLGRMK 950 anti-HBV ALLGRMKG 951 anti-HBV LDPAFR 952 anti-HBV CXXRGDC 953 Inhibition of platelet aggregation RPLPPLP 954 Src antagonist PPVPPR 955 Src antagonist XFXDXWXXLXX 956 Anti-cancer (particularly for sarcomas) KACRRLFGPVDSEQLSRDCD 957 p16-mimetic RERWNFDFVTETPLEGDFAW 958 p16-mimetic KRRQTSMTDFYHSKRRLIFS 959 p16-mimetic TSMTDFYHSKRRLIFSKRKP 960 p16-mimetic RRLIF 961 p16-mimetic KRRQTSATDFYHSKRRLIFSRQIKIWFQNRRMKWKK 962 p16-mimetic WO 2006/010057 PCT/US2005/024373 51 KRRLIFSKRQIKIWFQNRRMKWKK 963 p16-mimetic Asn Gin Gly Arg His Phe Cys Gly Gly Ala Leu lie His Ala Arg 964 CAP37 mimetic/LPS Phe Val Met Thr Ala Ala Ser Cys Phe Gln binding Arg His Phe Cys Gly Gly Ala Leu Ile His Ala Arg Phe Val Met 965 CAP37 mimetic/LPS Thr Ala Ala Ser Cys binding Gly Thr Arg Cys Gin Val Ala Gly Trp Gly Ser Gln Arg Ser Gly 966 CAP37 mimetic/LPS Gly Arg Leu Ser Arg Phe Pro Arg Phe Val Asn Val binding WHWRHRIPLQLAAGR 967 carbohydrate (GD I alpha) mimetic LKTPRV 968 B2GPI Ab binding NTLKTPRV 969 J32GPI Ab binding NTLKTPRVGGC 970 .52GPI Ab binding KDKATF 971 f2GPI Ab binding KDKATFGCHD 972 p2GPI Ab binding KDKATFGCHDGC 973 p2GPI Ab binding TLRVYK 974 02GPI Ab binding ATLRVYKGG 975 p2GPI Ab binding CATLRVYKGG 976 p2GPI Ab binding INLKALAALAKKIL 977 Membrane transporting GWT NR Membrane transporting GWTLNSAGYLLG 978 Membrane transporting GWTLNSAGYLLGKINLKALAALAKKIL 979 Membrane transporting CVHAYRS 980 Antiproliferative, antiviral CVHAYRA 981 Antiproliferative, antiviral CVHAPRS 982 Antiproliferative, antiviral CVHAPRA 983 Antiproliferative, antiviral CVHSYRS 984 Antiproliferative, antiviral CVHSYRA 985 Antiproliferative, antiviral CVHSPRS 986 Antiproliferative, antiviral CVHSPRA 987 Antiproliferative, antiviral CVHTYRS 988 Antiproliferative, antiviral CVHTYRA 989 Antiproliferative, antiviral CVHTPRS 990 Antiproliferative, antiviral CVHTPRA 991 Antiproliferative, antiviral HWAWFK 992 anti-ischemic, growth hormone-liberating WO 2006/010057 PCT/US2005/024373 52 TABLE 21 MYOSTATIN INHIBITOR PEPTIDES PEPTIBODY NAME SEQ ID PEPTIDE SEQUENCE Myostatin-TN8-Con1 1036 KDKCKMWHWMCKPP Myostatin-TN8-Con2 1037 KDLCAMWHWMCKPP Myostatin-TNS-Con3 1038 KDLCKMWKWMCKPP Myostatin-TN8-Con4 1039 KDLCKMWHWMCKPK Myostatin-TN8-Con5 1040 WYPCYEFHFWCYDL Myostatin-TN8-Con6 1041 WYPCYEGHFWCYDL Myostatin-TN8-Con7 1042 IFGCKWWDVQCYQF Myostatin-TN8-Con8 1043 IFGCKWWDVDCYQF Myostatin-TN8-Con9 1044 ADWCVSPNWFCMVM Myostatin-TN8-ConlO 1045 HKFCPWWALFCWDF Myostatin-TN8-1 1046 KDLCKMWHWMCKPP Myostatin-TN8-2 1047 IDKCAIWGWMCPPL Myostatin-TN8-3 1048 WYPCGEFGMWCLNV Myostatin-TN8-4 1049 WFTCLWNCDNE Myostatin-TN8-5 1050 HTPCPWFAPLCVEW Myostatin-TN8-6 1051 KEWCWRWKWMCKPE Myostatin-TN8-7 1052 FETCPSWAYFCLDI Myostatin-TN8-8 1053 AYKCEANDWGCWWL Myostatin-TN8-9 1054 NSWCEDQWHRCWWL Myostatin-TN8-10 1055 WSACYAGHFWCYDL Myostatin-TN8-11 1056 ANWCVSPNWFCMVM Myostatin-TN8-12 1057 WTECYQQEFWCWNL Myostatin-TN8-13 1058 ENTCERWKWMCPPK Myostatin-TN8-14 1059 WLPCHQEGFWCMNF Myostatin-TN8-15 1060 STMCSQWHWMCNPF Myostatin-TN8-16 1061 IFGCHWWDVDCYQF Myostatin-TN8-17 1062 IYGCKWWDIQCYDI Myostatin-TN8-18 1063 PDWCIDPDWWCKFW Myostatin-TN8-19 1064 QGHCTRWPWMCPPY Myostatin-TN8-20 1065 WQECYREGFWCLQT Myostatin-TN8-21 1066 WFDCYGPGFKCWSP Myostatin-TN8-22 1067 GVRCPKGHLWCLYP Myostatin-TN8-23 1068 HWACGYWPWSCKWV Myostatin-TN8-24 1069 GPACHSPWWWCVFG Myostatin-TN8-25 1070 TTWCISPMWFCSQQ Myostatin-TN8-26 1071 HKFCPPWAIFCWDF Myostatin-TN8-27 1072 PDWCVSPRWYCNMW Myostatin-TN8-28 1073 VWKCHWFGMDCEPT Myostatin-TN8-29 1074 KKHCQIWTWMCAPK Myostatin-TN8-30 1075 WFQCGSTLFWCYNL Myostatin-TN8-31 1076 WSPCYDHYFYCYTI Myostatin-TN8-32 1077 SWMCGFFKEVCMWV Myostatin-TN8-33 1078 EMLCMIHPVFCNPH Myostatin-TN8-34 1079 LKTCNLWPWMCPPL Myostatin-TN8-35 1080 VVGCKWYEAWCYNK Myostatin-TN8-36 1081 PIHCTQWAWMCPPT Myostatin-TN8-37 1082 DSNCPWYFLSCVIF Myostatin-TN8-38 1083 HIWCNLAMMKCVEM Myostatin-TN8-39 1084 NLQCIYFLGKCIYF Myostatin-TN8-40 1085 AWRCMWFSDVCTPG Myostatin-TN8-41 1086 WFRCFLDADWCTSV Myostatin-TN8-42 1087 EKICQMWSWMCAPP WO 2006/010057 PCT/US2005/024373 53 Myostatin-TN8-43 1088 WFYCHLNKSECTEP Myostatin-TN8-44 1089 FWRCAIGIDKCKRV Myostatin-TN8-45 1090 NLGCKWYEVWCFTY Myostatin-TNS-46 1091 IDLCNMWDGMCYPP Myostatin-TN8-47 1092 EMPCNIWGWMCPPV Myostatin-TN12-1 1093 WFRCVLTGIVDWSECFGL Myostatin-TN12-2 1094 GFSCTFGLDEFYVDCSPF Myostatin-TN12-3 1095 LPWCHDQVNADWGFCMLW Myostatin-TN12-4 1096 YPTCSEKFWIYGQTCVLW Myostatin-TN12-5 1097 LGPCPIH-HGPWPQYCVYW Myostatin-TN12-6 1098 PFPCETHQISWLGHCLSF Myostatin-TN12-7 1099 HWGCEDLMWSWHPLCRRP Myostatin-TN12-8 1100 LPLCDADMMPTIGFCVAY Myostatin-TN12-9 1101 SHWCETTFWMNYAKCVHA Myostatin-TN12-10 1102 LPKCTHVPFDQGGFCLWY Myostatin-TN12-11 1103 FSSCWSPVSRQDMFCVFY Myostatin-TN12-13 1104 SHKCEYSGWLQPLCYRP Myostatin-TN12-14 1105 PWWCQDNYVQHMLHCDSP Myostatin-TN12-15 1106 WFRCMLMNSFDAFQCVSY Myostatin-TN12-16 1107 PDACRDQPWYMFMGCMLG Myostatin-TN12-17 1108 FLACFVEFELCFDS Myostatin-TN12-18 1109 SAYCIITESDPYVLCVPL Myostatin-TN12-19 1110 PSICESYSTMWLPMCQHN Myostatin-TN12-20 1111 WLDCHDDSWAWTKMCRSH Myostatin-TN12-21 1112 YLNCVMMNTSPFVECVFN Myostatin-TN12-22 1113 YPWCDGFMIQQGITCMFY Myostatin-TN12-23 1114 FDYCTWLNGFKDWKCWSR Myostatin-TN12-24 1115 LPLCNLKEISHVQACVLF Myostatin-TN12-25 1116 SPECAFARWLGIEQCQRD Myostatin-TN12-26 1117 YPQCFNLHLLEWTECDWF Myostatin-TN12-27 1118 RWRCELYDSEFLPKCWFF Myostatin-TN12-28 1119 LVGCDNVWHRCKLF Myostatin-TN12-29 1120 AGWCHVWGEMFGMGCSAL Myostatin-TN12-30 1121 HHECEWMARWMSLDCVGL Myostatin-TN12-31 1122 FPMCGIAGMKDFDFCVWY Myostatin-TN12-32 1123 RDDCTFWPEWLWKLCERP Myostatin-TN12-33 1124 YNFCSYLFGVSKEACQLP Myostatin-TN12-34 1125 AHWCEQGPWRYGNICMAY Myostatin-TN12-35 1126 NLVCGKISAWGDEACARA Myostatin-TN12-36 1127 HNVCTIMGPSMKWFCWND Myostatin-TN12-37 1128 NDLCAMWGWRNTIWCQNS Myostatin-TN12-38 1129 PPFCQNDNDMLQSLCKLL Myostatin-TN12-39 1130 WYDCNVPNELLSGLCRLF Myostatin-TN12-40 1131 YGDCDQNHWMWPFTCLSL Myostatin-TN12-41 1132 GWMCHFDLHDWGATCQPD Myostatin-TN12-42 1133 YFHCMFGGHEFEVHCESF Myostatin-TN12-43 1134 AYWCWHGQCVRF Myostatin-Linear- 1 1135 SEHWTFTDWDGNEWWVRPF Myostatin-Linear-2 1136 MEMLDSLFELLKDMVPISKA Myostatin-Linear-3 1137 SPPEEALMEWLGWQYGKFT Myostatin-Linear-4 1138 SPENLLNDLYILMTKQEWYG Myostatin-Linear-5 1139 FHWEEGIPFHVVTPYSYDRM Myostatin-Linear-6 1140 KRLLEQFMNDLAELVSGHS Myostatin-Linear-7 1141 DTRDALFQEFYEFVRSRLVI Myostatin-Linear-8 1142 RMSAAPRPLTYRDIMDQYWH WO 2006/010057 PCT/US2005/024373 54 Myostatin-Linear-9 1143 NDKAHFFEvIFMFDVHNFVES Myostatin-Linear-lo 1144 QTQAQKIDGLWELLQSIRNQ Myostatin-Linear-1 1 1145 MLSEFEEFLGNLVHRQEA Myostatin-Linear-12 1146 YTPKMGSEWTSFWHNRIHYL Myostatin-Linear-13 1147 LNDTLLRELKMVLNSLSDMK Myostatin-Linear-14 1148 FDVERDLMRWLEGFMQSAAT Myostatin-Linear-15 1149 HHGWNYLRKGSAPQWFEAWV Myostatin-Linear-16 1150 VESLHQLQMWLDQKLASGPH Myostatin-Linear-17 1151 RATLLKDFWQLVEGYGDN Myostatin-Linear-18 1152 EELLREFYRFVSAFDY Myostatin-Linear-19 1153 GLLDEFSHIAEQFYQWMGG Myostatin-Linear-20 1154 YREMSMLEGLLDVLERLQHY Myostatin-Linear-21 1155 HNSSQMLLSELILVGSMMQ Myostatin-Linear-22 1156 WREHFLNSDYIRDKLIAIDG Myostatin-Linear-23 1157 QFPFYVFDDLPAQLEYWIA Myostatin-Linear-24 1158 EFFHWLHNHRSEVNHWLDM4N Myostatin-Linear-25 1159 EALFQNFFRDVLTLSEREY Myostatin-Linear-26 1160 QYWEQQWMTYFRENGLHVQY Myostatin-Linear-27 1161 NQRMMLEDLWRIMTPMFGRS Myostatin-Linear-29 1162 FLDELKAELSRHYALDDLDE Myostatin-Linear-30 1163 GKLIEGLLNELMQLETFMPD Myostatin-Linear-31 1164 ILLLDEYKKDWKSWF Myostatin-2xTN8-19 kc 1165 QGHCTRWPWMCPPYGSGSATGGSGSTASSGSGSATG QGHCTRWPWMCPPY Myostatin-2xTN8-con6 1166 WYPCYEGHFWCYDLGSGSTASSGSGSATGWYPCYEG HFWCYDL Myostatin-2xTN8-5 kc 1167 HTPCPWFAPLCVEWGSGSATGGSGSTASSGSGSATGH TPCPWFAPLCVEW Myostatin-2xTN8-18 kc 1168 PDWCIDPDWWCKFWGSGSATGGSGSTASSGSGSATG PDWCIDPDWWCKFW Myostatin-2xTN8-11 kc 1169 ANWCVSPNWFCMVMGSGSATGGSGSTASSGSGSAT GANWCVSPNWFCMVM Myostatin-2xTN8-25 kc 1170 PDWCIDPDWWCKFWGSGSATGGSGSTASSGSGSATG PDWCIDPDWWCKFW Myostatin-2xTN8-23 kc 1171 HWACGYWPWSCKWVGSGSATGGSGSTASSGSGSAT GHWACGYWPWSCKWV Myostatin-TN8-29-19 kc 1172 KKHCQIWTWMCAPKGSGSATGGSGSTASSGSGSATG QGHCTRWPWMCPPY Myostatin-TN8-19-29 kc 1173 QGHCTRWPWMCPPYGSGSATGGSGSTASSGSGSATG KKHCQIWTWMCAPK Myostatin-TN8-29-19 kn 1174 KKHCQIWTWMCAPKGSGSATGGSGSTASSGSGSATG QGHCTRWPWMCPPY Myostatin-TN8-29-19-8g 1175 KKHCQ1WTWMCAPKGGGGGGGGQGHCTRWPWMCP PY Myostatin-TN8-19-29-6gc 1176 QGHCTRWPWMCPPYGGGGGGKKHCQIWTWMCAPK WO 2006/010057 PCT/US2005/024373 55 TABLE 22 MYOSTATIN INHIBITOR PEPTIDES Affinity- matured SEQ ID peptibody NO: Peptide sequence mTN8-19-1 1177 VALHGQCTRWPWMCPPQREG mTN8-19-2 1178 YPEQGLCTRWPWMCPPQTLA mTN8-19-3 1179 GLNQGHCTRWPWMCPPQDSN mTN8-19-4 1180 MITQGQCTRWPWMCPPQPSG mTN8-19-5 1181 AGAQEHCTRWPWMCAPNDWI mTN8-19-6 1182 GVNQGQCTRWRWMCPPNGWE mTN8-19-7 1183 LADHGQCIRWPWMCPPEGWE mTN8-19-8 1184 ILEQAQCTRWPWMCPPQRGG mTN8-19-9 1185 TQTHAQCTRWPWMCPPQWEG mTN8-19-10 1186 VVTQGHCTLWPWMCPPQRWR mTN8-19-11 1187 IYPHDQCTRWPWMCPPQPYP mTN8-19-12 1188 SYWQGQCTRWPWMCPPQWRG mTN8-19-13 1189 MWQQGHCTRWPWMCPPQGWG mTN8-19-14 1190 EFTQWHCTRWPWMCPPQRSQ mTN8-19-15 1191 LDDQWQCTRWPWMCPPQGFS mTN8-19-16 1192 YQTQGLCTRWPWMCPPQSQR mTN8-19-17 1193 ESNQGQCTRWPWMCPPQGGW mTN8-19-18 1194 WTDRGPCTRWPWMCPPQANG mTN8-19-19 1195 VGTQGQCTRWPWMCPPYETG mTN8-19-20 1196 PYEQGKCTRWPWMCPPYEVE mTN8-19-21 1197 SEYQGLCTRWPWMCPPQGWK mTN8-19-22 1198 TFSQGHCTRWPWMCPPQGWG mTN8-19-23 1199 PGAHDHCTRWPWMCPPQSRY mTN8-19-24 1200 VAEBWHCRRWPWMCPPQDWR mTN8-19-25 1201 VGTQGHCTRWPWMCPPQPAG mTN8-19-26 1202 EEDQAHCRSWPWMCPPQGWV mTN8-19-27 1203 ADTQGHCTRWPWMCPPQHWF mTN8-19-28 1204 SGPQGHCTRWPWMCAPQGWF mTN8-19-29 1205 TLVQGHCTRWPWMCPPQRWV mTN8-19-30 1206 GMAHGKCTRWAWMCPPQSWK mTN8-19-31 1207 ELYHGQCTRWPWMCPPQSWA mTN8-19-32 1208 VADHGHCTRWPWMCPPQGWG mTN8-19-33 1209 PESQGHCTRWPWMCPPQGWG mTN8-19-34 1210 IPAHGHCTRWPWMCPPQRWR mTN8-19-35 1211 FTVHGHCTRWPWMCPPYGWV mTN8-19-36 1212 PDFPGHCTRWRWMCPPQGWE mTN8-19-37 1213 QLWQGPCTQWPWMCPPKGRY mTN8-19-38 1214 HANDGHCTRWQWMCPPQWGG mTN8-19-39 1215 ETDHGLCTRWPWMCPPYGAR mTN8-19-40 1216 GTWQGLCTRWPWMCPPQGWQ WO 2006/010057 PCT/US2005/024373 56 mTN8-19 con1 1217 VATQGQCTRWPWMCPPQGWG mTN8-19 con2 1218 VATQGQCTRWPWMCPPQRWG mTN8 con6-1 1219 QREWYPCYGGHLWCYDLHKA mTN8 con6-2 1220 ISAWYSCYAGHFWCWDLKQK mTN8 con6-3 1221 WTGWYQCYGGHLWCYDLRRK mTN8 con6-4 1222 KTFWYPCYDGHFWCYNLKSS mTN8 con6-5 1223 ESRWYPCYEGHLWCFDLTET TABLE 23 MYOSTATIN INHIBITOR PEPTIDES Affinity matured SEQID peptibody NO: Peptide Sequence L2 1224 MEMLDSLFELLKDMVPISKA mL2-Con1 1225 RMEMLESLLELLKEIVPMSKAG iL2-Con2 1226 RMEMLESLLELLKEIVPMSKAR mL2-1 1227 RMEMLESLLELLKDIVPMSKPS mL2-2 1228 GMEMLESLFELLQEIVPMSKAP mL2-3 1229 RMEMLESLLELLKDIVPISNPP mL2-4 1230 RIEMLESLLELLQEIVPISKAE mL2-5 1231 RMEMLQSLLELLKDIVPMSNAR mL2-6 1232 RMEMLESLLELLKEIVPTSNGT mL2-7 1233 RMEMLESLFELLKEIVPMSKAG mL2-8 1234 RMEMLGSLLELLKEIVPMSKAR niL2-9 1235 QMELLDSLFELLKEIVPKSQPA mL2-10 1236 RMEMLDSLLELLKEIVPMSNAR mL2-11 1237 RMEMLESLLELLHEIVPMSQAG mL2-12 1238 QMEMLESLLQLLKEIVPMSKAS mL2-13 1239 RMEMLDSLLELLKDMVPMTTGA mL2-14 1240 RIEMLESLLELLKDMVPMANAS mL2-15 1241 RMEMLESLLQLLNEIVPMSRAR mL2-16 1242 RMEMLESLFDLLKELVPMSKGV mL2-17 1243 RIEMLESLLELLKDIVPIQKAR mL2-18 1244 RMELLESLFELLKDMVPMSDSS mL2-19 1245 RMEMLESLLEVLQEIVPRAKGA mL2-20 1246 RMEMLDSLLQLLNEIVPMSHAR mL2-21 1247 RMEMLESLLELLKDIVPMSNAG mL2-22 1248 RMMLQSLFELLKGMVPISKAG mL2-23 1249 RMMLESLLELLKEIVPNSTAA mL2-24 1250 RMMLQSLLELLKEIVPISKAG niL2-25 1251 RIEMLDSLLELLNELVPMSKAR 1252 L-15 HHGWNYLRKGSAPQWFEAWV mL15-con1 1253 QVESLQQLLMWLDQKLASGPQG WO 2006/010057 PCT/US2005/024373 57 mL15-1 1254 RMELLESLFELLKEMVPRSKAV mL15-2 1255 QAVSLQIHLLMWLDQKLASGPQH mL15-3 1256 DEDSLQQLLMWLDQKLASGPQL mL15-4 1257 PVASLQQLLIWLDQKLAQGPHA mL15-5 1258. EVDELQQLLNWLDHKLASGPLQ mL15-6 1259 DVESLEQLLMWLDHQLASGPHG mL15-7 1260 QVDSLQQVLLWLEHIKLALGPQV mL15-8 1261 GDESLQIILLMWLEQKLALGPHG mL15-9 1262 QIEMLESLLDLLRDMVPMSNAF mL15-10 1263 VDSLQQLLMWLDQKLASGPQA mL15-11 1264 EDESLQQLLIYLDKMLSSGPQV mL15-12 1265 AMDQLHQLLIWLDHKLASGPQA mL15-13 1266 RIEMLESLLELLDEIALIPKAW mL15-14 1267 EVVSLQHLLMWLEHKLASGPDG mL15-15 1268 GGESLQQLLMWLDQQLASGPQR mL15-16 1269 GVESLQQLLIFLDIMLVSGpHD mL15-17 1270 NVESLEHLMMWLERLLASGPYA mL15-18 1271 QVDSLQQLLIWLDHQLASGPKR mL15-19 1272 EVESLQQLLMWLEHKLAQGPQG mL15-20 1273 EVDSLQQLLMWLDQKLASGPHA mL15-21 1274 EVDSLQQLLMWLDQQLASGPQK mL15-22 1275 GVEQLPQLLMWLEQKLASGPQR mL15-23 1276 GEDSLQQLLMWLDQQLAAGPQV mL15-24 1277 ADDSLQQLLMWLDRKLASGPHV mL15-25 1278 PVDSLQQLLIWLDQKLASGPQG L-17 1279 RATLLKDFWQLVEGYGDN nL17-conl 1280 DWRATLLKEFWQLVEGLGDNLV mL17-con2 1281 QSRATLLKEFWQLVEGLGDKQA mnL17-1 1282 DGRATLLTEFWQLVQGLGQKEA mL17-2 1283 LARATLLKEFWQLVEGLGEKVV mL17-3 1284 GSRDTLLKEFWQLVVGLGDMQT mL17-4 1285 DARATLLKEFWQLVDAYGDRMV mL17-5 1286 NDRAQLLRDFWQLVDGLGVKSW mL17-6 1287 GVRETLLYELWYLLKGLGANQG mL17-7 1288 QARATLLKEFCQLVGCQGDKLS mL17-8 1289 QERATLLKEFWQLVAGLGQNMR mL17-9 1290 SGRATLLKEFWQLVQGLGEYRW mL17-10 1291 TMRATLLKEFWLFVDGQREMQW mL17-11 1292 GERATLLNDFWQLVDGQGDNTG mL17-12 1293 DERETLLKEFWQLVHGWGDNVA mL17-13 1294 GGRATLLKELWQLLEGQGANLV mL17-14 1295 TARATLLNELVQLVKGYGDKLV mL17-15 1295 GMRATLLQEFWQLVGGQGDNWM mL17-16 1297 STRATLLNDLWQLMKGWAEDRG mL17-17 1298 SERATLLKELWQLVGGWGDNFG mL17-18 1299 VGRATLLKEFWQLVEGLVGQSR mL17-19 1300 EIRATLLKEFWQLVDEWREQPN mL17-20 1301 QLRATLLKEFLQLVHGLGETDS WO 2006/010057 PCT/US2005/024373 58 mL17-21 1302 TQRATLLKEFWQLIEGLGGKHV mL17-22 1303 HYRATLLKEFWQLVDGLREQGV mL17-23 1304 QSRVTLLREFWQLVESYRPIVN mL17-24 1305 LSRATLLNEFWQFVDGQRDKRM mL17-25 1306 WDRATLLNDFWHLMEELSQKPG niL17-26 1307 QERATLLKEFWRMVEGLGKNRG niL17-27 1308 NERATLLREFWQLVGGYGVNQR L-20 1309 YREMSMLEGLLDVLERLQHY mL20-1 1310 HQRDMSMLWELLDVLDGLRQYS mL20-2 1311 TQRDMSMLDGLLEVLDQLRQQR niL20-3 1312 TSRDMSLLWELLEELDRLGHQR mL20-4 1313 MQHDMSMLYGLVELLESLGHQI mL20-5 1314 WNRDMRMLESLFEVLDCiLRQQV mL20-6 1315 GYRDMSMLEGLLAVLDRLGPQL mL20 con1 1316 TQRDMSMLEGLLEVLDRLGQQR niL20 con2 1317 WYRDMSMLEGLLEVLDRLGQQR L-21 1318 HNSSQMLLSELIMLVGSMMQ mL21-1 1319 TQNSRQMLLSDFMMLVGSMIQG mL21-2 1320 MQTSRHILLSEFMMLVGSIMHlG mL21-3 1321 HDNSRQMLLSDLLHLVGTMIQG mL21-4 1322 MENSRQNLLRELIMLVGNMSHQ mL21-5 1323 QDTSRHMLLREFMMLVGEMIQG mL21 con1 1324 DQNSRQMLLSDLMILVGSMIQG L-24 1325 EFFHWLHNHRSEVNHWLDMN mL24-1 1326 NVFFQWVQKHGRVVYQWLDINV mL24-2 1327 FDFLQWLQNHRSEVEHWLVMDV TABLE 24 MYOSTATIN INHIBITOR PEPTIDES Peptibody Name Peptide 2x nTN8-Con6- M-GAQ-WYPCYEGHFWCYDL (N)-1K GSGSATGGSGSTASSGSGSATG-WYPCYEGHFWCYDL LE-5G-FC (SEQ ID NO: 1328) 2x mTN8-Con6- FC-5G-AQ-WYPCYEGHFWCYDL (C)-1K GSGSATGGSGSTASSGSGSATG- WYPCYEGHFWCYDL LE (SEQ ID NO: 1329) WO 2006/010057 PCT/US2005/024373 59 2x nTN8-Con7- M-GAQ-IFGCKWWDVQCYQF (N)-1K GSGSATGGSGSTASSGSGSATG-IFGCKWWDVQCYQF LE-5G-FC (SEQ ID NO: 1330) 2x mTN8-Con7- FC-5G-AQ-IFGCKWWDVQCYQF (C)-IK GSGSATGGSGSTASSGSGSATG-IFGCKWVDVQCYQF LE (SEQ ID NO: 1331) 2x mTN8-Con8- M-GAQ-IFGCKWWDVDCYQF (N)-lK GSGSATGGSGSTASSGSGSATG-IFGCKWWDVDCYQF LE-5G-FC (SEQ ID NO: 1332) 2x mTN8-Con8- FC-5G-AQ-IFGCKWWDVDCYQF (C)-IK GSGSATGGSGSTASSGSGSATG- IFGCKWWDVDCYQF LE (SEQ ID NO: 1333) 2X mTN8-19-7 FC-5G-AQ LADHGQCIRWPWMCPPEGWELEGSGSATGGSGSTASSG SGSATGLADHGQCIRWPWMCPPEGWE-LE (SEQ ID NO: 1334) 2X mTN8-19-7 FC-5G-AQ ST-GG del2x LADHGQCIRWPWMCPPEGWEGSGSATGGSGGGASSGSG LE SATGLADHGQCIRWPWMCPPEGWE (SEQ ID NO: 1335) 2X mTN8-19-21 FC-5G-AQ SEYQGLCTRWPWMCPPQGWKLEGSGSATGGSGSTASSG SGSATGSEYQGLCTRWPWMCPPQGWK -LE (SEQ ID NO: 1336) WO 2006/010057 PCT/US2005/024373 60 2X mTN8-19-21 FC-5G-AQ ST-GG del2x SEYQGLCTRWPWMCPPQGWKGSGSATGGSGGGASSGS LE GSATGSEYQGLCTRWPWMCPPQGWK (SEQ ID NO: 1337) 2X mTN8-19-22 FC-50-AQ TFSQGHCTRWPWMCPPQGWGLEGSGSATGGSGSTASSG SGSATGTFSQGHCTRWPWMCPPQGWG -L E (SEQ ID NO: 1338) 2X mTN8-19-32 FC-5G-AQ VADHGHCTRWPWMCPPQGWGLEGSGSATGGSGSTASS GSGSATGVADHGHCTRWPWMCPPQGWG-LE (SEQ ID NO: 1339) 2X mTN8-19-32 FC-5G-AQ ST-GG del2x VADHGHCTRWPWMCPPQGWGGSGSATGGSGGGASSGS LE GSATGVADHGHCTRWPWVCPPQGWG (SEQ ID NO: 1340) 2X mTN8-19-33 FC-5G-AQ PESQGHCTRWPWMCPPQGWGLEGSGSATGGSGSTASSG SGSATGPESQGHCTRWPWMCPPQGWGLE (SEQ ID NO: 1341) 2X mTN8-19-33 FC-5G-AQ ST-GG del2x PESQGHCTRWPWMCPPQGWGGSGSATGGSGGGASSGS LE GSATGPESQGHCTRWPWMCP PQGWG (SEQ ID NO: 1342) WO 2006/010057 PCT/US2005/024373 61 Table 25 Integrin-antagonist peptide sequences Sequence/structure SEQ. ID NO: CLCRGDCIC 1344 CWDDGWLC 1345 CWDDLWWLC 1346 CWDDGLMC 1347 CWDDGWMC 1348 CSWDDGWLC 1349 CPDDLWWLC 1350 NGR 1351 GSL 1352 RGD 1353 CGRECPRLCQSSC 1354 CNGRCVSGCAGRC 1355 CLSGSLSC 1356 GSL 1357 NGRAHA 1358 CNGRC 1359 CDCRGDCFC 1360 CGSLVRC 1361 DLXXL 1362 RTDLDSLRTYTL 1363 RTDLDSLRTY 1364 RTDLDSLRT 1365 RTDLDSLR 1366 GDLDLLKLRLTL 1367 GDLHSLRQLLSR 1368 RDDLHMLRLQLW 1369 SSDLHALKKRYG 1370 RGDLKQLSELTW 1371 CXXRGDC 1372 STGGFDDVYDWARGVSSALTTTLVATR 1373 STGGFDDVYDWARRVSSALTTTLVATR 1374 SRGVNFSEWLYDMSAAMKEASNVFPSRRSR 1375 SSQNWDMEAGVEDLTAAMLGLLSTIISSSR 1376 SSPSLYTQFLVNYESAATRIQDLLIASRPSR 1377 SSTGWVDLLGALQRAADATRTSIPPSLQNSR 1378 DVYTKKELIECARRVSEK 1379 RGDGX 1380 CRGDGXC 1381 CARRLDAPC 1382 CPSRLDSPC 1383 CDCRGDCFC 1384 CDCRGDCLC 1385 RGDLAALSAPPV 1386 WO 2006/010057 PCT/US2005/024373 62 Table 26 Selectin antagonist peptide sequences Sequence/structure SEQ ID NO: DITWDQLWDLMK 1387 DITWDELWKIMN 1388 DYTWFELWDMMQ 1389 QITWAQLWNMMK 1390 DMTWHDLWTLMS 1391 DYSWHDLWEMMS 1392 EITWDQLWEVMN 1393 HVSWEQLWDIMN 1394 HITWDQLWRIMT 1395 RNMSWLELWEHMK 1396 AEWTWDQLWHVMNPAESQ 1397 HRAEWLALWEQMSP 1398 KKEDWLALWRIMSV 1399 ITWDQLWDLMK 1400 DITWDQLWDLMK 1401 DITWDQLWDLMK 1402 DITWDQLWDLMK 1403 CQNRYTDLVAIQNKNE 1404 AENWADNEPNNKRNNED 1405 RKNNKTWTWVGTKKALTNE 1406 KKALTNEAENWAD 1407 CQXRYTDLVAIQNKXE 1408 AENWADGEPNNKXNXED 1409 Table 27 Vinculin binding peptides Sequence/structure SEQ ID NO: SSQNWDMEAGVEDLTAAMLGLLSTIHSSSR 1410 SSPSLYTQFLVNYESAATRIQDLLIASRPSR 1411 SSTGWVDLLGALQRAADATRTSIPPSLQNSR 1412 DVYTKKELIECARRVSEK 1413 STGGFDDVYDWARGVSSALTTTLVATR 1414 STGGFDDVYDWARRVSSALTTTLVATR 1415 SRGVNFSEWLYDMSAAMKEASNVFPSRRSR 1416 WO 2006/010057 PCT/US2005/024373 63 Table 28 Laminin-related peptide sequences Sequence/structure SEQ ID NO: YIGSRYIGSR [i.e., (YIGSR)2] 1417 YIGSRYIGSRYIGSR [i.e., (YIGSR) 3 ] 1418 YIGSRYIGSRYIGSRYIGSR [i.e., (YIGSR) 4 ] 1419 YIGSRYIGSRYIGSRYIGSRYIGSR [i.e., (YIGSR) 5 1 1420 IPCNNKGAHSVGLMWWMLAR 1421 YIGSRREDVEILDVPDSGR 1422 RGDRGDYIGSRRGD 1423 YIGSRYIGSRYIGSRYIGSRYIGSR 1424 REDVEILDVYIGSRPDSGR 1425 YIGSRREDVEILDVPDSGR 1426 Table 29 NGF Modulating Peptides SEQ ID NO: Sequence of Peptide Portion of Fc-Peptide Fusion Product 1427 TGYTEYTEEWPMGFGYQWSF 1428 TDWLSDFPFYEQYFGLMPPG 1429 FMRFPNPWKLVEPPQGWYYG 1430 VVKAPHFEFLAPPHFHEFPF 1431 FSYTWIDETPSNIDRYMLWL 1432 VNFPKVPEDVEPWPWSLKLY 1433 TWHPKTYEEFALPFFVPEAP t434 WHFGTPYIQQQPGVYWLQAP 1435 VWNYGPFFMNFPDSTYFLHE 1436 WRIHSKPLDYSHVWFFPADF 1437 FWDGNQPPDILVDWPWNPPV 1438 FYSLEWLKDHSEFFQTVTEW 1439 QFMELLKFFNSPGDSSHHFL 1440 TNVDWISNNWEHMKSFFTED 1441 PNEKPYQMQSWFPPDWPVPY 1442 WSHTEWVPQVWWKPPNHFYV 1443 WGEWINDAQVHMBEGFISES 1444 VPWEHDHDLWEIISQDWHIA 1445 VLHLQDPRGWSNFPPGVLEL 1446 IHGCWFTEEGCVWQ 1447 YMQCQFARDGCPQW 1448 KLQCQYSESGCPTI 1449 FLQCEISGGACPAP 1450 KLQCEFSTSGCPDL 1451 KLQCEFSTQGCPDL 1452 KLQCEFSTSGCPWL 1453 IQGCWFTEEGCPWQ 1454 SFDCDNPWGHVLQSCFGF 1455 SFDCDNPWGHKLQSCFGF WO 2006/010057 PCT/US2005/024373 64 Table 30 TALL MODULATING PEPTIDES Sequence/structure SEQ ID NO: LPGCKWDLLIKQWVCDPL-A-V' 1456 V'- A - LPGCKWDLLIKQWVCDPL 1457 LPGCKWDLLIKQWVCDPL - A - 1458 LPGCKWDLLIKQWVCDPL - A -V' V'- A - LPGCKWDLLIKQWVCDPL - A - 1459 LPGCKWDLLIKQWVCDPL SADCYFDILTKSDVCTSS- A -V 1 1460 V'- A - SADCYFDILTKSDVCTSS 1461 SADCYFDILTKSDVTSS- A - 1462 SADCYFDILTKSDVTSS - A -V'

V

1 - A - SADCYFDILTKSDVTSS - A - 1463 SADCYFDILTKSDVTSS FHDCKWDLLTKQWVCHGL- A -V' 1464 V'- A - FHDCKWDLLTKQWVCHGL 1465 FHDCKWDLLTKQWVCHGL - A - 1466 FHDCKWDLLTKQWVCHGL - A -V 1

V

1 - A - FHDCKWDLLTKQWVCHGL - A - 1467 FHDCKWDLLTKQWVCHGL Table 31 TALL-1 inhibitory peptibodies. Peptibody Peptibody Peptide Sequence SEQ ID NO TALL-1-8-1-a 1468 MPGTCFPFPW ECTHAGGGGG VDKTHTCPPC PAPELLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYNST YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY TLPPSRDELT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK TALL-1-8-2-a 1469 MWGACWPFPW ECFKEGGGGG VDKTHTCPPC PAPELLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYNST YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY TLPPSRDELT KNQVSLTCLV KGFYPSDLAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK TALL-1-8-4-a 1470 MVPFCDLLTK HCFEAGGGGG VDKTHTCPPC PAPELLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYNST YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY TLPPSRDELT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK TALL-1-12-4- 1471 MGSRCKYKWD VLTKQCFHHG GGGGVDKTHT CPPCPAPELL a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR WO 2006/010057 PCT/US2005/024373 65 DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12-3- 1472 MLPGCKWDLL IKQWVCDPLG GGGGVDKTHT CPPCPAPELL a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12-5- 1473 MSADCYFDIIL TKSDVCTSSG GGGG VDKTHT CPPCPAPELL a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAP]EKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12-8- 1474 MSDDCMYDQL TRMFICSNLG GGGGVDKTHT CPPCPAPELL GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12-9- 1475 MDLNCKYDEL TYKEWCQFNG GGGGVDKTHT CPPCPAPELL a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12- 1476 MFHDCKYDLL TRQMVCHGLG GGGGVDKTHT CPPCPAPELL 10-a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12- 1477 MRNHCFWDHL LKQDICPSPG GGGGVDKTHT CPPCPAPELL 11-a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1-12- 1478 MANQCWWDSL TKKNVCEFFG GGGGVDKTHT CPPCPAPELL 14-a GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEALHNH YTQKSLSLSP GK TALL-1- 1479 MFHDCKWDLL TKQWVCHGLG GGGGVDKTHT CPPCPAPELL consensus GGPSVFLFPP KPKDTLMISR TPEVTCVVVD VSBEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRVVSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SVMHEfALHNH YTQKSLSLSP GK TALL-1 12-3 1480 MLPGCKWDLL IKQWVCDPLG SGSATGGSGS TASSGSGSAT WO 2006/010057 PCT/US2005/024373 66 tandem dimer HMLPGCKWDL LIKQWVCDPL GGGGGVDKTH TCPPCPAPEL LGGPSVFLFP PKPKDTLMIS RTPEVTCVVV DVSHEDPEVK FNWYVDGVEV HNAKTKPREE QYNSTYRVVS VLTVLHQDWL NGKEYKCKVS NKALPAPIEK TISKAKGQPR EPQVYTLPPS RDELTKNQVS LTCLVKGFYP SDIAVEWESN GQPENNYKTT PPVLDSDGSF FLYSKLTVDK SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PGK TALL-1 1481 MFHDCKWDLL TKQWVCHGLG SGSATGGSGS TASSGSGSAT consensus HMIFHDCKWDL LTKQWVCHGL GGGGGVDKTH TCPPCPAPEL tandem dimer LGGPSVFLFP PKPKDTLMIS RTPEVTCVVV DVSHEDPEVK FNWYVDGVEV HNAKTKPREE QYNSTYRVVS VLTVLHQDWL NGKEYKCKVS NKALPAPIEK TISKAKGQPR EPQVYTLPPS RDELTKNQVS LTCLVKGFYP SDIAVEWESN GQPENNYKTT PPVLDSDGSF FLYSKLTVDK SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PGK TABLE 32 ANG-2 INHIBITOR PEPTIDES PEPTIDE SEQ ID NO. PEPTIDE SEQUENCE Con4-44 1482 PIRQEECDWDPWTCEHMWEV Con4-40 1483 TNIQEECEWDPWTCDHMPGK Con4-4 1484 WYEQDACEWDPWTCEHMAEV Con4-31 1485 NRLQEVCEWDPWTCEHMENV Con4-C5 1486 AATQEECEWDPWTCEHMPRS Con4-42 1487 LRHQEGCEWDPWTCEHMFDW Con4-35 1488 VPRQKDCEWDPWTCEHMYVG Con4-43 1489 SISHEECEWDPWTCEHMQVG Con4-49 1490 WAAQEECEWDPWTCEHMGRM Con4-27 1491 TWPQDKCEWDPWTCEHMGST Con4-48 1492 GHSQEECGWDPWTCEIHMGTS Con4-46 1493 QHWQEECEWDPWTCDHMPSK Con4-41 1494 NVRQEKCEWDPWTCEHMPVR Con4-36 1495 KSGQVECNWDPWTCEHMPRN Con4-34 1496 VKTQEHCDWDPWTCEHMREW Con4-28 1497 AWGQEGCDWDPWTCEHMILPM Con4-39 1498 PVNQEDCEWDPWTCEHMPPM Con4-25 1499 RAPQEDCEWDPWTCAHMDIK Con4-50 1500 HGQNMECEWDPWTCEHMFRY Con4-38 1501 PRLQEECVWDPWTCEHMPLR Con4-29 1502 RTTQEKCEWDPWTCEHMESQ Con4-47 1503 QTSQEDCVWDPWTCDHMVSS WO 2006/010057 PCT/US2005/024373 67 Con4-20 1504 QVIGRPCEWDPWTCEHLEGL Con4-45 1505 WAQQEECAWDPWTCDHMVGL Con4-37 1506 LPGQEDCEWDPWTCEHMVRS Con4-33 1507 PMNQVECDWDPWTCEHNVRS AC2-Con4 1508 FGWSHGCEWDPWTCEHMGST Con4-32 1509 KSTQDDCDWDPWTCEHMVGP Con4-17 1510 UPRISTCQWDPWTCEEEMDQL Con4-8 1511 STIGDMCEWDPWTGAIMQVD AC4-Con4 1512 VLGGQGCEWDPWTCRLLQGW Con4-1 1513 VLGGQGCQWDPWTCSHLEDG Con4-C1 1514 TTIGSMCEWDPWTCAHMQGG Con4-21 1515 TKGKSVCQWDPWTCSHMQSG Con4-C2 1516 TTIGSMCQWDPWTCAHMQGG Con4-18 1517 WVNEVVCEWDPWTCNI{WDTP Con4-19 1518 VVQVGMCQWDPWTCKHMRLQ Con4-16 1519 AVGSQTCEWDPWTCAHLYEV Con4-11 1520 QGMKMFCEWDPWTCAIIIVYR Con4-C4 1521 TTIGSMCQWDPWTCEHMQGG Con4-23 1522 TSQRVGCEWDPWTCQHLTYT Con4-15 1523 QWSWPPCEWDPWTCQTVWPS Con4-9 1524 GTSPSFCQWDPWTCSHMVQG TN8-Con4* 1525 QEECEWDPWTCEHM TABLE 33 ANG-2 INHIBITOR PEPTIDES Peptide SEQ ID NO. Peptide Sequence Li-i 1526 QNYKPLDELDATLYEHFIFHYT L1-2 1527 LNFTPLDELEQTLYEQWTLQQS LI-3 1528 TKFNPLDELEQTLYEQWTLQHQ LI-4 1529 VKFKPLDALEQTLYEIIWMFQQA Li-5 1530 VKYKPLDELDELLYEQQTFQER U1-7 1531 TNFMPIVIDDLEQRLYEQFLQQG L1-9 1532 SKFKPLDELEQTLYEQWTLQHA Li- 10 1533 QKFQPLDELEQTLYEQFMLQQA WO 2006/010057 PCT/US2005/024373 68 Li-11 1534 QNFKPMDELEDTLYKQFLFQHS L1-12 1535 YKFTPLDDLEQTLYEQWTLQHV L1-13 1536 QEYEPLDELDETLYNQWMFHQR L1-14 1537 SNFMPLDELEQTLYEQFMLQHQ Li-15 1538 QKYQPLDELDKTLYDQFMLQQG LI-16 1539 QKFQPLDELEETLYKQWTLQQR LI-17 1540 VKYKPLDELDEWLYHQFrLHHQ L1-18 1541 QKFMPLDELDEILYEQFMFQQS L1-19 1542 QTFQPLDDLEEYLYEQWIRRYH L1-20 1543 EDYMPLDALDAQLYEQFILLHG L1-21 1544 HTFQPLDELEETLYYQWLYDQL L1-22 1545 YKFNPMDELEQTLYEEFLFQHA AC6-L1 1546 TNYKPLDELDATLYEHWILQHS Li-Cl 1547 QKFKPLDELEQTLYEQWTLQQR LI-C2 1548 TKFQPLDELDQTLYEQWTLQQR L1-C3 1549 TNFQPLDELDQTLYEQWTLQQR LI 1550 KFNPLDELEETLYEQFTFQQ TABLE 34 ANG-2 INHIBITOR PEPTIDES Peptide SEQ ID NO. Sequence Conl-l 1551 AGGMRPYDGMLGWPNYDVQA Con1-2 1552 QTWDDPCMHILGPVTWRRCI Conl-3 1553 APGQRPYDGMLGWPTYQRIV Conl-4 1554 SGQLRPCEEIFGCGTQNLAL Con1-5 1555 FGDKRPLECMFGGPIQLCPR Con1-6 1556 GQDLRPCEDMFGCGTKDWYG Con1 1557 KRPCEEIFGGCTYQ WO 2006/010057 PCT/US2005/024373 69 TABLE 35 ANG-2 INHIBITOR PEPTIDES Peptide SEQ ID NO: Sequence 12-9-1 1558 GFEYCDGMEDPFTFGCDKQT 12-9-2 1559 KLEYCDGMEDPFTQGCDNQS 12-9-3 1560 LQEWCEGVEDPFTFGCEKQR 12-9-4 1561 AQDYCEGMEDPFTFGCEMQK 12-9-5 1562 LLDYCEGVQDPFTFGCENLD 12-9-6 1563 HQEYCEGMEDPFTFGCEYQG 12-9-7 1564 MLDYCEGMDDPFTFGCDKQM 12-9-C2 1565 LQDYCEGVEDPFTFGCENQR 12-9-Cl 1566 LQDYCEGVEDPFTFGCEKQR 12-9 1567 FDYCEGVEDPFTFGCDNH Table 36 Ang-2 Binding Peptides Peptide Seq Id No. Sequence TN8-8 1568 KRPCEEMWGGCNYD TN8-14 1569 HQICKWDPWTCKHW TN8-Conl 1570 KRPCEEIFGGCTYQ TN8-Con4 1571 QEECEWDPWTCEHM TN12-9 1572 FDYCEGVEDPFTFGCDNH L1 1573 KFNPLDELEETLYEQFTFQQ C17 1574 QYGCDGFLYGCMIN WO 2006/010057 PCT/US2005/024373 70 Table 37 Ang-2 Binding Peptides Peptibody Peptibody Sequence Li (N) MGAQKFNPLDELEETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 1575) LI (N) WT MKFNPLDELEETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 1576) LI (N) 1K WT MKFNPLDELEETLYEQFTFQQGSGSATGGSGSTASSGSGSAT HLEGGGGG-Fc (SEQ ID NO: 1577) 2xL1 (N) MGAQKFNPLDELEETLYEQFrFQQGGGGGGGGKFNPLDELE ETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 1578) 2xL1 (N) WT MKFNPLDELEETLYEQFTFQQGGGGGGGKFNPLDELEETLYE QFTFQQLEGGGGG-Fc (SEQ ID NO:1579) Con4 (N) MGAQQEECEWDPWTCEHMLEGGGGG-Fc (SEQ ID NO:1580) Con4 (N) 1K-WT MQEECEWDPWTCEHMGSGSATGGSGSTASSGSGSATHLEGG GGG-Fc (SEQ ID NO:1581) 2xCon4 (N) 1K MGAQQEECEWDPWTCEHMGSGSATGGSGSTASSGSGSATH QEECEWDPWTCEHMLEGGGGG-Fc (SEQ ID NO:1582) LI (C) M-Fc-GGGGGAQKFNPLDELEETLYEQFTFQQLE (SEQ ID NO:1583) Li (C) 1K M-Fc GGGGGAQGSGSATGGSGSTASSGSGSATHKFNPLDELEETLY EQFTFQQLE (SEQ ID NO:1584) 2xL1 (C) M-Fc GGGGGAQKFNPLDELEETLYEQFTFQQGGGGGGGGKFNPLD ELEETLYEQFTFQQLE (SEQ ID NO: 1585) Con4 (C) M-Fc-GGGGGAQQEECEWDPWTCEHMLE (SEQ ID NO:1586) Con4 (C) 1K M-Fc GGGGGAQGSGSATGGSGSTASSGSGSATHQEECEWDPWTCE HMLE (SEQIDNO:1587) M-Fc 2xCon4 (C) 1K GGGGGAQQEECEWDPWTCEHMGSGSATGGSGSTASSGSGS ATHQEECEWDPWTCEHMLE (SEQ ID NO:1588) Con4-L1 (N) MGAQEECEWDPWTCEHMGGGGGGGGKFNPLDELEETLYEQ FTFQQGSGSATGGSGSTASSGSGSATHLEGGGGG-Fc (SEQ .D NO: 1589) Con4-L1 (C) M-Fc GGGGGAQGSGSATGGSGSTASSGSGSATHKFNPLDELEETLY EQFTFQQGGGGGQEECEWDPWTCEHMLE (SEQ ID NO:1590) WO 2006/010057 PCT/US2005/024373 71 TN-12-9 (N) MGAQ-FDYCEGVEDPFTFGCDNHLE-GGGGG-Fc (SEQ ID NO:1591) MGAQ-QYGCDGFLYGCMINLE-GGGGG-Fc (SEQ ID C17 (N) NO:1592) MGAQ-KRPCEEMWGGCNYDLEGGGGG-Fc (SEQ ID TN8-8 (N) NO:1593) MGAQ-HQICKWDPWTCKHWLEGGGGG-Fc (SEQ ID TN8-14 (N) NO:1594) Con1 (N) MGAQ-KRPCEEIFGGCTYQLEGGGGG-Fc (SEQ ID NO: 1595) Table 38 Ang-2 Binding Peptides Con4 Derived Affinity-Matured Peptibody Sequence (Seq Id No:) Pbs M-Fe-GGGGGAQ-PIRQEECDWDPWTCEHMWEV-LE Con4-44 (C) (SEQ ID NO: 1596) M-Fc-GGGGGAQ-TNIQEECEWDPWTCDHMPGK-LE Con4-40 (C) (SEQ ID NO: 1597) M-Fc-GGGGGAQ-WYEQDACEWDPWTCEHMAEV-LE Con4-4 (C) (SEQ ID NO: 1598) M-Fc-GGGGGAQ-NRLQEVCEWDPWTCEHMENV-LE Con4-31 (C) (SEQ ID NO: 1599) M-Fc-GGGGGAQ-AATQEECEWDPWTCEHMPRS-LE Con4-C5 (C) (SEQ ID NO: 1600) M-Fe-GGGGGAQ-LRHQEGCEWDPWTCEHMFDW-LE Con4-42 (C) (SEQ ID NO: 1602) M-Fc-GGGGGAQ-VPRQKDCEWDPWTCEHMYVG-LE Con4-35 (C) (SEQ ID NO: 1602) M-Fc-GGGGGAQ-SISHEECEWDPWTCEHMQVG-LE Con4-43 (C) (SEQ ID NO: 1603) M-Fc-GGGGGAQ-WAAQEECEWDPWTCEHMGRM-LE Con4-49 (C) (SEQ ID NO: 1604) M-Fc-GGGGGAQ-TWPQDKCEWDPWTCEHMGST-LE Con4-27 (C) (SEQ ID NO: 1605) M-Fc-GGGGGAQ-GHSQEECGWDPWTCEHMGTS-LE Con4-48 (C) (SEQ ID NO: 1606) M-Fc-GGGGGAQ-QHWQEECEWDPWTCDHMPSK-LE Con4-46 (C) (SEQ ID NO: 1607) M-Fc-GGGGGAQ-NVRQEKCEWDPWTCEHMPVR-LE Con4-41 (C) (SEQ ID NO: 1608) M-Fc-GGGGGAQ-KSGQVECNWDPWTCEHMPRN-LE Con4-36 (C) (SEQ ID NO: 1609) M-Fc-GGGGGAQ-VKTQEHCDWDPWTCEHMREW-LE Con4-34 (C) (SEQ ID NO: 1610) M-Fc-GGGGGAQ-AWGQEGCDWDPWTCEHMLPM-LE Con4-28 (C) (SEQ ID NO: 1611) M-Fc-GGGGGAQ-PVNQEDCEWDPWTCEHMPPM-LE Con4-39 (C) (SEQ ID NO: 1612) WO 2006/010057 PCT/US2005/024373 72 M-Fc-GGGGGAQ-RAPQEDCEWDPWTCAHMDIK-LE Cou4-25 (C) (SEQ ID NO: 1613) M-Fc-GGGGGAQ-HGQNMECEWDPWTCEHMFRY-LE Con4-50 (C) (SEQ ID NO: 1614) M-Fc-GGGGGAQ-PRLQEECVWDPWTCEHMPLR-LE Con4-38 (C) (SEQ ID NO: 1615) M-Fc-GGGGGAQ-RTTQEKCEWDPWTCEHMESQ-LE Con4-29 (C) (SEQ ID NO: 1616) M-Fc-GGGGGAQ-QTSQEDCVWDPWTCDHMVSS-LE Con4-47 (C) (SEQ ID NO: 1617) M-Fc-GGGGGAQ-QVIGRPCEWDPWTCEHLEGL-LE Con4-20 (C) (SEQ ID NO: 1618) M-Fc-GGGGGAQ-WAQQEECAWDPWTCDHMVGL-LE Con4-45 (C) (SEQ ID NO: 1619) M-Fc-GGGGGAQ-LPGQEDCEWDPWTCEHMVRS-LE Con4-37 (C) (SEQ ID NO: 1620) M-Fc-GGGGGAQ-PMNQVECDWDPWTCEHMPRS-LE Con4-33 (C) (SEQ ID NO: 1621) M-Fc-GGGGGAQ-FGWSHGCEWDPWTCEHMGST-LE AC2-Con4 (C) (SEQ ID NO: 1622) M-Fc-GGGGGAQ-KSTQDDCDWDPWTCEHMVGP-LE Con4-32 (C) (SEQ ID NO: 1623) M-Fc-GGGGGAQ-GPRISTCQWDPWTCEHMDQL-LE Con4-17 (C) (SEQ ID NO: 1624) M-Fc-GGGGGAQ-STIGDMCEWDPWTCAHMQVD-LE Con4-8 (C) (SEQ ID NO: 1625) M-Fc-GGGGGAQ-VLGGQGCEWDPWTCRLLQGW-LE AC4-Con4 (C) (SEQ ID NO: 1626) M-Fc-GGGGGAQ-VLGGQGCQWDPWTCSILEDG-LE Con4-1 (C) (SEQ ID NO: 1627) M-Fc-GGGGGAQ-TTIGSMCEWDPWTCAHMQGG-LE Con4-C1 (C) (SEQ ID NO: 1628) M-Fc-GGGGGAQ-TKGKSVCQWDPWTCSHMQSG-LE Con4-21 (C) (SEQ ID NO: 1629) M-Fc-GGGGGAQ-TTIGSMCQWDPWTCAHMQGG-LE Con4-C2 (C) (SEQ ID NO: 1630) M-Fc-GGGGGAQ-WVNEVVCEWDPWTCNHWDTP-LE Con4-18 (C) (SEQ ID NO: 1631) M-Fc-GGGGGAQ-VVQVGMCQWDPWTCKHMRLQ-LE Con4-19 (C) (SEQ ID NO: 1632) M-Fc-GGGGGAQ-AVGSQTCEWDPWTCAHLVEV-LE Con4-16 (C) (SEQ ID NO: 1633) M-Fc-GGGGGAQ-QGMKMFCEWDPWTCAHIVYR-LE Con4-11 (C) (SEQ ID NO: 1634) M-Fc-GGGGGAQ-TTIGSMCQWDPWTCEHMQGG-LE Con4-C4 (C) (SEQ ID NO: 1635) M-Fc-GGGGGAQ-TSQRVGCEWDPWTCQHLTYT-LE Con4-23 (C) (SEQ ID NO: 1636) M-Fc-GGGGGAQ-QWSWPPCEWDPWTCQTVWPS-LE Con4-15 (C) (SEQ ID NO: 1637) M-Fc-GGGGGAQ-GTSPSFCQWDPWTCSHMVQG-LE Con4-9 (C) (SEQ ID NO: 1638) M-FC-GGGGGAQ-TQGLHQCEWDPWTCKVLWPS-LE Con4-10 (C) (SEQ ID NO: 1639) M-Fc-GGGGGAQ-VWRSQVCQWDPWTCNLGGDW-LE Con4-22 (C) (SEQ ID NO: 1640) WO 2006/010057 PCT/US2005/024373 73 M-Fc-GGGGGAQ-DKILEECQWDPWTCQFFYGA-LE Con4-3 (C) (SEQ ID NO: 1641) M-Fc-GGGGGAQ-ATFARQCQWDPWTCALGGNW-LE Con4-5 (C) (SEQ ID NO: 1642) M-Fc-GGGGGAQ-GPAQEECEWDPWTCEPLPLM-LE Con4-30 (C) (SEQ ID NO: 1643) M-Fc-GGGGGAQ-RPEDMCSQWDPWTWHLQGYC-LE Con4-26 (C) (SEQ ID NO: 1644) M-Fc-GGGGGAQ-LWQLAVCQWDPQTCDHMGAL-LE Con4-7 (C) (SEQ ID NO: 1645) M-Fc-GGGGGAQ-TQLVSLCEWDPWTCRLLDGW-LE Con4-12 (C) (SEQ ID NO: 1646) M-Fc-GGGGGAQ-MGGAGRCEWDPWTCQLLQGW-LE Con4-13 (C) (SEQ ID NO: 1647) M-Fc-GGGGGAQ-MFLPNECQWDPWTCSNLPEA-LE Con4-14 (C) (SEQ ID NO: 1648) M-Fc-GGGGGAQ-FGWSHGCEWDPWTCRLLQGW-LE Con4-2 (C) (SEQ ID NO: 1649) M-Fc-GGGGGAQ-WPQTEGCQWDPWTCRLLHGW-LE Con4-6 (C) (SEQ ID NO: 1650) M-Fc-GGGGGAQ-PDTRQGCQWDPWTCRLYGMW-LE Con4-24 (C) (SEQ ID NO: 1651) M-Fc-GGGGGAQ-TWPQDKCEWDPWTCRLLQGW-LE ACl-Con4 (C) (SEQ ID NO: 1652) M-Fc-GGGGGAQ-DKILEECEWDPWTCRLLQGW-LE AC3-Con4 (C) (SEQ ID NO: 1653) M-Fe-GGGGGAQ-AATQEECEWDPWTCRLLQGW-LE AC5-Con4 (C) (SEQ ID NO: 1654) L1 Derived Affinity- Peptibody Sequence (Seq Id No:) Matured Pbs MGAQ-TNFMPMDDLEQRLYEQFILQQG-LEGGGGG-Fc L1-7 (N) (SEQ ID NO: 1655) MGAQ-TNYKPLDELDATLYEHWILQHS LEGGGGG-Fc AC6-L1 (N) (SEQ ID NO: 1656) MGAQ-QKYQPLDELDKTLYDQFMLQQG LEGGGGG-Fc L1-15 (N) (SEQ ID NO: 1657) MGAQ-LNFTPLDELEQTLYEQWTLQQS LEGGGGG-Fc L1-2 (N) (SEQ ID NO: 1658) MGAQ-QKFQPLDELEQTLYEQFMLQQA LEGGGGG-Fc L1-10 (N) (SEQ ID NO: 1659) MGAQ-QEYEPLDELDETLYNQWMFHQR LEGGGGG-Fc L1-13 (N) (SEQ ID NO: 1660) MGAQ-VKYKPLDELDEILYEQQTFQER LEGGGGG-Fc L1-5 (N) (SEQ ID NO: 1661) MGAQ-TKFQPLDELDQTLYEQWTLQQR LEGGGGG-Fc L1-C2 (N) (SEQ ID NO: 1662) MGAQ-TNFQPLDELDQTLYEQWTLQQR LEGGGGG-Fc L1-C3 (N) (SEQ ID NO: 1663) MGAQ-QNFKPMDELEDTLYKQFLFQHS LEGGGGG-Fc L1-11 (N) (SEQ ID NO: 1664) MGAQ-VKYKPLDELDEWLYHQFTLHHQ LEGGGGG-Fc L1-17 (N) (SEQ ID NO: 1665) MGAQ-YKFTPLDDLEQTLYEQWTLQHV LEGGGGG-Fc L1-12 (N) (SEQ ID NO: 1666) MGAQ-QNYKPLDELDATLYEHFIFHYT LEGGGGG-Fc L1-1 (N) (SEQ ID NO: 1667) WO 2006/010057 PCT/US2005/024373 74 MGAQ-VKFKPLDALEQTLYEHWMFQQA LEGGGGG-Fc L1-4 (N) (SEQ ID NO: 1668) MGAQ-EDYMLDALDAQLYEQFILLHG LEGGGGG-FC L1-20 (N) (SEQ ID NO: 1669) MGAQ-YKFNPMDELEQTLYEEFLFQHA LEGGGGG-Fc L1-22 (N) (SEQ ID NO: 1670) MGAQ-SNFMPLDELEQTLYEQFMLQHQ LEGGGGG-Fc L1-14 (N) (SEQ ID NO: 1671) MGAQ-QKFQPLDELEETLYKQWTLQQR LEGGGGG-F L1-16 (N) (SEQ ID NO: 1672) MGAQ-QKFMPLDELDEILYEQFWQQS LEGGGGG-Fc L1-18 (N) (SEQ ID NO: 1673) MGAQ-TKFNPLDELEQTLYEQWTLQHQ LEGGGGG-Fc L1-3 (N) (SEQ ID NO: 1674) MGAQ-HTFQPLDELEETLYYQWLYDQL LEGGGGG-Fc L1-21 (N) (SEQ ID NO: 1675) MGAQ-QKFKPLDELEQTLYEQWTLQQR LEGGGGG-Fc L1-C1 (N) (SEQ ID NO: 1676) MGAQ-QTFQPL-DDLEEYLYEQWIRRYH LEGGGGG-Fc L1-19 (N) (SEQ ID NO: 1677) MGAQ-SKFKPLDELEQTLYEQWTLQHA LEGGGGG-Fc L1-9 (N) (SEQ ID NO: 1678) Con1 Derived Affinity- Peptibody Sequence (Seq Id No:) Matured Pbs M-FC-GGGGGAQ-SGQLRPCEEIFGCGTQNLAL-LE Conl-4 (C) (SEQ ID NO: 1679) M-Fe-GGGGGAQ-AGGMRPYDGMLGWPNYDVQA-LE Conl-1 (C) (SEQ ID NO: 1680) M-Fc-GGGGGAQ-GQDLRPCEDMFGCGTKDWYG-LE Conl-6 (C) (SEQ ID NO: 1681) M-Fc-GGGGGAQ-APGQRPYDGMLGWPTYQRIV-LE Conl-3 (C) (SEQ ID NO: 1682) M-Fc-GGGGGAQ-QTWDDPCMHILGPVTWRRCI-LE Con1-2 (C) (SEQ ID NO: 1683) M-Fc-GGGGGAQ-FGDKRPLECMFGGPIQLCPR-LE Conl-5 (C) (SEQ ID NO: 1684) M-Fe-GGGGGAQ-KRPCEEIFGGCTYQ-LE Parent: Con1 (C) (SEQ ID NO: 1685) 12-9 Derived Affinity- Peptibody Sequence (Seq Id No:) Matured Pbs M-Fc-GGGGGAQ-LQEWCEGVEDPFTFGCEKQR-LE 12-9-3 (C) (SEQ ID NO: 1686) M-FC-GGGGGAQ-MLDYCEGMDDPFTFGCDKQM-LE 12-9-7 (C) (SEQ ID NO: 1687) M-Fc-GGGGGAQ-HQEYCEGMEDPFTFGCEYQG-LE 12-9-6 (C) (SEQ ID NO: 1688) M-Fc-GGGGGAQ-LQDYCEGVEDPFTFGCENQR-LE 12-9-C2 (C) (SEQ ID NO: 1689) M-Fc-GGGGGAQ-LLDYCEGVQDPFTFGCENLD-LE 12-9-5 (C) (SEQ ID NO: 1690) M-Fc-GGGGGAQ-GFEYCDGMEDPFTFGCDKQT-LE 12-9-1 (C) (SEQ ID NO: 1691) M-Fc-GGGGGAQ-AQDYCEGMEDPFTFGCEMQK-LE 12-9-4 (C) (SEQ ID NO: 1692) M-Fc-GGGGGAQ-LQDYCEGVEDPFTFGCEKQR-LE 12-9-Cl (C) (SEQ ID NO: 1693) WO 2006/010057 PCT/US2005/024373 75 M-Fc-GGGGGAQ-KLEYCDGMEDPFTQGCDNQS-LE 12-9-2 (C) (SEQ ID NO: 1694) M-Fc-GGGGGAQ-FDYCEGVEDPFTFGCDNH-LE Parent: 12-9 (C) (SEQ ID NO: 1695) [0079] In addition to the TMP compounds set out in Table 6, the invention provides numerous other TMP compounds. In one aspect, TMP compounds comprise the following general structure: TMP1.(Ll)n-TMP 2 wherein TMPI and TMP 2 are each independently selected from the group of compounds comprising the core structure:

X

2

-X

3

-X

4

-X

5

-X

6

-X

7

-X

8

-X

9

-X

1 o, wherein,

X

2 is selected from the group consisting of Glu, Asp, Lys, and Val;

X

3 is selected from the group consisting of Gly and Ala;

X

4 is Pro;

X

5 is selected from the group consisting of Thr and Ser;

X

6 is selected from the group consisting of Leu, Ile, Val, Ala, and Phe;

X

7 is selected from the group consisting of Arg and Lys;

X

8 is selected from the group consisting of Gln, Asn, and Glu;

X

9 is selected from the group consisting of Trp, Tyr, and Phe;

X

10 is selected from the group consisting of Leu, Ile, Val, Ala, Phe, Met, and Lys; Li is a linker as described herein; and n is 0 or 1; and physiologically acceptable salts thereof. [0080] In one embodiment, L 1 comprises (Gly)n, wherein n is 1 through 20, and when n is greater than 1, up to half of the Gly residues may be substituted by another amino acid selected from the remaining 19 natural amino acids or a stereoisomer thereof.

WO 2006/010057 PCT/US2005/024373 76 [0081] In addition to the core structure X2-X 10 set forth above for TMP 1 and TMP2, other related structures are also possible wherein one or more of the following is added to the

TMP

1 and/or TMP 2 core structure: X 1 is attached to the N-terminus and/or X 1 1 , X 1 2, X 13 , and/or X 14 are attached to the C-terminus, wherein X 1 , X 12 , X 13 , and X14 are as follows:

X

1 is selected from the group consisting of Ile, Ala, Val, Leu, Ser, and Arg; XIi is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Ser, Thr, Lys, His, and Glu;

X

12 is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Gly, Ser, and Gln;

X

13 is selected from the group consisting of Arg, Lys, Thr, Val, Asn, Gln, and Gly; and

X

14 is selected from the group consisting of Ala, Ile, Val, Leu, Phe, Thr, Arg, Glu, and Gly. [0082] TMP compounds of the invention are made up of, i.e., comprising, at least 9 subunits (X 2

-X

10 ), wherein X 2

-X

1 0 comprise the core structure. The X 2 -X14 subunits are amino acids independently selected from among the 20 naturally-occurring amino acids, however, the invention embraces compounds where X 2

-X

14 are independently selected from the group of atypical, non-naturally occurring amino acids well known in the art. Specific amino acids are identified for each position. For example, X 2 may be Glu, Asp, Lys, or Val. Both three-letter and single letter abbreviations for amino acids are used herein; in each case, the abbreviations are the standard ones used for the 20 naturally-occurring amino acids or well-known variations thereof. These amino acids may have either L or D stereochemistry (except for Gly, which is neither L nor D), and the TMPs (as well as all other compounds of the invention) may comprise a combination of stereochemistries. The invention also provides reverse TMP molecules (as well as for all other peptides disclosed herein) wherein the amino terminal to carboxy terminal sequence of the amino acids is reversed. For example, the reverse of a molecule having the normal sequence Xi-X 2

-X

3 would be X 3

-X

2

-X

1 . The invention also provides retro-reverse TMP molecules (as well as for all other molecules of the invention described herein) wherein, like a reverse TMP, the amino terminal to carboxy WO 2006/010057 PCT/US2005/024373 77 terminal sequence of amino acids is reversed and residues that are normally "L" enatiomers in TMP are altered to the "D" stereoisomer form. [00831 Exemplary TMP compounds of the invention therefore include without limitation the following compounds. IEGPTLRQWLAARA-GPNG-IEGPTLRQWLAARA (SEQ. ID NO: 993) IEGPTLRQCLAARA-GGGGGGGG-IEGPTLRQCLAARA (cyclic) (SEQ. ID NO: 994) IEGPTLRQCLAARA-GGGGGGGG-IEGPTLRQCLAARA (linear) (SEQ. ID NO: 995) IEGPTLRQALAARA-GGGGGGGG-IEGPTLRQALAARA (SEQ. ID NO: 996) IEGPTLRQWLAARA-GGGKGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 997) IEGPTLRQWLAARA-GGGK(BrAc)GGGG-IEGPTLRQWLAARA (SEQ. ID NO: 998) IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 999) IEGPTLRQWLAARA-GGGK(PEG)GGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1000) IEGPTLRQWLAARA-GGGC(PEG)GGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1001) IEGPTLRQWLAARA-GGGNGSGG-IEGPTLRQWLAARA (SEQ. ID NO: 1002) IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA |IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1003) IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1004) Fc-IEGPTLRQWLAARA-GPNG-IEGPTLRQWLAARA (SEQ. ID NO: 1005) Fc-IEGPTLRQWLAARA-GPNG-IEGPTLRQWLAARA-Fe (SEQ. ID NO: 1006) IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA-Fc (SEQ. ID NO: 1007) Fc-GG-IEGPTLRQWLAARA-GPNG-IEGPTLRQWLAARA (SEQ. ID NO: 1008) Fc-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1009) WO 2006/010057 PCT/US2005/024373 78 Fc-IEGPTLRQCLAARA-GGGGGGGG-IEGPTLRQCLAARA (cyclic) I (SEQ. ID NO: 1010) Fc-IEGPTLRQCLAARA-GGGGGGGG-IEGPTLRQCLAARA (linear) (SEQ. ID NO: 1011) Fc-IEGPTLRQALAARA-GGGGGGGG-IEGPTLRQALAARA (SEQ. ID NO: 1012) Fc-IEGPTLRQWLAARA-GGGKGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1013) Fc-IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1014) Fc-IEGPTLRQWLAARA-GGGNGSGG-IEGPTLRQWLAARA (SEQ. ID NO: 1015) Fc-IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA Fc-IEGPTLRQWLAARA-GGGCGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1016) Fc-GGGGG-IEGPTLRQWLAARA-GGGGGGGG-IEGPTLRQWLAARA (SEQ. ID NO: 1017) [0084] The present invention is also particularly useful with peptides having activity in treatment of: e cancer, wherein the peptide is a VEGF-mirnetic or a VEGF receptor antagonist, a HER2 agonist or antagonist, a CD20 antagonist and the like; - asthma, wherein the protein of interest is a CKR3 antagonist, an IL-5 receptor antagonist, and the like; - thrombosis, wherein the protein of interest is a GPIIb antagonist, a GPIIIa antagonist, and the like; e autoimmune diseases and other conditions involving immune modulation, wherein the protein of interest is an IL-2 receptor antagonist, a CD40 agonist or antagonist, a CD40L agonist or antagonist, a thymopoietin mimetic and the like. [0085] Derivatives. The invention also contemplates derivatizing the peptide and/or vehicle portion (as discussed below) of the compounds. Such derivatives may improve the solubility, absorption, biological half life, and the like of the compounds. The moieties may alternatively eliminate or attenuate any undesirable side-effect of the compounds and the like. Exemplary derivatives include compounds in which: WO 2006/010057 PCT/US2005/024373 79 1. The compound or some portion thereof is cyclic. For example, the peptide portion may be modified to contain two or more Cys residues (e.g., in the linker), which could cyclize by disulfide bond formation. For citations to references on preparation of cyclized derivatives, see Table 2. 2. The compound is cross-linked or is rendered capable of cross-linking between molecules. For example, the peptide portion may be modified to contain one Cys residue and thereby be able to form an intermolecular disulfide bond with a like molecule. The compound may also be cross-linked through its C-terminus, as in the molecule shown below. HO

F

1 -(X')b-CO-N

NH

2

F-(X

1 H)b-CON NH 0 3. One or more peptidyl [-C(O)NR-] linkages (bonds) is replaced by a non peptidyl linkage. Exemplary non-peptidyl linkages are -CH2-carbamate [-CH2-OC(O)NR-], phosphonate, -CH2-sulfonamide [-CH2-S(O)2NR-], urea [-NHC(O)NH-], -CH2-secondary amine, and alkylated peptide [-C(O)NR6- wherein R6 is lower alkyl]. 4. The N-terminus is derivatized. Typically, the N-terminus may be acylated or modified to a substituted amine. Exemplary N-terminal derivative groups include -NRR1 (other than -NH2), -NRC(O)R1, -NRC(O)OR1, -NRS(O)2R1, -NHC(O)NHR1, succinimide, or benzyloxycarbonyl-NH- (CBZ-NH-), wherein R and RI are each independently hydrogen or lower alkyl and wherein the phenyl ring may be substituted with 1 to 3 substituents selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, chloro, and bromo. 5. The free C-terminus is derivatized. Typically, the C-terminus is esterified or amidated. For example, one may use methods described in the art to add (NH-CH2-CH2 NH2)2 to compounds of this invention. Likewise, one may use methods described in the art to add -NH2 to compounds of this invention. Exemplary C-terminal derivative groups include, for example, -C(O)R2 wherein R2 is lower alkoxy or -NR3R4 wherein R3 and R4 are independently hydrogen or C1-C8 alkyl (preferably CI-C4 alkyl). 6. A disulfide bond is replaced with another, preferably more stable, cross linking moiety (e.g., an alkylene). See, e.g., Bhatnagar et al. (1996), J. Med. Chem. 39: 3814 9; Alberts et al. (1993) Thirteenth Am. Pep. Symp., 357-9.

WO 2006/010057 PCT/US2005/024373 80 7. One or more individual amino acid residues is modified. Various derivatizing agents are known to react specifically with selected sidechains or terminal residues, as described in detail below. [0086] Lysinyl residues and amino terminal residues may be reacted with succinic or other carboxylic acid anhydrides, which reverse the charge of the lysinyl residues. Other suitable reagents for derivatizing alpha-amino-containing residues include imidoesters such as methyl picolinimidate; pyridoxal phosphate; pyridoxal; chloroborohydride; trinitrobenzenesulfonic acid; O-methylisourea; 2,4 pentanedione; and transaminase-catalyzed reaction with glyoxylate. [0087] Arginyl residues may be modified by reaction with any one or combination of several conventional reagents, including phenylglyoxal, 2,3-butanedione, 1,2 cyclohexanedione, and ninhydrin. Derivatization of arginyl residues requires that the reaction be performed in alkaline conditions because of the high pKa of the guanidine functional group. Furthermore, these reagents may react with the groups of lysine as well as the arginine epsilon-amino group. [00881 Specific modification of tyrosyl residues has been studied extensively, with particular interest in introducing spectral labels into tyrosyl residues by reaction with aromatic diazonium compounds or tetranitromethane. Most commonly, N-acetylimidizole and tetranitromethane are used to form O-acetyl tyrosyl species and 3-nitro derivatives, respectively. [00891 Carboxyl sidechain groups (aspartyl or glutamyl) may be selectively modified by reaction with carbodiimides (R'-N=C=N-R') such as 1-cyclohexyl-3-(2-morpholinyl-(4 ethyl) carbodiimide or 1-ethyl-3-(4-azonia-4,4-dimethylpentyl) carbodiimide. Furthermore, aspartyl and glutamyl residues may be converted to asparaginyl and glutaminyl residues by reaction with ammonium ions. [0090] Glutaminyl and asparaginyl residues may be deamidated to the corresponding glutamyl and aspartyl residues. Alternatively, these residues are deamidated under mildly acidic conditions. Either form of these residues falls within the scope of this invention. [0091] Cysteinyl residues can be replaced by amino acid residues or other moieties either to eliminate disulfide bonding or, conversely, to stabilize cross-linking. See, e.g., Bhatnagar et al. (1996), J. Med. Chem. 39: 3814-9.

WO 2006/010057 PCT/US2005/024373 81 [0092] Derivatization with bifunctional agents is useful for cross-linking the peptides or their functional derivatives to a water-insoluble support matrix or to other macromolecular vehicles. Commonly used cross-linking agents include, e.g., 1,1-bis(diazoacetyl)-2 phenylethane, glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with 4 azidosalicylic acid, homobifunctional imidoesters, including disuccinimidyl esters such as 3,3'-dithiobis(succinimidylpropionate), and bifunctional maleimides such as bis-N maleimido-1,8-octane. Derivatizing agents such as methyl-3-[(p azidophenyl)dithio]propioimidate yield photoactivatable intermediates that are capable of forming crosslinks in the presence of light. Alternatively, reactive water-insoluble matrices such as cyanogen bromide-activated carbohydrates and the reactive substrates described in U.S. Pat. Nos. 3,969,287; 3,691,016; 4,195,128; 4,247,642; 4,229,537; and 4,330,440 are employed for protein immobilization. [0093] Carbohydrate (oligosaccharide) groups may conveniently be attached to sites that are known to be glycosylation sites in proteins. Generally, O-linked oligosaccharides are attached to serine (Ser) or threonine (Thr) residues while N-linked oligosaccharides are attached to asparagine (Asn) residues when they are part of the sequence Asn-X-Ser/Thr, where X can be any amino acid except proline. X is preferably one of the 19 naturally occurring amino acids other than proline. The structures of N-linked and 0-linked oligosaccharides and the sugar residues found in each type are different. One type of sugar that is commonly found on both is N-acetylneuraminic acid (referred to as sialic acid). Sialic acid is usually the terminal residue of both N-linked and O-linked oligosaccharides and, by virtue of its negative charge, may confer acidic properties to the glycosylated compound. Such site(s) may be incorporated in the linker of the compounds of this invention and are preferably glycosylated by a cell during recombinant production of the polypeptide compounds (e.g., in mammalian cells such as CHO, BHK, COS). However, such sites may further be glycosylated by synthetic or semi-synthetic procedures known in the art. [0094] Other possible modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of seryl or threonyl residues, oxidation of the sulfur atom in Cys, methylation of the alpha-amino groups of lysine, arginine, and histidine side chains. Creighton, Proteins: Structure and Molecule Properties (W. H. Freeman & Co., San Francisco), pp. 79-86 (1983). [0095] Compounds of the present invention may be changed at the DNA level, as well. The DNA sequence of any portion of the compound may be changed to codons more WO 2006/010057 PCT/US2005/024373 82 compatible with the chosen host cell. For E. coli, which is the preferred host cell, optimized codons are known in the art. Codons may be substituted to eliminate restriction sites or to include silent restriction sites, which may aid in processing of the DNA in the selected host cell. The vehicle, linker and peptide DNA sequences may be modified to include any of the foregoing sequence changes. [0096] Isotope- and toxin-conjugated derivatives. Another set of useful derivatives are the above-described molecules conjugated to toxins, tracers, or radioisotopes. Such conjugation is especially useful for molecules comprising peptide sequences that bind to tumor cells or pathogens. Such molecules may be used as therapeutic agents or as an aid to surgery (e.g., radioimmunoguided surgery or RIGS) or as diagnostic agents (e.g., radioimmunodiagnostics or RID). [0097] As therapeutic agents, these conjugated derivatives possess a number of advantages. They facilitate use of toxins and radioisotopes that would be toxic if administered without the specific binding provided by the peptide sequence. They also can reduce the side effects that attend the use of radiation and chemotherapy by facilitating lower effective doses of the conjugation partner. [0098] Useful conjugation partners include: - radioisotopes, such as 90Yttrium, 131Iodine, 225Actinium, and 213Bismuth; - ricin A toxin, microbially derived toxins such as Pseudomonas endotoxin (e.g., PE38, PE40), and the like; - partner molecules in capture systems (see below); - biotin, streptavidin (useful as either partner molecules in capture systems or as tracers, especially for diagnostic use); and - cytotoxic agents (e.g., doxorubicin). [0099] One useful adaptation of these conjugated derivatives is use in a capture system. In such a system, the molecule of the present invention would comprise a benign capture molecule. This capture molecule would be able to specifically bind to a separate effector molecule comprising, for example, a toxin or radioisotope. Both the vehicle conjugated molecule and the effector molecule would be administered to the patient. In such a system, the effector molecule would have a short half-life except when bound to the vehicle-conjugated capture molecule, thus minimizing any toxic side-effects. The vehicle- WO 2006/010057 PCT/US2005/024373 83 conjugated molecule would have a relatively long half-life but would be benign and non toxic. The specific binding portions of both molecules can be part of a known specific binding pair (e.g., biotin, streptavidin) or can result from peptide generation methods such as those described herein. [00100] Such conjugated derivatives may be prepared by methods known in the art. In the case of protein effector molecules (e.g., Pseudomonas endotoxin), such molecules can be expressed as fusion proteins from correlative DNA constructs. Radioisotope conjugated derivatives may be prepared, for example, as described for the BEXA antibody (Coulter). Derivatives comprising cytotoxic agents or microbial toxins may be prepared, for example, as described for the BR96 antibody (Bristol-Myers Squibb). Molecules employed in capture systems may be prepared, for example, as described by the patents, patent applications, and publications from NeoRx. Molecules employed for RIGS and RID may be prepared, for example, by the patents, patent applications, and publications from NeoProbe. [00101] Vehicles. The invention requires the presence of at least one vehicle attached to a peptide through the N-terminus, C-terminus or a sidechain of one of the amino acid residues. Multiple vehicles may also be used. In one aspect, an Fe domain is the vehicle. The Fc domain may be fused to the N or C termini of the peptides or at both the N and C termini. [00102] In various embodiments of the invention, the Fc component is either a native Fc or an Fe variant. The immunoglobulin source of the native Fe is, in one aspect, of human origin and may, in alternative embodiments, be of any class of immunoglobulin. Native Fe domains are made up of monomeric polypeptides that may be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and/or non-covalent association. The number of intermolecular disulfide bonds between monomeric subunits of native Fe molecules ranges from one to four depending on class (e.g., IgG, IgA, IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgAl, IgGA2). One example of a native Fe is a disulfide-bonded dimer resulting from papain digestion of an IgG (see Ellison et al. (1982), Nucleic Acids Res. 10: 4071-9). [00103] It should be noted that Fe monomers will spontaneously dimerize when the appropriate cysteine residues are present, unless particular conditions are present that prevent dimerization through disulfide bond formation. Even if the cysteine residues that normally form disulfide bonds in the Fe dimer are removed or replaced by other residues, the WO 2006/010057 PCT/US2005/024373 84 monomeric chains will generally form a dimer through non-covalent interactions. The term "Fe" herein is used to mean any of these forms: the native monomer, the native dimer (disulfide bond linked), modified dimers (disulfide and/or non-covalently linked), and modified monomers (i.e., derivatives). [00104] As noted, Fc variants are suitable vehicles within the scope of this invention. A native Fc may be extensively modified to form an Fc variant, provided binding to the salvage receptor is maintained; see, for example WO 97/34631 and WO 96/32478. In such Fe variants, one may remove one or more sites of a native Fc that provide structural features or functional activity not required by the fusion molecules of this invention. One may remove these sites by, for example, substituting or deleting residues, inserting residues into the site, or truncating portions containing the site. The inserted or substituted residues may also be altered amino acids, such as peptidomimetics or D-amino acids. Fc variants may be desirable for a number of reasons, several of which are described below. Exemplary Fe variants include molecules and sequences in which: 1. Sites involved in disulfide bond formation are removed. Such removal may avoid reaction with other cysteine-containing proteins present in the host cell used to produce the molecules of the invention. For this purpose, the cysteine-containing segment at the N terminus may be truncated or cysteine residues may be deleted or substituted with other amino acids (e.g., alanyl, seryl). Even when cysteine residues are removed, the single chain Fe domains can still form a dimeric Fe domain that is held together non-covalently. 2. A native Fe is modified to make it more compatible with a selected host cell. For example, one may remove the PA sequence near the N-terminus of a typical native Fe, which may be recognized by a digestive enzyme in E. coli such as proline iminopeptidase. One may also add an N-terminal methionine residue, especially when the molecule is expressed recombinantly in a bacterial cell such as E. coli. 3. A portion of the N-terminus of a native Fc is removed to prevent N-terminal heterogeneity when expressed in a selected host cell. For this purpose, one may delete any of the first 20 amino acid residues at the N-terminus, particularly those at positions 1, 2, 3, 4 and 5. 4. One or more glycosylation sites are removed. Residues that are typically glycosylated (e.g., asparagine) may confer cytolytic response. Such residues may be deleted or substituted with unglycosylated residues (e.g., alanine).

WO 2006/010057 PCT/US2005/024373 85 5. Sites involved in interaction with complement, such as the Clq binding site, are removed. For example, one may delete or substitute the EKK sequence of human IgG1. Complement recruitment may not be advantageous for the molecules of this invention and so may be avoided with such an Fc variant. 6. Sites are removed that affect binding to Fe receptors other than a salvage receptor. A native Fe may have sites for interaction with certain white blood cells that are not required for the fusion molecules of the present invention and so may be removed. 7. The ADCC site is removed. ADCC sites are known in the art; see, for example, Molec. Immunol. 29 (5): 633-9 (1992) with regard to ADCC sites in IgG1. These sites, as well, are not required for the fusion molecules of the present invention and so may be removed. 8. When the native Fe is derived from a non-human antibody, the native Fc may be humanized. Typically, to humanize a native Fe, one will substitute selected residues in the non-human native Fc with residues that are normally found in human native Fc. Techniques for antibody humanization are well known in the art. [00105] An alternative vehicle would be a protein, polypeptide, peptide, antibody, antibody fragment, or small molecule (e.g., a peptidomimetic compound) capable of binding to a salvage receptor. For example, one could use as a vehicle a polypeptide as described in U.S. Pat. No. 5,739,277, issued April 14, 1998 to Presta et al. Peptides could also be selected by phage display for binding to the FcRn salvage receptor. Such salvage receptor-binding compounds are also included within the meaning of "vehicle" and are within the scope of this invention. Such vehicles should be selected for increased half-life (e.g., by avoiding sequences recognized by proteases) and decreased immunogenicity (e.g., by favoring non-immunogenic sequences, as discovered in antibody humanization). [00106] Variants, analogs or derivatives of the Fe portion may be constructed by, for example, making various substitutions of residues or sequences. [001071 Variant (or analog) polypeptides include insertion variants, wherein one or more amino acid residues supplement an Fe amino acid sequence. Insertions may be located at either or both termini of the protein, or may be positioned within internal regions of the Fe amino acid sequence. Insertion variants, with additional residues at either or both termini, can include for example, fusion proteins and proteins including amino acid tags or WO 2006/010057 PCT/US2005/024373 86 labels. For example, the Fc molecule may optionally contain an N-terminal Met, especially when the molecule is expressed recombinantly in a bacterial cell such as E. coli. [00108] In Fc deletion variants, one or more amino acid residues in an Fc polypeptide are removed. Deletions can be effected at one or both termini of the Fc polypeptide, or with removal of one or more residues within the Fe amino acid sequence. Deletion variants, therefore, include all fragments of an Fc polypeptide sequence. [00109] In Fe substitution variants, one or more amino acid residues of an Fe polypeptide are removed and replaced with alternative residues. In one aspect, the substitutions are conservative in nature and conservative substitutions of this type are well known in the art. Alternatively, the invention embraces substitutions that are also non conservative. [001101 For example, cysteine residues can be deleted or replaced with other amino acids to prevent formation of some or all disulfide crosslinks of the Fc sequences. Each cysteine residue can be removed and/or substituted with other amino acids, such as Ala or Ser. As another example, modifications may also be made to introduce amino acid substitutions to (1) ablate the Fc receptor binding site; (2) ablate the complement (Cq) binding site; and/or to (3) ablate the antibody dependent cell-mediated cytotoxicity (ADCC) site. Such sites are known in the art, and any known substitutions are within the scope of Fc as used herein. For example, see Molecular Immunology, Vol. 29, No. 5, 633-639 (1992) with regard to ADCC sites in IgGL. [001111 Likewise, one or more tyrosine residues can be replaced by phenylalanine residues. In addition, other variant amino acid insertions, deletions and/or substitutions are also contemplated and are within the scope of the present invention. Conservative amino acid substitutions will generally be preferred. Furthermore, alterations may be in the form of altered amino acids, such as peptidomimetics or D-amino acids. [001121 Fe sequences of the compound may also be derivatized as described herein for peptides, i.e., bearing modifications other than insertion, deletion, or substitution of amino acid residues. Preferably, the modifications are covalent in nature, and include for example, chemical bonding with polymers, lipids, other organic, and inorganic moieties. Derivatives of the invention may be prepared to increase circulating half-life, or may be designed to improve targeting capacity for the polypeptide to desired cells, tissues, or organs.

WO 2006/010057 PCT/US2005/024373 87 [00113] It is also possible to use the salvage receptor binding domain of the intact Fc molecule as the Fc part of a compound of the invention, such as described in WO 96/32478, entitled "Altered Polypeptides with Increased Half-Life." Additional members of the class of molecules designated as Fe herein are those that are described in WO 97/34631, entitled "Immunoglobulin-Like Domains with Increased Half-Lives." Both of the published PCT applications cited in this paragraph are hereby incorporated by reference. [00114] WSP components. Compounds of the invention further include at least one WSP. The WPS moiety of the molecule may be branched or unbranched. For therapeutic use of the end-product preparation, the polymer is pharmaceutically acceptable. In general, a desired polymer is selected based on such considerations as whether the polymer conjugate will be used therapeutically, and if so, the desired dosage, circulation time, resistance to proteolysis, and other considerations. In various aspects, the average molecular weight of each water soluble polymer is between about 2 kDa and about 100 kDa, between about 5 kDa and about 50 kDa, between about 12 kDa and about 40 kDa and between about 20 kDa and about 35 kDa. In yet another aspect the molecular weight of each polymer is between about 6 kDa and about 25 kDa. The term "about" as used herein and throughout, indicates that in preparations of a water soluble polymer, some molecules will weigh more, some less, than the stated molecular weight. Generally, the higher the molecular weight or the more branches, the higher the polymer/protein ratio. Other sizes may be used, depending on the desired therapeutic profile including for example, the duration of sustained release; the effects, if any, on biological activity; the ease in handling; the degree or lack of antigenicity and other known effects of a water soluble polymer on a therapeutic protein. [00115] The WSP should be attached to a peptide or protein with consideration given to effects on functional or antigenic domains of the peptide or protein. In general, chemical derivatization may be performed under any suitable condition used to react a protein with an activated polymer molecule. Activating groups which can be used to link the water soluble polymer to one or more proteins include without limitation sulfone, maleimide, sulfhydryl, thiol, triflate, tresylate, azidirine, oxirane and 5-pyridyl. If attached to the peptide by reductive alkylation, the polymer selected should have a single reactive aldehyde so that the degree of polymerization is controlled. [001161 Suitable, clinically acceptable, water soluble polymers include without limitation, PEG, polyethylene glycol propionaldehyde, copolymers of ethylene glycol/propylene glycol, monomethoxy-polyethylene glycol, carboxymethylcellulose, WO 2006/010057 PCT/US2005/024373 88 polyacetals, polyvinyl alcohol (PVA), polyvinyl pyrrolidone, poly-1, 3-dioxolane, poly-1,3,6 trioxane, ethylene/maleic anhydride copolymer, poly (.beta.-amino acids) (either homopolymers or random copolymers), poly(n-vinyl pyrrolidone)polyethylene glycol, propropylene glycol homopolymers (PPG) and other polyakylene oxides, polypropylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (POG) (e.g., glycerol) and other polyoxyethylated polyols, polyoxyethylated sorbitol, or polyoxyethylated glucose, colonic acids or other carbohydrate polymers, Ficoll or dextran and mixtures thereof. [00117] Polysaccharide polymers are another type of water soluble polymer which may be used for protein modification. Dextrans are polysaccharide polymers comprised of individual subunits of glucose predominantly linked by axl-6 linkages. The dextran itself is available in many molecular weight ranges, and is readily available in molecular weights from about 1 kD to about 70 kD. Dextran is a suitable water soluble polymer for use in the present invention as a vehicle by itself or in combination with another vehicle (e.g., Fc). See, for example, WO 96/11953 and WO 96/05309. The use of dextran conjugated to therapeutic or diagnostic immunoglobulins has been reported; see, for example, European Patent Publication No. 0 315 456, which is hereby incorporated by reference. Dextran of about 1 kD to about 20 kD is preferred when dextran is used as a vehicle in accordance with the present invention. [00118] In one embodiment, the WSP is PEG and the invention contemplates preparations wherein a compound is modified to include any of the forms of PEG that have been used to derivatize other proteins, such as and without limitation mono-(C1-C10) alkoxy or aryloxy-polyethylene glycol. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The PEG group may be of any convenient molecular weight and may be linear or branched. The average molecular weight of PEG contemplated for use in the invention ranges from about 2 kDa to about 100 kDa, from about 5 kDa to about 50 kDa, from about 5 kDa to about 10 kDa. In another aspect, the PEG moiety has a molecular weight from about 6 kDa to about 25 kDa. PEG groups generally are attached to peptides or proteins via acylation or reductive alkylation through a reactive group on the PEG moiety (e.g., an aldehyde, amino, thiol, or ester group) to a reactive group on the target peptide or protein (e.g., an aldehyde, amino, or ester group). Using methods described herein, a mixture of polymer/peptide conjugate molecules can be prepared, and the advantage provided herein is the ability to select the proportion of polymer/peptide conjugate to include in the mixture. Thus, if desired, a mixture of peptides with various numbers of polymer WO 2006/010057 PCT/US2005/024373 89 moieties attached (i.e., zero, one or two) can be prepared with a predetermined proportion of polymer/protein conjugate. [00119] A useful strategy for the PEGylation (other methods are discussed in more detail herein) of synthetic peptides consists of combining, through forming a conjugate linkage in solution, a peptide and a PEG moiety, each bearing a special functionality that is mutually reactive toward the other. The peptides can be easily prepared with conventional solid phase synthesis. The peptides are "preactivated" with an appropriate functional group at a specific site. The precursors are purified and fully characterized prior to reacting with the PEG moiety. Ligation of the peptide with PEG usually takes place in aqueous phase and can be easily monitored by reverse phase analytical HPLC. The PEGylated peptides can be easily purified by preparative HPLC and characterized by analytical HPLC, amino acid analysis and laser desorption mass spectrometry. [00120] Linkers. Any linkedr" group is optional, whether positioned between peptides, peptide and vehicle or vehicle and WSP. When present, its chemical structure is not critical, since it serves primarily as a spacer. The linker is preferably made up of amino acids linked together by peptide bonds. Thus, in preferred embodiments, the linker is made up of from 1 to 20 amino acids linked by peptide bonds, wherein the amino acids are selected from the 20 naturally occurring amino acids. Some of these amino acids may be glycosylated, as is well understood by those in the art. In a more preferred embodiment, the 1 to 20 amino acids are selected from glycine, alanine, proline, asparagine, glutamine, and lysine. Even more preferably, a linker is made up of a majority of amino acids that are sterically unhindered, such as glycine and alanine. Thus, preferred linkers are polyglycines (particularly (Gly)4, (Gly)5), poly(Gly-Ala), and polyalanines. Other specific examples of linkers are: (Gly)3Lys(Gly)4 (SEQ ID NO: 1018); (Gly)3AsnGlySer(Gly)2 (SEQ ID NO: 1019); (Gly)3Cys(Gly)4 (SEQ ID NO: 1020); and GlyProAsnGlyGly (SEQ ID NO: 1021). [00121] To explain the above nomenclature, for example, (Gly)3Lys(Gly)4 means Gly-Gly-Gly-Lys-Gly-Gly-Gly-Gly. Combinations of Gly and Ala are also preferred. The linkers shown here are exemplary; linkers within the scope of this invention may be much longer and may include other residues.

WO 2006/010057 PCT/US2005/024373 90 [00122] Non-peptide linkers are also possible. For example, alkyl linkers such as -NH (CH2)s-C(O)-, wherein s = 2-20 could be used. These alkyl linkers may further be substituted by any non-sterically hindering group such as lower alkyl (e.g., C1-C6) lower acyl, halogen (e.g., Cl, Br), CN, NH2, phenyl, etc. An exemplary non-peptide linker is a PEG linker, 0 H wherein n is such that the linker has a molecular weight of 100 to 5000 kD, preferably 100 to 500 kD. The peptide linkers may be altered to form derivatives in the same manner as described above. [00123] Peptide production. A peptide having been identified may be made in transformed host cells using recombinant DNA techniques. If the vehicle component is a polypeptide, the peptide-vehicle fusion product may be expressed as one. To do so, a recombinant DNA molecule encoding the peptide is first prepared using methods well known in the art. For instance, sequences coding for the peptides could be excised from DNA using suitable restriction enzymes. Alternatively, the DNA molecule could be synthesized using chemical synthesis techniques, such as the phosphoramidate method. Also, a combination of these techniques could be used. The invention therefore provides polynucleotides encoding a compound of the invention. [00124] The invention also provides vectors encoding compounds of the invention in an appropriate host. The vector comprises the polynucleotide that encodes the compound operatively linked to appropriate expression control sequences. Methods of effecting this operative linking, either before or after the polynucleotide is inserted into the vector, are well known. Expression control sequences include promoters, activators, enhancers, operators, ribosomal binding sites, start signals, stop signals, cap signals, polyadenylation signals, and other signals involved with the control of transcription or translation. [00125] The resulting vector having the polynucleotide therein is used to transform an appropriate host. This transformation may be performed using methods well known in the art.

WO 2006/010057 PCT/US2005/024373 91 [00126] Any of a large number of available and well-known host cells may be used in the practice of this invention. The selection of a particular host is dependent upon a number of factors recognized by the art. These include, for example, compatibility with the chosen expression vector, toxicity of the peptides encoded by the DNA molecule, rate of transformation, ease of recovery of the peptides, expression characteristics, bio-safety and costs. A balance of these factors must be struck with the understanding that not all hosts may be equally effective for the expression of a particular DNA sequence. Within these general guidelines, useful microbial hosts include bacteria (such as E. coli), yeast (such as Saccharomyces) and other fungi, insects, plants, mammalian (including human) cells in culture, or other hosts known in the art. [00127] Next, the transformed host is cultured and purified. Host cells may be cultured under conventional fermentation conditions so that the desired compounds are expressed. Such fermentation conditions are well known in the art. Finally, the peptides are purified from culture by methods well known in the art. [00128] Depending on the host cell utilized to express a compound of the invention, carbohydrate (oligosaccharide) groups may conveniently be attached to sites that are known to be glycosylation sites in proteins. Generally, 0-linked oligosaccharides are attached to serine (Ser) or threonine (Thr) residues while N-linked oligosaccharides are attached to asparagine (Asn) residues when they are part of the sequence Asn-X-Ser/Thr, where X can be any amino acid except proline. X is preferably one of the 19 naturally occurring amino acids not counting proline. The structures of N-linked and O-linked oligosaccharides and the sugar residues found in each type are different. One type of sugar that is commonly found on both is N-acetylneuraminic acid (referred to as sialic acid). Sialic acid is usually the terminal residue of both N-linked and O-linked oligosaccharides and, by virtue of its negative charge, may confer acidic properties to the glycosylated compound. Such site(s) may be incorporated in the linker of the compounds of this invention and are preferably glycosylated by a cell during recombinant production of the polypeptide compounds (e.g., in mammalian cells such as CHO, BHK, COS). However, such sites may further be glycosylated by synthetic or semi synthetic procedures known in the art. [00129] Alternatively, the compounds may be made by synthetic methods. For example, solid phase synthesis techniques may be used. Suitable techniques are well known in the art, and include those described in Merrifield (1973), Chem. Polypeptides, pp. 335-61 (Katsoyannis and Panayotis eds.); Merrifield (1963), J. Am. Chem. Soc. 85: 2149; Davis et WO 2006/010057 PCT/US2005/024373 92 al. (1985), Biochem. Intl. 10: 394-414; Stewart and Young (1969), Solid Phase Peptide Synthesis; U.S. Pat. No. 3,941,763; Finn et al. (1976), The Proteins (3rd ed.) 2: 105-253; and Erickson et al. (1976), The Proteins (3rd ed.) 2: 257-527. Solid phase synthesis is the preferred technique of making individual peptides since it is the most cost-effective method of making small peptides. [00130] Compounds that contain derivatized peptides or which contain non-peptide groups are particularly amendable to synthesis by well-known organic chemistry techniques. [00131] WSP modification. For obtaining a compound covalently attached to a WSP, any method described herein or otherwise known in the art is employed. Methods for preparing chemical derivatives of polypeptides will generally comprise the steps of (a) reacting the polypeptide with the activated polymer molecule (such as a reactive ester or aldehyde derivative of the polymer molecule) under conditions whereby the polypeptide becomes attached to one or more polymer molecules, and (b) obtaining the reaction product(s). The optimal reaction conditions will be determined based on known parameters and the desired result. For example, the larger the ratio of polymer molecules:protein, the greater the percentage of attached polymer molecule. [00132] A biologically active molecule can be linked to a polymer through any available functional group using standard methods well known in the art. Examples of functional groups on either the polymer or biologically active molecule which can be used to form such linkages include amine and carboxy groups, thiol groups such as in cysteine resides, aldehydes and ketones, and hydroxy groups as can be found in serine, threonine, tyrosine, hydroxyproline and hydroxylysine residues. [00133] The polymer can be activated by coupling a reactive group such as trichloro-s triazine [Abuchowski, et al., (1977), J. Biol. Chem. 252:3582-3586, incorporated herein by reference in its entirety], carbonylimidazole [Beauchamp, et al., (1983), Anal. Biochem. 131:25-33, incorporated herein by reference in its entirety], or succinimidyl succinate [Abuchowski, et al., (1984), Cancer Biochem. Biophys. 7:175-186, incorporated herein by reference in its entirety] in order to react with an amine functionality on the biologically active molecule. Another coupling method involves formation of a glyoxylyl group on one molecule and an arninooxy, hydrazide or semicarbazide group on the other molecule to be conjugated [Fields and Dixon, (1968), Biochem. J. 108:883-887; Gaertner, et al., (1992), Bioconjugate Chem. 3:262-268; Geoghegan and Stroh, (1992), Bioconjugate Chem. 3:138- WO 2006/010057 PCT/US2005/024373 93 146; Gaertner, et al., (1994), J. Biol. Chem. 269:7224-7230, each of which is incorporated herein by reference in its entirety]. Other methods involve formation of an active ester at a free alcohol group of the first molecule to be conjugated using chloroformate or disuccinimidylcarbonate, which can then be conjugated to an amine group on the other molecule to be coupled [Veronese, et al., (1985), Biochem. and Biotech. 11:141-152; Nitecki, et al., U.S. Patent No. 5,089,261; Nitecki, U.S. Patent No. 5,281,698, each of which is incorporated herein by reference in its entirety]. Other reactive groups which may be attached via free alcohol groups are set forth in Wright, EP 0539167A2 (incorporated herein by reference in its entirety), which also describes the use of imidates for coupling via free amine groups. [00134] Another chemistry involves acylation of the primary amines of a target using the NHS-ester of methoxy-PEG (O-[(N-succinimidyloxycarbonyl)-methyl]-O' methylpolyethylene glycol). Acylation with methoxy-PEG-NHS results in an amide linkage which will eliminate the charge from the original primary amine. Other methods utilize mild oxidation of a target under conditions selected to target the pendant diol of the penultimate glycosyl unit sialic acid for oxidation to an aldehyde. The resultant glycoaldehyde was then reacted with a methoxy-PEG-hydrazide (O-(Hydrazinocarbonylmethyl)-O' methylpolyethylene glycol) to form a semi-stable hydrazone between PEG and target. The hydrazone is subsequently reduced by sodium cyanoborohydride to produce a stable PEG conjugate. See for example, U.S. Patent No. 6,586,398 (Kinstler, et al., July 1, 2003), incorporated herein by reference in its entirety. [001351 In specific applications of techniques for chemical modification, for example, U.S. Patent No. 4,002,531 ( incorporated herein by reference in its entirety) states that reductive alkylation was used for attachment of polyethylene glycol molecules to an enzyme. U.S. Patent No. 4,179,337 (incorporated herein by reference in its entirety) discloses PEG:protein conjugates involving, for example, enzymes and insulin. U.S. Patent No. 4,904,584 (incorporated herein by reference in its entirety) discloses the modification of the number of lysine residues in proteins for the attachment of polyethylene glycol molecules via reactive amine groups. U.S. Patent No. 5,834,594 (incorporated herein by reference in its entirety) discloses substantially non-immunogenic water soluble PEG:protein conjugates, involving for example, the proteins IL-2, interferon alpha, and IL-lra. The methods of Hakimi et al. involve the utilization of unique linkers to connect the various free amino groups in the protein to PEG. U.S. Patent Nos. 5,824,784 and 5,985,265 (each of which is WO 2006/010057 PCT/US2005/024373 94 incorporated herein by reference in its entirety) teach methods allowing for selectively N terminally chemically modified proteins and analogs thereof, including G-CSF and consensus interferon. Importantly, these modified proteins have advantages as relates to protein stability, as well as providing for processing advantages. [00136] WSP modification is also described in Francis et al., In: Stability of protein pharmaceuticals: in vivo pathways of degradation and strategies for protein stabilization (Eds. Ahern., T. and Manning, M. C.) Plenum, N. Y., 1991 (incorporated herein by reference in its entirety), is used. In still another aspect, the method described in Delgado et al., "Coupling of PEG to Protein By Activation With Tresyl Chloride, Applications In Immunoaffinity Cell Preparation", In: Fisher et al., eds., Separations Using Aqueous Phase Systems, Applications In Cell Biology and Biotechnology, Plenum Press, N.Y.N.Y., 1989 pp. 211-213 (incorporated herein by reference in its entirety), which involves the use of tresyl chloride, which results in no linkage group between the WSP moiety and the polypeptide moiety. In other aspects, attachment of a WSP is effected through use of N-hydroxy succinimidyl esters of carboxymethyl methoxy polyethylene glycol, as well known in the art. [00137] For other descriptions of modification of a target with a WSP, see, for example, U.S patent application No. 20030096400; EP 0 442724A2; EP 0154316; EP 0401384; WO 94/13322; U.S. Patent Nos. 5,362,852; 5,089,261; 5,281,698; 6,423,685; 6,635,646; 6,433,135; International application WO 90/07938; Gaertner and Offord, (1996), Bioconjugate Chem. 7:38-44; Greenwald et al., Crit Rev Therap Drug Carrier Syst. 2000;17:101-161; Kopecek et al., J Controlled Release., 74:147-158, 2001; Harris et al., Clin Pharmacokinet. 2001;40(7):539-51; Zalipsky et al., Bioconjug Chem. 1997;8:111-118; Nathan et al., Macromolecules. 1992;25:4476-4484; Nathan et al., Bioconj Chem. 1993;4:54 62; and Francis et al., Focus on Growth Factors, 3:4-10 (1992), the disclosures of which are incorporated herein by reference in their entirety. [00138] Reductive alkylation. In one aspect, covalent attachment of a WSP is carried out by reductive alkylation chemical modification procedures as provided herein to selectively modify the N-terminal a-amino group, and testing the resultant product for the desired biological characteristic, such as the biological activity assays provided herein. [00139] Reductive alkylation for attachment of a WSP to a protein or peptide exploits differential reactivity of different types of primary amino groups (e.g., lysine versus the N terminal) available for derivatization in a particular protein. Under the appropriate reaction WO 2006/010057 PCT/US2005/024373 95 conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved. [00140] For reductive alkylation, the polymer(s) selected could have a single reactive aldehyde group. A reactive aldehyde is, for example, polyethylene glycol propionaldehyde, which is water stable, or mono C-C 10 alkoxy or aryloxy derivatives thereof (see U.S. Patent No. 5,252,714, incorporated herein by reference in its entirety). In one approach, reductive alkylation is employed to conjugate a PEG-aldehyde (O-( 3 -Oxopropyl)-O' methylpolyethylene glycol) to a primary amine. Under appropriate conditions, this approach has been demonstrated to yield PEG conjugates predominately modified through the a-amine at the protein N-terminus. [00141] An aldehyde functionality useful for conjugating the biologically active molecule can be generated from a functionality having adjacent amino and alcohol groups. In a polypeptide, for example, an N-terminal serine, threonine or hydroxylysine can be used to generate an aldehyde functionality via oxidative cleavage under mild conditions using periodate. These residues, or their equivalents, can be normally present, for example at the N terminus of a polypeptide, may be exposed via chemical or enzymatic digestion, or may be introduced via recombinant or chemical methods. The reaction conditions for generating the aldehyde typically involve addition of a molar excess of sodium meta periodate and under mild conditions to avoid oxidation at other positions in the protein. The pH is preferably about 7.0. A typical reaction involves the addition of a 1.5 fold molar excess of sodium meta periodate, followed by incubation for 10 minutes at room temperature in the dark. [00142] The aldehyde functional group can be coupled to an activated polymer containing a hydrazide or semicarbazide functionality to form a hydrazone or sernicarbazone linkage. Hydrazide-containing polymers are commercially available, and can be synthesized, if necessary, using standard techniques. PEG hydrazides for use in the invention can be obtained from Shearwater Polymers, Inc., 2307 Spring Branch Road, Huntsville, Ala. 35801 (now part of Nektar Therapeutics,150 Industrial Road, San Carlos, CA 94070-6256). The aldehyde is coupled to the polymer by mixing the solution of the two components together and heating to about 37*C until the reaction is substantially complete. An excess of the polymer hydrazide is typically used to increase the amount of conjugate obtained. A typical reaction time is 26 hours. Depending on the thermal stability of the reactants, the reaction temperature and time can be altered to provide suitable results. Detailed determination of reaction conditions for both oxidation and coupling is set forth in Geoghegan and Stroh, WO 2006/010057 PCT/US2005/024373 96 (1992), Bioconjugate Chem. 3:138-146, and in Geoghegan, U.S. Patent No. 5,362,852, each of which is incorporated herein by reference in its entirety. [00143] Using reductive alkylation, the reducing agent should be stable in aqueous solution and preferably be able to reduce only the Schiff base formed in the initial process of reductive alkylation. Reducing agents are selected from, and without limitation, sodium borohydride, sodium cyanoborohydride, dimethylamine borate, timethylamine borate and pyridine borate. [00144] The reaction pH affects the ratio of polymer to protein to be used. In general, if the reaction pH is lower than the pKa of a target reactive group, a larger excess of polymer to protein will be desired. If the pH is higher than the target pKa, the polymer:protein ratio need not be as large (i.e., more reactive groups are available, so fewer polymer molecules are needed). [00145] Accordingly, the reaction is performed in one aspect at a pH which allows one to take advantage of the pKa differences between the -- amino groups of the lysine residues and that of the a-amino group of the N-terminal residue of the protein. By such selective derivatization, attachment of a water soluble polymer to a protein is controlled; the conjugation with the polymer takes place predominantly at the N-terminus of the protein and no significant modification of other reactive groups, such as the lysine side chain amino groups, occurs. [001461 In one aspect, therefore, methods are provided for covalent attachment of a WSP to a target compound and which provide a substantially homogenous preparation of WSP/protein conjugate molecules, in the absence of further extensive purification as is required using other chemical modification chemistries. More specifically, if polyethylene glycol is used, methods described allow for production of an N-terminally PEGylated protein lacking possibly antigenic linkage groups, i.e., the polyethylene glycol moiety is directly coupled to the protein moiety without potentially toxic by-products. [00147] Depending on the method of WSP attachment chosen, the proportion of WSP molecules attached to the target peptide or protein molecule will vary, as will their concentrations in the reaction mixture. In general, the optimum ratio (in terms of efficiency of reaction in that there is no excess unreacted protein or polymer) is determined by the molecular weight of the WSP selected. In addition, when using methods that involve non- WO 2006/010057 PCT/US2005/024373 97 specific attachment and later purification of a desired species, the ratio may depend on the number of reactive groups (typically amino groups) available. [00148] Purification. The method of obtaining a substantially homogeneous WSP modified preparation is, in one aspect, by purification of a predominantly single species of modified compound from a mixture of species. By way of example, a substantially homogeneous species is first separated by ion exchange chromatography to obtain material having a charge characteristic of a single species (even though other species having the same apparent charge may be present), and then the desired species is separated using size exclusion chromatography. Other methods are reported and contemplated by the invention, includes for example, PCT WO 90/04606, published May 3, 1990, which describes a process for fractionating a mixture of PEG-protein adducts comprising partitioning the PEG/protein adducts in a PEG-containing aqueous biphasic system. [00149] Thus, one aspect of the present invention is a method for preparing a WSP modified compound conjugate comprised of (a) reacting a compound having more than one amino group with a water soluble polymer moiety under reducing alkylation conditions, at a pH suitable to selectively activate the a-amino group at the amino terminus of the protein moiety so that said water soluble polymer selectively attaches to said a-amino group; and (b) obtaining the reaction product. Optionally, and particularly for a therapeutic product, the reaction products are separated from unreacted moieties. [00150] As ascertained by peptide mapping and N-terminal sequencing, a preparation is provided which comprises at least 50% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide. In other embodiments, preparations are provided which comprises at least 75% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide; at least 85% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide; at least 90% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide; at least 95% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide; and at least 99% PEGylated peptide in a mixture of PEGylated peptide and unreacted peptide. [00151] Pharmaceutical Compositions. The present invention further provides pharmaceutical compositions comprising a preparation of the invention. Such pharmaceutical compositions may be for administration for injection, or for oral, nasal, transdermal or other forms of administration, including, e.g., by intravenous, intradermal, intramuscular, WO 2006/010057 PCT/US2005/024373 98 intramammary, intraperitoneal, intrathecal, intraocular, retrobulbar, intrapulmonary (e.g., aerosolized drugs) or subcutaneous injection (including depot administration for long term release); by sublingual, anal, vaginal, or by surgical implantation, e.g., embedded under the splenic capsule, brain, or in the cornea. The treatment may consist of a single dose or a plurality of doses over a period of time. In general, comprehended by the invention are pharmaceutical compositions comprising effective amounts of a compound of the invention together with pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions include diluents of various buffer content (e.g., Tris-HCl, acetate, phosphate), pH and ionic strength; additives such as detergents and solubilizing agents (e.g., Tween 80, Polysorbate 80), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., Thimersol, benzyl alcohol) and bulking substances (e.g., lactose, mannitol); incorporation of the material into particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc. or into liposomes. Hyaluronic acid may also be used, and this may have the effect of promoting sustained duration in the circulation. The pharmaceutical compositions optionally may include still other pharmaceutically acceptable liquid, semisolid, or solid diluents that serve as pharmaceutical vehicles, excipients, or media, including but are not limited to, polyoxyethylene sorbitan monolaurate, magnesium stearate, methyl- and propylhydroxybenzoate, starches, sucrose, dextrose, gum acacia, calcium phosphate, mineral oil, cocoa butter, and oil of theobroma. Such compositions may influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the present proteins and derivatives. See, e.g., Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) pages 1435-1712 which are herein incorporated by reference. The compositions may be prepared in liquid form, or may be in dried powder, such as lyophilized form. Implantable sustained release formulations are also contemplated, as are transdermal formulations. [00152] Oral Dosage. Contemplated for use herein are oral solid dosage forms, which are described generally in Chapter 89 of Remington's Pharmaceutical Sciences (1990), 18th Ed., Mack Publishing Co. Easton Pa. 18042, incorporated herein by reference. Solid dosage forms include tablets, capsules, pills, troches or lozenges, cachets or pellets. Alternatively, proteinoid encapsulation may be used (as, for example, proteinoid microspheres reported in U.S. Pat. No. 4,925,673), or liposomal encapsulation may be used, the liposomes optionally derivatized with various polymers (e.g., U.S. Pat. No. 5,013,556). A description of solid WO 2006/010057 PCT/US2005/024373 99 dosage forms for therapeutics in general is given in Chapter 10 of Marshall, K., Modem Pharmaceutics (1979), edited by G. S. Banker and C. T. Rhodes, incorporated herein by reference. In general, the formulation will include a preparation of the invention and inert ingredients which allow for protection against the stomach environment, and release of the biologically active material in the intestine. [00153] If necessary, the compounds may be chemically modified so that oral delivery is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the compound molecule itself, where said moiety permits (a) inhibition of proteolysis; and (b) uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the compound and increase in circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline (Abuchowski and Davis, Soluble Polymer-Enzyme Adducts, Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, (1981), pp 367-383; Newmark, et al., J. Appl. Biochem. 4:185-189 (1982)). Other polymers that could be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. [00154] For oral delivery dosage forms, it is also possible to use a salt of a modified aliphatic amino acid, such as sodium N-(8-[2-hydroxybenzoyl]amino) caprylate snack) , as a carrier to enhance absorption of the therapeutic compound. The clinical efficacy of a heparin formulation using SNAC has been demonstrated in a Phase II trial conducted by Emisphere Technologies. See US Pat. No. 5,792,451, "Oral drug delivery composition and methods." [00155] Preparations of the invention can be included in formulation as fine multiparticulates in the form of granules or pellets of particle size about, for example, one mm. The formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets. Compositions are optionally prepared by compression. [00156] Colorants and flavoring agents may be included. For example, the preparation may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents. [00157] Preparations of the invention are, in one aspect, diluted or increased in the volume with an inert material. Exemplary diluents include carbohydrates, especially WO 2006/010057 PCT/US2005/024373 100 mannitol, a-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Certain inorganic salts may also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell. [00158] Preparations including disintegrants are further contemplated in solid dosage form compositions. Materials used as disintegrants include, but are not limited to, starch (including the commercial disintegrant based on starch, Explotab), sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite. Another form of disintegrant is an insoluble cationic exchange resin. Powdered gums may also be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants. [00159] Pharmaceutical compositions including binders are further contemplated to hold the therapeutic agent together to form a hard tablet and exemplary binders include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic. [00160] An antifrictional agent in a pharmaceutical composition is further contemplated to prevent sticking during the formulation process. Lubricants include, but are not limited to, stearic acid, including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000. [00161] Glidants that might improve the flow properties of a pharmaceutical composition during formulation and to aid rearrangement during compression are also provided. Exemplary glidants include starch, talc, pyrogenic silica and hydrated silicoaluminate. [00162] To aid dissolution of a composition into the aqueous environment, incorporation of a surfactant as a wetting agent is contemplated. Exemplary surfactants include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents are contemplated, including for example and WO 2006/010057 PCT/US2005/024373 101 without limitation, benzalkonium chloride or benzethonium chloride. In another aspect, compositions using as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. Compositions comprising these surfactants, either alone or as a mixture in different ratios, are therefore further provided. [00163] Optionally, additives are included in a pharmaceutical composition to enhance uptake of the compound, such additives including, for example and without limitation, fatty acids oleic acid, linoleic acid and linolenic acid. [00164] Controlled release compositions. In another aspect, controlled release formulation are provided. A preparation of the invention is incorporated into an inert matrix which permits release by either diffusion or leaching mechanisms e.g., gums. Slowly degenerating matrices, e.g., alginates, polysaccharides, may also be incorporated into the formulation. Another form of a controlled release is by a method based on the Oros therapeutic system (Alza Corp.), i.e., the drug is enclosed in a semipermeable membrane which allows water to enter and push drug out through a single small opening due to osmotic effects. Some enteric coatings also have a delayed release effect. [00165] Other coatings may be used in compositions of the invention, including for example, a variety of sugars which could be applied in a coating pan. The compositions also include a film coated tablet and the materials used in this instance are divided into two groups. The first includes the nonenteric materials, such as and without limitation methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methylhydroxy-ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl-methyl cellulose, sodium carboxy-methyl cellulose, providone and the polyethylene glycols. The second group consists of the enteric materials that are commonly esters of phthalic acid. [00166] A mix of materials is also contemplated to provide the optimum film coating. Film coating may be carried out in a pan coater or in a fluidized bed or by compression coating. [001671 Pulmonary delivery. Also contemplated herein is pulmonary delivery of a preparation of the invention. The compound is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. Pulmonary delivery is described in Adjei et al., Pharma. Res. (1990) 7: 565-9; Adjei et al. (1990), WO 2006/010057 PCT/US2005/024373 102 Internat. J. Pharmaceutics 63: 135-44; Braquet et al. (1989), J. Cardiovasc. Pharmacol. 13 (suppl.

5 ): s.143-146; Hubbard et al. (1989), Annals Int. Med. 3: 206-12; Smith et al. (1989), J. Clin. Invest. 84: 1145-6; Oswein et al. (March 1990), "Aerosolization of Proteins", Proc. Symp. Resp. Drug Delivery II, Keystone, Colo.; Debs et al. (1988), J. Immunol. 140: 3482-8 and Platz et al., U.S. Pat. No. 5,284,656, the disclosures of which are incorporated herein by reference. [00168] Mechanical devices. Also contemplated for practice of the invention is a wide range of mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art. Specific examples of commercially available devices suitable for the practice of this invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II nebulizer, manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, N.C.; and the Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Mass. [00169] All such devices require the use of formulations suitable for the dispensing a preparation of the invention. Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material, in addition to diluents, adjuvants and/or carriers useful in therapy. [00170] For effective delivery to distal lung, the composition is prepared in particulate form with an average particle size in one aspect of less than 10 pm (or microns), and in an alternative aspect 0.5 to 5 pm. [00171] Formulations suitable for use with a nebulizer, either jet or ultrasonic, will typically comprise the inventive compound dissolved in water at a concentration of about 0.1 to 25 mg of biologically active protein per mL of solution. The formulation may also include a buffer and a simple sugar (e.g., for protein stabilization and regulation of osmotic pressure). The nebulizer formulation may also contain a surfactant, to reduce or prevent surface induced aggregation of the protein caused by atomization of the solution in forming the aerosol. [001721 Formulations for use with a metered-dose inhaler device will generally comprise a finely divided powder containing the inventive compound suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a WO 2006/010057 PCT/US2005/024373 103 hydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be useful as a surfactant. [00173] Formulations for dispensing from a powder inhaler device will comprise a finely divided dry powder containing the inventive compound and may also include a bulking agent, such as lactose, sorbitol, sucrose, mannitol, trehalose, or xylitol in amounts which facilitate dispersal of the powder from the device, e.g., 50 to 90% by weight of the formulation. [00174] Nasal delivery. Nasal delivery of preparations of the invention is also contemplated. Nasal delivery allows the passage of the protein to the blood stream directly after administering the therapeutic product to the nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with dextran or cyclodextran. Delivery via transport across other mucous membranes is also contemplated. [00175] Buccal delivery forms. Buccal delivery of the inventive compound is also contemplated. Buccal delivery formulations are known in the art for use with peptides. [00176] Carriers. Pharmaceutically acceptable carriers include carbohydrates such as trehalose, mannitol, xylitol, sucrose, lactose, and sorbitol. Other ingredients for use in formulations may include DPPC, DOPE, DSPC and DOPC. Natural or synthetic surfactants may be used. PEG may be used (even apart from its use in derivatizing a compound of the invention). Dextrans, such as cyclodextran, may be used. Bile salts and other related enhancers may be used. Cellulose and cellulose derivatives may be used. Amino acids may be used, such as use in a buffer formulation. [00177] Other formulations. The use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is also contemplated. [00178] Dosages. The dosage regimen involved in a method for treating a condition described herein will be determined by the attending physician, considering various factors which modify the action of drugs, e.g. the age, condition, body weight, sex and diet of the patient, the severity of any infection, time of administration and other clinical factors. In various aspects, the daily regimen is in the range of 0.1-1000 pg of a preparation per kilogram of body weight (calculating the mass of the protein alone, without chemical modification) or 0.1-150 pig/kg.

WO 2006/010057 PCT/US2005/024373 104 [00179] Preparations of the invention may be administered by an initial bolus followed by a continuous infusion to maintain therapeutic circulating levels of drug product. As another example, the inventive compound may be administered as a one-time dose. Those of ordinary skill in the art will readily optimize effective dosages and administration regimens as determined by good medical practice and the clinical condition of the individual patient. The frequency of dosing will depend on the pharmacokinetic parameters of the agents and the route of administration. The optimal pharmaceutical formulation will be determined by one skilled in the art depending upon the route of administration and desired dosage. See for example, Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) pages 1435-1712, the disclosure of which is hereby incorporated by reference. Such formulations may influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the administered agents. Depending on the route of administration, a suitable dose may be calculated according to body weight, body surface area or organ size. Further refinement of the calculations necessary to determine the appropriate dosage for treatment involving each of the above mentioned formulations is routinely made by those of ordinary skill in the art without undue experimentation, especially in light of the dosage information and assays disclosed herein, as well as the pharmacokinetic data observed in the human clinical trials discussed above. Appropriate dosages may be ascertained through use of established assays for determining blood levels dosages in conjunction with appropriate dose-response data. The final dosage regimen will be determined by the attending physician, considering various factors which modify the action of drugs, e.g. the drug's specific activity, the severity of the damage and the responsiveness of the patient, the age, condition, body weight, sex and diet of the patient, the severity of any infection, time of administration and other clinical factors. As studies are conducted, further information will emerge regarding the appropriate dosage levels and duration of treatment for various diseases and conditions. [00180] The therapeutic methods, compositions and compounds of the present invention may also be employed, alone or in combination with other cytokines, soluble c-Mpl receptor, hematopoietic factors, interleukins, growth factors or antibodies in the treatment of disease states characterized by other symptoms as well as platelet deficiencies. It is anticipated that the preparations of the invention will prove useful in treating some forms of thrombocytopenia in combination with general stimulators of hematopoiesis, such as IL-3 or GM-CSF. Other megakaryocytic stimulatory factors, i.e., meg-CSF, stem cell factor (SCF), WO 2006/010057 PCT/US2005/024373 105 leukemia inhibitory factor (LIF), oncostatin M (OSM), or other molecules with megakaryocyte stimulating activity may also be employed with Mpl ligand. Additional exemplary cytokines or hematopoietic factors for such co-administration include IL-i alpha, IL-I beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-11, colony stimulating factor-1 (CSF-1), M-CSF, SCF, GM-CSF, granulocyte colony stimulating factor (G-CSF), EPO, interferon-alpha (TFN alpha), consensus interferon, IFN-beta, IFN-gamma, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-14, IL-15, IL- 16, IL-17, IL-18, thrombopoietin (TPO), angiopoietins, for example Ang-1, Ang-2, Ang-4, Ang-Y, the human angiopoictin-like polypeptide, vascular endothelial growth factor (VEGF), angiogenin, bone morphogenic protein-1, bone morphogenic protein-2, bone morphogenic protein-3, bone morphogenic protein-4, bone morphogenic protein-5, bone morphogenic protein-6, bone morphogenic protein-7, bone morphogenic protein-8, bone morphogenic protein-9, bone morphogenic protein- 10, bone morphogenic protein-11, bone morphogenic protein-12, bone morphogenic protein-13, bone morphogenic protein-14, bone morphogenic protein-15, bone morphogenic protein receptor IA, bone morphogenic protein receptor IB, brain derived neurotrophic factor, ciliary neutrophic factor, ciliary neutrophic factor receptor, cytokine-induced neutrophil chemotactic factor 1, cytokine-induced neutrophil, chemotactic factor 2a, cytokine-induced neutrophil chemotactic factor 2P, p endothelial cell growth factor, endothelin 1, epidermal growth factor, epithelial-derived neutrophil attractant, fibroblast growth factor 4, fibroblast growth factor 5, fibroblast growth factor 6, fibroblast growth factor 7, fibroblast growth factor 8, fibroblast growth factor 8b, fibroblast growth factor 8c, fibroblast growth factor 9, fibroblast growth factor 10, fibroblast growth factor acidic, fibroblast growth factor basic, glial cell line-derived neutrophic factor receptor al, glial cell line-derived neutrophic factor receptor a2, growth related protein, growth related protein a, growth related protein P, growth related protein y, heparin binding epidermal growth factor, hepatocyte growth factor, hepatocyte growth factor receptor, insulin-like growth factor I, insulin-like growth factor receptor, insulin-like growth factor II, insulin-like growth factor binding protein, keratinocyte growth factor, leukemia inhibitory factor, leukemia inhibitory factor receptor a, nerve growth factor nerve growth factor receptor, neurotrophin-3, neurotrophin-4, placenta growth factor, placenta growth factor 2, platelet-derived endothelial cell growth factor, platelet derived growth factor, platelet derived growth factor A chain, platelet derived growth factor AA, platelet derived growth factor AB, platelet derived growth factor B chain, platelet derived growth factor BB, platelet derived growth factor receptor a, platelet derived growth factor receptor p, pre-B cell growth stimulating factor, stem cell factor receptor, TNF, including TNFO, TNF1, TNF2, WO 2006/010057 PCT/US2005/024373 106 transforming growth factor a, transforming growth factor P, transforming growth factor $31, transforming growth factor p1.2, transforming growth factor P2, transforming growth factor P3, transforming growth factor $5, latent transforming growth factor $1, transforming growth factor [ binding protein I, transforming growth factor P binding protein II, transforming growth factor p binding protein III, tumor necrosis factor receptor type I, tumor necrosis factor receptor type II, urokinase-type plasminogen activator receptor, vascular endothelial growth factor, and chimeric proteins and biologically or immunologically active fragments thereof. It may further be useful to administer, either simultaneously or sequentially, an effective amount of a soluble mammalian c-Mpl, which appears to have an effect of causing megakaryocytes to fragment into platelets once the megakaryocytes have reached mature form. Thus, administration of a preparation of the invention (to enhance the number of mature megakaryocytes) followed by administration of the soluble c-Mpl (to inactivate the ligand and allow the mature megakaryocytes to produce platelets) is expected to be a particularly effective means of stimulating platelet production. The dosage recited above would be adjusted to compensate for such additional components in the therapeutic composition. Progress of the treated patient can be monitored by conventional methods. [00181] Conditions in general. The compounds of this invention have pharmacological activity resulting from their ability to bind to proteins of interest as agonists, mimetics or antagonists of the native ligands of such proteins of interest. The utility of specific compounds is shown in Table 2. The activity of these compounds can be measured by assays known in the art. For the TPO-mimetic compounds, an in vivo assay is further described in the Examples section herein. [00182] In addition to therapeutic uses, the compounds of the present invention are useful in diagnosing diseases characterized by dysfunction of their associated protein of interest. In one embodiment, a method of detecting in a biological sample a protein of interest (e.g., a receptor) that is capable of being activated comprising the steps of: (a) contacting the sample with a compound of this invention; and (b) detecting activation of the protein of interest by the compound. The biological samples include tissue specimens, intact cells, or extracts thereof. The compounds of this invention may be used as part of a diagnostic kit to detect the presence of their associated proteins of interest in a biological sample. Such kits employ the compounds of the invention having an attached label to allow for detection. The compounds are useful for identifying normal or abnormal proteins of interest. For the EPO mimetic compounds, for example, presence of abnormal protein of interest in a biological WO 2006/010057 PCT/US2005/024373 107 sample may be indicative of such disorders as Diamond Blackfan anemia, where it is believed that the EPO receptor is dysfunctional. [00183] Therapeutic uses of EPO-mimetic compounds. The EPO-mimetic compounds of the invention are useful for treating disorders characterized by low red blood cell levels. Included in the invention are methods of modulating the endogenous activity of an EPO receptor in a mammal, preferably methods of increasing the activity of an EPO receptor. In general, any condition treatable by erythropoietin, such as anemia, may also be treated by the EPO-mimetic compounds of the invention. These compounds are administered by an amount and route of delivery that is appropriate for the nature and severity of the condition being treated and may be ascertained by one skilled in the art. Preferably, administration is by injection, either subcutaneous, intramuscular, or intravenous. [00184] Therapeutic uses of TPO-mimetic compounds. For the TPO-mimetic compounds, one can utilize such standard assays as those described in W095/26746 entitled "Compositions and Methods for Stimulating Megakaryocyte Growth and Differentiation". In vivo assays also appear in the Examples hereinafter. [00185] The conditions to be treated are generally those that involve an existing megakaryocyte/platelet deficiency or an expected megakaryocyte/platelet deficiency (e.g., because of planned surgery or platelet donation). Such conditions will usually be the result of a deficiency (temporary or permanent) of active Mpl ligand in vivo. The generic term for platelet deficiency is thrombocytopenia, and hence the methods and compositions of the present invention are generally available for treating thrombocytopenia in patients in need thereof. [00186] Thrombocytopenia may be present for various reasons, including chemotherapy and other therapy with a variety of drugs, radiation therapy, surgery, accidental blood loss, and other specific disease conditions. Exemplary specific disease conditions that involve thrombocytopenia and may be treated in accordance with this invention are: aplastic anemia, idiopathic or immune thrombocytopenia (ITP), including idiopathic thrombocytopenic purpura associated with breast cancer; HIV associated ITP and HIV related thrombotic thrombocytopenic purpura; metastatic tumors which result in thrombocytopenia, systemic lupus erythematosus, including neonatal lupus syndrome, splenomegaly, Fanconi's syndrome, vitamin B 12 deficiency, folic acid deficiency, May Hegglin anomaly, Wiskott-Aldrich syndrome, chronic liver disease; myelodysplastic WO 2006/010057 PCT/US2005/024373 108 syndrome associated with thrombocytopenia; paroxysmal nocturnal hemoglobinuria, acute profound thrombocytopenia following C7E3 Fab (Abciximab) therapy; alloinmune thrombocytopenia, including maternal alloimmune thrombocytopenia; thrombocytopenia associated with antiphospholipid antibodies and thrombosis; autoimmune thrombocytopenia; drug-induced immune thrombocytopenia, including carboplatin-induced thrombocytopenia, heparin-induced thrombocytopenia; fetal thrombocytopenia; gestational thrombocytopenia; Hughes' syndrome; lupoid thrombocytopenia; accidental and/or massive blood loss; myeloproliferative disorders; thrombocytopenia in patients with malignancies; thrombotic thrombocytopenia purpura, including thrombotic microangiopathy manifesting as thrombotic thrombocytopenic purpura/hemolytic uremic syndrome in cancer patients; autoimmune hemolytic anemia; occult jejunal diverticulum perforation; pure red cell aplasia; autoimmune thrombocytopenia; nephropathia epidemica; rifampicin-associated acute renal failure; Paris Trousseau thrombocytopenia; neonatal alloimmune thrombocytopenia; paroxysmal nocturnal hemoglobinuria; hematologic changes in stomach cancer; hemolytic uremic syndromes in childhood; hematologic manifestations related to viral infection including hepatitis A virus and CMV-associated thrombocytopenia. Also, certain treatments for AIDS result in thrombocytopenia (e.g., AZT). Certain wound healing disorders might also benefit from an increase in platelet numbers. Also, certain treatments for AIDS result in thrombocytopenia (e.g., AZT). Certain wound healing disorders might also benefit from an increase in platelet numbers. [001871 The TPO-mimetic compounds of this invention may be used in any situation in which production of platelets or platelet precursor cells is desired, or in which stimulation of the c-Mpl receptor is desired. Thus, for example, the compounds of this invention may be used to treat any condition in a mammal wherein there is a need of platelets, megakaryocytes, and the like. Such conditions are described in detail in the following exemplary sources: W095/26746; W095/21919; W095/18858; W095/21920 and are incorporated herein. [00188] The TPO-mimetic compounds of this invention may also be useful in maintaining the viability or storage life of platelets and/or megakaryocytes and related cells. Accordingly, it could be useful to include an effective amount of one or more such compounds in a composition containing such cells. [00189] The therapeutic methods, compositions and compounds of the present invention may also be employed, alone or in combination with other cytokines, soluble Mpl receptor, hematopoietic factors, interleukins, growth factors or antibodies in the treatment of WO 2006/010057 PCT/US2005/024373 109 disease states characterized by other symptoms as well as platelet deficiencies. It is anticipated that the inventive compound will prove useful in treating some forms of thrombocytopenia in combination with general stimulators of hematopoiesis, such as IL-3 or GM-CSF. Other megakaryocytic stimulatory factors, i.e., meg-CSF, stem cell factor (SCF), leukemia inhibitory factor (LIF), oncostatin M (OSM), or other molecules with megakaryocyte stimulating activity may also be employed with Mpl ligand. Additional exemplary cytokines or hematopoietic factors for such co-administration include IL-I alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-11, colony stimulating factor-I (CSF-1), SCF, GM CSF, granulocyte colony stimulating factor (G-CSF), EPO, interferon-alpha (IFN-alpha), consensus interferon, IFN-beta, or IFN-gamma. It may further be useful to administer, either simultaneously or sequentially, an effective amount of a soluble mammalian Mpl receptor, which appears to have an effect of causing megakaryocytes to fragment into platelets once the megakaryocytes have reached mature form. Thus, administration of an inventive compound (to enhance the number of mature megakaryocytes) followed by administration of the soluble Mpl receptor (to inactivate the ligand and allow the mature megakaryocytes to produce platelets) is expected to be a particularly effective means of stimulating platelet production. The dosage recited above would be adjusted to compensate for such additional components in the therapeutic composition. Progress of the treated patient can be monitored by conventional methods. [00190] In cases where the inventive compounds are added to compositions of platelets and/or megakaryocytes and related cells, the amount to be included will generally be ascertained experimentally by techniques and assays known in the art. An exemplary range of amounts is 0.1 pg to 1 mg inventive compound per 106 cells. [00191] The conditions to be treated are generally those that involve an existing megakaryocyte/platelet deficiency or an expected megakaryocyte/platelet deficiency (e.g., because of planned surgery or platelet donation). Such conditions will usually be the result of a deficiency (temporary or permanent) of active thrombopoietin in vivo. The generic term for platelet deficiency is thrombocytopenia, and the methods and preparations of the present invention are generally available for treating thrombocytopenia in patients in need thereof. [00192] With regard to anticipated platelet deficiencies, e.g., due to future surgery, a compound of the present invention could be administered several days to several hours prior to the need for platelets. With regard to acute situations, e.g., accidental and massive blood WO 2006/010057 PCT/US2005/024373 110 loss, a preparation or composition of the invention is optionally administered along with blood or purified platelets. [00193] The TPO-mimetic compounds of the invention are useful in stimulating certain cell types other than megakaryocytes if such cells are found to express c-Mpl. Conditions associated with such cells that express the c-Mpl, which are responsive to stimulation by a preparation or composition described herein are also within the scope of this invention. [00194] The following examples are not intended to be limiting but only exemplary of specific embodiments of the invention. Example 1 Expression Construct Assembly [00195] A polynucleotide encoding a TMP fusion protein comprising a murine Fe region (mFc-TMP) was constructed by combining nucleotide sequences individually encoding murine Fc and a TMP (described in EP01124961A2). In the first round of PCR, the murine Fc-encoding component was amplified with PCR primers 3155-58 (SEQ ID NO: 1022) and 1388-00 (SEQ ID NO: 1023). 3155-58: CCGGGTAAAGGTGGAGGTGGTGGTATCGA (SEQ ID NO: 1024) 3155-59: CCACCTCCACCTTTACCCGGAGAGTGGGAG (SEQ ID NO: 1025) [00196] In a separate reaction, an TMP-encoding polynucleotide was amplified with primers 1209-85 (SEQ ID NO: 1026) and 3155-59 (SEQ ID NO: 1027). 1209-85: CGTACAGGTTTACGCAAGAAAATGG (SEQ ID NO: 1028) 1388-00: CTAGTTATTGCTCAGCGG (SEQ ID NO: 1029) [00197] The resulting PCR fragments were gel purified and combined in a single tube for a second round of PCR with primers 1209-85 (SEQ ID NO: 1030) and 1388-00 (SEQ ID NO: 1031). The PCR product from this second round of amplification was gel purified and digested with restriction enzymes NdeI and XhoL The digestion fragment was purified and ligated into the vector pAMG21, previously digested with the same enzymes. This ligation mix was transformed via electroporation into the E. coli strain Amgen 393 and plated onto LB + Kanamycin media. Colonies were screened via PCR and DNA sequencing. A positive WO 2006/010057 PCT/US2005/024373 111 clone with a nucleotide sequence (SEQ ID NO: 1032) encoding the mFc-TMP fusion protein (SEQ ID NO: 1033) was identified and designated 6397. [00198] Murine Fc-TMP fusion protein-encoding polynucleotide (SEQ ID NO: 1034) 1 GATTTGATTC TAGATTTGTT TTAACTAATT AAAGGAGGAA TAACAT Open RF: ATGGTCGACGGTTG TAAGCCATGC ATTTGTACAG TCCCAGAAGT ATCATCTGTC 101 TTCATCTTCC CCCCAAAGCC CAAGGATGTG CTCACCATTA CTCTGACTCC 151 TAAGGTCACG TGTGTTGTGG TAGACATCAG CAAGGATGAT CCCGAGGTCC 201 AGTTCAGCTG GTTTGTAGAT GATGTGGAGG TGCACACAGC TCAGACGCAA 251 CCCCGGGAGG AGCAGTTCAA CAGCACTTTC CGCTCAGTCA GTGAACTTCC 301 CATCATGCAC CAGGACTGGC TCAATGGCAA GGAGTTCAAA TGCAGGGTCA 351 ACAGTGCAGC TTTCCCTGCC CCCATCGAGA AAACCATCTC CAAAACCAAA 401 GGCAGACCGA AGGCTCCACA GGTGTACACC ATTCCACCTC CCAAGGAGCA 451 GATGGCCAAG GATAAAGTCA GTCTGACCTG CATGATAACA GACTTCTTCC 501 CTGAAGACAT TACTGTGGAG TGGCAGTGGA ATGGGCAGCC AGCGGAGAAC 551 TACAAGAACA CTCAGCCCAT CATGGACACA GATGGCTCTT ACTTCGTCTA 601 CAGCAAGCTC AATGTGCAGA AGAGCAACTG GGAGGCAGGA AATACTTTCA 651 CCTGCTCTGT GTTACATGAG GGCCTGCACA ACCACCATAC TGAGAAGAGC 701 CTCTCCCACT CTCCGGGTAA AGGTGGAGGT GGTGGTATCG AAGGTCCGAC 751 TCTGCGTCAG TGGCTGGCTG CTCGTGCTGG TGGTGGAGGT GGCGGCGGAG 801 GTATTGAGGG CCCAACCCTT CGCCAATGGC TTGCAGCACG CCCATAA 3' Sequence: TCTCGAGGATCCG CGGAAAGAAG AAGAAGAAGA AGAAAGCCCG AAAGG [00199] Murine Fc-TMP protein sequence (SEQ ID NO: 1035) 1 MVDGCKPCIC TVPEVSSVFI FPPKPKDVLT ITLTPKVTCV VVDISKDDPE 51 VQFSWFVDDV EVHTAQTQPR EEQFNSTFRS VSELPIMHQD WLNGKEFKCR 101 VNSAAFPAPI EKTISKTKGR PKAPQVYTIP PPKEQMAKDK VSLTCMITDF 151 FPEDITVEWQ WNGQPAENYK NTQPIMDTDG SYFVYSKLNV QKSNWEAGNT 201 FTCSVLHEGL HNHHTEKSLS HSPGKGGGGG IEGPTLRQWL AARAGGGGGG 251 GGIEGPTLRQ WLAARA* WO 2006/010057 PCT/US2005/024373 112 Example 2 Fermentation of Strain 6397 [00200] Fermentation of strain 6397 was initiated by inoculation of 500 mL of sterilized Luria broth with a seed culture of the strain in a shake flask. When cell density reached 0.9 at 600 nm, the contents were used to inoculate a 15 L fermentor containing 10 L of complex based growth medium (800 g glycerol, 500 g trypticase, 3 g sodium citrate, 40 g

KH

2

PO

4 , 20 g (NH 4

)

2

SO

4 , 5 ml Fluka P-2000 antifoam, 10 ml trace metals (ferric chloride 27.0 g/L, zinc chloride 2.00 g/L, cobalt chloride 2.00 g/L, sodium molybdate 2.00 g/L, calcium chloride 1.00 g/L, cupric sulfate 1.90 g/L, boric acid 0.50 g/L, manganese chloride 1.60 g/L, sodium citrate dihydrate 73.5 g/L), 10 ml vitamins (biotin 0.060 g/L, folic acid 0.040 g/L, riboflavin 0.42 g/L, pyridoxine HCl 1.40 g/L, niacin 6.10 g/L, pantothenic acid 5.40 g/L, sodium hydroxide 5.30 ml/L), add water to bring to 10 L). The fermenter was maintained at 37*C and pH 7 with dissolved oxygen levels kept at a minimum of 30% saturation. When the cell density reached 13.1 OD units at 600 nm, the culture was induced by the addition 10 ml of 0.5 mg/ml N-(3-oxo-hexanoyl) homoserine lactone. At 6 hours post induction, the broth was chilled to 10'C, and the cells were harvested by centrifugation at 4550g for 60 min at 5'C. The cell paste was then stored at -80*C. Example 3 Protein Refolding [002011 E. coli paste (300 g) from strain 6397 expressing mc-TMP was dissolved in 2250 ml lysis buffer (50 mM Tris HCl, 5 mM EDTA, pH 8.0) and passed through a chilled microfluidizer two times at 13,000 PSI. The homogenate was then centrifuged at 11,300g for 60 minutes at 4*C. The supernatant was discarded, and the pellet was resuspended in 2400 ml of water using a tissue grinder. The homogenate was then centrifuged at 11,300g for 60 minutes at 4'C. The supernatant was discarded, and the pellet was resuspended in 200 ml volumes of water using a tissue grinder. The homogenate was centrifuged at 27,200g for 30 minutes at 4 'C, and the supernatant was discarded. About 12.5% of the pellet was resuspended in 28 ml 20 mM Tris HCl, pH 8.0, with 35 mg hen egg white lysozyme (Sigma, St Louis, MO) using a tissue grinder and incubated at 37'C for 20 min. Following incubation, the suspension was centrifuged at 27,200g for 30 minutes at 22'C, and the supernatant was discarded. The pellet was resuspended in 35 ml 8 M guanidine HCl, 50 mM Tris HCI, pH 8.0, after which 350 R1 1 M DTT (Sigma, St Louis, MO) was added and WO 2006/010057 PCT/US2005/024373 113 material was incubated at 37 'C for 30 minutes. The solution was then centrifuged at 27,200g for 30 minutes at 22*C. The supernatant was then transferred to 3.5 L of refolding buffer (50 mM Tris base, 160 mM arginine HCl, 3 M urea, 20% glycerol, pH 9.5, 1 mM cysteine, 1 mM cystamine HCI) at 1 ml/min with gentle stirring at 4'C. Example 4 Construct Purification [00202] After about 40 hours incubation at 4C with gentle agitation, the refold solution described in Example 3 was concentrated to 500 pl using a tangential flow ultrafiltration apparatus with a 30 kDa cartridge (Satorius,Goettingen, Germany) followed by diafiltration against 3 L of Q-Buffer A (20 mM Tris HCI, pH 8.0). The concentrated material was filtered through a Whatman GF/A filter and loaded on to an 86 ml Q-Sepharose fast flow column (2.6 cm ID) (Amersham Biosciences, Piscataway, NJ) at 15 ml/min. After washing the resin with several column volumes of Q-Buffer A, the protein was eluted using a 20 column volume linear gradient to 60% Q-Buffer B (20 mM Tris HClI, 1 M NaCl, pH 8.0) at 10 mi/min. The peak fractions were pooled, and the pool was passed through a Mustang E syringe filter (Pall Corporation, East Hills, NY) at 1 ml/min. The filtered material was filtered a second time through a 0.22 gm cellulose acetate filter and stored at -80'C. Example 5 Protein PEGylation [00203] To a cooled (4'C), stirred solution of mFc-TMP (3.5 ml, 0.8 mg/ml) in a 100 mM sodium acetate buffer, pH 5, containing 20 mM NaCNBH 3 , was added a 3.8-fold molar excess of methoxypolyethylene glycol aldehyde (MPEG) (average molecular weight, 20 kDa) (Nektar). The stirring of the reaction mixture was continued at the same temperature. The extent of the protein modification during the course of the reaction was monitored by SEC HPLC using a Superose 6 HR 10/30 column (Amersham Biosciences) eluted with a 0.05 M phosphate buffer with 0.15 M NaCl, pH 7.0 at 0.4 ml/min. After 16 hours the SEC HPLC analysis indicated that the majority of the protein has been conjugated to MPEG. At this time the reaction mixture was buffer-exchanged into a 20 mM Tris/HCl buffer, pH 8.12. The MPEG-mFc-AMP2 conjugates were isolated by ion exchange chromatography using a 1 ml Hi Trap HP Q column (Amersham Biosciences) equilibrated with a 20 mM Tris/HCl buffer, pH 8.12. The reaction mixture was loaded on the column at a flow rate of 0.5 ml/min and the unreacted MPEG aldehyde was eluted with three column volumes of the starting buffer. A WO 2006/010057 PCT/US2005/024373 114 linear 20-column-volume gradient from 0% to 100 % 20 mM Tris/HCl buffer, pH 8.12, containing 0.5 M NaCL was used to the elute the protein-polymer conjugates. Fractions (2 ml) collected during ion exchange chromatography separation were analyzed by HPLC SEC as described above. A fraction containing the mono- and di-MPEG-nFc-TMP conjugates in an approximate ratio of 2.3 to 1 (as determined by SEC HPLC) was concentrated, and sterile filtered. Example 6 In vivo Testing [00204] BDF1 mice (Charles River Laboratories, Wilmington, Massachusetts) were divided into groups of 10 and injected on days 0, 21, and 42 subcutaneously with either diluting agent (Dulbecco's PBS with 0.1% bovine serum albumin) or diluting agent with 50 pg test mono- and di-MPEG-mFc-TMP conjugate protein (as described above) per kg animal. Each group was divided in half and bled (140 pl) from the retro-orbital sinus on alternate time points (days 0, 3, 5, 7, 10, 12, 14,19, 24, 26, 28,31, 33, 40, 45, 47, 49, 52 and 59). On day 59, mice were anesthetized with isoflurane prior to bleeding. The collected blood was analyzed for a complete and differential count using an ADVIA 120 automated blood analyzer with murine software (Bayer Diagnostics, New York, NY). [00205] Results showed that administration of mono- and di-MPEG-mFc-TMP conjugates on days 0, 21 and 41 resulted in subsequent increases in platelet levels to essentially the same degree with each dose. See FIG. 1. In comparison, administration of mFc-TMP (lacking a PEG moiety) on the same days (i.e., days 0, 21 and 41) also resulted in increased levels of platelets, but the increase following administration on day 21 was significantly lower (i.e., attenuated) than the increase after administration on day 0, and the increase following administration on day 41 was less (i.e., further attenuated) than that observed following administration on day 21. Overall, these results indicate that the Fc-TMP modified to include a WSP (e.g., PEG) moiety was able to induce platelet production that did not decrease when administered in a multiple dosage regimen. [00206] The present invention has been described in terms of particular embodiments found or proposed to comprise preferred modes for the practice of then invention. It will be appreciated by those of ordinary skill in the art that, in light of the present disclosure, numerous- modifications and changes can be made in the particular embodiments exemplified without departing from the intended scope of the invention.

Claims (27)

1. A substantially homogenous compound comprising a structure set out in Formula I, Formula I: [(X)a-F-(X 2 )b]-(L)c-WSPd wherein: F1 is a vehicle; X1 is selected from P -(L2)e P 2 -(L)f -P 1 -(L 2 )e P3(L 4 )_p 2 _(L)f p'_1 2 )e- and p4-L_ h -(L )gp2-(L2) 1 -pI -(L2)e P(L5)h-P3 2_3pl X 2 is selected from: -(L 2e.P1 -(L2)e_p1-(L)f -P2 -(L 2 )eP1-(L )rP2-(L4 )P3, and -(L 2 )e_p 1 -(L 3 )fp 2 -(L 4 ); -P3-(L 5 )h-p 4 wherein P1, P 2 ' p 3 , and P 4 are each independently sequences of pharmacologically active peptides; L', L2, L3, L , and L5 are each independently linkers; a, b, c, e, f, g, and h are each independently 0 or 1, provided that at least one of a and b is 1; d is at least 1; and WSP is a water soluble polymer, the attachment of which is effected at any reactive moiety in F 1 ; said compound having a property of improved bioefficacy when administered in a multidose regimen WO 2006/010057 PCT/US2005/024373 116

2. The compound of claim 1 which is a multimer.

3. The compound of claim 2 which is a dimer.

4. The compound of claim 1 comprising a structure set out in Formula II Formula II: [X 1 -F']-(L)c-WSP wherein F' is an Fe domain and is attached at the C-terminus of X 1 , and one or more WSP is attached to the Fc domain, optionally through linker Ll;

5. The compound of claim 4 which is a multimer.

6. The compound of claim 5 which is a dimer.

7. The compound of claim 1 comprising a structure set out in Formula III Formula HI: [F1-X 2 ]-(L')c-WSPd wherein F' is an Fc domain and is attached at the N-terminus of X 2 , and one or more WSP is attached to the Fc domain, optionally through linker L';

8. The compound of claim 7 which is a multimer.

9. The compound of claim 8 which is a dimer.

10. The compound of claim 1 comprising a structure set out in Formula IV Formula IV: [F'-(L)e-PL]-(L')c-WSPd wherein F1 is an Fc domain and is attached at the N-terminus of -(L')c-Pl and one or more WSP is attached to the Fc domain, optionally through linker L'.

11. The compound of claim 10 which is a multimer.

12. The compound of claim 11 which is a dimer.

13. The compound of claim 1 comprising a structure set out in Formula V Formula V: [F 1 -(L')e-P -(L 2 )rP 2 ]-(L1)c-WSPd wherein F1 is an Fc domain and is attached at the N-terminus of -L'-P'-L 2 _p 2 and one or more WSP is attached to the Fc domain, optionally through linker L'.

14. The compound of claim 10 which is a multimer. WO 2006/010057 PCT/US2005/024373 117

15. The compound of claim 11 which is a dimer.

16. The compound of any one of claims 1 through 15 wherein P1 and/or P 2 are independently selected from a peptide set out in any one of Tables 4 through 20.

17. The compound of claim 16 wherein P1 and/or P 2 have the same amino acid sequence.

18. The compound of any one of claims 1 through 15 wherein F 1 is an Fc domain.

19. The compound of any one of claims 1 through 15 wherein WSP is PEG.

20. The compound of any one of claims 1 through 15 wherein F1 is an Fc domain and WSP is PEG.

21. The compound of claim 19 wherein PEG has a molecular weight of between about 2 kDa and 100 kDa.

22. The compound of claim 21 wherein said PEG has a molecular weight of between about 6 kDa and 25 kDa.

23. A composition comprising the compound of claim 19, wherein said composition comprises at least 50% PEGylated compound.

24. The composition of claim 23 comprising at least 75% PEGylated compound.

25. The composition of claim 23 comprising at least 85% PEGylated compound.

26. The composition of claim 23 comprising at least 90% PEGylated compound..

27. The composition of claim 23 comprising at least 95% PEGylated compound.